Professional Documents
Culture Documents
Prosidingpih2015 PDF
Prosidingpih2015 PDF
PROSIDING
PERSIDANGAN
INDUSTRI HERBA
PUTRAJAYA
Editor
O Asiah
HF Lim
BJ Chee
M Nik Musaadah
M Mastura
K Getha
MGH Khoo
P Mazura
S Vimala
FRIM Proceedings No. 7
Editor
O Asiah
HF Lim
BJ Chee
M Nik Musaadah
M Mastura
K Getha
MGH Khoo
P Mazura
S Vimala
2015
© Institut Penyelidikan Perhutanan Malaysia 2015
Ketua Pengarah
Institut Penyelidikan Perhutanan Malaysia
52109 Kepong
Selangor Darul Ehsan
Malaysia
Tel: 603-62797000
Faks: 603-62731314
http://www.frim.gov.my
ISBN 978-967-0622-47-7
MS ISO 9001:2008
Diset dalam Calibri 11/12
Dihasilkan di Malaysia oleh Institut Penyelidikan Perhutanan Malaysia, Kepong
KANDUNGAN
Prakata ix
Pembentangan Pleno 1
Pembentangan Lisan 5
iii
Mengangkat Tongkat Ali ke Mata Dunia: Pengalaman 51
Pengkomersialan
MY Awang Ahmad
Pembentangan Poster 71
iv
Pemilihan dan Penghasilan Baka Limau Purut (Citrus hystrix) 106
Bermutu Tinggi
MA Farah Fazwa, J Mailina, MA Nor Azah, M Nur Nazihah, A Mohd.
Zaki, SB Syafiqah Nabilah et al.
Pembiakan Aksesi Terpilih Kacip Fatimah (Labisia pumila var. alata) 118
Melalui Kaedah Keratan dan Penilaian Pengeluaran Pucuk Pada
Peringkat Tapak Semaian
M Nur Nazihah, MA Farah Fazwa, S Norhayati, SB Syafiqah Nabilah, L
Mohd Asri & Z Mohd Zaini
Serangan Atteva sciodoxa (Ulat Harimau) Pada Tanaman Tongkat Ali 125
di Hutan dan Ladang Semenanjung Malaysia
WA Wan Muhd Azrul, A Mohd Farid, AS Sajap, S Tosiah & M Patahayah
v
Trend Penggunaan 18 Spesies Tumbuhan Ubatan di Bawah Program 170
NKEA di Kalangan Pengamal Perubatan Tradisional Melayu di
Semenanjung Malaysia
M Dionysia, MS Abdul Hayat, M Nik Musaadah, B Intan Nurulhani, MN
Madihah, Z Nurul Husna et al.
vi
Asam Gelugur Powder Rich in HCA (Hydroxycitric Acid): a Potential 228
Crop for Weight Management
HZ Umi Kalsum, HA Hashimah, A Sharizan, A Mohamed Nazim, M
Aida, S Nor Fadhilah et al.
vii
Effect of Organic-Based Fertilizer Rate and Planting Distance on 289
Biomass Yield of Belalai Gajah (Clinacanthus nutans)
AG Rosnani & M Syahida
Medicinal Plants Used for Women’s Healthcare Among the Jakun 309
Community in Kg. Peta: a Preliminary Study
I Nur Amalina, M Maryati & AB Mohd. Fadzelly
viii
PRAKATA
Prosiding yang mengandungi kompilasi kertas kerja mengenai dasar, akta dan
strategipengetahuan tradisi, penyelidikan dan pembangunan serta pasaran
herba dapat memberi maklumat berguna dan menjadi rujukan penting bagi
pihak yang berkepentingan dalam penyelidikan mahupun industri herba. Saya
berharap usaha pengumpulan maklumat mengenai herba dan hasilan semula
jadi daripada segala aspek akan diteruskan agar hasrat dan aspirasi Kerajaan
Malaysia yang mengenal pasti industri herba sebagai sumber pertumbuhan
ekonomi Negara di bawah NKEA dalam program transformasi ekonomi (ETP)
akan terlaksana.
ix
PEMBENTANGAN
PLENO
1
MEMARTABATKAN PENGETAHUAN TRADISI: ANTARA KHAZANAH
WARISAN BANGSA DAN SUMBER KEKAYAAN BAHARU NEGARA
H Norini
Institut Penyelidikan Perhutanan Malaysia (FRIM), 52109 Kepong, Selangor
Tel: 03-6279 7540 E-mel: norini@frim.gov.my
ABSTRAK
2
consent digunakan bagi memastikan sokongan padu setiap subetnik. Ternyata
pendekatan ini membuahkan hasil yang amat lumayan. Usaha
mendokumenkan TK berupaya mengekalkan khazanah warisan sementara
menghasilkan prototaip dan kemudian mengkomersialkan pula boleh
menjadikan herba/tumbuhan ubatan sebagai sumber kekayaan baharu negara.
Paling penting, hasil mahsul mengkomersialkan produk dikongsi kembali
dengan empunya ilmu. Dengan tekad membara kami terus melangkah, kali ini
dengan mendokumentasikan TK Orang Melayu pula pada pertengahan 2013.
Kami juga sedang berusaha membangunkan kerangka bagi perkongsian faedah.
Sebenarnya, terlalu banyak yang ingin dilaksanakan. Dengan dana yang terhad
kami cuba melangkah setapak demi setapak, tetapi kami yakin langkah kami
mampu menggegarkan dunia kerana hingga kini tidak ada mana-mana institusi
yang mengutarakan pakej penyelidikan yang sedemikian.
3
TONGKAT ALI: FROM LAB TO MARKET
ABSTRACT
The paper shares the experience of Biotropics Malaysia Berhad, as one of the
many vehicles in Malaysia, in the country’s collective effort to establish the
presence of Tongkat Ali, as a new health product in the international market.
The paper discusses the research works that have been undertaken not only to
meet the different requirements and restrictions by the health regulators in
the various countries, but also to meet the more sophisticated demands of the
modern and educated consumers of health products worldwide. The paper
looks at the commercialisation initiatives undertaken to introduce this
wondrous Malaysian herb in global platforms, and educating the consumers
on its numerous health benefits. The paper also shares some challenges faced
by Biotropics in this long, arduous but fulfilling journey, and tries to extract
some learning from the experiences that may be useful to other players who
may wish to tread the same journey.
4
PEMBENTANGAN
LISAN
5
HALA TUJU INDUSTRI HERBA DI BAWAH NKEA
ABSTRAK
6
Pembangunan Herba (HDD) merupakan badan penggerak utama dalam
pelaksanaan program EPP1 ini.
PENGENALAN
Fokus utama industri herba adalah untuk memperkasa kualiti produk bagi
menembusi pasaran eksport global dalam produk nutraseutikal dan perubatan
botaniKal. Di dalam NKEA Pertanian, EPP1 Produk Herbal Bernilai Tinggi
dijangka memberi nilai pasaran yang besar iaitu sebanyak AS$93 bilion pada
tahun 2015 dan dijangka meningkat kepada AS$107 bilion pada tahun 2017.
Faktor utama pertumbuhan positif nilai pasaran herba ini adalah berdasarkan
manfaat dan khasiat tersendiri herba yang diamalkan sejak dahulu lagi yang
secara tidak langsung mencetuskan kesedaran ke atas penggunaan herba di
kalangan rakyat Malaysia.
7
Rajah 1. 18 Fokus Herba di Bawah NKEA EPP#1 Produk Herba Bernilai Tinggi
Klaster Herba
8
maksimum sebanyak RM14,500/ha diberikan kepada peserta yang melibatkan
pembangunan kawasan dan infrastruktur ladang iaitu pembersihan kawasan,
sistem pengairan dan saliran, jalan dan parit, ladang serta pagar. Projek ini
dilihat berpotensi dalam memenuhi keperluan dan permintaan semasa bahan
mentah yang berkualiti dan konsisten kepada pengusaha-pengusaha tempatan
yang terlibat dengan program NKEA Pertanian untuk sub kumpulan projek
herba negara di bawah pengurusan sistem rantaian bekalan yang konsisten.
Objektif penubuhan Taman Penanaman Herba atau Herbal Park ini adalah
untuk penanaman bahan mentah secara komersial yang diterajui oleh Majlis
Pembangunan Koridor Ekonomi Pantai Timur (ECERDC) di Pasir Raja, Chegar
Perah dan Durian Mentangau. Ianya dijangka dapat memenuhi keperluan
industri herba memandangkan tanaman herba yang ditanam secara
berkelompok dan sebagai hab bekalan kepada pengusaha-pengusaha
mendapatkan bahan mentah. Sebanyak tiga syarikat peneraju telah dilantik
bagi mengusaha penanaman ini melalui pemilihan penilaian teknikal yang ketat
di peringkat HDD untuk memastikan projek ini dapat dilaksanakan mengikut
jadual. Model Pelaksanaan Taman Penanaman Herba ini adalah diuruskan
sepenuhnya oleh syarikat peneraju yang dilantik dengan dipantau secara mikro
oleh pihak ECERDC. Selain daripada itu, kaedah pelaksananaan penanaman
secara Satellite Farm di Herbal Park ini juga dibuka kepada petani-petani kecil
di kawasan tersebut. Satelliet Farm ini merupakan salah satu platform bantuan
kepada pengusaha-pengusaha kecil menambah pendapatan dan seterusnya
memangkin ekonomi Industri Kecil dan Sederhana (IKS). Mesyuarat
pemantauan berkala dan lawatan tapak sentiasa dilaksanakan bagi memastikan
projek ini dilaksanakan dengan lancar dan mencapai sasaran yang dikehendaki
oleh Kerajaan.
9
I. Penemuan (Discovery)
Penyelidikan melalui kajian kimia dan penilaian terapeutik ke atas
tumbuhan ubatan dan beraroma bagi mengenalpasti sebatian bioaktif
dan mekanisma tindak balas bahan aktif dalam penghasilan produk
yang berkualiti dan selamat.
10
satu inisiatif utama HDD. Sebanyak 14 syarikat telah berjaya melalui tatacara
penilaian yang ketat untuk dipilih sebagai syarikat peneraju. Syarikat-syarikat
tersebut akan menggunakan geran yang diperuntukkan untuk melaksanakan
kajian pra-klinikal dan klinikal bagi produk yang terpilih yang dikeluarkan oleh
syarikat di makmal-makmal di dalam dan di luar negara. Produk nutraseutikal
bernilai tinggi ini dengan kenyataan-kenyataan kesihatan yang telah disahkan
dengan hasil ujian, akan dipasarkan dan dilancarkan dalam pasaran global yang
dijangka berpotensi untuk dijual pada harga 10 kali ganda daripada harga asal.
Geran kategori ini disediakan bagi melaksanakan projek yang boleh membantu
merancakkan lagi pembangunan industri herba negara. Beberapa projek khas
yang telah dilaksanakan dengan kerjasama beberapa institusi/agensi antaranya
adalah Herbal Extraction Centre oleh Institute of Bio product Development
(IBD), Herbal Trading House oleh Marditech dan Monograph yang dilaksanakan
oleh Institut Penyelidikan Perubatan (IMR). Monograph merupakan laporan
ilmiah berkaitan herba tempatan yang digunakan sebagai bahan rujukan
penerbitan penting mengenai kualiti pokok herba yang terdapat di Negara.
Website Globinmed.com merupakan aplikasi digital yang menjadikan tempat
rujukan monograf ini. Sepanjang tempoh pelaksanaan projek monograph di
bawah geran ini sebanyak 27 tanaman herba dikenal pasti iaitu merungai,
lempoyang, gelenggang, belalai gajah, halia bara, ketum, cengkih, kayu manis,
cekur, karas, kari, kunyit, lada hitam, cucur atap, peria katak, capa, temu hitam,
lengkuas, bongelai, lemuni, pandan, kantan, limau purut, pecah beling, serai
makan, kenanga dan buah pala.
Selain daripada itu, HDD juga telah menyalurkan geran kepada pihak
FRIM untuk melaksanakan Projek Pengetahuan Tradisi Melayu (TK Malayu) dan
Kajian Rantaian Nilai Industri Herba Terpilih di Semenanjung Malaysia yang
merupakan pangkalan data industri herba di peringkat huluan dan hiliran yang
bertujuan bagi mengenal pasti permintaan dan penawaran bahan mentah dan
penghasilan produk herba. Sistem MyTKDL yang dibangunkan oleh MYIPO
merupakan pangkalan data pengetahuan tradisi dan sumber genetik dari
Malaysia dalam bentuk digital untuk kegunaan semasa meluluskan
permohonan antara geran di bawah projek khas ini.
11
dan juga ianya dibawa ke peringkat antarabangsa. Di peringkat hiliran, HDD
telah merangka halatuju yang lebih progresif bagi memastikan produk-produk
herba yang dihasilkan memberi pulangan berganda kepada negara.
RUJUKAN
12
PERANAN FRIM DALAM MEMPERKASA INDUSTRI HERBA DI MALAYSIA
MA Rasadah
Institut Penyeldikan Perhutanan Malaysia (FRIM), 52109 Kepong, Selangor
Tel: 03-6279 7330 E-mel: rasadah@frim.gov.my
ABSTRAK
Bahagian Hasilan Semula Jadi (BHS), FRIM telah ditubuhkan pada tahun 1995
telah diberi mandat secara langsung untuk melaksanakan beberapa projek
penyelidikan yang berteraskan penemuan sebatian semula jadi (Natural
Products Discovery) serta pembangunan produk herba & penjagaan kesihatan
daripada tumbuhan ubatan dan beraroma yang berorientasikan pasaran.
Terdapat dua pendekatan yang dilaksanakan oleh Bahagian Hasilan Semula Jadi
iaitu program jangka pendek (1-3 tahun) dan jangka panjang (3-5 tahun).
Program jangka pendek adalah untuk penghasilan data-data saintifik yang
diperlukan bagi menyokong dakwaan dan kegunaan herba untuk kesihatan &
penjagaan diri, sementara jangka panjang pula adalah penghasilan prototaip
produk dan proses komersialisasi. Penyelidikan yang terperinci adalah
diperlukan untuk menghasilkan produk yang berkualiti, berkesan dan selamat
digunakan. Sehingga kini lebih daripada 30 prototiap produk herba telah
dapat dihasilkan.
Berikut adalah usaha-usaha yang sedang dijalankan oleh FRIM dalam usaha
untuk memantapkan industri herba ke peringkat yang boleh dibanggakan:
Domestikasi dan penanaman tumbuhan herba
Bioprospek sebatian semula jadi dari sumber hutan
Kawalan kualiti produk herba
Formulasi dan pembangunan produk
Teknologi lepas tuai dan pemprosesan
Penemuan sebatian perintis dari sumber semula jadi
FRIM juga terlibat dengan pelbagai program dan latihan teknologi herba
dengan pelbagai kementerian seperti dengan Kementerian Luar Bandar dan
Wilayah (KKLW), MOA, KKM, MARA, MTDC dan MITI untuk melatih usahawan
herba dalam usaha untuk meningkatkan mutu penghasilan produk herba yang
mampu menembusi pasaran antarabangsa. Selain dari melaksanakan
penyelidikan, BHS juga menawarkan perkhidmatan dan berkongsi teknologi
terkini dalam pemprosesan & penghasilan produk herba. Sehingga kini lebih
daripada 200 usahawan herba telah mendapat manafaat dari program dan
latihan yang dikendalikan.
13
Kata kunci: FRIM, Bahagian Hasilan Semula Jadi, perintis, peranan, industri
herba
LATAR BELAKANG
Langkah baru dunia dalam era ekonomi global menuntut satu transformasi
pembangunan industri herba yang berteknologi tinggi di negara kita, Malaysia.
Penggunaan tumbuhan ubatan dan herba dalam perubatan tradisonal telah
melalui pelbagai transformasi sejak beberapa dekad yang lalu. Ianya dapat
menjana ekonomi negara sekiranya dieksplotasi dengan baik tanpa
menjejaskan eko-sistem terutamanya hutan semula jadi. Untuk itu, potensi
tumbuhan ubatan dilihat begitu besar sehingga dijadikan sebagai projek
pemulaan yang pertama (EPP1) dalam Bidang Keberhasilan Utama Ekonomi
(NKEA) pertanian di bawah seliaan Kementerian Pertanian dan Industri Asas
Tani Malaysia (MOA). Tumbuhan ubatan kini telah dinilai daripada segi aspek
farmaseutikal dan klinikal serta permintaan pasaran global yang meluas telah
meningkatkan nilai ekonominya setaraf dengan ubatan moden. EPP1 ini
memfokuskan ke arah memperkasakan kualiti produk dan usaha mempromosi
untuk menembusi pasaran global untuk produk nutraseutikal dan ubatan
botanikal.
14
PROGRAM-PROGRAM PENYELIDIKAN
15
spesies tumbuhan yang terpilih. Pemiawaian memainkan peranan yang penting
dalam penambah nilai. Pemiawaian ekstrak menjadi norma bagi produk yang
berkualiti.
2. Pemiawaian herba
Ekstrak piawai memainkan peranan penting dalam mempengaruhi penerimaan
pengguna terhadap produk berasaskan herba. Walau bagaimanapun, kajian ke
atas aspek pemiawaian bagi pemprosesan dan pengeluaran ekstrak herba di
Malaysia masih diperingkat awal. Untuk itu pihak FRIM mengorak langkah
16
dalam menjalankan kajian ini. Terkini lebih daripada 35 jenis ekstrak piawai
yang terdiri dari metabolit primer dan sekunder telah dapat dihasilkan. Selain
daripada itu, lebih daripada 12 ekstrak piawai untuk aktiviti antioksidan telah
dibangunkan dan dilesenkan kepada pihak industri herba. Beberapa Prosedur
Operasi Piawai (SOP) yang mematuhi garis panduan perundangan pasaran
telah dibangunkan dan sedia untuk dikomersialkan. Antaranya ialah mengkudu,
kacip fatimah, cucur atap, gelenggang, misai kucing, karas, pokok kapalterbang,
minyak pati dan sebagainya.
3. Polisakarida
FRIM merupakan satu-satunya badan penyelidikan di Malaysia yang
menjalankan R&D secara intensif terhadap polisakarida daripada tumbuhan
ubatan sebagai sumber produk farmaseutikal dan makanan berfungsi. Hasil
penyelidikan polisakarida daripada buah mengkudu telah dipaten dan
dikomersialkan kepada Syarikat Poly-Xtract Sdn. Bhd. untuk menghasilkan
fraksi polisakarida pada skala pandu (pilot scale) dan fraksi polisakarida ini
telah digunakan oleh Syarikat Nourish Care Sdn. Bhd. sebagai bahan aktif untuk
penghasilan produk makanan jenama Le’Natura iaitu biskut dan kopi putih
mesra diabetes. Produk-produk ini telah pun berada di rangkaian-rangkaian
pasar raya seperti Aeon Jusco, OTK, Mydin, Tesco dan Giant. Hasil penyelidikan
ini juga telah dipilih di antara 42 projek penyelidikan daripada seluruh negara
oleh pihak Agensi Inovasi Malaysia (AIM) sebagai peluang perniagaan inovatif
(IBO) yang berpotensi menjana pendapatan RM 1 billion selepas 3 tahun.
Inovasi daripada projek penyelidikan ini juga telah terpilih ke peringkat akhir
Anugerah Inovasi Negara 2012.
4. Pengredan gaharu
Pemintaan terhadap kayu gaharu dan hasilan produk berasaskan gaharu sangat
tinggi. Penentuan harga dan gred bagi kayu dan minyak gaharu adalah tidak
seragam. Untuk itu, FRIM telah memperkenalkan satu sistem pengklasifikasi
pintas (gaharu sense) yang berkonsepkan teknologi alat pengesanan bauan
(hidung elektronik). Sistem pengredan ini telah berjaya dipatenkan dan sedia
untuk ditawarkan kepada industri gaharu di negara ini.
17
membangunkan asai Cap-E untuk menganalisa kebolehdapatan bio antioksidan
pada peringkat sel dalam hasilan semula jadi dan ekstrak tumbuhan. Ujian
antioksidan/pemutih kulit dan Cap-E telah ditawarkan kepada pihak industri
herba.
Pada masa yang sama, Bahagian Hasilan Semula Jadi juga menjalankan
penilaian bioefikasi dan potensi bio dan mewujudkan 25 bioasai berkaitan
dengan bioterapeutik, kegagalan metabolik dan sistem pengenalpastian
mikrobial. Buat masa ini , Bahagian mempunyai pangkalan data bagi 564
spesies bakteria aerobik gram-negatif, 480 spesies bakteria anerobik, 619
spesies filamen kulat dan 267 species yis serta 19 Methicilin superbug
Staphyloccocus aureus untuk penyelidikan.
7. Pengiktirafan
Kejayaan Bahagian Hasilan Semula Jadi dalam tempoh yang singkat iaitu sejak
tahun 1995 dengan kepakaran 50 pegawai penyelidik dalam menjalankan
kajian herba dan hasilan semula jadi banyak dibantu oleh kemudahan makmal
berteknologi tinggi. Pusat Teknologi Herba (HTC) yang dimiliki FRIM, dilengkapi
dengan peralatan sejajar dengan matlamat dan peranannya sebagai pentas
yang menghubungkan agensi-agensi kerajaan termasuk agensi penyelidikan &
penguatkuasaan dengan pengusaha herba bagi penghasilan produk yang
berkualiti tinggi. Di samping itu, HTC telah menerima persijilan GMP untuk
pemprosesan ubatan berbentuk kapsul dan tablet daripada Kementerian
Kesihatan Malaysia (KKM) pada tahun 2007.
18
bumi, kunyit, limau nipis, serai wangi, lengundi, kalampayan, asam gelugor, dan
karas.
8. Pemindahan teknologi
Setiap tahun, bahagian ini telah melatih lebih daripada 250 orang usahawan
herba dan pemakanan melalui khidmat nasihat dan pemindahan teknologi
serta menawarkan antara 100 hingga 150 perkhidmatan pemprosesan herba.
Kebanyakan program latihan yang dianjurkan setiap tahun membantu industri
herba dalam penghasilan produk yang berkualiti, berkesan dan selamat
digunakan. Program-program ini melibatkan usahawan dari MARA, KKLW,
MTDC dalam usaha untuk meningkatkan mutu penghasilan produk herba dan
penanaman tumbuhan herba yang berkualiti.
19
herba dan spa. Program ini boleh menjadi contoh kepada institusi-institusi
pengajian tinggi tempatan untuk memulakan langkah yang sama.
KESIMPULAN
20
MENYUSUN STRATEGI BERPANDUKAN FAKTA RANTAIAN NILAI
INDUSTRI HERBA
ABSTRAK
Kata kunci: strategi, rantaian nilai, industri herba, hab herba Malaysia
PENGENALAN
21
kekayaan baharu negara sekiranya negara kita benar-benar serius untuk
menjadikan Malaysia sebagai salah satu hab herba dunia.
KAEDAH
Kertas kerja ini dibangunkan berdasar input bancian, survei dan siri bengkel
yang dijalankan oleh FRIM melalui kajian rantaian nilai industri herba terpilih di
Semenanjung Malaysia. Temu bual melibatkan pelbagai pihak berkepentingan
serta pemerhatian kumpulan penyelidikan terhadap senario semasa industri
herba dan rujukan pelbagai sumber turut dilakukan.
22
komersialisasi spesies turut penting bagi menentukan hab herba Malaysia
sentiasa di tahap optimum.
Selain itu, suatu pelan strategik yang memfokuskan matlamat utama untuk
pembangunan hab herba Malaysia beridentiti tersendiri perlu dirangka segera.
Kami mencadangkan pemilihan spesies berpotensi untuk dikembangkan adalah
berdasarkan beberapa kriteria seperti berikut:
23
2. Spesies yang digunakan secara tradisional, disokong secara saintifik
serta mempunyai potensi untuk bersaing di peringkat ASEAN - contoh:
Kacip fatimah.
3. Spesies yang turut dikaji serta popular disebabkan trend/keperluan
semasa - contoh: Belalai gajah
RUMUSAN
RUJUKAN
Gerard, B., Hood, S., Ruzy Suliza, H., Christof, J., Joerg, G., & Zurinawati, Z.A.
(Eds.). (2009). Health and Beauty from the Rainforest ─ Malaysian
Traditions of Ramuan. Kuala Lumpur: Editions Didier Millet Pte Ltd.
24
Jabatan Pertanian Semenanjung Malaysia. (2013). Statistik Tanaman (Sub
Sektor Tanaman Makanan). Jabatan Pertanian Semenanjung Malaysia.
Putrajaya: Jabatan Pertanian Semenanjung Malaysia.
Rohana, A. R., Nur Fazreen, Z., Ariff Fahmi, A. B., Nur Syazni, A., Siti Zubaidah,
S., Lim, H. F., et al. (2015). Directory of Herbal Cultivators in Peninsular
Malaysia. Kuala Lumpur: Forest Research Institute Malaysia.
Rohana, A. R., Nur Fazreen, Z., Ariff Fahmi, A. B., Nur Syazni, A., Siti Zubaidah,
S., Lim, H. F., et al. (2015). Value Chain Study for Holistic Development
of Herbal Industry. Proceeding 2015 3rd International Conference on
Rural Development & Entrepreneurship, (pp. 211─219). Hebei Province,
China.
25
PENDOKUMENAN PENGETAHUAN TRADISI MELAYU BERKAITAN
TUMBUHAN UBATAN DI SEMENANJUNG MALAYSIA
ABSTRAK
PENGENALAN
26
Pertanian dan Industri Asas Tani (MOA). Pelaksanaan aktiviti projek ini diterajui
oleh Institut Penyelidikan Perhutanan Malaysia (FRIM) dengan kerjasama
Herbwalk Consultancy; Institut Penyelidikan Perubatan (IMR) dan Bahagian
Perubatan Tradisional dan Komplementari, Kementerian Kesihatan Malaysia;
Perbadanan Harta Intelek Malaysia (MyIPO); Perpustakaan Negara Malaysia
dan Dr Harun Mat Piah (pakar filologi).
KAEDAH
27
Transliterasi daripada Jawi klasik kepada Rumi telah dilaksanakan oleh ahli
filologi.
Bancian Pengamal
28
Transliterasi Manuskrip Perubatan Melayu/Kitab Tib
Projek ini telah berjaya mentransliterasikan tiga buah kitab tib iaitu MSS 2999,
Kitab Tib Muzium Terengganu dan juga MSS 1292. Hasil transliterasi ini
sebanyak dua buah buku diterbitkan.
Output Projek
Hasil projek ini pada keseluruhannya adalah seperti Rajah 1. Secara ringkasnya
output yang dihasilkan adalah dalam bentuk penerbitan, pangkalan data,
koleksi spesimen baucer/herbarium dan germplasma untuk tujuan konservasi,
dan juga pengumpulan peralatan bagi tujuan penubuhan muzium.
KESIMPULAN
PENGHARGAAN
Kami sangat berterima kasih kepada semua pengamal yang terlibat dalam
projek ini dan juga pegawai pengumpul data yang telah melaksanakan bancian
dan juga survei. Projek ini dibiayai oleh MOA.
RUJUKAN
29
Kitab Tib/ Manuskrip Bancian pengamal Buku statistik pengamal
perubatan tradisional
Melayu di Sem. Malaysia
& PIC 1
Buku rujukan berkaitan
kriteria pengamal (BPTK)
Buku MSS2999:
pandangan dan
tafsiran perubatan Koleksi artifak/ peralatan/
moden terhadap manuskrip (IMR)
manuskrip perubatan
Melayu
Buku Transliterasi Penubuhan Muzium
Kitab Tib Muzium Perubatan Tradisional
Terengganu Melayu (IMR)
Buku Peralatan
Perubatan Tradisional
Survei pengamal terpilih
(penggunaan tumbuhan
ubatan & sosio ekonomi) Buku kajian sosio ekonomi
& PIC 2 pengamal
Dokumentasi TK
• Buku spesies di bawah
NKEA
• Buku spesies yang
diguna dalam
perubatan tradisional
Melayu
• Pangkalan data (MyTKDL
& BRAHMS)
30
PERALATAN DALAM PERUBATAN TRADISIONAL MELAYU: KHAZANAH
ILMU DAN PERADABAN WARISAN BANGSA
ABSTRAK
31
memberi penghargaan khusus kepada warisan pengetahuan tradisi Melayu
melalui penggiatan aktiviti pendokumentasian sebelum ia hilang ditelan masa.
PENGENALAN
32
(Koharuddin 2003). Bahan asas seperti jenis batu, tanah dan besi bagi
menghasilkan alatan ini juga memainkan peranan berbeza dan memberikan
kesan terapeutik yang berlainan. Jenis penyakit yang ingin dirawat, jantina, usia
pesakit dan tahap keseriusan penyakit juga menentukan jenis alatan yang
sesuai digunakan untuk penyediaan ubatan dan merawat pesakit tersebut.
Kajian ini dijalankan khusus bagi pengumpulan maklumat dan kajian
alatan/artifak perubatan Melayu untuk mencapai 2 objektif iaitu:
Aktiviti temu bual ini dijalankan dari September 2014 hingga Ogos 2015
ke atas seramai 65 orang pengamal yang dipilih dari seluruh Semenanjung
Malaysia. Pengamal ditemubual berpandukan borang soal selidik khusus sama
ada di rumah atau di premis perkhidmatan mereka. Demonstrasi berkaitan
juga divideokan. Temu bual ini merekod dan mendokumentasikan sejarah
amalan dan pengetahuan, termasuk maklumat sumber pengetahuan, jangka
masa mengamal, dan penerangan berkaitan amalan khusus mereka.
Perolehan Peralatan
Kajian ini juga mengumpul alatan dan artifak berkaitan sebagai koleksi dan
artifak khusus Alatan Perubatan Tradisional Melayu untuk dipamerkan di
Muzium Bioperubatan, IMR. Pengumpulan alatan dilakukan serentak atau
selepas mendapat maklumat melalui temu bual dilakukan ke atas pengamal.
33
Hasil kajian ini adalah terbitan sebuah buku mewah dengan memuatkan
gambar dan penerangan peralatan terpilih yang bersesuaian.
Jenis-jenis Amalan
Seramai 65 orang pengamal terpilih telah dicerap jenis amalan mereka melalui
temu bual yang dijalankan. Hasil temu bual mendapati amalan mereka boleh
dibahagi kepada kumpulan seperti dinyatakan di Jadual 1.
Peralatan
34
berjaya diperolehi daripada pengamal akan dibeli di tempat lain, sama ada atas
cadangan daripada pengamal atau penyelidik mencari sendiri dengan bertanya
kepada orang yang berpengetahuan. Selain daripada itu, alatan yang tidak
boleh diperolehi dari mana-mana premis, alatan tersebut direplikasikan melalui
perkhidmatan pembuat replika profesional. Foto yang menunjukkan dimensi
dan saiz alatan dijadikan rujukan bagi menghasilkan replika.
KESIMPULAN
Pelbagai maklumat yang diperolehi melalui kajian ini amat penting dipelihara
dalam bentuk pendokumentasian kerana ini merupakan khazanah warisan
bangsa yang amat bernilai. Selain daripada memantapkan identiti Melayu
dalam bidang perubatan tradisional, kajian seperti ini dapat mencetuskan
penemuan baru dalam bidang perubatan moden.
PENGHARGAAN
RUJUKAN
Editor, DBP. (2013). Kamus Dewan Edisi Keempat. Cetakan Keempat. Dewan
Bahasa dan Pustaka. Malaysia Dewan Bahasa dan Pustaka 2013.
35
PENYELIDIKAN DAN PEMBANGUNAN SERTA PENGKOMERSIALAN
(R&D&C) PRODUK BERASASKAN TANAMAN UBATAN DAN BERAROMA
DI MARDI
ABSTRAK
Kemajuan pesat industri herba negara perlu dimanfaatkan oleh semua pihak
terlibat agar komoditi ini dapat dijadikan sumber baru ekonomi negara. Usaha
yang tidak mengenal jemu semua pihak perlu digalakkan untuk mengambil
peluang memasukkan sains ke dalam herba tempatan agar kualiti bahan
mentah dapat dikekalkan pada aras yang tinggi dan keberkesanan serta
keselamatan produk herba dapat dibuktikan secara saintifik. Sehubungan itu,
dalam menunaikan tanggungjawab yang dimandatkan, MARDI sebagai sebuah
agensi di bawah Kementerian Pertanian dan Industri Asas Tani telah dan akan
melaksanakan tanggungjawab tersebut melalui program penyelidikan dan
pembangunan serta pengkomersilan (R&D&C). Ini meliputi bidang
pembaikbakaan, agronomi, pengurusan serangga dan penyakit, pengendalian
lepas tuai, pemprosesan primer dan sekunder serta pembangunan produk
makanan dan bukan makanan berasaskan herba bagi menambah nilai produk
herba tempatan. Pembaikbakaan bertujuan untuk mengumpul dan memilih
germplasma herba yang berkualiti agar tidak pupus. Pengeluaran bahan
tanaman berkualiti yang dipilih melalui proses pemilihan yang intensif juga
termasuk dalam aktiviti pembaikbakaan. Agronomi pula merangkumi semua
aspek pengurusan tanaman termasuk penyelidikan jarak tanaman, keperluan
baja, serta lain-lain keperluan asas hidup tumbuhan seperti air dan cahaya.
Pengurusan serangga dan penyakit yang berkesan pula merupakan faktor
kritikal untuk pertumbuhan pokok yang baik yang boleh merangsang hasil
biomassa dan kandungan fitokimia yang tinggi. Bahan mentah yang banyak dari
peringkat ladang perlu dikendalikan dengan cekap di peringkat lepas tuai bagi
memastikan kualiti yang diperoleh semasa dikutip dapat dikekalkan sehingga
penyimpanan. Pemprosesan primer dan sekunder yang cekap pula dapat
memastikan hasil dan kualiti bahan mentah dapat dikekalkan sehinggalah
proses pengumpulan bahan mentah untuk pembuatan produk. Apabila hasil
dan kualiti bahan mentah yang dipulihara sejak dari pengutipan dapat
digunakan sehingga proses pembangunan produk dapat memastikan produk
36
yang dikeluarkan sentiasa mempunyai aras kualiti yang tinggi. Namun begitu,
program penyelidikan yang dirangka ini hanya boleh berjaya apabila diberi
sokongan teguh oleh agensi utama sama ada dengan mewujudkan dan
memperkemaskan polisi, akta dan peraturan yang menyokong perkembangan
industri herba berserta peruntukan bajet yang mencukupi.
PENGENALAN
Industri herba telah menjadi satu fenomena sejak dua dekad lalu berikutan
peningkatan kesedaran dalam konsep pengurusan kesihatan, penambahbaikan
kualiti produk serta bukti-bukti saintifik terhadap keberkesanan dan
keselamatan produk. Lantaran itu, industri herba tempatan perlu dimajukan
untuk mengambil peluang yang terbuka itu. Warisan biologi negara yang kaya
dengan spesies (t)anaman (u)batan dan (b)er(a)roma (TUbA = medicinal and
aromatic plants, MAP = herba) telah membolehkan Malaysia menjadi negara
penyumbang yang siginifikan kepada industri herba.
37
MARDI sebagai satu agensi di bawah Kementerian Pertanian dan
Industri Asas Tani telah dipertanggungjawabkan untuk melakukan penyelidikan
dan pembangunan serta pengkomersialan (R&D&C) untuk meningkatkan nilai
sepanjang rantaian pengeluaran TUbA. Usaha MARDI tidak akan berjaya tanpa
sokongan daripada jabatan dan agensi lain yang turut terlibat dalam industri
ini.
Pembaikbakaan
38
MAB), penemuan dan pengesahan penanda molekul, pengenotipan dan
latihan.
Agronomi
Serangan serangga dan penyakit yang serius boleh mendatangkan kesan yang
sangat negatif kepada pertumbuhan tanaman yang seterusnya berupaya
menjejaskan hasil biomassa serta kandungan fitokimia dalam bahan mentah
TUbA. Penyelidikan terbaru di MARDI terhadap tanaman dukung anak telah
mengenalpasti penyakit utama bagi tanaman ini. Kaedah menentukan aras
infestasi dan keterukan penyakit telah dibangunkan. Penggunaan biopestisid
juga telah dikenal pasti berpotensi untuk mengawal perebakan insiden
penyakit pada tanaman ini. Penyelidikan terhadap perosak tanaman tongkat ali
pula telah mengenalpasti ulat harimau sebagai perosak penting yang berupaya
menjejaskan pertumbuhan pokok tongkat ali.
39
produk siap boleh berlaku disebabkan spesies yang tidak sesuai, kutipan pada
tahap tumbesaran yang tidak optimum, kontaminasi oleh agen mikrob dan
kimia serta pengekstrakan yang tidak optimum.
Selepas dikutip, bahan mentah perlu diproses untuk memastikan kualiti dapat
dikekalkan. Apabila pemprosesan melibatkan TUbA jenis minyak pati, bahan
yang dikutip perlu diproses dengan mengerat atau mericik daun/batang
menjadi saiz yang lebih kecil untuk diproses ke peringkat seterusnya. Dalam
pemprosesan sekunder, bahan mentah tanaman minyak pati akan diproses di
dalam tempat pemprosesan yang sesuai seperti penyulingan hidro agar hasil
minyak pati dapat diperoleh pada kadar pengekstrakan yang tinggi.
Pembangunan Produk
Produk TUbA boleh dibahagikan kepada dua kategori iaitu makanan dan bukan
makanan. Produk makanan boleh berupa makanan tambahan atau
nutraseutikal untuk tujuan penjagaan kesihatan. Kategori kedua ialah produk
bukan makanan seperti aromaterapi, toiletries, biopestisida berasaskan TUbA.
Minyak pati misalnya, boleh diniagakan dalam bentuk 100% tulen tanpa perlu
diproses lagi. Minyak pati juga boleh digunakan sebagai satu daripada ramuan
dalam produk aromaterapi dan boleh juga digunakan dalam pembuatan
produk penjagaan diri seperti sabun dan gel mandian berasaskan minyak
tersebut. Minyak serai, serai wangi dan jeremin telah dibangunkan sebagai
biopestisida bagi menangani masalah perosak tanaman kekupu belakang intan
(Plutella xylostella) dan ulat ratus (Spodoptera spp) yang banyak menyerang
sayuran seperti kobis dan sawi. Produk biopestisida ini sedang diuji di peringkat
kajian lapangan. Beberapa minyak pati utama yang lain seperti nilam, gelam
dan gajus (Anacardium occidentale) juga telah digunakan untuk
membangunkan produk nilai tambah seperti sabun, syampu dan formulasi anti-
kuman.
Pengkomersialan
Usaha telah dilakukan oleh MARDI untuk menggunakan teknologi moden bagi
menghasikan produk yang boleh dikomersialkan. Dalam RMKe-10, usaha ini
telah berjaya menghasilan beberapa produk yang berjaya dikomersialkan.
Contoh produk tersebut ialah NutrimaTM – Just Great (Roowi et al. 2013, Roowi
et al. 2012, Roowi & Crozier 2011, Roowi et al. 2010, Roowi et al. 2009 dan
Roowi 2008) dan ekstrak dukung anak (Lee et al. 2013). Produk-produk
40
tersebut telah berjaya memenangi beberapa anugerah dari dalam dan luar
negara.
41
KESIMPULAN
RUJUKAN
Gimlette, J. D. (1971). Malay Poisons and Charm Cures. Oxford University Press,
New York.
Lee, S.H., Tang ,Y.Q., Jaganath, I.B., Sekaran, S.D. et al (2013). Effects of cocktail
of four local Malaysian medicinal plants (Phyllanthus spp.) against
dengue virus 2. BMC Complement Altern. Med. 26;13:192. doi:
10.1186/1472─6882─13─192.
42
Roowi, S., William, M. and Crozier, A. (2012). Free phenolic acids in human
urine after drinking coffee rich with chlorogenic acids. J. Trop. Agric. And
Fd. Agric. (402):221-232.
Roowi, S., Hussin, Z., Othman, R., Muhammad, S. A., and Jusoh, A.Z. (2013)
Phenolic acid in selected tropical citrus. Asian Journal of Plant Biology
1:1─5.
Roowi, S., Mullen, W., Edwards, C. A. and Crozier, A. (2009). Yoghurt impacts
on the excretion of phenolic acids derived from colonic breakdown of
orange juice flavanones in humans. Mol. Nutr. And Food Res. 53 (S1):68 –
75.
Roowi, S., Stalmach, A., Mullen, W., Lean, M. E.J., Edwards, C. A. and Crozier, A.
(2010).Green Tea Flavan-3-ols: Colonic Degradation and Urinary
Excretion of Catabolites. J. Trop. Agric. And Fd. Chem. 58 (2)─1296–1304.
43
KAJIAN RANTAIAN NILAI INDUSTRI HERBA TERPILIH DI SEMENANJUNG
MALAYSIA
ABSTRAK
Industri herba telah dikenal pasti sebagai salah satu pemacu kegiatan ekonomi
yang berpotensi memberi sumbangan penting kepada pertumbuhan ekonomi
Malaysia bagi mencapai status negara maju menjelang tahun 2020 di bawah
Program Transformasi Ekonomi (ETP). Sehingga kini, maklumat berkaitan
industri herba kurang diselaras dan dikoordinasi. Sehubungan itu, masih tiada
satu pangkalan data yang mengandungi maklumat komprehensif industri
herba. Bagi memenuhi aspirasi ini, kajian berkaitan rantaian nilai industri herba
adalah penting dan meliputi aspek permintaan dan penawaran. Rantaian nilai
industri herba merangkumi pembekal bahan tanaman, penanam, pengusaha,
pemborong dan peruncit. Kajian ini dijalankan bagi membangunkan pangkalan
data industri herba di peringkat huluan dan hiliran melalui bancian
komprehensif di seluruh Semenanjung Malaysia. Konsep rantaian nilai oleh
Micheal Porter menjadi asas kajian ini. Hasil bancian mendapati sebanyak
6,570 individu dan syarikat terlibat dalam rantaian nilai industri herba. Hasil
kajian ini dapat membantu dalam pembuatan keputusan bagi merencana hala
tuju industri herba berasaskan maklum balas langsung dari pihak industri
herba.
PENGENALAN
44
menyokong pembuatan keputusan telah digunakan dalam strategi daya saing
yang dibangunkan oleh Porter pada awal tahun 1979.
45
berkaitan 5 aktiviti utama dan 4 aktiviti sokongan berdasarkan Teori
Michael Porter seperti berikut:
Aktiviti utama:
Aktiviti sokongan:
46
xi. Pembangunan laman sesawang kajian
Bagi tujuan promosi serta penyebaran maklumat kajian, pautan laman
sesawang projek melalui laman sesawang FRIM telah dibangunkan.
Kesemua informasi terkini berkaitan kajian dikemaskini dari masa ke
semasa di laman tersebut.
Sumber: Sekaran (2000), Wagner et al. (2012) & Rohana et al. (2015)
47
sasar yang sukar untuk memberi maklumat sewaktu bancian, dengan masing-
masing 14%, 10% dan 9% menolak untuk bekerjasama sewaktu ditemu bual
berbanding penanam dan pembekal bahan tanaman yang hanya 4% dan 3%
menolak sewaktu ditemu bual.
48
Jadual 3. Bilangan responden yang bekerjasama, tidak bekerjasama dan
individu tiada di lokasi perniagaan sewaktu bancian
Individu
Tidak tiada/kedai
Bekerjasama Jumlah
bekerjasama ditutup semasa
bancian
Pembekal
86 3 1 90
bahan tanaman
Penanam 435 17 10 462
Pengusaha 451 50 33 534
RUMUSAN
RUJUKAN
49
Rohana, A.R., Nur Fazreen, Z., Ariff Fahmi, A.B., Nur Syazni, A., Siti Zubaidah, S.,
Lim, H.F., et al. (2015). Directory of Herbal Cultivators in Peninsular
Malaysia. Kuala Lumpur: Forest Research Institute Malaysia.
Rohana, A.R., Nur Fazreen, Z., Ariff Fahmi, A.B., Nur Syazni, A., Siti Zubaidah,
S., Lim, H.F., et al. (2015). Pp. 211–219 in Value Chain Study for Holistic
Development of Herbal Industry. Proceeding 2015 3rd International
Conference on Rural Development & Entrepreneurship, Hebei Province,
China.
Wagner, C., Kawulich, B. & Garner, M. (2012). Doing social research: a global
context. McGraw-Hill Higher Education, UK.
50
MENGANGKAT TONGKAT ALI KE MATA DUNIA: PENGALAMAN
PENGKOMERSIALAN
MY Awang Ahmad
Kompleks Perhutanan, Fakulti Sains dan Sumber Alam, Universiti Malysia
Sabah, 88999, Kota Kinabalu, Sabah Malaysia
Tel: 088-320118 Faks: 088-320876 E-mel: agahmad@ums.edu.my
ABSTRAK
PENGENALAN
51
tongkat ali. Keadaan ini boleh mengurangkan keuntungan kepada industri
herba di Malaysia, sekiranya ingin menembusi pasaran luar negara. Kertas kerja
ini akan membincangkan kegunaan lain tongkat ali selain dari kegunaannya
sebagai afrodisiak, dari pengalaman penulis serta kajian literatur.
Bhat dan Karim (2010) melaporkan tongkat ali mempunyai pelbagai jenis
kegunaan dari bahagian akar hingga ke daun. Banyak kajian berkaitan sifat
pharmakologi tongkat ali telah dilakukan. Antaranya akar, batang, dan kulit
mempunyai aktiviti anti plasmodial (anti malaria), manakala daun
mengandungi aktiviti antitumor dan antimikrobial. Sebatian kimia juga telah
berjaya diipencilkan dari berbagai bahagian pokok tongkat ali. Namun demikian
bagi mendapatkan sumber mentah tongkat ali yang berkualiti, penghasilan
sumber yang boleh dipercayai dan mapan harus dititikberatkan. Sehubungan
dengan itu, penulis telah berjaya membangunkan satu teknologi penanaman
tongkat ali secara mapan iaitu EurypotTM.
Eurypot™
52
Tongkat Ali Sebagai Pokok Hiasan (Landskap)
Tanaman hiasan merupakan salah satu lagi cara untuk meningkatkan nilai
pokok tongkat ali. Pada masa ini, pokok tongkat ali belum dianggap sebagai
pokok hiasan dan belum disenaraikan dalam senarai tumbuhan hiasan di
bawah Jabatan Landskap Negara. Melalui kajian yang dijalankan oleh Program
Taman Alam dan Rekreasi di Universiti Malaysia Sabah (UMS), mendapati
bahawa, tongkat ali mempunyai bentuk monopodial sama seperti pokok
eksotik Brazilian Tower Tree yang sering digunakan sebagai pokok hiasan di
perbandaran di Malaysia. Walau bagaimanapun, kos pembelian pokok eksotik
ini adalah agak tinggi iaitu dalam lingkungan RM100-300 sepokok.
53
Tongkat Ali Sebagai Makanan Tambahan Haiwan
54
KESIMPULAN
Walaupun pokok tongkat ali dinobatkan sebagai raja herba di Malaysia, namun
usaha untuk mempromosi dan meninggikan nilai tongkat ali masih lagi rendah
jika dibandingkan dengan ginseng Korea. Pelaburan jutaan ringgit dalam
penyelidikan tongkat ali harus diterjemahkan dalam bentuk hasil bermanfaat
kepada pengguna. Sehubungan dengan itu, sudah tiba masanya untuk
menghasilkan pelbagai produk tongkat ali, daripada penghasilan bahan mentah
sehingga kepada produk ubatan dan kosmetik yang bernilai tinggi. Inovasi
produk berasaskan tongkat ali hasruslah disokong dengan pemberian dana
yang setimpal.
RUJUKAN
Bhat, R. And Karim, A.A. (2010). Tongkat Ali (Eurycoma longifolia Jack): A
review on its ethnobotany and pharmacological importance.
Fitoterapia 81(7): 669─679.
Wiart, C. (2002). Medicinal Plants of South East Asia. Prentice Hall, Pearson
Malaysia. Sdn.Bhd., Selangor Malaysia.
55
ANALYSING MALAY MEDICAL MANUSCRIPT MSS2999: THE FINDINGS
AND THE HOPE FOR THE FUTURE
H Abdul Ghani
INTRODUCTION
Traditional Malay medicine is not a new after thoughts but rather, it is at the
very least more than 6,000 years old. It stems out from the need to treat
diseases of the community. Like most other traditional medicine, it is based on
the understanding of what is normal and what comprise abnormality. It is this
that directed the practitioner towards concocting the appropriate remedies to
correct the abnormalities. The practice initially was based on instinctual
observations using simple remedies comprising of single materia medica, to
the eventual understanding of the varied forces in the materia medica which
could help in enhancing the effects of the remedies prescribed. This had
resulted in the cooking up of more complex remedies utilising varied materia
medica to render the treatment better directed. The old practitioners indeed
are people with great knowledge and great minds for without them they could
not possibly have craftily concocted remedies very well directed to the cure of
the diseases being addressed.
THE MANUSCRIPT
MSS 2999 is a small manuscript of 22 pages with 141 prescriptions for the
treatment of 42 different diseases in 12 different anatomical regions. It was
obtained from Pusat Manuskrip Melayu of the National Library of Malaysia.
Not much is known of the origin or the author of the manuscript. It is probably
notes of a student during his learning process with his ‘guru’. This is reflected
upon the lack of organisation of the prescriptions according to system of
disease.
56
THE DISEASES
Most of the diseases addressed were trivial and it can be assumed the most
common ailments of the period the manuscript was written. It covers diseases
of the eye, ear, nose and throat, dental problems, gastrointestinal tract,
respiratory tract, urinary tract, fevers and infective processes and diseases of
women. There was no mention of treatment of chronic diseases like diabetes,
hypertension, cardiac diseases nor of cancer. It may be assumed that either
these diseases were not common or the practitioner then has no knowledge of
these disorders. The approach to treatment at times are very similar to modern
medical therapy while at time the approach differs but is deemed more
appropriate than those of modern therapy.
CONCLUSION
For a nation with unwritten history of more than 6,000 years, one cannot deny
the advancement of their knowledge. With the discovery of the archaeological
site at Sungai Batu, Kedah, it further provides evidence of this long history and
puts the Malay nation a being far more matured than most other nations with
technological advancement in the field of iron smelting as being the
oldest.Traditional Malay medicine too may have evolved to an advanced stage
with knowledge being spread to wherever they used to trade via the sea route.
Exchange of knowledge is a continuous learning process of this nation as can
57
be noticed from the use of materia medica from China, India, the Middle East
and Europe.
58
COMMERCIALIZED INNOVATIVE HERBAL PRODUCTS FROM UPM
ABSTRACT
The product invention is an additive which can be incorporated into edible oil
composition especially cooking oil to provide therein antioxidant effect and
reduce oil adsorption into the food prepared. The additive is composed of
biophenolics formulated from plant extracts of the Rutaceae family. The
antioxidant characteristics of Rutaceae leaf alcohol extract were evaluated in
refined, bleached and deodorized (RBD) palm olein at 180oC using an
accelerated oxidation and deep frying studies for up to 40 hours. The extract,
at the optimum concentration 0.1%, retarded oil deterioration significantly
(P<0.05) in deep frying studies, comparable to 0.02% BHT (Butylated Hydroxy
Toluene) in tests such as peroxide value, anisidine value, free fatty acid,
Oxidative Stability Index (OSI), polar and polymer compounds content. The
sensory evaluation of french fries fried in oil containing the extract showed
significant (P<0.05) improvement in crispiness, taste and overall quality,
especially after the 40th hour of frying compared to those similarly fried in the
control oils and the oils containing BHT. The polyphenols content of the extract
were 103.2 mg/g gallic acid equivalent. Rutaceae leaf extract exhibited
excellent heat stable natural antioxidant properties that retarded oil
degradation and improved the quality of the fried product.
INTRODUCTION
59
antioxidants have been proven to be capable of retarding oxidation in frying
and in accelerated oxidation study. Phenolic compounds such as catechin and
catechin composites have been shown to significantly improve the stability of
peanut oil and palm superolein frying performances. The objective of this study
was to evaluate the antioxidative characteristics of Rutaceae leaf extract in
palm olein during accelerated oxidation and frying.
The cleaned Rutaceae leaves were dried in a hot air oven at 45oC for 24 hr,
ground to a fine powder before extracting with 10 times its weight in ethanol
for 8 hr at 50oC. The solvent was evaporated using rotary evaporator.
Frying Experiment
Deep frying was carried out in a stainless steel electrical open fryer. The
treatments conducted simultaneously were, (1) Afdhal-x, palm olein containing
0.1% Rutaceae leaf extract (10 kg oil was introduced into separate fryers, and
heated up to 60oC before adding 0.1% extract, and stirred until completely
dissolved) (2) palm olein containing 0.02% BHT, and (3) palm olein without any
additive as control. Approximately 400 g oil samples were collected from each
fryer to represent sample for day 0 before frying. The remaining oil was heated
at 180 2oC and was allowed to equilibrate at this temperature for 30 min.
About 14 batches at 200g per batch of french fries were fried for 2.5 min per
day at 30 min intervals for 8 hours daily. The fryers were turned off at the end
of the frying experiment each day and the oil cooled to 60oC, before filtering
using separate filters to remove debris. Accurately weighed 400g of the frying
oil from each fryer were sampled into amber bottles at the end of each day. All
oil samples were flushed with slow bubbles of nitrogen from the bottom of the
bottles and stored in at -20oC for physical and chemical analysis.
Sensory Evaluation
Sensory evaluation was conducted on the same day using the 5th and 6th
batches of fried French fries. The fryers were topped off up to 10 kg with oil
containing antioxidants (0.02% BHT or 0.1% Rutaceae leaf extract) depending
on the oil loss. The whole procedure was repeated consecutively for 5 days.
The French fries colour, flavour, oiliness, crispiness, taste and overall quality
were evaluated using a 9- point hedonic scale (1= very poor; 9= very good). The
sensory was done by 20 trained panellists on day 1, 3 and 5 equivalent to 8, 24
and 40 hours of frying in the same oil.
60
Statistical Analysis
Each experiment and analysis, including leaves sampling and extraction, were
conducted in triplicate. The MINITAB 14 software was used to analyse data for
determining ANOVA, standard deviation and Duncan’s multiple range test for
significance at a 5% level.
In deep frying studies the peroxides present in the oil containing the extract
(Afdhal oil) were lower than in the BHT, indicating that in the presence of food,
the compounds in the extract significantly (P<0.05) reduced peroxides
formation since less of the peroxides have changed to the secondary products
as confirmed by the anisidine values, when compared to the oils containing
BHT and the untreated control (Figure 1).
The Peroxide values (Figure 1) which measures the primary oxidation products
in the oil, is expected to initially increase and then decrease after certain hours
of heating, due to the formation of secondary oxidation products such as
ketones, aldehydes, hydrocarbons, epoxides etc. which were then gauged by
the anisidine values. Peroxide levels were evaluated in the frying oil from their
ability to liberate iodine from potassium iodide. Peroxides formed in palm oil
were relatively low (below 3m eq oxygen/kg oil after 30-40 hours at 180°C)
compared to other more unsaturated cooking oils because it contains about
50% monounsaturated fatty acids and about 45% saturated fatty acids with
very low levels of polyunsaturated fatty acids. Palm oil is also very rich in
tocotrienols, which act as endogenous antioxidants, making the oil fairly stable
to high temperatures use.
Sensory Evaluation
Sensory evaluation of French fries fried on the 40th hours in the various oils
showed significantly better overall quality, for samples fried in oils containing
Rutaceae leaf extract (Afdhal-x), unlike the control and BHT samples. However,
no significant difference (P<0.05) were found in scores for oiliness, crispiness
and taste observed between samples throughout the frying experiment (Figure
1). Evaluation on the colour of French fries shows that Rutaceae and control
samples were not significantly different (P<0.05) from one another during the
40 hours of frying. Although the colour of oil and French fries were darker in
the presence Rutaceae extract, and increases significantly during frying, the
colour sensory scores of French fries were the reverse. French fries fried in oil
containing Rutaceae extract were preferred as the number of frying hours
61
increased to the 40th hour (Figure 1). Samples fried in the control oil showed a
slight increase in the colour intensity with increasing frying time, as expected.
BHT caused slight decrease in scores as the frying hours increased. Flavour
scores for all samples were not affected by increasing frying time, but were
slightly different from one another by the 40th hour of frying. The mean scores
were not significantly different (P<0.05) because of the high standard
deviations in the scores given by the sensory panellists. All samples fried in oil
containing Rutaceae extract were preferred even at the 40th hour of frying
(average score of 7.00); unlike those fried in oil containing BHT which scored
lower than 5. The leaf extract improved the fry’s characteristics especially for
flavour scores after using the oil for 40 hours of frying.
62
The Oil Content of the Sample (%)
Oil content (%) 5
4 3.54
Nugget
3.10
2.85
3 2.52 2.61 2.51
2.30 French
fries
2 1.74
1.38 1.22
1.35 1.13
1.26 1.12 Keropok
0.96 Lekor
1
0.38
Doughnut
0
control c.grandis c.aurantifolia c.microcarpa
Sample
63
CONCLUSION
This study shows that Rutaceae leaf alcoholic extracts are effective at retarding
oil oxidation. The leaf extract maintained the sensory qualities of French fries,
better than BHT, in reused oil, showing that Rutaceae leaf is a good antioxidant
for oils, besides having other beneficial health properties.
ACKNOWLEDGEMENTS
The authors would like to thank Ministry of Education for financially supporting
the commercialisation of this work
REFERENCES
Mohamed, S. and Nor, F.M. (2008) Additive for edible oil composition. US
Patent App. 12/745,635.
64
DEVELOPMENT OF HERBAL PRODUCT FOR CANCER THERAPY
ABSTRACT
INTRODUCTION
65
The resistance of metastatic cancerous cells to chemotherapy and its
adverse effects has become a serious challenge in cancer research. Despite the
intensive progress in chemotherapeutics in the last decades, the need to
discover and to develop new, alternative, or adjuvant therapeutic agents
remains.
Natural Products, especially plants, have been used for the treatment of
various diseases since antiquity to date. Secondary metabolites from plants
have such as flavonoids, alkaloids, terpenoids and glycosides have been prized
for their protective effect against different pathogens (e.g bacteria, fungi or
insects), pain-relieving and healing activity or growth regulatory molecules (e.g.
hormone-like substances). Also, Phytochemicals have a fundamental role in
prevention and treatment of cancer either by modulating the activity of
specific hormones and enzymes responsible for carcinogenesis or by direct
cytotoxicity against cancer cells (Kintzios & Barberaki 2004). Natural products
have been forefront in cancer treatment; recently pharmaceutical companies
have explored more than 25,000 plants for anti-cancer drugs. About 60% of the
anti-cancer drug over the globe have been discovered from plant origin
between 1983 and 1994 (Sakarkar & Deshmukh 2011). Phytoconstituents
derived from plants are paclitaxel from Taxus brevifolia Nutt, vinblastine
and vincristine from Catharanthus roseus, etoposide and teniposide which
are epimers of podophyllotoxin that isolated from the roots of different
podophyllum species, also camptothecin which is extracted from the Chinese
tree Camptotheca acuminata (Newman et al. 2000). The significance of
phytoconstituents in treatment of cancer is very obvious, for instance the
discovery of paclitaxel (Taxol®) evident the success of natural products in
combating cancers (Oberlies & Kroll 2004, Cragg & Newman 2005).
It’s well known that the growth of solid tumors depends mainly on
angiogenesis. Many natural compounds proved to have antiangiogenic effects
such as asisoliquiritin and combrestastin were isolated from Glycyrrhiza glabra
and Combretum caffhum respectively were found to inhibit MMPs and tubules
formation, in turn interrupt angiogenesis (Pettit et al. 1989, Kang et al. 2010).
Other natural compounds showed potent antiangiogenic activity including
resveratrol from grapes (Cao et al. 2005) quercetin (Tan et al. 2003),
rosmarinic acid (Huang & Zheng 2006), genistein (Su et al. 2005) and curcumin
(Gururaj et al. 2002) with various mechanism of actin. Some compounds
modulates VEGF signalling pathway either by decreasing the production of
VEGF or interfering with binding of VEGF to its receptors such as Vincristine,
Ginsenosides, Taxol and Triptolide extracted from V. rosea, P. ginseng, T.
brevifolia and T. wilfordii respectively (Attardo & Sartori 2002). Several flavones
66
and flavonols isolated from different plants reported to inhibit endothelial cell
proliferation and migration the key steps in angiogenesis (Fotsis et al. 1997).
Plant as sources of antiangiogenic agents could be of great value, as they can
be produced commercially in large quantities (Al-Suede et al). Although, there
is no plant based antiangiogenic compounds available commercially at present.
An herb is a plant or plant part used for its scent, flavor, or therapeutic
properties. Herbal medicines are dietary supplement used to treat various
diseases. Recently, traditional medicine such as Chinese medicine, Kampo
medicine and Ayurveda are used in Asian countries and most of them are
accepted as popular complementary and alternative medicine therapies for
cancer worldwide (Elujoba et al. 2005). In some countries, herbal medicines are
in some forms of drugs listed in Chinese and Japanese pharmacopoeias now
used in daily cancer clinical practice. It is common approach in herbal medicine
studies nowadays to find active herb in a composite formula for certain disease
and further explore active ingredients in the herb, thereafter composite
formula can be standardized for biological activity and chemical composition.
FDA classify the drug products from herbal medicine as dietary supplements
and botanical drug products, with regard to the experiences and problems in
herbal medicines for its use as a single and as combination (Feng et al. 2011).
Botanical products from herbal medicines are extracts/fractions containing
several active components which can be determined precisely using chemical
analysis technologies such as gas chromatography (GC), high-performance
67
liquid chromatography (HPLC) and mass spectrophotometer (MS)(Choi et al.
2009)
CONCLUSION
REFERENCES
Bray, F., Ren, J. S., Masuyer, E. & Ferlay, J. (2013). Global Estimates of Cancer
Prevalence for 27 Sites In The Adult Population In 2008. International Journal
of Cancer 132(5): 1133─1145.
Cao, Y., Fu, Z.D., Wang, F., Liu, H.Y. & Han, R. (2005). Anti-Angiogenic Activity Of
Resveratrol, A Natural Compound From Medicinal Plants. Journal of Asian
natural products research 7(3): 205─213.
Choi, W.C., Kwon, S.P. & Park, S.J. (2009). Process for Preparation of Rhus Verniciflua
Extracts Having Excellent Anti-Cancer Activity and Anti-Cancer Pharmaceutical
Composition Containing The Same. Google Patents.
Chorawala, M., Oza, P. & Shah, G. (2012). Mechanisms of anticancer drugs resistance:
an overview. International Journal of Pharmaceutical Sciences and Drug
Research 4(1): 1─9.
Feng, Y., Wang, N., Zhu, M., Feng, Y., Li, H. & Tsao, S. (2011). Recent Progress on
Anticancer Candidates in Patents of Herbal Medicinal Products. Recent patents
on food, nutrition & agriculture 3(1): 30─48.
68
Fotsis, T., Pepper, M. S., Aktas, E., Breit S., Rasku, S., Adlercreutz, H., Wähälä, K.,
Montesano, R. & Schweigerer, L. (1997). Flavonoids, Dietary-Derived
Inhibitors of Cell Proliferation And in Vitro Angiogenesis. Cancer Research
57(14): 2916─2921.
Gupta, V. K. & Sharma S. K. (2006). Plants as Natural Antioxidants. Nat. Prod. Rad 5(4):
326─334.
Gururaj, A. E., Belakavadi, M., Venkatesh, D. A., Marme, D. & Salimath, B. P. (2002).
Molecular Mechanisms of Anti-Angiogenic Effect of Curcumin. Biochem
Biophys Res Commun 297(4): 934─942.
Huang, S.S. & Zheng R.l. (2006). Rosmarinic Acid Inhibits Angiogenesis and Its
Mechanism of Action In Vitro. Cancer letters 239(2): 271─280.
Kang, S. W., Choi, J. S., Choi, Y. J., Bae, J. Y., Li, J., Kim, D. S., Kim, J. L., Shin, S. Y., Lee, Y.
J., Kwun, I. S. & Kang, Y. H. (2010). Licorice Isoliquiritigenin Dampens
Angiogenic Activity via Inhibition of Mapk-Responsive Signaling Pathways
Leading to Induction of Matrix Metalloproteinases. J Nutr Biochem 21(1):
55─65.
Kintzios, S. E. & Barberaki, M. G. (2004). Plants That Fight Cancer, CRC Press.
Newman, D. J., Cragg, G. M. & Snader, K. M. (2000). The Influence of Natural Products
upon Drug Discovery. Natural product reports 17(3): 215─234.
Pal, S., Bhattacharjee A., Ali, A., Mandal, N. C., Mandal, S. C. & M. Pal (2014). Chronic
Inflammation And Cancer: Potential Chemoprevention Through Nuclear Factor
Kappa B and P53 Mutual Antagonism. J. Inflamm.(Lond.) 11: 23.
Pettit, G., Singh, S., Hamel, E., Lin, C. M., Alberts, D. & Garcia-Kendal, D. (1989).
Isolation and Structure of The Strong Cell Growth and Tubulin Inhibitor
Combretastatin A-4. Experientia 45(2): 209─211.
Pham-Huy, L. A., He, H. &Pham-Huy, C. (2008). Free Radicals, Antioxidants in Disease
and Health. International journal of biomedical science: IJBS 4(2): 89.
69
Su, S.J., Yeh, T.M. , Chuang, W.J., Ho, C.L., Chang, K.L.,. Cheng, H.L, Liu, H.S., Cheng,
H.L., Hsu P.Y. & Chow, N.H. (2005). The Novel Targets for Anti-Angiogenesis of
Genistein on Human Cancer Cells. Biochemical pharmacology 69(2): 307─318.
Tan, W.F., Lin, L.P., Li, M.H., . Zhang, Y.X , Tong, Y.H., Xiao, D. & Ding, J. (2003).
Quercetin, a Dietary-Derived Flavonoid, Possesses Antiangiogenic Potential.
European journal of pharmacology 459(2): 255─262.
70
PEMBENTANGAN
POSTER
71
SARINGAN FITOKIMIA DAN AKTIVITI ANTI-RADANG TUMBUHAN
UBATAN TERPILIH WARISAN ORANG ASLI SEMELAI, POS ISKANDAR,
BERA, PAHANG
ABSTRAK
PENGENALAN
72
pengetahuan tradisi kaum Semelai di Pos Iskandar, Bera, Pahang. Kaedah
penyaringan fitokimia telah digunakan untuk mengenalpasti metabolit
sekunder yang dihasilkan oleh tumbuhan. Anti-radang atau dikenali anti-
inflamasi dilaksanakan melalui penentuan nilai perencatan enzim iaitu enzim
lipoksigenase dan enzim xanthine oksidase serta pembengkakan telinga mencit
yang diaruh dengan TPA. Perencatan aktiviti enzim tersebut boleh
mengurangkan kesan buruk akibat keradangan. Profil bioaktiviti dan fitokimia
tumbuhan terpilih ini akan menjadi asas kajian lanjutan R&D ke arah
pembangunan produk dari hasilan semula jadi.
Penyaringan fitokimia
Saponin: Larutan methanol dan air digoncang untuk mengesan kehadiran buih.
(Simes 1995)
73
suhu 25°C. Tindakbalas enzim kemudiannya dimulakan dengan penambahan
asid natrium linoleik (substrat) sebanyak 10 µL sebelum pengeraman dilakukan
sekali lagi selama 10 minit pada suhu 25°C. Aktiviti perencatan lipoksigenase
diukur dengan menggunakan spektrofotometer pada panjang gelombang 234
nm. Nordihidroguairetik asid (NDGA) digunakan sebagai kawalan positif. Ujian
diulang sebanyak tiga kali.
Kajian ke atas model haiwan yang biasa digunakan untuk melihat aktiviti
antiradang bagi sampel yang diuji adalah dengan menggunakan aplikasi topikal
TPA (12-0-tetradekanoilforbol-13-asetat) untuk mengaruh bengkak atau
edema. Dalam kajian ini, aruhan bengkak pada telinga mencit dilakukan
dengan menyapu larutan TPA pada kedua-dua belah telinga mencit 40 minit
selepas larutan ekstrak/sampel disapu pada telinga kanan manakala aseton
disapu pada sebelah kiri sebagai kawalan. Selepas 6 jam pemberian TPA,
mencit akan dibunuh dan ditebuk kedua belah telinga pada diameter 7 mm
dan ditimbang. Kesan perencatan bagi setiap sample dikira dengan
membandingkan perbezaan berat antara telinga kiri dan kanan.
Penyaringan fitokimia
74
Jadual 1. Kehadiran kumpulan sebatian kimia. Sa=Saponin, Fla=Flavonoid,
Ta=Tannin, Tri= Triterpenoid, dan Ste= Steroid
Kod spesies Bahagian Penyaringan fitokimia
tumbuhan Sa Fla Ta Tri Ste
1 TD 01/10 daun 1+ 0 C2 3+ 3+
batang 1+ 0 C2 2+ 2+
rizom 1+ 1+ C1 1+ 0
2 TD 02/10 daun 2+ 2+ C1 1+ 2+
batang 2+ 0 C1 0 0
dahan 3+ 2+ C2 0 1+
3 TD 03/10 daun 1+ 0 C2 3+ 3+
batang 2+ 1+ C1 0 0
4 TD 04/10 daun 2+ 0 H2 3+ 3+
batang 1+ 0 H2 0 1+
5 TD 05/10 daun 1+ 0 C2 3+ 2+
6 TD 06/10 daun 1+ 0 C3 3+ 3+
batang 0 0 H1 1+ 1+
dahan 0 2+ H2 0 1+
akar 0 0 0 0 0
7 TD 07/10 daun 0 0 C1 3+ 3+
batang 0 2+ H1 0 0
8 TD 08/10 daun 1+ 0 H2 0 3+
batang 1+ 3+ C1 1+ 0
9 TD 09/10 batang 3+ 1+ 0 0 0
kulit luar 1+ 1+ C1 0 0
10 TD 10/10 daun 0 0 C2 0 1+
batang 1+ 1+ H3 0 1+
75
Jadual 2. Aktiviti perencatan dari pelbagai bahagian tumbuhan untuk 10
spesies
Kod Bahagian Aktiviti anti-radang
tumbuhan LOX (%)a XO (%)a TPA (%)
1 TD 01/10 daun 20.17± 4.44 41.79 ± 6.11 43%
batang NA 24.74± 1.75 2%
rizom 10.63±7.97 56.42± 2.67 26%
2 TD 02/10 daun 50.33± 1.80 78.66± 5.32 41%
batang 42.81±2.12 57.75 ± 2.07 34%
dahan 62.04± 1.22 77.92 ± 4.06 26%
3 TD 03/10 daun NA 63.20 ± 0.69 34%
batang 72.3± 6.28 61.40 ± 7.96 33%
4 TD 04/10 daun 95.02± 4.76 ND 26%
batang 84.92± 1.50 86.30 ± 5.08 29%
5 TD 05/10 daun 28.23±2.33 84.98 ± 7.21 49%
6 TD 06/10 daun 12.78± 6.44 41.47 ± 5.88 14%
batang 7.49±5.92 52.88 ± 7.07 7%
dahan 83.47± 5.33 83.18 ± 2.27 17%
akar 14.28± 6.36 46.97 ± 8.11 14%
7 TD 07/10 daun 10.55±5.86 10.35 ± 3.45 51%
batang 55.33±6.33 68.46 ± 1.81 37%
8 TD 08/10 daun 77.83± 74.33 ± 16%
14.33
batang 75.91± 4.67 81.40 ± 9.22 40%
9 TD 09/10 batang 12.12±3.03 NA 40%
Kulit luar 86.18±4.39 53.72±2.26 27%
10 TD 10/10 daun 27.33±1.99 27.13±3.79 34%
batang 33.27±1.28 50.44±3.15 26%
a
Semua nilai adalah min ± S.E.M., kepekatan akhir adalah 100 µg/mL.
Nota: H- Tinggi (71-100%) M-Sederhana (41-70%) L-rendah (0-40%) ND- Tidak
ditentukan
RUMUSAN
76
RUJUKAN
Azhar, U.H., Abdul, M., Itrat, A., Sher Bahadar, K., Ezaz, A., Sarfraz, A.N., &
Muhammad, I. (2004). Enzymes Inhibiting Lignin from Vitex negundo.
Chemical and Pharmaceutical Bulletin: 52(11): 1269─1272.
Simes, J.J.H., Tracey J.G.,Webb, I.J. & Dunstaon, W.J. (1995). Australian
Phytochemicals Survey III. Saponins in the Eastern Australian Flowering
Plants. Bulletin No. 281., CSIRO, Melbourne Australia.
77
KANDUNGAN FITOKIMIA MAS COTEK BERLAINAN AKSESI
ABSTRAK
Mas cotek (Ficus deltoidea) digolongkan dalam famili Moraceae. Pokok ini
mempunyai pelbagai jenis bentuk daun, terdapat bentuk daun yang elips,
obovate atau spatulate. Daun berbentuk spatulate paling banyak ditanam dan
dijual. Daun-daun ini biasanya berwarna hijau, tebal dan keras dan hampir
berair. Ciri daun mas cotek adalah daun yang mempunyai bintik berwarna
emas pada permukaan daun. Daun mas cotek juga dikategorikan kepada
spesies jantan dan betina. Perbezaan daun mas cotek jantan dan betina adalah
berdasarkan bentuk daun dan warna bintik di bahagian bawah daun. Daun
pokok ini dikatakan mampu merawat pelbagai jenis penyakit. Kajian ini
dilaksanakan untuk menentukan kandungan vitexin (kandungan aktif) dalam
daun mas cotek berlainan aksesi. Kandungan aktif dikenal pasti dengan
menggunakan peralatan kromatografi cecair berprestasi tinggi (KCPT/HPLC).
Sepuluh aksesi mas cotek telah dianalisis dan dua aksesi, V10 dan V11
menunjukkan kandungan vitexin yang tinggi iaitu masing-masing pada 44.14
dan 20.46 µg/ml.
PENGENALAN
Mas cotek (Ficus deltoidea) dari famili Moraceae adalah pokok renek dengan
akar serabut yang biasanya bermula sebagai epifit. Pokok ini biasanya dilihat
sebagai pokok renek epifit yang melata pada pokok besar. Bentuk daun pokok
ini sangat berbeza-beza biarpun tergolong dalam genus yang sama. Bentuknya
sama ada eliptikal, lanseolat (seperti pisau pembedah) kepada obovat atau
spatulat. Mat-Salleh & Latiff (2002) menyatakan Ficus deltoidea mempunyai
enam varieti iaitu F. deltoidea var. angustifolia, F. deltoidea var. bilobata, F.
78
deltoidea var. trengganuensis, F. deltoidea var. kunstleri, F. deltoidea var.
intermedia dan F. deltoidea var. motleyana. Varieti-varieti ini dibeza
berdasarkan kelebaran buah, tulang daun (bercabang atau sebaliknya), bentuk
daun apeks (membulat, trunkat atau bentuk lain) serta saiz daun. F. deltoidea
var. deltoidea pula telah dilaporkan oleh Zunoliza et al. (2009). Sebanyak 30
aksesi mas cotek telah dikenal pasti berdasarkan morfologi bagi daun, batang
dan tabiat pertumbuhannya (Musa 2005).
Bahan Tanaman
Sepuluh aksesi sampel mas cotek (MC) telah ditanam dan dikutip pada tiga
masa tuaian yang berbeza. Tuaian dilaksanakan pada 15 inci dari pucuk.
Sampel segar dikeringkan menggunakan pengering solar selama seminggu dan
kemudian dikisar untuk analisis kandungan sebatian vitexin.
79
µm) Lichrocart (Merck). Aliran pelarut, 1.0 ml/min dilakukan selama 25 minit.
Puncak bagi vitexin dikesan pada gelombang 254 nm. Pelarut bergerak yang
terdiri daripada 0.1% H3PO4 dalam ACN (A) dan 0.05% H3PO4 dalam air yang
diionkan (B). Program bagi pelarut bergerak adalah seperti berikut:
Sepuluh aksesi MC yang ditanam di Bachok, Kelantan telah dituai pada tiga
masa tuaian berbeza. Semua sampel ini telah dianalisa menggunakan HPLC dan
menunjukkan data kandungan vitexin yang baik. Sebatian vitexin (Rajah 1)
telah digunakan sebagai sebatian standard dalam MC. Perbezaan bentuk daun
MC ditunjukkan dalam Rajah 2.
OH
HO
OH
O OH
HO
HO O
OH O
Rajah 1. Struktur kimia bagi sebatian utama MC
80
V1 V4 V6 V 10
V 11 V 14 V 19 V 23
V 28 V 39
Rajah 2. Perbezaan bentuk daun mas cotek
Rajah 3. Nilai peratus ekstrak mentah daun mas cotek berlainan aksesi
81
Jadual 1. Kandungan vitexin dalam aksesi berlainan daun mas cotek
Aksesi Tuaian pertama Tuaian kedua Tuaian ketiga
(µg/ml) (µg/ml) (µg/ml)
V1 3.6650 5.2235 5.5729
V4 5.0100 1.8567 2.9464
V6 2.3338 3.9667 4.9244
V10 20.4572 6.5406 12.1605
V11 44.1429 21.8074 40.1726
V14 4.1802 2.4409 3.4023
V19 4.2593 4.0829 4.6634
V23 9.4406 3.4575 7.4782
V28 4.3228 2.3226 2.6882
V39 6.0560 7.5688 6.2472
RUMUSAN
Sebanyak sepuluh aksesi mas cotek yang dituai pada tiga masa tuaian yang
berbeza telah dianalisis. Aksesi V1 merupakan aksesi terbaik yang
menunjukkan jumlah kandungan vitexin paling tinggi dengan nilai 44.14 µg/ml.
PENGHARGAAN
RUJUKAN
Burkill, I.H. & Haniff, M. (1930). Malay Village Medicine. Garden’s Bulletin 6(2):
67─332.
Choo, C.Y., Sulong, N.Y., Man, F. & Wong, T.W. (2012). Vitexin and Isovitexin
from the Leaves of Ficus deltoidea with In-Vivo -Glucosidase
Inhibition. Journal of Ethnopharmacology 142: 776–781.
Fasihuddin, B.A. & Din, L.B. (2002). Medicinal Plants Used by Ethnic Groups in
Sabah. Proceedings of French Malaysian-Symposium on Natural
Products. Universiti Malaya Kuala Lumpur. 85pp.
82
Mat-Salleh, K. & Latiff, M.A. (2002). Tumbuhan Ubatan Malaysia. Universiti
Kebangsaan Malaysia, Bangi. 184pp.
Zunoliza, A., Khalid, H., Zhari, I. & Rasadah, M.A. (2009). Anti-Inflammatory
Activity of Standardized Extracts of Leaves of Three Varieties of Ficus
deltoidea. International Journal of Pharmaceutical and Clinical
Research 1(3): 100─105.
83
PENENTUAN JULAT EURYCOMANONE DALAM PRODUK TONGKAT ALI
MELALUI KAEDAH KROMATOGRAFI CECAIR ULTRAPRESTASI (UPLC)
ABSTRAK
PENGENALAN
Tongkat ali terkenal sebagai minuman herba yang paling popular di kalangan
pengusaha industri herba. Terdapat pelbagai produk minuman seperti kopi
campuran, teh tongkat ali dan juga minuman tin yang dicampurkan dengan
ekstrak tongkat ali. Hampir kesemua produk di pasaran dikatakan mempunyai
khasiat yang boleh meningkatkan tenaga. Berdasarkan laporan sebelum ini,
ekstrak tongkat ali mengandungi beberapa sebatian kimia dan eurycomanone
84
merupakan satu daripadanya (Zhari et al. 1999). Beberapa kajian yang
dijalankan oleh penyelidik melapurkan eurycomanone adalah bahan aktif bagi
aktiviti anti-malaria dan anti-ulcer (Chan et al. 2004). Eurycomanone turut
dijadikan sebatian penanda di dalam ekstrak organik dan akuas pada larutan
ekstrak piawai tongkat ali (Mohd Radzi et al. 2004). Analisa kandungan
eurycomanone telah dijalankan di Makmal Fitokimia (MF), Bahagian Hasilan
Semulajadi sejak tahun 2006. Analisa dijalankan dengan menggunakan
peralatan kromatografi cecair prestasi tinggi (HPLC) sehingga tahun 2009.
Bermula tahun 2010 hingga kini analisa dijalankan dengan menggunakan
peralatan kromatografi cecair ultraprestasi (UPLC).
Sampel untuk kajian ini adalah sampel yang diterima oleh Makmal Fitokimia,
sejak tahun 2010 hingga 2014 (Jadual 1).
Penyediaan Sampel
Sampel yang diterima adalah berbentuk serbuk akar, ekstrak, serbuk daun atau
buah tongkat ali. Kesemua serbuk bahan mentah yang diterima akan ditapis
bagi mendapatkan sebuk yang bersaiz 1-5mm. Hasil tapisan kemudiannya akan
dicampurkan dengan larutan akuas pada kadar 1 nisbah 10 iaitu 1 gram sampel
dengan 10 mililiter akuas. Larutan kemudiannya diekstrak mengggunakan
sonicator pada suhu 50oC selama 90 minit. Larutan ditapis dengan kertas turas
dan hasil larutan dikeringkan menggunakan rotary evaporator. Berat ekstrak
yang dihasilkan ditimbang dan kemudiannya ditambahkan semula larutan
akuas dengan kepekatan akhir larutan mestilah tidak melebihi 3 mg/ml.
Larutan ekstrak mesti ditapis terlebih dahulu menggunakan penapis bersaiz
0.45 µ sebelum analisa UPLC dijalankan.
85
Bahan Kimia
Bahan kimia yang digunakan adalah acetonitril gred HPLC (MERCK) dan asid
formik (0.1%) (Fluka Chemica) sebagai pelarut fasa gerak. Eurycomanone (P)
(Chromadex) digunakan sebagai bahan piawai.
Analisa Kimia
Kelinearan (linearity)
86
Rajah 1. Contoh graf kalibrasi untuk pengiraan eurycomanone
87
Rajah 2. Profile UPLC untuk analisa sampel
RUMUSAN
88
campuran serbuk tongkat ali yang dikisar halus di dalam kapsul, berdasarkan
terdapatnya mendakan yang tidak larut di dalam larutan ujikaji. Mendakan
tidak larut ini boleh membahayakan kesihatan pengguna jika kapsul ini
dimakan dalam kuantiti yang melebihi had yang dibenarkan. Daun tongkat ali
juga mempunyai kadar peratusan eyrucomanone yang dapat dikesan. Satu
kajian menyeluruh juga harus dijalankan bagi mengkaji kesesuaian daun
sebagai bahan produk sebagai alternatif pengunaan akar.
RUJUKAN
Chan K.L, Choo C.Y., Noor Ain A. & Zakiah I. (2004). Antimalarial and
Toxicological Studies of Eurycomanone from Eurycoma Longifolia Jack
and Its Synthesized Derivatives. 25─28 Mac 2004 Herbal Asia, Kuala
Lumpur.
Mohd Radzi A., Mohd Ilham A., Norijas A.A., Mohd Noh J., & Mohd Ghawas M.
(2004). Penentuan Kualiti Berdasarkan Kandungan Kimia dan Usia
Penanaman Tongkat Ali Secara Perladangan. Pp 279-285. dlm Chang, Y.
S., Vimala, S., Mazura, P. & Ong, B. K. (eds). Proceedings of the Seminar
on Medicinal and Aromatic Plants: Current Trends and Perspectives.
Forest Research Institute Malaysia (FRIM), Kuala Lumpur.
Mohd Radzi, A., Ong, B. K., Mohd Ilham, A. & Mohd Hafidz, H.A. (2008)
Pembangunan Kaedah Penilaian Kawalan Mutu Minuman Kopi Tongkat
Ali Berasaskan Penanda Kimia Eurycomanone. Pp 272─279. Dlm Chang,
Y. S., Mazura, P. & Nik Musaadah, M. (eds). Proceedings Plant, Health
and Man-Past, Present and Future, FRIM, Kepong.
Zhari, I., Norhayati, I. & Jaafar. L. (1999). Radix Eurycomas. Malaysian Herbal
Monograph Vol. 1.: 29–32.
89
PENILAIAN KANDUNGAN FITOKIMIA DAN AKTIVITI SITOTOKSITI
SPESIES TERPILIH TUMBUHAN UBATAN BERASASKAN PENGETAHUAN
TRADISIONAL SUBETNIK JAHAI/TEMIAR DI KAMPUNG AIR BANUN,
GERIK, PERAK
ABSTRAK
PENGENALAN
Persepsi bahawa penggunaan ubatan herba adalah dijamin selamat
bersandarkan fakta ia berasal dari sumber semula jadi adalah kurang tepat.
Produk yang berasaskan semula jadi tidak semestinya selamat. Penggunaan
turun-temurun juga tidak boleh dijadikan bukti bahawasanya ia tidak akan
memberi risiko dan membahayakan pengguna dengan kesan-kesan sampingan
90
(Haq 2004). Bahan persediaan herba, ekstrak dan sebatian fitokimia mungkin
mempunyai risiko yang sama seperti sebatian sintetik.
Penyaringan Fitokimia
Flavonoid: Larutan klorofom dikeringkan dan kemudian diekstrak dengan
kombinasi ether dan larutan ammonia,
Saponin: Larutan metanol dan air digoncang untuk mengesan kehadiran buih.
(Simes, 1995)
91
Saringan Sitotoksiti Titisan Sel Normal
Dua jenis titisan sel normal telah digunakan di dalam penyelidikan ini iaitu
WRL-68 (human liver cells) dan Vero (African green monkey kidney). Kedua-
dua jenis titisan sel normal diperolehi daripada ATCC. Ia telah dikulturkan di
dalam Medium Dulbecco’s Modified Eagle medium (Sigma, USA) dan
dicampurkan dengan serum 5% foetal bovine (PAA Laboratories), amphotericin
B 1 % dan penisilin-streptomisin (PAA Laboratories) dan gentamycin 0.5% (PAA
Laboratories). Sel disimpan di dalam inkubator yang mengandungi 5%
CO2/95% air pada suhu 37°C.
Sel yang yang telah tumbuh pada kadar eksponential telah diletakkan
di dalam piring 96-well untuk mencapai jumlah 10,000 sel/telaga. Selepas 24
jam tempoh pengeraman, sel telah didedahkan dengan ekstrak tumbuhan
dengan kepekatan berjulat 0.1 hingga 1000 µg/ml untuk tempoh 72 jam. Untuk
tujuan perbandingan, Paclitaxel telah digunakan dan setiap eksperimen
dijalankan di dalam triplikat.
92
Ekstrak selebihnya didapati sederhana sitotoksik (19 ekstrak) dan lemah
sitotoksik (tujuh ekstrak). Hanya ekstrak yang kurang toksik akan dipilih untuk
kajian lanjutan.
93
Jadual 2. Kesan sitotoksiti 10 spesies tumbuhan herba berasaskan pengetahuan
tradisional sub etnik Jahai/Temiar di Kampung Air Banun, Gerik, Perak
94
RUMUSAN
Data yang diperolehi daripada kajian ini memberikan informasi yang berguna
untuk tujuan penyelidikan yang lebih mendalam ke atas spesies-spesies
tersebut yang mempunyai potensi biologi.
RUJUKAN
Haq, I. (2004). Safety of medicinal plants. Pakistan Journal of Medical Research.
43. 203─210.
Skehan P., Storeng R., Scuidero D., Monks A., McMahon J., Vistica D., Warren
J.T., Bokesch H., Kenney S. & Boyd M.R. (1990). New Calorimetric
cytotoxicity assay for anti-cancer-drug screening. Journal of the
National Cancer Institute. 82: 1107─1102.
95
ASAS MOLEKUL AKTIVITI ANTI-MALARIA KURKUMIN
ABSTRAK
PENGENALAN
96
penglibatan kurkumin dalam merencat pengaktifan GSK3 dalam kajian malaria
masih belum jelas.
Sel hepar Chang, diperoleh daripada American Type Culture Collection (ATCC)
dan dikultur dalam Dulbecco Modified Eagle Medium (DMEM) (Gibco, USA)
berdasarkan kaedah Mossmann (1983).
Mencit jantan ICR (25±5 g; berusia 6-8 minggu) diperoleh daripada Kompleks
Rumah Haiwan, UKM. Kelulusan untuk kajian haiwan diperoleh daripada
97
Jawatankuasa Etika Haiwan Universiti Kebangsaan Malaysia (UKMAEC). Strain
Plasmodium berghei NK65 (klorokuin-sensitif) daripada Malaria Research and
Reference Reagent Resource Center (MR4, USA). Ujian penekanan terapeutik
dan profilaksis berdasarkan kaedah Peters (1975).
Pada dos 30 mg/kg berat tubuh, kurkumin menindas lebih 50–60% populasi P.
berghei NK65 melalui ujian terapeutik dan profilaksis dan mampu melanjutkan
tempoh hayat mencit terinfeksi-malaria. Hasil serupa telah dilaporkan oleh
(Reddy et al. 2005) tetapi melibatkan strain yang berbeza.
Daripada analisis protein bagi ujian terapeutik dan profilaksis, didapati bahawa
kurkumin mempunyai ciri perencatan enzim GSK3 berbanding haiwan yang
tidak diberi rawatan (data tidak ditunjukkan). Walaupun sebelum ini, kurkumin
pernah dilaporkan mempunyai kesan perencatan terhadap molekul pengatur
sistem inflamasi sel (NF-kB), penemuan kami bahawa kurkumin menyebabkan
penindasan aktiviti GSK3 merupakan laporan buat pertama kali. Hasil kajian ini
selari dengan peranan GSK3 dalam mengawal pengaktifan NF-kB.
98
KESIMPULAN
Hasil kajian ini merupakan satu penemuan baru mengenai kefahaman asas
molekul aktiviti anti-malaria kurkumin yang berasaskan eksperimen pada
haiwan (in vivo) dan kultur parasit (in vitro).
PENGHARGAAN
RUJUKAN
Bradford, M.M. (1976). A Rapid and Sensitive Method for the Quantitation of
Microgram Quantities of Protein Utilizing the Principle of Protein-Dye
Binding. Analytical Biochemistry: 72(1─2): 248─254.
Bustanji, Y., Mutasem, O.T., Ihab, M.A., Mohamed, A.S.A., Mohammad, K.M. &
Hatim, S.A. (2009). Inhibition of Glycogen Synthase Kinase By
Curcumin: Investigation by Simulated Molecular Docking and
Subsequent In Vitro/In Vivo Evaluation. Journal of Enzyme Inhibition
and Medicinal Chemistry: 24 (3):771─778.
Cortés-Vieyra, R., Alejandro, B.P., Juan, J.V.A., Marcos, C.J., Finlay, B.B. &Víctor,
M. B.A. (2012). Role of Glycogen Synthase Kinase-3 Beta in The
Inflammatory Response Caused by Bacterial Pathogens. Journal of
Inflammation (Lond): 9 (1):23─23.
Dondorp, A.M., François, N., Poravuth, Y., Debashish, D., Aung, P.P., Joel, T.,
Khin, M.L., Frederic, A., Warunee, H. & Sue, J.L. (2009). Artemisinin
Resistance in Plasmodium Falciparum Malaria. New England Journal of
Medicine: 361 (5):455─467.
Jain, K., Sood, S. & Gowthamarajan, K. (2013). Modulation of Cerebral Malaria
by Curcumin as An Adjunctive Therapy. The Brazilian Journal of
Infectious Diseases: 17(5): 579─591.
Kizhakkayil, J., Faisal, T., Shahanas, C., Abdulkader, H., Mahendra, P.&
Sehamuddin, G. (2012). Glutathione Regulates Caspase-Dependent
Ceramide Production and Curcumin-Induced Apoptosis in Human
Leukemic Cells. Free Radical Biology and Medicine: 52 (9):1854─1864.
Kohli, K., Ali, J., Ansari, M.J.& Raheman, Z. (2005). Curcumin: A Natural
Antiinflammatory Agent. Indian Journal of Pharmacology: 37 (3):141.
99
Leammi, U.K. (1970). Cleavage of Structural Proteins During The Assembly of
The Head of Bacterial Phage T4. Nature: 227( 5259):680─685.
Lee, C. (2007). Protein Extraction from Mammalian Tissues. Methods in
Molecular Biology: 362: 385─389.
Mimche, P.N., Donatella, T. & Livia, V. (2011). The Plant-Based
Immunomodulator Curcumin as A Potential Candidate for The
Development of an Adjunctive Therapy for Cerebral Malaria. Malaria
Journal: 10 (1):10.
Mosmann, T. (1983). Rapid Colorimetric Assay for Cellular Growth and Survival:
Application to Proliferation and Cytotoxicity Assays. Journal of
Immunological Methods: 65(1): 55─63.
Peters, W. (1975). The Chemotherapy of Rodent malaria, xxii. The Value of
Drug-Resistant Strains of in Screening for Blood Schizontocidal Activity.
Annals of Tropical Medicine and Parasitology: 69 (2): 155─171.
Reddy, R.C., Vatsala, P.G., Keshamouni, V.G., Padmanaban, G., & Rangarajan,
P.N. (2005). Curcumin for Malaria Therapy. Biochemical and
Biophysical Research Communications: 326(2), 472─474.
Sharma, S., Kulkarni, S.K., & Chopra, K. (2006). Curcumin, The Active Principle
of Turmeric (Curcuma Longa), Ameliorates Diabetic Nephropathy in
Rats. Clinical and Experimental Pharmacology and Physiology: 33(10):
940─945.
Trager, W. & Jensen, J.B. (1976). Human Malaria Parasites in Continuous
Culture. Science: 193(4254): 673─675.
100
MEKANISME TINDAKAN SEBATIAN BIOAKTIF ANTI-MALARIA DALAM
PAKU PAKIS UBATAN, GLEICHENIA TRUNCATA
ABSTRAK
PENGENALAN
101
Usaha penyelidikan berterusan dijalankan melalui eksploitasi fungsi
protein kinase yang berpotensi dijadikan sebagai sasaran drug anti-malaria.
Salah satu protein kinase yang diberi perhatian dalam kajian sasaran drug anti-
malaria adalah glikogen sintase kinase-3 (GSK3), iaitu kinase serin/treonin yang
berfungsi sebagai kunci utama dalam penyakit berkait-inflamasi penyakit
infeksi virus, bakteria serta parasit menjadikan kinase ini sebagai sasaran
berpotensi bagi drug anti-malaria (Dai et al. 2012). Perencatan aktiviti GSK3
dilihat mampu memberi perlindungan kepada hos kerana enzim ini
merangsang penghasilan sitokin pro-inflamasi secara berlebihan semasa infeksi
malaria (Dai et al. 2012). Perencat GSK3, LiCl dilaporkan berupaya merencat
pertumbuhan Plasmodium falciparum dan P. berghei masing-masing melalui
kajian in vitro dan in vivo (Marhalim et al. 2014; Nurul Aiezzah et al. 2010).
Justeru itu, perencatan aktiviti GSK3 bukan sahaja dilihat mampu
mengurangkan kesan patologi malaria dalam hos, malahan merencat
perkembangan parasit semasa infeksi malaria. Dalam kajian ini kesan
perlakuan metil-4-hidroksisinamat (M4H), iaitu sebatian bioaktif G. truncata
diuji menggunakan model haiwan malaria.
102
PENEMUAN DAN PERBINCANGAN
Hasil kajian in vivo dalam model infeksi-P. berghei NK65 (strain sensitif-
klorokuin) menunjukkan pemberian 30 mg/kg BT M4H merencat
perkembangan parasit P. berghei peringkat eritrosit dengan aktif
(kemopenindasan melebihi 60% pada H4 selepas infeksi) berdasarkan
pengkelasan Rasoanaivo et al. (2004). Pemberian 30 mg/kg BT M4H juga
dicerap mampu memanjangkan tempoh kemandirian haiwan terinfeksi-P.
berghei berbanding kumpulan kawalan tanpa perlakuan (data tidak
ditunjukkan). Pemerhatian ini mungkin berkait dengan pengurangan kesan
patologi malaria (Mota & Rodriguez 2002).
KESIMPULAN
PENGHARGAAN
RUJUKAN
103
Chai, T.T., Elamparuthi, S., Yong, A.L., Quah, Y., Ong, H.C. & Wong, F.C. (2013).
Antibacterial, anti-glucosidase, and antioxidant activities of selected
highland ferns of Malaysia. Botanical Studies: 1─7.
Dai, M., Freeman, B., Shikani, H. J., Bruno, F. P., Collado, J. E., Macias, R.,
Reznik, S. E., Davies, P., Spray, D. C., Tanowitz, H. B., Weiss, L. M. &
Desruisseaux, M. S. (2012). Altered regulation of Akt signaling with
murine cerebral malaria, effects on long-term neuro-cognitive
function, restoration with lithium treatment. PLoS One: 1─15.
Ho, R., Teai, T., Bianchini, J.-P., Lafont, R. & Raharivelomanana, P. (2010).
Working with Ferns. Springer New York.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head
of bacteriophage T4. Nature: 680─685.
Marhalim, L., Embi, N. & Sidek, H.M. (2014). Synergistic effects of GSK3 and
p38 mapk inhibitors on growth of Plasmodium falciparum ex vivo.
Malaysian Applied Biology: 65─71.
Suhaini, S., Liew, S.Z., Norhaniza, J., Lee, P.C., Jualang, G., Embi, N. & Hasidah,
M.S. (2015). Antimalarial and anti-inflammatory effects of Gleichenia
truncata are mediated through inhibition of GSK3β. Tropical
Biomedicine: 1─15
104
Towbin, H., Staehelin, T. & Gordon, J. (1979). Electrophoretic transfer of
proteins from polyacrylamide gels to nitrocellulose sheets: procedure
and some applications. Proceedings of National Academy of Science
USA: 4350─4354.
105
PEMILIHAN DAN PENGHASILAN BAKA LIMAU PURUT (CITRUS HYSTRIX)
BERMUTU TINGGI
ABSTRAK
Pokok limau purut tergolong di dalam keluarga Rutaceae. Walaupun pokok ini
berasal dari benua Asia tetapi pada hari ini ia banyak ditanam di Malaysia dan
Thailand. Minyak pati dari daun limau purut (Citrus hystrix) sering digunakan di
dalam industri pewangi, aromaterapi, perasa, makanan dan perubatan
tradisional. Melihat kepada kepentingan spesies ini di dalam industri herba
negara, satu kajian bagi mendapatkan baka-baka berkualiti dari segi hasil
minyak pati dan kandungan sitronellalnya telah di jalankan oleh Program
Membaikbiak Tumbuhan dari Institut Penyelidikan Perhutanan Malaysia
(FRIM). Sebanyak 220 pokok ibu dari 11 populasi limau purut di Semenanjung
Malaysia telah disaring kuantiti dan kualiti minyak patinya. Secara
keseluruhannya, kertas kerja ini membincangkan mengenai i) kaedah
pengumpulan sampel, ii) pengekstrakan minyak pati secara mobil, iii)
penyaringan kandungan kimia minyak pati, iv) pemilihan baka berkualiti dan
seterusnya iv) kaedah pembiakan baka terpilih dan v) penubuhan bank klonal.
Hasil daripada kajian ini ialah baka-baka limau purut yang berkualiti dapat
diperolehi dan dijadikan sebagai sumber stok tanaman untuk aktiviti
pembiakbakaan terhadap spesies ini pada masa hadapan.
Kata kunci: minyak pati, sitronellal, kuantiti dan kualiti, stok tanaman,
pembiakbakaan
PENGENALAN
Citrus hystrix, lebih dikenali nama tempatannya sebagai limau purut adalah
satu daripada spesies liar di Malaysia. Ia juga dikenali dengan nama “Wild
Lime” dalam bahasa Inggeris, “Kaffir Lime” dalam bahasa Danish dan “Som
Makrut” di Thailand. Tanaman saka ini berasal dari rantau Asia Tenggara dan
telah banyak ditanam di Indonesia, Thailand dan Malaysia. Spesies ini tumbuh
liar di tanah kering dan terdedah kepada cahaya matahari. Pertumbuhan limau
adalah sekitar 3-5 meter dan mencapai sehingga ketinggian 30 meter, tetapi
jika dibandingkan dengan spesies limau lain, pertumbuhannya adalah perlahan.
106
Daun limau purut mempunyai kepanjangan antara 7.5–10.0 cm dan
mempunyai tangkai bersayap yang memberi gambaran dua helai daun yang
dicantumkan hujung ke hujung (Yahaya & Ahmad Puat 2005). Buahnya
berbentuk “pear” dan mencapai kepanjangan sehingga 10 cm dan mempunyai
diameter antara 5.0 cm hingga 7.5 cm. Buahnya berwarna hijau dan bertukar
kepada kekuningan apabila telah masak, berkedut dan berpermukaaan kasar
(Chin & Yong 1980).
107
KAEDAH KAJIAN
Sebanyak 180 pokok ibu dari 9 populasi limau purut di Semenanjung Malaysia
telah disaring daunnya bagi mendapatkan kuantiti dan kualiti minyak pati.
Populasi tersebut adalah dari 1) Yan, Kedah 2) Balik Pulau, Pulau Pinang 3)
Banting, Selangor; 4) Teluk Intan, Perak; 5) Kuala Pilah, Negeri Sembilan; 6)
Jasin, Melaka; 7) Raub, Pahang 8) Kuala Berang, Terengganu, dan 9) Ketereh,
Kelantan. Setiap populasi dinamakan mengikut kod-kod yang berbeza. Pokok-
pokok ibu yang dipilih mempunyai sifat-sifat fenotip yang baik seperti jumlah
daun dan buah yang lebat serta bebas dari serangan penyakit dan perosak
(Rajah 1). Bagi mengelakkan berlakunya pengurangan pada hasil minyak pati
dalam daun yang mungkin berlaku semasa dalam perjalanan, semua daun
dikutip dan disaring secara langsung di lapangan atau dinamakan “mobile lab”.
Rajah 1. Contoh pokok ibu yang dipilih untuk tujuan pemilihan baka berkualiti
terletak di Yan, Kedah.
Penyulingan minyak pati dijalankan sebaik sahaja sampel daun dikutip dari
lapangan. Daun matang yang sesuai diperolehi dengan memotong daun-daun
tersebut dari ranting pokok. Proses penyulingan bagi setiap sampel dijalankan
selama 6 jam menggunakan peralatan jenis Clavenger. Sebanyak 3 replikasi
digunakan bagi setiap sampel pokok ibu (Rajah 2).
108
Rajah 2. Gambar menunjukkan proses penyulingan minyak pati yang
dijalankan secara “mobile lab” di setiap lokasi kajian.
Analisa kualitatif kromatografi gas (GC) dan kromatografi gas jisim spektrometri
(GCMS) dijalankan menggunakan alat Shimadzu GC2010 Plus dan Agilent
Technologies 7890A/5975C MSD (Rajah 3) dengan masing-masing
menggunakan kolum kapilari BP-5 (30 m x 0.25 mm, 0.25 mm tebal filem) dan
HP-5MS (30 m x 0.25 mm, 0.25 mm tebal filem). Kromatografi gas dilengkapi
dengan FID dan menggunakan suntikan split mode. Helium digunakan sebagai
gas pembawa dengan kadar tetapnya 1ml/min. Suhu pengesan ditetapkan
pada 250 °C. Peralatan GC diprogramkan pada awalnya dengan suhu 60°C
selama 10 minit dan kemudian ditingkatkan ke suhu 230 °C selama 1 minit
dengan kadar aliran 3°C/min. Puncak komponen dan masa penahanan diukur
menggunakan integrasi elektronik. Bagi GC/MS, program suhu yang sama
ditetapkan seperti pada peralatan GC. Pengenalpastian komponen-komponen
kimia adalah dengan membandingkan nilai kovat indek komponen di dalam
sampel dengan nilai rujukan (Adams 2005) dan juga perbandingan spektrum
jisim sampel dengan spektrum di pangkalan data.
Keputusan hasil minyak pati yang diperolehi dari 9 populasi adalah seperti di
dalam Jadual 1. Hasil menunjukkan kesemua pokok ibu yang disaring
mempunyai julat di antara 0.9 hingga 4.7%. Jadual 2 menunjukkan purata
minyak pati yang dihasilkan dari 9 populasi di mana populasi Kuala Berang
mencatatkan jumlah minyak pati tertinggi (4.2%).
109
Jadual 1. Julat hasil minyak pati dari yang disaring dari 9 populasi kajian
Populasi Julat minyak pati (%)
Yan 1.70-4.6
Balik Pulau 1.25-2.2
Teluk Intan 1.30-2.7
Banting 3.10-4.5
Kuala Pilah 1.30-4.7
Jasin 0.90-2.2
Raub 2.20-4.2
Kuala Berang 1.40-2.7
Ketereh 2.20-3.6
Jadual 2. Purata jumlah minyak pati limau purut dari 9 populasi kajian
Populasi Purata jumlah minyak pati (%)
Yan 3.2%
Balik Pulau 3.4%
Teluk Intan 3.7%
Banting 3.6%
Kuala Pilah 3.2%
Jasin 2.8%
Kuala Berang 4.2%
Raub 3.2%
Ketereh 3.0%
110
Jadual 3. Kandungan kimia utama (%) yang dijumpai dalam minyak pati limau
purut yang disaring dari 9 populasi di Semenanjung Malaysia
Populasi Yan Balik Teluk Banting Kuala Jasin Kuala Raub Ketereh
Pulau Intan Pilah Berang
Sebatian
Sitronellal 68.1 74.4 82.0 72.7 54.4 76.9 83.7 63.6 77.5
Sitronellol 10.0 4.0 5.8 9.6 10.7 8.6 4.8 9.4 9.0
Linalool 2.2 2.2 2.5 2.5 1.5 2.2 2.7 1.9 2.3
Sitronellil
4.2 3.4 2.9 3.7 3.8 3.7 2.1 2.7 2.9
asetat
Rajah 3. Klon bank limau purut yang ditubuhkan di FRIM bagi menempatkan
baka-baka limau purut yang berkualiti
KESIMPULAN
111
industri ini akan dapat diperolehi dengan mudah dan produk yang dikeluarkan
adalah dari bahan yang berkualiti tinggi.
RUJUKAN
Chin, H.F. & Yong, H.S. (1980). Malaysian Fruits in Color. Tropical Press Sdn.
Bhd, Kuala Lumpur, Malaysia. Pp 52.
Nor Azah, M.A., Mastura, M., Zaridah, M.Z., Mailina, J., Mohamad Shahidan.,
Norseha, A., Zainon, A.S., Norhanan, M.Y., Abdul Majid, J. & Abu Said, A.
(2004). Potential application of Citrus hystrix essential oils for personal
care products. Prosiding Hasil Kajian IRPA RMK8 2004 FRIM. Pp :
273─277.
Takeshi, Y., Akemi, S., Tatsuya, O., Hiroyuki, M., Miharu, O. & Tsuneyoshi, K.
(2004). Flavour and Fragrance Journal Volume 19, Issue 2. Pp : 121─133.
112
PENGELUARAN DAN PENILAIAN MINYAK DAUN KAYU MANIS
MALAYSIA
ABSTRAK
Kandungan kimia minyak dedaun kayu manis biasanya adalah eugenol dan
sinnamik aldehid. Bagi penyulingan secara stim skala industri, bahan campuran
dedaun dan cabang ranting kayu manis memberikan 0.4–0.5% hasil minyak
pati, manakala skala makmal penyulingan secara hidro dedaun kayu manis
sahaja memberikan hasil penyulingan sebanyak 1.6%. Minyak dedaun kayu
manis bewarna kekuningan cerah dan berkilau. Komponen utama dalam
minyak pati daun kayu manis adalah kumpulan eugenol dan sinnamik aldehid.
Aroma minyak pati daun kayu manis adalah sangat segar, manis serta sedikit
tajam. Bahan ini merupakan bahan perisa dan wangian (Flavour & Fragrance
(F&F)) bagi ramuan makanan, minuman, minyak wangi dan kosmetik.
PENGENALAN
113
bagi penilaian penyelidikan dan pembangunan (R&D) minyak pati dedaun kayu
manis.
Bahan mentah dedaun kayu manis dituai di MARDI Kuala Linggi dan disuling
menggunakan penyulingan stim berskala 100 kg. Kandungan bahan mentah
yang disuling secara industri adalah merangkumi dahan, ranting dan dedaun.
Manakala bagi hasil sebenar kandungan minyak dedaun adalah secara
penyulingan berskala makmal dengan menggunakan apparatus Clavenger (2 L).
Analisis GC-MS-Tof (1D) digunakan untuk pengenalpastian kandungan kimia
beberapa aroma kimia makanan dan bahan wangian. Pengesan ToF ini adalah
singkatan bagi time-of-flight mass spectrometer menggunakan alat GC-MS
Pegasus 4D, Leco Corporation, St. Joseph, MI, USA. Parameter penggunaan
kolum bagi GC-MS-Tof adalah 1D column model ZB-5MS, panjang 30 m,
diameter 0.25 mm, ketebalan filem dalaman 0.25 μm, fasa utama column 5%
Polysilarylene dan 95% Polydimethylsiloxane (Zebron, Phenomenex
Corporation).
114
PENEMUAN DAN PERBINCANGAN
Kandungan minyak pati daun kayu manis yang diperolehi secara skala makmal
adalah sehingga 1.6%, manakala bahan campuran dedaun, dahan dan ranting
memberikan hasil minyak yang rendah dengan hanya 0.4–0.5%. Namun
operasi penyulingan berskala besar boleh menghasilkan sehingga 500 ml
minyak daun kayu manis premium bagi penyulingan 1 vessel/batch. Rajah 1
menunjukkan produk minyak pati kayu manis bergred premium iaitu campuran
dahan dan ranting bagi pengeluaran produk secara pukal menggunakan
penyulingan skala kilang di MARDI.
Rajah 1. Satu liter minyak pati kayu manis Malaysia gred premium daripada
campuran dahan dan ranting)
Rajah 2. Komposisi kimia utama (eugenol dan iso-eugenol) dalam minyak daun
kayu manis
115
Daripada analisis ini kandungan kumpulan eugenol dan benzyl benzoate
masing-masing adalah sebanyak 56.0% dan 14.2% (Rajah 3). Di samping itu
komponen kimia terpilih dan penting dalam industri minyak wangi juga telah
dikenal pasti iaitu eugenol asetat (6.5%), E-sinnamaldehid (sinnamil asetat
(2.4%) dan hidrosinnamil asetat (0.2%).
Bahan aroma sinnamaldehid ini juga berfungsi dan efektif sebagai bahan
anti-mikrob menurut laporan kajian serapan gas terhadap Escherichia coli,
Staphylococcus aureus, Streptococcus pyogenes, S. pneumoniae dan
Haemohilus influenza (Inouye 2001). El-Baroty et al. (2010) pula melaporkan
penggunaan minyak kayu manis di antara 20–100 µg/ml boleh bertindak
sebagai bahan pengawet asli dan menghalang pengoksidaan bahan (lipid
peroxidation). Secara tidak langsung eugenol dan sinnamik aldehid yang
terkandung dalam minyak dedaun dan ranting kayu manis ini berpotensi
sebagai agen anti-oksida dan anti-bakteria.
Rajah 3. Profil 1D GC-MS Tof bagi minyak daun kayu manis Malaysia
KESIMPULAN
Minyak pati daun kayu manis yang dikeluarkan di Malaysia mempunyai potensi
sebagai bahan perisa kerana kandungan kumpulan aktif eugenol yang tinggi
dan bahan sinnamaldehid. Produk berasaskan minyak pati boleh menggantikan
produk oleoresin kayu manis yang banyak menggunakan pelarut dan
mengandungi residu pelarut yang mungkin boleh menjejaskan kesihatan
116
pengguna. Kajian ini melengkapkan lagi data minyak pati daun kayu manis yang
berpotensi bagi industri minyak pati di Malaysia, serta boleh memberikan data
sokongan kepada cadangan Teknologi Penanaman dan Pengeluaran Bahan
herba F&F dalam Rancangan Malaysia kesebelas (RMK11), sekiranya tumbuhan
beraroma terpilih sebagai herba baharu dalam EPP1.
PENGHARGAAN
RUJUKAN
Culinary Herbs and Spices, the Seasoning and Spice Association. 2015.
http://www.seasoningandspice.org.uk/ssa/background_culinary-herbs-
spices.aspx. [23 September 2015].
El-Baroty, G.S., Abd El-Bakri H.H., Farag, R.S. & Saleh, M.A. (2010).
Characterization of Antioxidant and Antimicrobial Compounds of
Cinnamon and Ginger Essential Oils. African J. of Biochem. Research
Vol. 4(6): 167─174.
117
PEMBIAKAN AKSESI KACIP FATIMAH (LABISIA PUMILA VAR. ALATA)
TERPILIH MELALUI KAEDAH KERATAN DAN PENILAIAN PENGELUARAN
PUCUK PADA PERINGKAT TAPAK SEMAIAN
ABSTRAK
Sejumlah tiga belas aksesi kacip fatimah (Labisia pumila var. alata) yang
mempunyai kandungan fenolik yang tinggi telah dibiakkan melalui kaedah
keratan daun. Tujuan kajian ini dijalankan adalah untuk menilai keupayaan
pengakaran dan menganalisa prestasi pertumbuhan aksesi kacip fatimah yang
dihasilkan daripada kaedah keratan daun. Melalui kaedah ini, daun kacip
fatimah dirawat menggunakan hormon pengakaran (Seradix 1) dan dibiakkan
di dalam rumah keratan dalam sistem renjisan selama 17 minggu. Setelah
dirawat dengan hormon, keratan yang berakar telah dipindahkan ke dalam
polibeg yang mengandungi media tanaman campuran tanah: kompos: sabut:
pasir (3: 1: 1: 1) dan dipindahkan ke tapak semaian dengan 70% teduhan. Data
seperti kadar kelangsungan hidup, jumlah keratan berakar dan jumlah pucuk
yang terhasil diperhatikan sehingga minggu ke 30. Daripada keputusan yang
diperolehi, aksesi LP7 mencatatkan peratus pengakaran tertinggi (91%) diikuti
oleh LP15 dan LP28 (87%), LP17 (85%) dan LP12 (84%). Manakala, LP15, LP17
dan LP28 menghasilkan bilangan keratan berakar tertinggi dengan purata
masing-masing 13.00 ± 1.00, 13.00 ± 0.58 dan 13.00 ± 1.00. Dari segi
pengeluaran pucuk, LP15 dan LP21 masing-masing menghasilkan bilangan
pucuk yang tinggi iaitu 2.43 ± 0.271 dan 2.43 ± 0.386. Hasil kajian ini dapat
membantu para pembiakbaka dan pengusaha herba mengenal pasti aksesi-
aksesi yang terbaik dari segi pertumbuhan bermula dari peringkat awal
pertumbuhan (peringkat nurseri) sebelum ianya ditanam di lapangan.
Pemilihan baka tanaman yang baik dapat menjamin pengeluaran stok bahan
tanaman secara berterusan.
118
PENGENALAN
Malaysia terkenal dengan pelbagai jenis herba yang selalu digunakan dalam
perubatan tradisional. Labisia pumila atau lebih dikenali sebagai kacip fatimah
merupakan salah satu herba tradisi yang terkenal dalam kalangan wanita
Melayu terutamanya bagi wanita mengandung. Air rebusan kacip fatimah
mempunyai banyak khasiat terutamanya dapat mengurangkan kesakitan
semasa bersalin dan mempercepat proses bersalin (Ibrahim & Hawa 2011). Air
kacip fatimah juga dapat memberi tenaga kepada ibu hamil sepanjang proses
bersalin (Wan Ezumi et al. 2007). Selain itu, kacip fatimah juga dikatakan dapat
menyembuhkan penyakit seperti disentri, senggugut, kembung perut dan
gonorea.
Bahan Tanaman
Sebanyak 13 aksesi kacip fatimah telah diambil dari tiga lokasi hutan simpan
yang berbeza di Semenanjung Malaysia iaitu HS Tembat, Kuala Berang,
Terengganu (LP5, LP6, LP11, LP21); HS Bukit Larut, Taiping, Perak (LP7, LP9,
LP12, LP10, LP13, LP15, LP30); dan HS Batu Papan, Gua Musang, Kelantan
(LP17, LP28). Kesemua pokok ibu ini ditanam sebagai koleksi germplasma yang
telah ditubuhkan di FRIM. Pokok-pokok ibu tersebut dipilih berdasarkan jumlah
119
daun iaitu lebih daripada lima dan juga daun yang sihat tanpa sebarang
penyakit.
Daun diambil dari pokok-pokok ibu terpilih yang ada di dalam koleksi
germplasma kacip fatimah. Daun dipotong mengikut saiz 30 cm2. Hormon
pengakaran (Seradix 1) telah diletakkan pada pangkal keratan. Keratan
dibiakkan di dalam rumah keratan sistem renjisan selama 17 minggu. Pasir
sungai steril digunakan sebagai media tanaman. Sebanyak 585 keratan telah
dihasilkan daripada 13 aksesi. Keratan tersebut ditanam mengikut kaedah
Rekabentuk Rawak Blok Lengkap (RCBD) dengan tiga replikasi (Rajah 1).
120
Pengumpulan Data
Data prestasi pertumbuhan akar telah dicatat pada minggu ke 6 hingga minggu
17. Data yang dikumpul tertakluk kepada analisis varians (ANOVA) untuk
menentukan kesan aksesi terhadap jumlah pertumbuhan akar dalam 17
minggu menggunakan perisian SPSS versi 16.0.
Data prestasi pertumbuhan pucuk telah dicatat pada minggu 22 hingga minggu
30. Data telah direkod setiap dua bulan sehingga minggu 30. Parameter yang
diperhatikan ialah bilangan daun. Data telah dianalisa menggunakan perisian
SPSS versi 16.0 untuk menilai dan membandingkan prestasi pertumbuhan
antara aksesi.
121
Jadual 1. Data pertumbuhan akar tiga belas aksesi kacip fatimah sehingga
minggu ke 17
Aksesi Bilangan akar Panjang akar (cm) Peratus pengakaran (%)
LP5 2.78ab ± 0.364 3.06a ± 0.584 27.0d
LP6 2.56b ± 0.412 4.64a ± 0.708 73.0abc
LP7 4.11ab ± 0.992 4.31a ± 0.972 91.0a
LP9 4.33ab ± 0.553 4.77a ± 0.424 78.0abc
LP10 3.67ab ± 0.373 4.00a ± 0.367 71.0abc
LP11 3.78ab ± 0.465 3.80a ± 0.439 56.0c
LP12 3.44ab ± 0.648 4.20a ± 0.745 84.0ab
LP13 3.78ab ± 0.703 4.04a ± 0.701 71.0abc
LP15 4.22ab ± 0.465 4.68a ± 0.659 87.0ab
LP17 4.78ab ± 0.572 4.40a ± 0.283 85.0ab
LP21 3.89ab ± 0.389 4.91a ± 0.312 78.0abc
LP28 3.67ab ± 0.471 4.60a ± 0.336 87.0ab
LP30 4.56ab ± 1.132 3.86a ± 0.870 60.0bc
122
Jadual 2. Data pertumbuhan daun tiga belas aksesi kacip fatimah sehingga
minggu 30
Aksesi Bilangan pucuk
LP5 1.16b ± 0.254
LP6 1.13b ± 0.152
LP7 1.50b ± 0.184
LP9 2.41a ± 0.281
LP10 1.55b ± 0.152
LP11 1.54b ± 0.346
LP12 1.39b ± 0.189
LP13 1.90ab ± 0.269
LP15 2.43a ± 0.271
LP17 1.07b ± 0.219
LP21 2.43a ± 0.386
LP28 1.46b ±0.187
LP30 1.84ab ± 0.465
*Purata diikuti oleh huruf yang sama tidak mempunyai perbezaan yang signifikan
p<0.05
Jadual 3. Data persekitaran di tapak semaian dari jam 8.30 pagi hingga 4.00
petang
KESIMPULAN
PENGHARGAAN
123
RUJUKAN
Aminah, H., Naimah, C.L., Mohd Zaki, A., & Lokmal, N. (2008). Rooted Leaf
Cuttings of Labisia pumila. J. Tropical Med. Plants, 9(1): 593─599.
Farah Fazwa, M.A., Norhayati, S., Syafiqah Nabilah, S.B. & Mohd. Adi Faiz, A.F.
(2013). Evaluation of Rooting Ability of Five Superior Genotypes of
Labisia pumila var. alata on sand media. Proceedings of Soils Science
Conference of Malaysia 2013, 329─333.
Mohd Noh, J., Rezuwan, K., & Md Akhir, H. (2006). Performance of Kacip
Fatimah (Labisia pumila) Production Under Shade House. ISHS Acta
Horticulturae 710: International Symposium on Greenhouse,
Environmental Controls and In-house Mechanization for Crop
Production in the Tropics and Sub-tropics: 399─403.
Wan Ezumi, M.F., Amrah, S.S., Suhaimi, A. & Mohsin, S. (2007). Evaluation of
The Female Reproductive Toxicity 0f Aqueous Extract of Labisia pumila
var. alata in Rats. Indian J Pharmacol 2007; 39:30─2.
124
SERANGAN ATTEVA SCIODOXA (ULAT HARIMAU) PADA TANAMAN
TONGKAT ALI DI HUTAN DAN LADANG SEMENANJUNG MALAYSIA
ABSTRAK
125
ladang. Di ladang, penjagaan dan pengurusan yang baik dapat membantu
dalam mengawal serangan ulat harimau ini.
PENDAHULUAN
Eurycoma longifolia Jack, atau nama tempatannya tongkat ali merupakan salah
satu tumbuhan herba yang terkenal di Malaysia. Tumbuhan ini mempunyai
nilai perubatan dalam meningkatkan tahap kesihatan manusia. Permintaan
bahan mentah tongkat ali sangat menggalakkan namun kebanyakannya diambil
daripada hutan asli. Bagi memenuhi permintaan ini, penanaman pokok tongkat
ali secara ladang telah diperkenalkan. Teknik penanaman yang sering
diamalkan oleh para pengusaha adalah tanaman secara monokultur dan juga
secara selingan dengan pokok-pokok pertanian. Penanaman secara begini
boleh mengelakkan kepupusan pokok tongkat ali di dalam hutan akibat
penuaian yang tidak terkawal. Walau bagaimanapun serangan penyakit dan
perosak telah memberi masalah dalam usaha ini. Patahayah et al. (2011) dan
Mohd. Noh (2004) melaporkan beberapa jenis penyakit dan perosak berbahaya
kerap ditemui di ladang tongkat ali. Antaranya adalah sindrom mati mengejut
atau Sudden Death Syndrom (SDS), serangan serangga Atteva sciodoxa (Ulat
Harimau), serangga teritip dan ulat penebuk batang (Zeuzera sp.). Serangan
ulat harimau merupakan perosak utama pokok tongkat ali (Mohd. Noh et al.
2004; Mohd Farid et al. 2014). Perosak ini diperhatikan hadir sepanjang masa
dan merupakan pemakan daun yang agresif. Serangan yang serius mampu
menyebabkan kematian atau pokok menjadi renggeh. Namun masih belum ada
kajian secara menyeluruh dilakukan bagi mengetahui tahap kerosakan akibat
serangan ulat harimau ini di Semenanjung Malaysia. Satu kajian telah
dijalankan bagi menilai status kehadiran dan tahap keterukan serangan ulat
harimau pada tongkat ali di ladang dan di hutan.
126
Kehadiran ulat harimau (%) = Bilangan pokok diserang ulat harimau x 100
Jumlah bilangan pokok yang dinilai
Kajian ini melibatkan sejumlah 28 lokasi tanaman tongkat ali yang dipantau.
Daripada jumlah ini, 11 lokasi merupakan petak tongkat ali yang ditubuhkan di
dalam hutan, 9 lokasi adalah ladang tongkat ali dengan selingan tanaman
pertanian atau pokok hutan dan 6 lokasi adalah ladang monokultur tongkat ali
(Rajah 1). Kesemua petak tongkat ali di dalam hutan adalah di bawah
pengurusan Jabatan Hutan Negeri bertujuan untuk pemuliharaan daripada
kepupusan spesies tersebut. Spesies yang ditanam secara selingan dengan
tongkat ali adalah jati, karas, akasia dan herba seperti serai wangi.
Pemerhatian juga mendapati bahawa, secara purata pokok tongkat ali di dalam
hutan mempunyai ketinggian kurang daripada 3 m dengan usia 1 hingga 5
tahun. Manakala, ketinggian pokok tongkat ali di ladang monokultur dan
selingan adalah di antara 1 m hingga 10 m dengan usia 5-10 tahun. Ladang
monokultur dan selingan menunjukkan saiz yang lebih besar di antara 2 hektar
ke 12.5 hektar berbanding tanaman di kawasan hutan iaitu 0.5 hektar hingga 2
hektar. Kebanyakan ladang tongkat ali yang dipantau terbiar dan tidak urus
dengan baik. Kebanyakan pemilik menyatakan bahawa aktiviti-aktiviti
penyelenggaraan seperti membaja, merumput dan meracun hanya dijalankan
pada peringkat awal penubuhan ladang sahaja.
127
Petunjuk: Tanaman di kawasan hutan
Tanaman secara selingan
Tanaman secara monokultur
Rajah 1. Lokasi kawasan tanaman tongkat ali yang terlibat di dalam survei
serangan ulat harimau
128
petak di Pengkalan Hulu dan Bukit Hijau. Di Pengkalan Hulu, kehadiran ulat
harimau adalah 30% manakala, di Bukit Hijau mencatatkan sehingga 83.33%
kehadiran perosak tersebut. Kerosakan pada tanaman tongkat ali di Bukit
Hijau amat serius dengan purata kerosakan pada bahagian silara tongkat ali
adalah melebihi 75% . Bilangan tongkat ali yang masih hidup juga berkurangan
kepada 90% dari bilangan anak benih yang ditanam.
129
menyebabkan serangan ulat harimau tidak menunjukkan kesan yang ketara.
Sebaliknya, ladang monokultur menunjukkan nasib yang berbeza berbanding
petak tongkat ali di kawasan hutan. Kajian mendapati hanya satu ladang
monokultur tongkat ali merekodkan kehadiran ulat harimau yang rendah iaitu
di Felda Jengka 16 (16.67%). Ladang-ladang tongkat ali yang lain seperti di
Felda Sungai Tekam dan Sungai Menyala masing-masing mencatatkan
kehadiran ulat harimau sebanyak 86.7% dan 83.33 %. Kedua-dua kawasan ini
juga menunjukkan tahap kerosakan pada silara pokok (DSI) yang tinggi iaitu
melebihi 75%.
RUMUSAN
Serangan ulat harimau ke atas tongkat ali berlaku di semua kawasan tanaman
tongkat ali termasuk di hutan dan di ladang (selingan dan monokultur). Walau
bagaimanapun, ladang tongkat ali monokultur adalah lebih terdedah kepada
serangan ulat harimau berbanding ladang selingan dan petak yang ditubuhkan
di kawasan hutan. Langkah terbaik dalam mengatasi masalah ini adalah dengan
membuat pemantauan berkala supaya serangan pada peringkat awal dapat
kesan dan langkah pengawalan dan pencegahan yang sesuai dapat diambil.
130
RUJUKAN
Mohd Anuar, A. 2003. Some Notes On The Control of The Tiger Moth, Atteva
sciodoxa (Lepidoptera: Yponomeutidae) and Scale Insects on Tongkat
Ali (Eurycoma Longifolia). The 6th International Conference on Plant
Protection in the Tropics; 11─14th. August 2003; Kuala Lumpur. P 92.
Mohd Farid, A., Wan Muhd Azrul, W.A., Ong S.P., & Lee S.S. 2014. Destructive
Leaf Cutter (Tiger Moth) of Eurycoma longifolia. FRIM Technical
Information No. 73.
Mohd. Noh Hj., Mohd Ilham, A. & Fauziah, I. 2004. Ancaman-Ancaman Utama
Dalam Penanaman Tongkat Ali Di Semenanjung Malaysia. Pp 42─46 in
Chang Y.S, Vimala, S., Mazura, M.P. & Ong, B.K. (Eds.). Proceeding of
the Seminar on Medicinal Plants: Tongkat Ali, Kacip Fatimah and
Pegaga - New Dimension in Complementary Healthcare. Kuala Lumpur.
Patahayah, M., Lee, S.S. & Mohd Farid, A. 2011. Penyakit, Perosak Dan
Gangguan Tanaman Tongkat Ali. FRIM Technical Information Handbook
No.41.
131
PRODUK HERBA DAN TREND PENCEMARAN MIKROORGANISMA
ABSTRAK
132
PENGENALAN
133
PENEMUAN DAN PERBINCANGAN
Sebanyak 184 sampel bahan mentah, 66 produk separa siap dan 75 produk siap
telah dijalankan ujian pencemaran mikroorganisma. Dalam kategori bahan
mentah 64% adalah sampel daun, 21% adalah rizom/akar, 10% buah dan 5%
batang. Bagi kategori produk separa siap, 76% sampel adalah ekstrak dan 24%
adalah bahan persediaan herba. Manakala produk siap pula 65% adalah produk
untuk penggunaan oral dan 35% adalah produk untuk penggunaan topikal.
Hasil daripada ujian paras kandungan mikroorganisma didapati kategori bahan
mentah memberikan bacaan CFU/g@ml paling tinggi iaitu sehingga 1012
CFU/g@ml, berbanding kategori separa siap bacaan CFU/g@ml tertinggi adalah
sehingga 1010 CFU/g@ml dan produk siap sehingga 106 CFU/g@ml. Hasil
pemerhatian menunjukkan terdapat sampel herba dengan paras pencemaran
mikrooganisma melebihi had spesifikasi sehingga 10 x 103 kali ganda. Nilai
CFU/g@ml tertinggi iaitu 5.2 x 1011 CFU/g@ml adalah sampel kategori bahan
mentah daripada bahagian rizom. Bagi kategori separa siap; ekstrak herba
daripada bahagian rizom/akar dan buah mencatatkan nilai CFU/g@ml paling
tinggi iaitu 2.5 x 1010 CFU/g@ml. Manakala produk penggunaan topikal
memberikan bacaan yang paling tinggi berbanding produk penggunaan oral
iaitu 2.1 x 107 CFU/g@ml untuk kategori produk siap.
Peratusan sampel yang memenuhi had spesifikasi dapat dilihat seperti jadual 1.
Hampir 80% daripada sampel untuk semua kategori yang diuji memenuhi had
spesifikasi yang dietapkan untuk TAMC. Manakala hanya 50% memenuhi had
spesifikasi untuk TYMC.
134
Meneliti trend pencemaran mikroorganisma secara umum, jelas
bahawa paras pencemaran mikroorganisma pada produk kategori bahan
mentah herba adalah paling tinggi, diikuti produk kategori separa siap. Paras
pencemaran mikroorganisma paling rendah pada produk kategori siap (Rajah
1). Pola pencemaran ini dipercayai berlaku kerana sampel bahan mentah herba
yang dikutip dari ladang mempunyai hubungan langsung dengan sumber
pencemaran seperti bahagian rizom/akar dengan tanah dan bahagian
buah/daun yang dicemari oleh mikroorganisma. Oleh itu, adalah penting
proses pemilihan dan sanitasi bahan mentah herba dijalankan dengan segera
agar kontaminasi mikroorganisma tidak merebak dan semakin meningkat.
Proses sanitasi bahan mentah perlu dibuat dengan lebih rapi untuk bahan
mentah daripada bahagian rizom/akar dengan memastikan tanah dan kotoran
dibasuh bersih. Bahagian buah atau daun yang tercemar dengan
mikroorganisma juga perlu diasingkan.
135
melibatkan penggunaan pelarut dalam penyediaan ekstrak dan juga haba
dalam penyediaan ekstrak air turut membantu mengurangkan paras
pencemaran mikroorganisma. Seterusnya proses pengeringan ekstrak seperti
kering beku (freeze dry) juga dapat mengelakkan pembiakan mikroorganisma
terutamanya kulat yang mudah membiak dalam keadaan yang lembap.
Walaubagaimanapun pencemaran mikroorganisma untuk kategori separa siap
dan produk siap ini boleh berlaku semula semasa peringkat pengendalian,
pembungkusan dan penyimpanan produk.
RUMUSAN
136
PENGHARGAAN
RUJUKAN
137
RUMPAI MIANG MEXICO, ANCAMAN MELEBIHI MANFAAT
ABSTRAK
PENGENALAN
138
menyenangkan (Rezaul 2010). Rumpai ini berasal dari Mexico, Amerika tengah
dan Amerika Selatan. Rumpai daripada keluarga Asteraceae ini boleh tumbuh
di pelbagai jenis tanah termasuk tanah yang kurang subur.
Rumpai miang mexico seakan ulam raja pada peringkat muda, daunnya
berbulu halus dan tersusun membulat. Bunganya kecil bewarna putih krim
muncul berjambak di hujung pokok dan bertukar menjadi coklat apabila
matang. Batang biasanya bercabang dan diselaputi bulu halus atau dipanggil
trikom dengan ketinggian mencapai sehingga 1.5 hingga 2 m. Biji benih
berbentuk baji dengan warna perang kehitaman. Satu pokok yang sihat boleh
menghasilkan 10,000 hingga 15,000 biji benih yang mampu bercambah dalam
satu kitaran yang lengkap (Patel 2011).
Biji benihnya boleh merebak melalui air mengalir, angin, makanan ternakan
yang dicemari, kenderaan dan jentera, haiwan ternakan yang diimport dan
manusia. Seluruh pokok termasuk debunga dan trikom mempunyai bahan
kimia toksin yang dipanggil sesquiterpen lakton dan pelbagai jenis bahan
alelokimia (Maishi et al., 1998). Bahan kimia ini boleh menyebabkan keburukan
kepada manusia, haiwan, tumbuhan dan ekosistem. Antaranya ia boleh
menyebabkan asma, bronkitis dan dermatitis kepada manusia. Kadar 10-50%
parthenium yang tercemar di dalam makanan ternakan boleh menyebabkan
kesakitan akut dan membawa kepada kematian. Anjing yang terdedah dengan
tanaman ini turut menyebabkan aneroksia, diarrhea dan sakit mata. Kehadiran
bahan kimia seperti parthenin, hysterin, hymenin dan ambrosin telah memberi
kesan alelopati atau perencatan terhadap tumbuhan lain yang tumbuh di
sekitarnya. Kajian telah membuktikan bahawa parthenin adalah bahan kimia
yang boleh merencat pertumbuhan akar pelbagai jenis tumbuhan dikot dan
monokot. Keistimewaan ini membolehkannya mudah menguasai sesuatu
kawasan dan menyesarkan tanaman lain. Apabila rumpai agresif ini menempati
sesuatu kawasan, maka biodiversiti kawasan tersebut akan berubah.
Perubahan drastik ini pasti menyebabkan ketidakseimbangan ekosistem
berlaku dan perlu ditangani dengan segera.
139
Oleh kerana telah tersebar luas tentang keburukan rumpai ini kepada
orang awam, maka kajian telah dijalankan untuk mengenalpasti taburan
rumpai miang mexico di beberapa tempat yang telah dilaporkan terdapat di
Malaysia. Di samping itu, kajian turut menekankan jenis tanah bagaimanakah
rumpai ini paling senang ditemui.
Kajian dijalankan di tiga tempat yang telah dilaporkan merebak seperti di Ulu
Yam, Selangor; Sungai Siput, Perak dan Sungai Pasir, Kedah. Pemantauan yang
dijalankan meliputi beberapa aspek termasuk keluasan kawasan yang telah
ditumbuhi, jenis tanah, faktor persekitaran yang memungkinkan berlakunya
penyebaran dan jenis aktiviti kawasan tersebut.
Kajian ini bertujuan untuk mengenal pasti keupayaan biji benih bercambah
pada tanah yang berbeza. Biji benih dan tanah telah diambil dan ujian
percambahan telah dilakukan di atas piring petri dan juga di atas tanah. Biji
benih telah direndam dengan bahan kimia kalium nitrate (0.2%) dan juga air
yang telah ditapis untuk memecahkan dormansi selama 24 jam. Kemudian biji
benih tersebut telah ditos selama 12 jam sebelum disemai di atas piring petri
dan tiga jenis tanah yang berbeza. Selepas dua minggu, pemerhatian dan
keputusan percambahan telah diambil.
Soal selidik telah dijalankan dengan menemu ramah beberapa orang pegawai
pengembangan di kawasan tersebut. Soalan merangkumi kes terhadap
kesihatan manusia, haiwan ternakan dan tanaman. Kes adalah berdasarkan
kepada laporan yang disampaikan kepada pegawai terlibat.
140
PENEMUAN DAN PERBINCANGAN
Analisis tekstur tanah (Jadual 1) menunjukkan bahawa tanah dari kawasan Ulu
yam adalah berpasir manakala tanah dari Sg. Pasir adalah lempung liat dan Sg.
Siput adalah lempung.
Sg Lempung
Siput*
Nota: * Ditentukan berdasarkan pemerhatian terhadap tanah bukan
berdasarkan analisis tanah
141
Rajah 1. Kadar percambahan biji benih di atas tanah dan piring petri selepas
rawatan dengan 0.2% KNO3
142
Rajah 3. Perbezaan kadar percambahan biji benih P. hysterophorus selepas
rawatan dengan KNO3 dan air suling
143
KESIMPULAN
Kajian ini membuktikan biji benih P. hysterophorus yang diambil dari tiga
kawasan yang berbeza di Semenanjung Malaysia mempunyai keupayaan yang
tinggi untuk bercambah walaupun pada tekstur tanah berbeza yang berbeza.
Walaupun kesan buruk tumbuhan terhadap manusia dan haiwan masih boleh
dikawal, namun kesannya terhadap tanaman dan ekosistem telah jelas berlaku.
Oleh kerana itu, pengawalan dan pemantauan yang serius perlu diambil oleh
semua pihak agar rumpai ini tidak terus tersebar.
PENGHARGAAN
RUJUKAN
Adkins, S. W., O’Donnell, C., Khan, N., Nguyen, T., Shabbir, A., Dhileepan, K.,
George, D. & Navie, S. (2010). Parthenium Weed (Parthenium
hysterophorus L.) Research in Australia: New Management Possibilities.
pp. 120-123 in Zydenbos, S.M.(ed) Proceedings 17th Australasian
Weeds Conference.
Kohli, R. K., Batish, D. R., Singh, H. P., & Dogra, K. S. (2006). Status, Invasiveness
and Environmental Threats of Three Tropical American Invasive Weeds
(Parthenium hysterophorus L., Ageratum conyzoides L., Lantana
camara L.) in India. Biological Invasions, 8(7), 1501─1510.
Maishi, A. I., Ali, P. S., Chaghtai, S. A., & Khan, G. (1998). A Proving of
Parthenium hysterophorus, L. British Homoeopathic Journal, 87(1),
17─21.
144
Patel, S. (2011). Harmful and Beneficial Aspects of Parthenium hysterophorus:
an Update. 3 Biotech, 1(1), 1─9.
Patel, V. S., Chitra, V., Prasanna, P. L., & Krishnaraju, V. (2008). Hypoglycemic
Effect of Aqueous Extract of Parthenium hysterophorus l. in Normal and
Alloxan Induced Diabetic Rats. Indian journal of pharmacology, 40(4),
183.
145
KONSERVASI SECARA EX SITU BAGI KACIP FATIMAH VARIETI
LANCEOLATA
ABSTRAK
Kacip fatimah atau nama saintifiknya Labisia pumila ialah sejenis tumbuhan
dalam famili Primulaceae yang digunakan oleh masyarakat Melayu sejak 400
tahun yang lalu. Pokok kacip fatimah adalah sejenis tumbuhan renek yang
tumbuh secara menegak dengan anggaran ketinggian 30–50 cm. Pokok ini
mempunyai akar yang unik, iaitu akar tunjang yang panjang dan dikelilingi oleh
anak akar yang halus. Malahan, anak pokok kacip fatimah boleh tumbuh keluar
di sepanjang akarnya itu. Hasil daripada kajian taksonomi botani, terdapat tiga
jenis varieti kacip fatimah yang biasa dijumpai di Malaysia iaitu Labisia pumila
varieti alata, Labisia pumila varieti pumila dan Labisia pumila varieti
lanceolata. Proses pengecaman bagi varieti lanceolata mudah dilakukan
berbanding varieti pumila dan varieti alata kerana tangkainya tidak
mempunyai sayap. Walau bagaimanapun, habitat bagi varieti lanceolata
tumbuh lebih berselerak berbanding dua varieti yang lain yang tumbuh secara
berumpun. Setakat ini, kajian saintifik mengenai kegunaan dan kelebihan
varieti lanceolata adalah masih kurang berbanding dua varieti yang lain.
Sebagai langkah awal, Program Membaikbiak Tumbuhan, Institut Penyelidikan
Perhutanan Malaysia (FRIM) telah mengambil inisiatif untuk melakukan
konservasi secara ex situ bagi varieti ini untuk tujuan penyelidikan dan koleksi
germplasma. Dalam kajian ini, pokok-pokok ibu varieti lanceolata telah dikutip
dari lima hutan simpan di lima negeri iaitu Perak, Negeri Sembilan, Johor,
Terengganu dan Pahang. Selain pokok, sampel tanah bagi setiap lokasi juga
turut diambil bagi mengenalpasti kandungan nutrien yang membantu
pertumbuhan pokok ini di habitat asalnya. Pokok-pokok ibu yang dibawa
pulang dibiarkan pulih di tapak semaian sebelum ianya ditanam ke plot
germplasma. Pembiakan tampang menggunakan kaedah keratan juga turut
dilakukan untuk memperbanyakkan anak pokok. Keupayaan pengakaran varieti
lanceolata bagi populasi Perak telah direkodkan di dalam kajian ini. Hasil kajian
menunjukkan 80% keratan dari 12 genotip bagi populasi Perak telah berjaya
berakar dalam tempoh 9 minggu. Kadar pertumbuhan pucuk pada keratan
akar juga didapati lebih cepat berbanding keratan daun, di mana keratan yang
berakar telah mula mengeluarkan pucuk pada minggu ketiga selepas keratan
berakar dipindahkan ke dalam polibeg. Anak-anak pokok yang terhasil dari
kaedah keratan kemudiannya dipindahkan ke tapak semaian bagi tujuan
146
pengikliman dan penyesuaian sebelum ditanam di germplasma. Kadar
pertumbuhan pokok di germplasma juga direkodkan setiap bulan. Sebagai
kesimpulannya, pokok-pokok kacip fatimah yang ada di germplasma ini
digunakan sebagai bahan penyelidikan (breeding materials) khususnya dalam
bidang pembiakbakaan dan juga sebagai pusat rujukan bagi spesies ini. Selain
itu, populasi kacip fatimah di hutan simpan semula jadi juga dapat dilindungi
dan dipelihara.
PENGENALAN
147
BAHAN DAN KAEDAH
Lebih dari 150 genotip kacip fatimah varieti lanceolata telah dikutip dari lima
hutan simpan (HS) di Semenanjung Malaysia iaitu HS Gunung Korbu, Perak; HS
Gunung Berembun, Negeri Sembilan; HS Gunung Berlumut, Johor; HS Pulau
Selimbar, Terengganu; dan HS Betung, Pahang. Setiap pokok dilabel dengan
kod yang berbeza dan dibawa pulang ke FRIM. Sampel tanah bagi setiap
populasi kacip fatimah yang dikutip turut diambil untuk dianalisa di Makmal
Kimia Tanah. Antara analisis yang dijalankan adalah penentuan jumlah nitrogen
(%), fosforus (ppm) dan potassium (cmol/kg).
Dalam kajian ini, keratan akar telah digunakan untuk memperbanyakkan anak
pokok kacip fatimah. Akar dikerat sepanjang 5 cm dan hormon penggalak akar
diletakkan di bahagian bawah keratan. Keratan akar ditanam di dalam media
pasir dan disiram menggunakan sistem renjisan automatik selama satu minit
setiap jam. Peratus pengakaran direkodkan dalam tempoh 9 minggu.
Penubuhan Germplasma
148
Bancian Pertumbuhan Pokok Di Germplasma
Bancian pertumbuhan seperti tinggi pokok (cm) dan diameter kolar (mm)
direkodkan setiap bulan. Analisis statistik telah dijalankan untuk mengenalpasti
perbezaan signifikan pertumbuhan pokok bagi setiap populasi.
Analisa sampel tanah dari tiga populasi iaitu HS Gunung Korbu, Perak; HS
Gunung Berembun, Negeri Sembilan; dan HS Gunung Berlumut, Johor
direkodkan. Keputusan analisa makronutrien tanah bagi ketiga-tiga populasi
tersebut direkodkan di dalam Jadual 1.
Jadual 1. Sifat kimia tanah di habitat asal kacip fatimah varieti lanceolata
Populasi Jumlah N % F K pH kering
(ppm) (cmol/kg)
HS Gunung
0.36a ± 0.03 12.57a ± 1.88 0.12a ± 0.01 4.05b ± 0.04
Korbu
HS Gunung
0.25b ± 0.01 2.42b ± 0.19 0.11a ± 0.01 4.23a ± 0.04
Berlumut
HS Gunung
0.33a ± 0.02 4.49b ± 0.31 0.12a ± 0.01 4.08b ± 0.05
Berembun
Bagi ciri-ciri fizikal tanah pula (Jadual 2), HS Gunung Berembun, Negeri
Sembilan mencatatkan peratusan pasir kasar yang tinggi (45.90%) berbanding
pasir halus (11.30%). Manakala HS Gunung Berlumut, Johor pula mengandungi
pasir halus yang tinggi dan kelodak dengan masing-masing berjumlah 23.81%
dan 19.00%. Hutan Simpan Gunung Korbu, Perak pula mencatatkan kandungan
149
tanah liat yang tinggi (41.53%) diikuti dengan HS Gunung Belumut, Johor
(38.38%). Ciri fizikal tanah memainkan peranan penting kerana ia memberikan
kesan terhadap pertumbuhan pokok terutamanya keupayaan memegang air,
pengudaraan dan pengekalan bahan organik (Fisher & Binkley 2000).
Jadual 2. Ciri-ciri fizikal tanah di habitat asal kacip fatimah varieti lanceolata
Populasi Pasir kasar Pasir halus Kelodak Tanah liat
(%) (%) (%) (%)
HS Gunung
36.06b ± 1.78 20.13ab ± 6.76 10.00b ± 0 .49 41.53a ± 1.38
Korbu
HS Gunung
37.5b ± 3.94 23.81a ± 2.38 19.00a ± 1.92 38.38a ± 16.96
Berlumut
HS Gunung
45.9a ± 1.94 11.13b ± 1.50 13.47b ± 0.69 31.40a ± 2.99
Berembun
Bagi analisis prestasi keratan akar pula, hanya 12 genotip kacip fatimah var.
lanceolata (AL8, AL10, AL22, AL26, AL27, AL32, AL34, AL36, AL41, AL44, AL48
dan AL52) dari HS Gunung Korbu, Perak direkodkan dalam kajian ini. Selepas
sembilan minggu keratan, kebanyakan genotip yang diuji menunjukkan 80%
pengakaran dan hanya tiga genotip yang berjaya memberikan 100%
pengakaran iaitu AL10, AL32 dan AL36. Berdasarkan Rajah 1, genotip AL34
menunjukkan pertumbuhan akar yang cepat pada minggu ketiga diikuti dengan
genotip dari AL8, AL10, AL32, AL36, AL41 dan AL44.
Peratus pertumbuhan pucuk (%)
60
Peratus pertumbuhan akar (%)
50
40 50
30 40
20 30
10 20
0 10
Genotip Genotip
150
mana kebanyakannya mula mengeluarkan pucuk pada minggu ketujuh. Aminah
(2005) dan Rozihawati (2008) melaporkan pertumbuhan pucuk pada keratan
akar adalah lebih cepat berbanding keratan daun. Maklumat ini adalah penting
untuk pengusaha ladang yang ingin membiakkan tanaman kacip fatimah secara
konvensional.
Prestasi pertumbuhan pokok-pokok ibu dari lima populasi kacip fatimah varieti
lanceolata yang telah ditanam di germplasma direkodkan setiap bulan. Hasil
analisis statistik (ANOVA) menunjukkan prestasi pertumbuhan lima populasi
kacip fatimah varieti lanceolata menunjukkan perbezaan yang signifikan pada
p<0.05. Min ketinggian dan diameter pokok bagi setiap populasi selepas tiga
bulan ditanam direkodkan dalam Jadual 3.
RUMUSAN
Maklumat dari hasil kajian ini boleh digunapakai untuk tujuan pemuliharaan
dan pemeliharaan spesies kacip fatimah khususnya dan juga bagi spesies-
spesies herba yang lain. Kesimpulannya, kacip fatimah varieti lanceolata juga
boleh diperbanyakkan menggunakan kaedah keratan sama seperti varieti alata
dan pumila. Para petani dan pengusaha herba boleh menggunakan teknik ini
untuk memperbanyakkan bahan tanaman di masa akan datang tanpa perlu
151
bergantung sepenuhnya daripada sumber hutan bagi mengawal dari
berlakunya kepupusan, dengan itu keselamatan diversiti spesies akan terjamin.
RUJUKAN
Aminah, H., Naimah, C.L., Mohd Zaki, A. & Lokmal, N. (20079. Rooted Leaf
Cuttings of Labisia pumila. Journal of Tropical Medicinal Plants: 9(1):
593─599.
Fathilah, S.N., Norazlina, M., Norliza, M., Isa, N.M., Ima, N.S. & Ahmad Nazrun,
S. (2013). Labisia pumila regulates bone-related genes expressions in
postmenopausal ostereoporosis model. BMC Complementary &
Alternative Medicine: 13:217.
Fisher, R.F. & Binkley, D. (2000). Ecology and Management of Forest Soil. 3rd ed.
John Willey & sons, New York. Pp489.
Hyun-kyung ,C., Dong-hyun, K., Jin Wook, K., Sulaiman, N., Mohamad Roji, S., &
Chang S. P. (2010). Labisia pumila Extract Protects Skin Cells from
Photoaging Caused by Uvb Irradiation. Journal of Bioscience and
Bioengineering: 109(3), 291–296.
Intan, I.H. & Nik, H.N.H. (2015). Kenapa Kacip Fatimah? Dewan Bahasa dan
Pustaka, Kuala Lumpur.
Saari, A & Noraini, O. (2013). Strategic Planning, Issues, Prospects and the
Future of the Malaysian Herbal Industry. International Journal of
Academic Research in Accounting, Finance and Management Sciences:
91─102.
152
TAPAK WARISAN FRIM: POTENSI SEBAGAI TAPAK KONSERVASI EX
SITU TERBESAR BAGI GERMPLASMA TUMBUHAN UBATAN DI
SEMENANJUNG MALAYSIA
ABSTRAK
LATAR BELAKANG
FRIM ditubuhkan dengan visi sebagai institusi penyelidikan yang terunggul dan
berautoritatif dalam penyelidikan perhutanan tropika di peringkat
antarabangsa. Kampus FRIM diwartakan sebagai Tapak Warisan Semula Jadi
Negara pada 2009 dan seterusnya pada 2012, diangkat sebagai Tapak Warisan
Kebangsaan. Kampus FRIM berkeluasan 544.31 ha merupakan satu-satunya
institusi penyelidikan di dunia diiktiraf sebagai tapak warisan. Justeru, pelbagai
usaha sedang dilaksanakan bagi menggerakkan FRIM sebagai Tapak Warisan
Dunia UNESCO menjelang tahun 2020. Tapak ini merupakan contoh terbaik
usaha sustainability science jangka panjang dan perlu diteruskan demi manfaat
sains sejagat serta bagi memastikan inovasi generasi terdahulu dapat dikongsi
dengan generasi akan datang.
153
Pembentukan Hutan Tropika Regenerasi Tertua di Dunia
Hutan tropika regenerasi yang berusia hampir 100 tahun di Kampus FRIM,
terbentuk menerusi konsep “on-going ecological and biological processes”
merupakan bukti penting pembentukan hutan buatan manusia dari kawasan
gersang. Inovasi dan pengurusan yang dilaksanakan sejak tahun 1920-an
berjaya memulihkan kawasan yang dahulunya bekas lombong, kebun sayur dan
belukar, bertukar menjadi hutan regenerasi tertua dunia dan kini menjadi
model pemulihan hutan serta mendapat pengiktirafan pakar hutan tropika.
Kawasan hutan asal sebenarnya telah dibalak beberapa kali. Kini terdapat
pelbagai jenis dirian hutan yang dibangunkan menerusi penanaman satu
spesies atau pelbagai spesies merangkumi spesies tempatan dan eksotik
bernilai komersial.
Taman Etnobotanikal
Taman Etnobotani ditubuhkan pada tahun 1995 sebagai pusat pemuliharaan ex
situ serta pusat domestikasi bagi tumbuhan ubatan dan beraroma yang
digunakan oleh golongan etnik di Semenanjung Malaysia. Berfungsi untuk
membangkitkan minat masyarakat umum dalam tumbuhan ubatan dan
beraroma, terdapat hampir 200 spesies tumbuhan mewakili 85 famili.
154
Denai Alam FRIM
Sehingga tahun 2009, terdapat enam buah denai alam dibangunkan dengan
tema berbeza bagi pelawat berekreasi sambil mempelajari sumber hutan yang
ditemui di sepanjang perjalanan. Antara denai alam yang terawal menyimpan
pelbagai spesies tumbuhan ubatan ialah Denai Keruing, manakala Denai
Sebasah yang dibina pada tahun 2008 menggambarkan pula kemandirian flora
dan fauna di kawasan hutan becah.
Arboretum FRIM
Semenjak 1926 penyelidikan botani dan perhutanan memerlukan sokongan
koleksi tumbuhan hidup yang terurus. Kini, Arboretum FRIM telah menjadi
pusat pengumpulan tumbuhan hidup terbesar dan mempunyai koleksi yang
mewakili keseluruhan flora di Malaysia. Terdapat tujuh arboretum di Kampus
FRIM dan kini menyimpan kira-kira 354 spesies dari 149 genera mewakili 48
famili tumbuhan yang ditanam.
155
BAHAN DAN KAEDAH
156
PENEMUAN DAN PERBINCANGAN
Berdasarkan 21,743 rekod dari pelombongan data (data mining) dan sistem
pangkalan data myBotani, dianggarkan terdapat 592 spesies mewakili 335
genera dari 109 famili tumbuhan ubatan di Kampus FRIM. Berdasarkan
anggaran awal keseluruhan 2,434 spesies tumbuhan yang ditemui, kira-kira
24.32% merupakan spesies ubatan. Manakala 10 famili utama tertera pada
Jadual 1.
2 Euphorbiaceae 13 17
3 Lamiaceae 7 10
4 Lauraceae 4 11
5 Leguminosae 18 26
6 Moraceae 7 16
7 Myrtaceae 8 17
8 Palmae 6 11
9 Rubiaceae 13 19
10 Zingiberaceae 9 12
Secara keseluruhan, terdapat enam buah tapak konservasi ex situ utama bagi
koleksi germplasma tumbuhan ubatan. Didapati Taman Etnobotani FRIM
merupakan tapak ex situ yang menyimpan paling banyak koleksi germplasma
tumbuhan ubatan iaitu sejumlah 152 spesies, diikuti oleh plot-plot
penyelidikan FRIM (128 spesies) dan Taman Etnoflora, KBG (28 spesies) seperti
ditunjukkan di Rajah 2.
157
Bilangan Spesies Tumbuhan Ubatan di Tapak Konservasi
Ex-Situ Kampus FRIM
10
12 Taman Etnobotani
Plot Penyelidikan
26
Taman Etnoflora KBG
28 Arboretum
128 Denai Alam
Rumah Tradisional
152
Melayu Melaka
0 50 100 150 200
Bilangan Koleksi ex-situ Spesies Tumbuhan Ubatan
Rajah 2. Bilangan spesies tumbuhan ubatan di enam tapak konservasi ex situ
Kampus FRIM
KESIMPULAN
RUJUKAN
158
PENGGUNAAN TUMBUHAN DALAM KEHIDUPAN ORANG ASLI
KUMPULAN SENOI DI SEMENANJUNG MALAYSIA: KHUSUSNYA
SUBETNIK TEMIAR DAN SEMAI
ABSTRAK
159
nyiru), perhiasan diri (tempok, rantai manik), alat muzik (tensol, genggong),
upacara sewang atau pemujaan roh (Hari Moyang, sakit, keraian), kematian,
kelahiran dan ukiran kayu (patung dan topeng). Masyarakat Orang Asli Senoi
didapati amat bergantung kepada spesies tumbuhan daripada jenis buluh,
palma (e.g. rotan, pinang, bertam, palas) pisang, mengkuang dan terap selain
daripada kutipan buah-buah dan pucuk tumbuhan dalam hutan.
PENGENALAN
160
BAHAN DAN KAEDAH
Secara umumnya, masyarakat Orang Asli pada zaman dahulu amat bergantung
kepada bahan berasaskan tumbuhan di sekeliling kediaman mereka untuk
pakaian dan keperluan kehidupan harian. Pakaian mereka diperbuat daripada
kulit kayu terap di mana lelaki akan memakainya sebagai cawat manakala
perempuan pula akan memakainya sebagai skirt pendek dengan membiarkan
bahagian atas tubuhnya terdedah. Walau bagaimanapun, Orang Asli pada
zaman sekarang telah memakai pakaian seperti masyarakat moden. Pakaian
daripada kulit kayu terap hanya akan digunakan semasa upacara tertentu atau
ketika berburu. Dalam arus kemodenan, beberapa makanan tradisi yang masih
diamalkan oleh masyarakat Orang Asli adalah seperti nasi dalam buluh, ayam
dalam buluh, pulut dalam periuk kera, ubi bakar, ikan pais, ubi lempak, ubi
kacau dan ikan dalam buluh. Peralatan dapur yang digunakan pula banyak
dihasilkan daripada buluh (cawan, bekas air, bekas nasi/lauk, peniup api,
sudip), tempurung kelapa (sudu, sengkalam,senduk, bekas nasi/lauk), upih
pinang (pinggan, timba), rotan (nyiru,pemarut) dan bertam (pemarut).
161
atapnya merupakan jalinan pelepah bertam atau nipah yang rapi. Antara jenis
rumah yang popular dibina oleh komuniti Temiar adalah seperti Dekreh -
pondok, Deklare -rumah dengan hiasan dinding motif corak kelawar dan bunga
kelarai, Dek Rendetek -rumah buluh, Dek Tagon dan Dek Ulau -rumah panjang
dan Dek Jalaq Juh - atas pokok (Nisra Nisran 1995).
Pelbagai jenis bubu daripada buluh dan rotan dihasilkan oleh komuniti
Temiar di mana penggunaan setiap jenis bubu adalah bergantung kepada lokasi
sungai di sekitar kediamannya seperti bubu panchok, bubu tempirai, bubu Che
O’, tangkul dan ce’ak. Peralatan seperti serempang buluh dan kail (bertam)
serta perahu dan rakit (buluh) turut digunakan dalam aktiviti menangkap ikan.
Sementelahan pula, komuniti Semai banyak melibatkan diri dalam aktiviti
bercucuk tanam. Antara peralatan yang digunakan adalah seperti beliung (kayu
dan rotan), tabung getah (buluh), parang (kayu dan anyaman rotan), raga galas
(rotan), lesung kaki, kepok padi (upih pinang), lesung tumbuk padi, keri, kuku
kambing, pisau raut dan nyiru.
162
Semasa upacara perkahwinan, bujam (pandan) akan digunakan untuk
meletak pinang, kapur, sireh, gambir, bunga cengkih dan rokok tembakau.
Perhiasan pengantin seperti rantai manik (biji keril- Temiar) atau rantai buluh
perindu (Semai), selimpar (nipah), simpai (rotan) dan tempok (rumput, nipah,
kelapa) serta lilitan pinggang (kelapa, nipah) turut digunakan. Dalam upacara
kelahiran, uri budak dari komuniti Semai akan disimpan di dalam bekas yang
dimasukkan tanah terlebih dahulu sebelum disembur dan dijampi
menggunakan batang bonglai dan dikambus dengan tanah sebelum ditutup
dan diletakkan di dapur agar sentiasa panas supaya bayi tidak diganggu oleh
makhluk halus. Sebaliknya, komuniti Temiar pula akan membungkus uri bayi
berserta pisau buluh yang digunakan semasa upacara kelahiran dengan kain
untuk diikat di dahan pokok besar.
RUMUSAN
RUJUKAN
Hood, S. (2006). Peoples and Traditions. The Encyclopedia of Malaysia Vol. 12.
Editions Didier Millet, Singapore. Pp. 148.
JHEOA (Jabatan Hal Ehwal Orang Asli) (2002). Kehidupan, Budaya dan Pantang
Larang Orang Asli. Kuala Lumpur: Perniagaan Rita. Pp. 154.
163
PENGGUNAAN HERBA DALAM PERBIDANAN TRADISIONAL MELAYU
SELEPAS BERSALIN DI ZON TENGAH, SEMENANJUNG MALAYSIA
ABSTRAK
164
PENGENALAN
Kajian ini menggunakan kaedah temu bual ke atas 30 orang bidan kampung.
Kawasan kajian pula meliputi kawasan dalam lingkungan 100 km daripada
Hospital Putrajaya. Senarai bidan ini dipilih daripada data kajian Study on
Traditional medicine: Midwives yang dijalankan oleh Pusat Penyelidikan
Perubatan Herba pada tahun 2006 hingga 2007 di Semenanjung Malaysia.
Calon responden dihubungi melalui telefon untuk membuat temu janji.
Responden akan menandatangani borang keizinan) (PIC) dan perkongsian
faedah (ABS) sebelum ditemu bual. Temu bual dilakukan dengan
berpandukan soalan kaji selidik yang telah dibina. Maklumat dicatat dan data
terkumpul dianalisis.
165
Jadual 1. Senarai spesies tumbuhan dan kegunaannya perbidanan tradisional
Melayu
Bil. Nama Penggunaan tradisional
tempatan
1. Asam gelugor Ramuan param, tangas kering dan mandi daun.
/keping
2. Asam jawa Ramuan pilis, ubat periuk, jamu, mandi daun.
3. Akar angin Ramuan param
4. Betik jantan Balut tungku (daun)
5. Buah keras Ramuan mandi daun
6. Bunga lawang Ramuan ubat periuk, jamu
7. Bawang putih Ramuan tangas kering (kulit), tangas basah (kulit),
ubat periuk (rizom), minyak urut (rizom)
8. Bawang merah Ramuan tangas kering (kulit), tangas basah (kulit)
166
25. Inai Ramuan ubat periuk, jamu, mandi daun (daun)
26. Jemuju Ramuan ubat periuk,
27. Jintan manis Ramuan pilis, ubat periuk, jamu, tangas basah, mandi
daun, isi tungku
28. Jintan putih Ramuan pilis, ubat periuk, jamu, tangas basah, mandi
daun, isi tungku
29. Jerangau Ramuan param, tangas basah (daun), mandi daun
(daun), minyak urut
30. Jarak pagar Balut tungku (daun)
31. Jering (kering) Ramuan jamu
32. Jambu batu Ramuan tangas basah (daun, pucuk), mandi daun
(daun)
33. Jarum emas Ramuan ubat periuk
34. Jintan hitam Ramuan pilis, jamu, tangas basah, isi tungku, minyak
urut
35. Ketumbar Ramuan ubat periuk, jamu, tangas basah, mandi daun
36. Kayu manis Ramuan param, pilis, ubat periuk, jamu, minyak urut
167
53. Lengkuas Ramuan jamu, tangas kering, tangas basah (daun),
mandi daun (daun), balut tungku (daun), isi tungku
(rizom), minyak urut
54. Lempuyang Ramuan jamu, tangas basah, mandi daun, minyak
urut
55. Lada ekor Ramuan pilis, ubat periuk, jamu
56. Lada sireh Ramuan pilis, ubat periuk, jamu
57. Manjakani Ramuan param, ubat periuk, jamu, tangas basah, isi
tungku
58. Mengkudu Ramuan tangas kering, mandi daun (daun), balut
tungku (daun)
59. Nilam Ramuan mandi daun
60. Pinang Ramuan ubat periuk
61. Padi (Beras) Ramuan param (buah), pilis (buah)
62. Pala Ramuan param
63. Putarwali Ramuan ubat periuk
64. Pandan wangi Ramuan param, tangas basah (daun), mandi daun
(daun), balut tungku (daun)
65. Pegaga Ramuan pilis, ubat periuk, mandi daun
66. Pelaga Ramuan jamu (buah)
67. Rambai Balut tungku (daun)
68. Rambutan Ramuan mandi daun (daun)
69. Serai wangi Ramuan param, tangas basah (daun), mandi daun
(daun), minyak urut, balut tungku (daun)
70. Seringan Ramuan mandi daun (daun)
71. Senduduk Ramuan ubat periuk
72. Sireh Ramuan pilis, ubat periuk, jamu, tangas basah, mandi
daun (daun, batang), balut tungku (daun)
168
RUMUSAN
PENGHARGAAN
RUJUKAN
Ibrahim, Y. (2006). Komuniti Melayu Pulau Tioman: Isu dan Cabaran dalam
Konteks Pembangunan Pelancongan. Jurnal Melayu, 2: 141–153.
Lin, K. W. (2005). Ethnobotanical Study of Medicinal Plants Used by The Jah Hut
Peoples in Malaysia. Indian Journal of Medical Sciences, 59(4): 156.
Zal, W. A., Salleh, H., Omar, M., & Halim, S. (2014). Kebolehcapaian dan
Keterancaman Modal Semula Jadi Orang Asli Lanoh di
Malaysia. Geografia: Malaysian Journal of Society and Space, 10(2): 178–
188.
169
TREND PENGGUNAAN 18 SPESIES TUMBUHAN UBATAN DI BAWAH
PROGRAM NKEA DI KALANGAN PENGAMAL PERUBATAN TRADISIONAL
MELAYU DI SEMENANJUNG MALAYSIA
ABSTRAK
Di bawah program National Key Economic Area (NKEA) Herba EPP#1, sebanyak
18 spesies tumbuhan ubatan telah dikenal pasti sebagai spesies yang
mempunyai potensi ekonomi. Justeru, pendekatan atau pengenalan spesies
tersebut kepada orang ramai adalah perlu untuk memartabatkan lagi
kepentingan dan penggunaan spesies tumbuhan ubatan tersebut dalam
menyumbang kepada peningkatan ekonomi negara. Analisis penggunaan
spesies tumbuhan ubatan di bawah program NKEA oleh 355 orang pengamal
perubatan tradisional Melayu di seluruh Semenanjung Malaysia telah
dilaksanakan hasil daripada pelaksanaan survei dalam projek
Pendokumentasian Pengetahuan Tradisi Melayu Berkaitan Tumbuhan Ubatan
di Semenanjung Malaysia. Empat spesies NKEA yang paling popular digunakan
oleh pengamal ialah mengkudu (33%), dukung anak (29%), halia (26%) dan
senduduk (17%) manakala spesies yang paling sedikit digunakan adalah kelo
(1%) dan rosel (0.3%). Amalan perubatan terutama sekali yang melibatkan
penggunaan tumbuhan dapat menjadi asas kepercayaan atau keyakinan orang
ramai untuk menggunakan tumbuhan tersebut sebagai sumber perubatan,
seterusnya menyumbang kepada usaha untuk memperkenalkan spesies
tumbuhan ubatan yang mempunyai nilai potensi ekonomi kepada masyarakat.
170
PENGENALAN
KAEDAH
171
tumbuhan di kalangan pengamal dilaksanakan bagi mendapatkan maklumat
lebih terperinci tentang trend pengunaan serta tujuan penggunaan tumbuhan
berkenaan.
172
tumbuhan asal di Malaysia. Kelo dipercayai berasal dari India dan rosel juga
dikatakan berasal dari India dan Afrika Barat. Tumbuhan yang diperkenalkan
dari negara luar tersebut mungkin merupakan salah satu faktor ianya tidak
kerap digunakan dan kelihatan sangat asing di kalangan pengamal perubatan
tradisional Melayu.
KESIMPULAN
173
membolehkan para penyelidik melakukan kajian saintifik untuk membuktikan
kesahihan tumbuhan ubatan tersebut dijadikan ubat kepada penyakit tertentu
tanpa memberi kesan mudarat kepada pengguna. Pengetahuan tradisi juga
turut menjadi asas penting untuk memartabatkan spesies komersial
berdasarkan kepentingan dan penggunaannya dalam masyarakat. Namun
demikian, pendekatan yang lebih meluas tentang penggunaan spesies
tumbuhan terutama tumbuhan yang mempunyai potensi ekonomi adalah perlu
supaya populariti penggunaan tumbuhan lebih tinggi seterusnya menyumbang
kepada permintaan yang tinggi terhadap produk herba dalam pasaran.
PENGHARGAAN
RUJUKAN
http://www.worldagroforestry.org/treedb/AFTPDFS/Hibiscus_sabdariffa.PDF
http://www.worldagroforestry.org/treedb/AFTPDFS/Moringa_oleifera.PDF
174
AMALAN PERUBATAN TRADISIONAL MELAYU DI SEMENANJUNG
MALAYSIA: ADAKAH MASIH RELEVAN PADA MASA KINI ?
ABSTRAK
PENGENALAN
175
KAEDAH
1. Profil Pengamal
Daripada 355 pengamal yang ditemu ramah, seramai 229 (64.5%) adalah
perempuan manakala selebihnya, iaitu 126 (35.5%) adalah lelaki. Ini
menunjukkan bahawa penglibatan dalam perubatan tradisional Melayu
dipelopori oleh kaum wanita. Keadaan ini boleh dikaitkan dengan permintaan
terhadap perkhidmatan perbidanan untuk tujuan kesihatan dan kecantikan di
kalangan wanita. Dari segi umur, 62% iaitu seramai 219 individu PTM berumur
lebih daripada 60 tahun, manakala 38% individu lagi (136 orang) terdiri
daripada golongan dalam lingkungan umur 40-60 tahun. Pengamal PTM yang
masih aktif adalah dalam lingkungan umur 51-80 tahun, iaitu melibatkan 321
orang (90%) daripada jumlah keseluruhan pengamal PTM.
176
7.3% (26 individu), campuran sebanyak 5.6% (20 individu) dan yang paling
sedikit ialah bidang perbidanan iaitu 4.8% (17 individu).
3. Pengkhususan Pengamal
4. Produk Pengamal
177
adalah dalam lingkungan 1-50 individu sebulan. Seramai 13 orang pengamal
PTM (4%) mampu merawat lebih daripada 200 pesakit dalam sebulan.
178
Jadual 2. Pendapatan dan purata pendapatan bulanan hasil daripada
perkhidmatan mengikut pengkhususan (n=289)
Pengkhususan Jumlah pendapatan Jumlah Purata
bulanan (RM) pengamal pendapatan
(RM)
Umum 112,183.40 141 796
Perbidanan 34,340 49 701
Khusus 39,850 51 781
Campuran 35,510 48 740
Jumlah 221,883.40 289 768
KESIMPULAN
PENGHARGAAN
179
PERALATAN PENYEDIAAN UBATAN DALAM KAEDAH RAWATAN
PERUBATAN TRADISIONAL MELAYU
AZ Wan Nurul Syafinaz, A Ida Farah, SM Ami Fazlin & BA Siti Khairul
Pusat Penyelidikan Perubatan Herba (HMRC), Institut Penyelidikan Perubatan,
Jalan Pahang, 50588 Kuala Lumpur, Malaysia
Tel: 03-2616 2666 Faks: 03-2693 9335 E-mel: wannurulsyafinaz@gmail.com
ABSTRAK
Salah satu elemen perawatan yang diamalkan oleh masyarakat Melayu ialah
rawatan herba yang mana kemujaraban ubat bergantung kepada bahan yang
digunakan serta cara penyediaan ramuan tersebut. . Peralatan yang sesuai di
dalam penyediaan bahan ramuan dari tumbuhan ubatan sangat penting bagi
mengoptimakan kemujaraban bahan ramuan tersebut. Kajian ini merekod
secara terperinci peralatan yang digunakan oleh pengamal perubatan
tradisional Melayu dalam penyediaan bahan ramuan untuk dijadikan ubatan
dalam merawat sesuatu penyakit. Pengamal dipilih melalui bancian yang
dijalankan melaui projek Pendokumentasian Secara Komprehensif
Pengetahuan Tradisional Melayu Berkaitan Tumbuhan Ubatan di Semenanjung
Malaysia berdasarkan jenis pengetahuan dan peralatan yang digunakan dalam
amalan mereka. Temu bual telah dijalankan dengan menggunakan borang kaji
selidik untuk mengumpul maklumat sejarah, pengalaman pengamal, kaedah
rawatan dan jenis peralatan yang digunakan dalam amalan rawatan mereka.
Peralatan diperolehi daripada pengamal semasa temu bual dijalankan atau
dibeli dari premis berkaitan. Seramai 65 orang pengamal ditemu bual bagi
kajian ini, dan 25 orang pengamal daripadanya menggunakan peralatan khusus
dalam penyediaan ubatan mereka. Sebanyak 21 jenis peralatan penyediaan
tumbuhan ubatan telah dikumpul sepanjang kajian ini. Penyediaan bahan
ramuan ubatan untuk merawat membabitkan proses termasuk menumbuk,
memipis, menggiling, mengasah, merendam, mencampur, meramas, dan
merendang. Peralatan yang dilaporkan dalam kajian ini terbahagi mengikut
kegunaannya untuk memproses bahan ramuan atau dalam proses
penyampaian rawatan itu sendiri. Pendokumentasian ini dapat merekod
amalan tradisional dalam penyediaan tumbuhan ubatan untuk digunakan
dalam perubatan tradisional Melayu. Kajian ini juga dapat memelihara ilmu
perubatan tradisional Melayu daripada lenyap ditelan zaman.
180
PENGENALAN
Daripada 65 orang yang telah ditemu bual bagi kajian ini, seramai 25 orang
pengamal menggunakan alatan khusus dalam penyediaan ubatan. Pengamal
menggunakan peralatan sama ada dalam proses penyediaan bahan ramuan
atau ketika rawatan dijalankan. Pengamal perubatan Melayu menggunakan
beberapa cara dalam proses penyediaan bahan ramuan daripada tumbuhan
181
ubatan. Sebagai contoh; rebus, celur, uli, asah, goreng, tumbuk, jemur, giling,
tanak, rendang, salai, bakar, keringkan, ramas dan rendam (Fatan 2003).
182
Jadual 3. Peralatan memproses
Peralatan Kegunaan
Lesung batu Menumbuk bahan ramuan supaya hancur atau
lumat dalam kuantiti yang sedikit.
Lesung kayu Menumbuk bahan ramuan supaya hancur atau
lumat untuk kuantiti yang besar.
Piring asah Mengasah bahagian tumbuhan ubatan untuk
ubat / piring mendapatkan hasil serbuk atau air daripada asahan
kayu asah ubat tersebut.
Pasu asah ubat Mengasah bahagian tumbuhan ubatan untuk
mendapatkan hasil serbuk atau air daripada asahan
tersebut
Batu giling Menggiling/memipis bahan ramuan sehingga
hancur/lumat.
Kacip Memotong/mengacip bahagian tumbuhan ubatan
mengikut ukuran yang ditetapkan.
183
KESIMPULAN
PENGHARGAAN
RUJUKAN
Abdul Razak, A.K. (2006). Analisis Bahasa Dalam Kitab Tib Pontianak.
Perpustakaan Negara Malaysia, Kuala Lumpur. 192pp.
184
PHYSICO─CHEMICAL PROPERTIES OF SPRAY DRIED CLINACANTHUS
NUTANS EXTRACT
ABSTRACT
Clinacanthus nutans or Sabah snake grass (SSG) powder was produced using an
Anhydro Lab S1 spray dryer. The powders were produced using three different
extract formulations as practised by You Dun Chao Healthcare Products.
Maltodextrin was blended with the extract and acts as an encapsulating agent.
The spray drying parameters were air inlet temperature of 1500C and outlet
temperature ranging from 75 to 850C, feed pump speed of 250 rpm and
compressed air pressure for atomization of 0.05 MPa. The SSG powders
produced were tested for water activity, bulk density, colour (L*, a*, b*),
moisture content, protein, oil content, ash and crude fibre. Results showed
that the ash, protein, oil and fibre contents were about the same as the
amount found in most salad herbs of Malaysia. Colorimetric analyses showed
that the L*, a* and b* values of the SSG powder showed significant differences
(p<0.05) among the formulations.
INTRODUCTION
185
several advantages over the liquid form for industrial purposes, such as high
stability, easy to be standardized, transported and stored (Oliveira et al. 2006).
It also facilitates the manufacturing of solid dosage forms in the production of
tablets or capsules, which represent most of the medicines used worldwide
(Leuenberger & Lanz 2005).
The spray drying process requires a drying aid to facilitate drying. The most
common drying aid used is maltodextrin (Bhandari et al. 1997). It helps to
reduce the agglomeration problems during storage, therefore improving
product stability (Bhandari et al. 1993; Silva et al. 2006). Maltodextrin is a
starch hydrolysate prepared by partial hydrolysis of corn starch or potato
starch using suitable acids and enzymes (USFDA). It is also classified as material
that has a dextrose equivalent of 3 to 20 with characteristics of no sweetness,
bland and not masking other flavours. Dextrose equivalent refers to the
percentage of reducing sugar in syrup calculated as dextrose on a dry weight
basis.
The aim of the study was to produce Sabah snake grass powder using a
spray dryer and evaluating its physico-chemical properties.
Raw Materials
The Sabah snake grass (SSG) leaves were obtained from You Dun Chao Herbs
Farm Sdn. Bhd while the extract was prepared by You Dun Chao Healthcare
Products Sdn. Bhd for spray drying. The maltodextrin was donated by ITS
Interscience Sdn. Bhd, Shah Alam, Selangor.
186
Spray Drying Procedures of SSG
The SSG extract was blended with 10-12 DE maltodextrin. The total soluble
solid of the mixture was measured using a hand held refractometer (Atago,
Master-alpha).The feed mixtures were stirred constantly to ensure feed
homogeneity during spray drying. The mixture was spray dried using Lab 1 APV
spray drier with inlet temperature at 150oC resulting with an outlet
temperature of 75oC to 85oC. The feed was pumped through the pressure
nozzle atomizer with a counter-current hot air movement produced by the
heater and the blower. The compressed air pressure for atomizing was set at
0.05 MPa.
1. Moisture content
The moisture content was determined by drying the SSG powder in an oven at
1050C overnight. Drying was continued until a constant reading was obtained.
The weight of the SSG powder before and after the drying process was
recorded. The moisture content of the powder was calculated.
2. Water activity
Water activity of SSG powder was measured using a water activity meter
(Aqualab Series 3, USA).
3. Bulk density
Bulk density of the powder was measured by weighing 10 g of sample and
placing it in a 100 ml graduated cylinder. A steady vibration was conducted on
a vibrator (Main et al. 1978). The volume was then recorded and used to
calculate bulk density as g/ml.
4. Colour (L, a, b)
Colour was measured using a colour reader (Minolta chromameter, model CR
300/colorimeter series). The values of L (Lightness), a (redness) and b
(yellowness) were determined.
5. Protein.
Crude protein was determined according to the Kjeldahl method using the
conversion factor of 6.25 (AOAC, 1990).
6. Oil content
Oil content was determined according to AOAC method 920.39C (AOAC, 1990).
187
7. Crude fiber
Crude fiber was determined using the fibretec system (according to Weende).
8. Ash
Ash content was determined according to AOAC method 923.03 (AOAC 1990).
Statistical analysis
Three formulations, SSG3 (addition of lemon juice), SSG5 (normal) and SSG8
(concentrated) were analysed in triplicate. Analysis of variance was carried out
to differentiate the means of the variables for the samples. The results were
expressed as mean values standard deviation (SD).
Physicochemical properties
Moisture content of SSG powder ranges from 0.51 to 0.91% (Table1) and no
differences (p>0.05) in moisture content were detected between formulations.
The least amount of moisture shows the rate of heat transferred to the
particles was high. The production of low moisture content powder is
important in spray drying where food powder with 10% moisture content helps
to retard mould growth (Hagan 2007).
Water activity for all powders were less than 0.5. According to Quek et
al. (2007), food that has water activity less than 0.6 is classified as
microbiologically stable. This indicates the powder might have longer shelf life.
The bulk density of the powder varied from 0.37 to 0.42 g/ml. A high
drying temperature would have caused higher surface ratio to volume for the
spray dried powder particles. In addition, the lesser amount of compressed air
for atomizing may produce larger particle size of the powder. This may lead to
lower bulk density of the powder. The larger the size of the particles would
have resulted in more trapped air between them. Thus, leading to a greater
possibility for oxygen degeneration of the pigment and reducing storage
stability.
The percentage of protein, fiber, ash and oil content in all the samples
were about the same as the amount found in most salad herbs of Malaysia as
reported by Wan Hassan and Mustaffa (2010).
188
Table 1. Physicochemical composition of spray dried SSG powder
Variables SSG 3 SSG 5 SSG 8
(Addition of (Normal) (Concentrated)
lemon juice)
Moisture content (%) 0.51 ± 0.32a 0.59 ± 0.06a 0.91 ± 0.52a
Water activity, aw 0.21 ± 0.01c 0.30 ± 0.01b 0.35 ± 0.02a
b a
Ash (%) 0.57 ± 0.01 0.84 ± 0.11 0.76 ± 0.05a
b a
Protein (%) 0.08 ± 0.00 0.16 ± 0.02 0.16 ± 0.01a
Oil content (%) 5.35 ± 2.74ab 10.75 ± 5.27a 2.11 ± 1.10b
a
Fibre (%) 0.51 ± 0.24 0.56 ± 0.29a 0.45 ± 0.12a
Bulk density (g/ml) 0.37 ± 0.14a 0.39 ± 0.05a 0.42 ± 0.01a
*Means with the same letter are not significantly different (for variables
Colour
The colour on the SSG powder was measured (Figure 1). The L*, a*, and b*
values showed significant differences (p<0.05) among the formulations. The
highest value for L* was given by SSG 3 indicating lighter product towards
whiteness. This might be due to the addition of lemon juice that impaired the
green pigment in the extract. SSG 5 had a* negative value corresponding to a
greenness although the extract is more diluted compared to SSG 8. SSG 5 had
the highest b* value. This indicates that the yellowness of the product was not
influenced by the lemon juice in SSG 3.
189
CONCLUSION
Sabah snake grass powder was obtained using 10% maltodextrin at inlet
temperature of 150oC. Addition of lemon juice in the extract helps to mask the
unpalatable taste in normal extract of SSG. Further study is required to look
into its potential as a health supplement.
ACKNOWLEDGEMENTS
The author thanks Mr. Hazmi bin Haron and Mr. L.E. Chong from You Dun Chao
Healthcare Products & You Dun Chao Herbs Farm Sdn. Bhd for the supply of
Sabah snake grass extract.
REFERENCES
Bhandari, B.R., Snoussi, A., Dumoulin, E.D. & Lebert, A. (1993). Spray Drying of
Concentrated Fruit Juices. Drying Technology 11(5):1081–1092.
Bhandari, B.R., Datta, N. & Howes, T. (1997). Problems Associated with Spray
Drying of Sugar-Rich Foods. Drying Technology 15 (2): 671–684.
Hagan, A.T. (2007). Part 1X of Food Storage FAQ. Shelf Reliance. http://
www.shelfreliance.com/library/view/57 (accessed April, 2009).
Main, J.H., Clydesdale, F.M. & Francis, F.J. (1978). Spray Drying Anthocyanin
Concentrates for Use as Food Colorant. J. Food Sci. 43: 1693–1694.
National Drug Committee. (2006). List of Herbal Medicinal Products A.D. 2006.
ISBN: 974-244-217-7, Chuoomnoom Sahakorn Karnkaset Publisher,
Bangkok, Thailand. Pp. 59–61.
Oliveira, W.P., Bott, R.B. & Souza, C.R.F. (2006). Manufacture of Standardized
Dried Extracts from Medicinal Brazilian Plants. Drying Technology
24(4): 523–533.
190
Quek, S.Y., Chok, N.K. & Swedlund, P. (2007). The Physicochemical Properties
of Spray Dried Watermelon Powder. Chemical Engineering and
Processing 46(5): 386–392.
Reineccius, G.A. & Risch, S.J. (1988). Spray Drying of Food Flavors. In Flavor
Encapsulation. Washington D.C: American Chemical Society. Pp. 55–66.
Silva, M.A., Sobral, P.J.A. & Kieckbusch, T.G. (2006). State Diagrams of Freeze-
Dried Camu-Camu (Myrciaria dubia (HBK) Mc Vaugh) Pulp With and
Without Maltodextrin Addition. Journal of Food Engineering 77(3):
426–432.
Teixeira, C.C.C., Teixeira, G.A. & Freitas, L.A.P. (2011). Spray Drying of Extracts
from Red Yeast fFrmentation Broth. Drying Technology 29: 342–350.
Wan Hassan,W.E. & Mustaffa, M. (2010). Ulam Species. In Ulam: Salad Herbs of
Malaysia. Masbe Publisher, Kuala Lumpur, Malaysia. Pp. 40–239.
191
EXTRACTION OF ANTIOXIDANTS FROM LEAVES OF CLINACANTHUS
NUTANS (BURM.F.) LINDAU: EFFECTS OF EXTRACTION METHOD AND
SOLVENT
NA Karim1 & II Muhamad 1,2
1
Department of Bioprocess Engineering, Faculty of Chemical Engineering,
Universiti Teknologi Malaysia (UTM), Johor Bahru, 81310 Johor
2
IJN-UTM Cardiovascular Engineering Centre, Faculty of Bioscience and
Medical Engineering, Universiti Teknologi Malaysia, Johor Bahru, 81310 Johor
Tel: 07-5535577/ 07-5558564 Fax: 07-5588166 E-mail: idayu@cheme.utm.my
ABSTRACT
The aim of this study was to determine the best method for extracting
antioxidants from Clinacanthus nutans (Burm.f.) Lindau, (C. nutans) leaves. The
extraction was done by four different methods: decoction (100ᵒC, 20 min),
soxhlet (8 hour), maceration (3 day) and ultrasound-assisted extraction (25ᵒC,
40 min) using different solvents: ethanol, methanol and water respectively.
Antioxidant activity, total phenol, and flavonoid content were determined from
the various extraction methods. Total phenolic and flavonoid contents were
determined by Folin–Ciocalteau and aluminum chloride methods,
respectively. Antioxidant activity was detected by 2,2-diphenyl-1-picrylhydrazyl
(DPPH) free radical scavenging activity and reducing capacity assessment
(FRAP). The results showed that the extraction method and solvent used
significantly affect the extraction of antioxidant from the leaves. Conventional
extraction method (water extracts) proved to be the best method with
maximum yield (31.60 ± 0.44 %, w/w), total phenolic (24.76 ± 3.39 mg gallic
acid equivalent per g dry weight sample) and flavonoid content (6.25 ± 0.42 mg
quercetin equivalent per g dry weight sample). It also showed strongest DPPH
free radical scavenging activity (16.29 ± 2.69 mg trolox equivalent antioxidant
capacities (TEAC)/g dry weight sample) and FRAP assay (19.40 ± 0.24 TEAC/g
dry weight sample). As a conclusion, C. nutans leaves are potential natural
antioxidant for the development of nutraceutical and functional food.
INTRODUCTION
192
potential in the treatment and prevention of cancer, cardiovascular and other
chronic diseases (Karabegovic et al. 2014). Clinacanthus nutans (Burm.f.)
Lindau (C. nutans), belonging to the family of Acanthaceae is native to the
Southeast Asia region and is cultivated in Thailand, Malaysia and Indonesia. It is
an edible erect herbal plant, characteristically described as a finely pubescent
scrub with a drooping and clambering trait (Quattrocchi 2012). It has been
reported to have biological activities and high medicinal values, including
antioxidant activity (Yong et al. 2013).
193
number SK2781/2015. The leaves and stems were separated and washed
thoroughly with tap water, oven dried at 50oC homogenized to fine powder.
They were kept in an airtight plastic 4°C until further extraction.
Antioxidant Assays
Total antioxidant activity was analysed using two assays: DPPH free radical
scavenging assay based on method by Abu Bakar et al. (2009) and ferric-
reducing antioxidant power assay (FRAP) according to the procedure of Benzie
and Strain (1996). Trolox (0.02 – 0.10 mg/ml) was used as reference standard.
The antioxidant activity was expressed as mg of Trolox equivalents antioxidant
capacity (TEAC) per gram of plant material on a dry weight as it showed
accurate and descriptive expression than assays that express antioxidant
194
activity as the percentage decrease in absorbance. The results provide direct
comparison of the antioxidant activity with Trolox.
The total phenolic content (TPC) and total flavonoid content (TFC) determined
in C. nutans leaves extracts obtained by different extraction methods and
solvents were depicted in Figure 2. The TPC varied from 3.88 to 44.76 mg of
GAE/g of dry mass sample and the TFC ranged from 1.33 to 12.22 mg of QE/g
of dry mass sample. The aqueous extracts of C. nutans obtained by all studied
methods had highest TPC values (ranging from 30.24 to 44.76), as compared to
195
alcoholic extracts (TPC: water > methanol > ethanol). From each different
extraction methods tested, soxhlet and decoction methods showed higher TPC
value with 6.51 to 45.19 and 9.48 to 44.76 mg GAE/g dry weight sample,
respectively. These results showed that phenolic contents were strongly
dependent on polarity of solvents used, which polar fractions (water) had more
phenolics in them than the less-polar fractions (methanol/ethanol) (Hayouni et
al. 2007). The TFC results showed a contradict trend as compared to TPC. The
ethanolic extracts exhibited a higher amount of TFC as compared to methanolic
and aqueous extracts in soxhlet method (TFC: ethanol > methanol > water).
These could be attributed to the solubility of flavonoid compounds which is
soluble in alcoholic solvents. The flavonoid content in C. nutans leaves could be
from flavones C-glycosides (shaftoside, isomollupentin 7-O-𝛽-glucopyranoside,
orientin, isoorientin, vitexin, and isovitexin) as previously reported by Teshima
et al. (1998). It was also reported that compounds such as flavonoid, which
contain hydroxyl functional groups, are responsible for antioxidant effects in
the plants (Jaberian et al. 2013).
Figure 2. Total phenolic (A) and total flavonoid contents (B) of C. nutans leaves
by different extraction methods and solvents
196
activity in sonication and maceration. Heat treatment applied in soxhlet and
decoction methods increased the concentration of antioxidant in C. nutans, in
which the heat treatment enhanced diffusivity, soften the plant tissues and
promoted elution of attached bioactive compounds (Cabana et al. 2013).
CONCLUSION
Decoction method (aqueous extracts) of C. nutans was found to be the best
extraction method and solvent as it showed high extraction efficiency and
strong antioxidants in term of high TPC and TFC values. In addition, C. nutans
leaves also showed significantly higher antioxidant contents for both DPPH and
FRAP assays. Therefore, the traditional decoction method was still a reliable
extraction process for and was considered as environmental-friendly, simple,
safe, reproducible and inexpensive method for extracting antioxidant.
ACKNOWLEDGEMENTS
REFERENCES
Abu Bakar, M.F.A., Mohamed, M., Rahmat, A. & Fry, J. (2009). Phytochemicals
and Antioxidant Activity of Different Parts of Bambangan (Mangifera
Pajang) and Tarap (Artocarpus Odoratissimus). Food Chemistry. 113:
479–483.
197
Azmir, J., Zaidul, I.S.M., Rahman, M.M., Sharif, K.M., Mohamed, A., Sahena, F.,
Jahurul, M.H.A., Ghafoor, K., Norulaini, N.A.N. & Omar, A.K.M. (2013).
Methods for Extraction of Bioactive Compounds from Plant Materials: A
Review. Journal of Food Engineering. 117: 426–436.
Bampouli, A., Kyriakopoulou, K., Papaefstathiou, G., Louli, V., Krokida, M. &
Magoulas, K. (2014). Comparison of Different Extraction Methods of
Pistacia lentiscus var. Chia Leaves: Yield, Antioxidant Activity and
Essential Oil Chemical Composition. Journal of Applied Research on
Medicinal and Aromatic Plants.1: 81–91.
Benzie, I.F. & Strain, J.J. (1996). The Ferric Reducing Ability Of Plasma (Frap) as
a Measure of "Antioxidant Power": The Frap Assay. Analytical
Biochemistry. 239(1): 70─76.
Cabana, R., Silva, L.R., Valentao, P., Viturro, C.I. & Andrade, P.B. (2013). Effect
of Different Extraction Methodologies on The Recovery of Bioactive
Metabolites from Satureja parvifolia (Phil.) Epling (Lamiaceace).
Industrial Crops and Products. 48: 49–56.
Hayouni, E.A., Abedrabba, M., Bouix, M. & Hamdi, M. (2007). The Effects of
Solvents and Extraction Method on The Phenolic Contents and Biological
Activities In Vitro of Tunisian Quercus coccifera l. and Juniperus
phoenicea l. Fruit Extracts. Food Chemistry. 105: 1126–1134.
Jaberian, H., Piri, K. & Nazari, J. (2013). Phytochemical Composition and In Vitro
Antimicrobial and Antioxidant Activities of Some Medicinal Plants. Food
Chemistry. 136: 237–244.
Karabegovic, I.T., Stojicevi, S.S., Velickovic, D.T., Todorovic, Z.B., Nikolic, N.C. &
Lazic, M.L. (2014). The Effect of Different Extraction Methods on The
Composition and Antioxidant Activity of Cherry Laurel (Prunus
laurocerasus) Leaves and Fruit Extracts. Industrial Crops and Products.
54: 142–148.
Sulaiman, N., Ida Idayu, M., Ramlan, A.Z., Nur Fashya, M., Nor, Farahiyah, A.N.,
Mailina, J. & Nor Azah, M.A. (2015). Effects of Extraction Methods on
Yield And Chemical Compounds Gaharu (Aquilaria malaccensis) Journal
of Tropical Forest Science. 27(3): 413─419
198
Teshima, K., Kaneko, T., Ohtani, K., Kasai, R., Lhieochaiphant, S.,
Picheansoonthon, C. & Yamasaki, K. (1998). Sulfur-Containing Glucosides
from Clinacanthus nutans. Phytochemistry. 48(5): 831─835.
Yong, Y.K., Tan, J.J., The, S.S., Mah, S.H., Ee, G.C.L., Chiong, H.S. & Ahmad, Z.
(2013). Clinacanthus nutans Extracts are Antioxidant with
Antiproliferative Effect on Cultured Human Cancer Cell Lines. Evidence-
Based Complementary and Alternative Medicine. 1─8.
199
ANTI-PROLIFERATIVE EFFECT OF CLINACANTHUS NUTANS ON
OVARIAN, BREAST AND COLORECTAL CANCER CELL LINES
ABSTRACT
INTRODUCTION
200
circulatory, respiratory and infectious diseases (Ministry of Health 2014).
Cancer starts from over proliferation of abnormal cells that can spread to other
parts of its origin and often diagnosed at late stage, thus may reduce chance of
survival. Often, the treatment used includes chemotherapy that is the use of
drug either alone or in combination with other drug or mode of treatment (i.e.
radiotherapy, immunotherapy, etc.). The role of chemotherapy is to kill the
cancer cells but sometimes this treatment is ineffective due to the toxic side
effects and drug resistance problems. Hence, the searches of new drug
candidate that can reduce these problems are in dire needs.
General Procedure
1
H and 13C NMR spectra were recorded in ppm (δ) in CDCl3, CD3OD and DMSO-
d6 employing a Bruker DRX 300 spectrometer operating at 300 MHz for 1H and
75 MHz for 13C, respectively. Column chromatographies were performed with
silica gel 60 (0.040-0.063 mm, Merck, Germany), MCI gel CHP 20P (75-150 µm,
Supelco, USA), Diaion HP-20 (250-850 µm, Supelco, USA), Diaion HP-20SS (75-
150 µm, Supelco, USA), Chromatorex ODS (100-200 mesh, Fuji Silysia Chemical,
Ltd.) and Sephadex LH-20 (GE Healthcare Bio-Science AB, Sweden). TLC was
performed on precoated Si gel 60 F254 plates (0.2 mm thick, Merck) and spots
were detected by UV illumination and by spraying with 10% H2SO4 solution
followed by heating.
201
Plant Materials
The fresh aerial part of C. nutans was purchased from Pusat Pertanian Pantai,
Jalan Pantai Batu 7, Pantai, Seremban. The leaves were separated from the
stems. Then they were dried in an oven at 40–45°C, followed by grinding into
powder.
Preparation of Extract
Ovarian (SKOV-3), breast (MCF-7) and colorectal (HT-29) cancer cell lines were
purchased from American Type Culture Collections, USA. The cell lines were
cultured and sub-cultured in Dulbecco’s Modified Eagle’s medium (DMEM)
supplemented with 5% fetal bovine serum (FBS), 1% penicillin-streptomycin,
0.25% amphotericin B and 1% gentamycin. Approximately 4000 to 6000 cells
were seeded in each well of the 96 well plates and incubated in a humidified
incubator at 37C and 5% carbon dioxide in air for 24 h. Each cell line was then
treated at five different concentrations of C. nutans’s extracts and fractions (1,
5, 25, 125, 625 µg/mL) and compounds (1, 10, 20, 50 100 µg/mL) in triplicate.
At least 24 wells were untreated with C. nutans in each plate. Cisplatin, a
known chemo-drug was also treated on these cell lines at different
concentrations (0.032, 0.16, 0.8, 4, 20 µg/mL) as for comparison studies. The
202
cells were then incubated in the same incubator with the mentioned
conditions for 72 h. The experiment was repeated at least three times.
The isolation and purification work on the hexane and water fractions of leaves
has resulted in the identification of 4 known compounds in the leaves as
shaftoside (1) (Osterdahl 1979, Kondo et al. 2000), stigmasterol (3), β-sitosterol
(4) (Vencata et al. 2012a) and lupenone (6) (Prachayasittikul et al. 2010).
Among these, lupenone was also obtained from the stems. Additionally,
structure analysis of three compounds, i.e., 2, 5 and 7, are still on-going.
CH3
CH3
HO HO HO
H3C CH3
203
Anti-proliferative Effects of Extracts, Fractions and Compounds from C.
nutans
204
Table 1. The IC50 values of the C. nutans samples (µg/mL) treated in ovarian
(SKOV-3), breast (MCF-7) and colorectal (HT-29) cancer cell lines
Cancer Cell Line SKOV-3 MCF-7 HT-29
C. nutans Name/ Codes
samples
Extract Methanol 292.63 ± 359.17 ± 330.40 ± 9.79
11.58 20.22
Hexane 311.38 ± 320.11 ± 341.39 ± 6.35
0.35 5.37
Ethyl Acetate 254.61 ± 268.93 ± 282.10 ± 11.65
33.69 16.11
Aqueous 284.58 ± 297.03 ± undetermined
18.02 4.07
205
CONCLUSION
ACKNOWLEDGEMENTS
REFERENCES
American Cancer Society. (2011). Global Cancer Facts & Figures 2nd Edition.
American Cancer Society Inc., Atlanta. 57pp.
Arullappan, S., Rajamanickam, P., Thevar, N. & Kodimani, C.C. (2014) In vitro
Screening of Cytotoxic, Antimicrobial and Antioxidant Activities of
Clinacanthus nutans (Acanthaceae) Leaf Extracts. Tropical Journal of
Pharmaceutical Research: 13(9): 1455─1461.
Awad, A.B. & Fink, C.S. (2000). Phytosterols as Anticancer Dietary Components:
Evidence and Mechanism of Action. The Journal of Nutrition: 130(9):
2127─2130.
Chelyn, J.L., Omar, M.H., Mohd Yousof, N.S.A., Ranggasamy, R., Wasiman, M.I.
& Ismail, Z. (2014). Analysis of Flavone C -Glycosides in The Leaves of
Clinacanthus nutans (Burm. f.) Lindau by HPTLC and HPLC-UV/DAD.
Scientific World Journal: 2014:724267. doi: 10.1155/2014/724267. Epub
2014 Oct 22.
206
Kongkaew, C. & Chaiyakunapruk, N. (2011). Efficacy of Clinacanthus nutans
Extracts in Patients with Herpes Infection: Systematic Review and Meta-
Analysis of Randomised Clinical Trials. Complementary Therapies in
Medicine: 19(1):47─53.
Ministry of Health. (2014). Health Facts 2014. Ministry of Health Virtual Library,
Malaysia. 18 pp.
Newman, D.J. & Cragg, GM. (2012). Natural Products as Sources of New Drugs
over the 30 Years from 1981 to 2010. Journal of Natural Products: 75(3):
311- 335.
Skehan, P., Storeng, R., Scudiero, D., Monks, A., McMahon, J., Vistica D.,
Warren, J.T., Bokesch, H., Kenne,y S., Boyd, M.R. (1990). New
Colorimetric Assay for Anticancer-Drug Screening. Journal of National
Cancer Institute: 82: 1107–1112.
207
Venkata, C., Prakash., S. & Prakash, I. (2012b). Isolation and Structural
Characterization of Lupine Triterpenes from Polypodium vulgare.
Research Journal of Pharmaceutical Sciences: 1(1): 23─27.
Wanikiat, P., Panthong, A., Sujayanon, P., Yoosook, C., Rossi, A.G., Reutrakul, V.
(2008). The Anti-Inflammatory Effects and The Inhibition of
Neutrophil Responsiveness by Barleria Lupulina and Clinacanthus nutans
Extracts. Journal of Ethnopharmacology: 116(2): 234─244.
Yang, H.S., Peng, T.W., Madhavan, P., Shukkoor, M.S.A., Akowuah, G.A. (2013).
Phytochemical Analysis and Antibacterial Activity of Methanolic Extract
of Clinacanthus nutans Leaf. International Journal of Drug Development
and Research: 5(3): 349─355.
Yong Y.K., Tan J.J., Teh S.S., Mah S.H., Ee G.C.L., Chiong H.S., Ahmad Z. (2013).
Clinacanthus nutans Extracts are Antioxidant with Antiproliferative Effect
on Cultured Human Cancer Cell Lines. Evidence-based Complementary
and Alternative Medicine. 2013:462751. doi: 10.1155/2013/462751.
Epub 2013 8pp.
208
DETECTION OF IRRADIATED HERBS USING PHOTO-STIMULATED
LUMINESCENCE TECHNIQUE (PSL)
ABSTRACT
INTRODUCTION
Food irradiation has been proven as one of the alternative techniques to the
existing methods for increasing the shelf-life, improving the hygienic quality,
and enhancing the functional properties of different food items (Akram et al.
2012). An important problem is to develop method for unambiguous
identification of irradiated foodstuff even in the absence of a non-irradiated
(control) sample.
209
probably due to variation of mineral grains accidentally being on the surface of
the sample layer (Bayram & Delincee 2004). The PSL technique has been
validated by the European Committee for Standardization (EN 13571, 2009) for
food such as spices and aromatic herbs. In view of the increasing use of
irradiation technology for decontamination of herbs, this study aims to
investigate whether PSL measurement can be employed to identify three
different types of Malaysian herbs.
Three types of herbs, namely mas cotek (Ficus deltoidea), misai kucing
(Orthosiphon stamineus) and hempedu bumi (Andrographis paniculata) were
purchased from local market and packed in black polyethylene bags (about 10
g each bag). The samples were exposed to γ-radiation at the dose of 5 kGy at
room temperature using a cobalt-60 gamma source (Gamma cell 220, at
Malaysian Nuclear Agency).
210
RESULTS AND DISCUSSION
The PSL photon counts (PCs) for all samples, measured as a function of
irradiation dose are presented in Figure 1. The PCs of all non-irradiated
samples were less than the lower threshold value (700 counts/60 s), indicating
them as negative (T1, non-irradiated). The PCs of the 5 kGy irradiated herbs
measured were higher than the upper threshold value
(T2 = 5000 counts/60 s) indicating irradiation treatment. Among the three
herbs, mas cotek recorded the highest PCs with values over three times more
compared to misai kucing and hempedu bumi.
Figure 1. Effect of irradiation dose on the signal intensity of mas cotek, misai
and hempedu bumi
211
quantity of silicate materials such as feldspar and quartz contaminating the
herbs, as reported by Bayram & Delincée (2004). Based on the results obtained,
it was clear that index sensitivity value is a better indicator than using the PSL
response to discriminate between non-irradiated and irradiated samples.
CONCLUSION
ACKNOWLEDGEMENTS
The authors wish to express their gratitude to the Malaysian Nuclear Agency
(MOSTI) for their support in this project.
REFERENCES
Akram, K., Ahn, J. & Kwon, J. (2012). Chapter 1. Analytical Methods for the
Identification of Irradiated Foods. Pp. 1─36 in Belotserkovsky, E. &
Ostaltsov, Z. (eds.) Ionizing Radiation.
212
European Standard EN 13751, (2009). Foodstuff - Detection of Irradiated Food
Using Photostimulated Luminescenc. Brussels, Belgium: European
Committee for Standardization.
Sanderson, D.C.W., Carmichael, L.A., Spencer, J.Q. & Naylor, J.D. (1996). Recent
Advances in Thermoluminescence and Photo-Stimulated Luminescence
Detection Methods for Irradiated Foods, International Meeting on
Analytical Detection Methods for Irradiation Treatment of Foods. Pp.
124–38 in McMurray, C.H., Stewart, E.M., Gray, R. & Pearce, J. (eds.).
Detection Methods for Irradiated Food—Current Status. Cambridge,
U.K.: Royal Society of Chemistry.
Zhang, L., Lin, T., Jiang, Y. & Bi F. (2013). A New Criterion of Photostimulated
Luminescence (PSL) Method to Detect Irradiated Traditional Chinese
Medicinal Herbs. Radiation Physics and Chemistry 92: 105–111.
213
OPTIMIZATION OF ANTIOXIDANT ACTIVITY IN CLINACANTHUS NUTANS
(BELALAI GAJAH/SABAH SNAKE GRASS)
ABSTRACT
This study focused on the effects of solvents, extraction time and repeated extraction
towards antioxidant properties of Clinacanthus nutans (Belalai Gajah/Sabah Snake
Grass). Freeze-dried stems were homogenized before testing its antioxidant activity:
folin–ciocalteu index (TPC), ferric reducing/antioxidant power (FRAP) and radical-
scavenging activity (DPPH) on the effect of different solvents (absolute methanol,
ethanol, acetone, and their aqueous solutions at 50%, 70%, and 100% concentrations),
st nd rd
time of extraction (15, 30 and 60 min) and repeated extraction (1 , 2 and 3 times).
The data obtained were statistically analyzed using SPSS Version 22 (Chicago, Inc.).
st
High antioxidant activity was shown on the 1 extraction session of 0.1 g of sample
immersed with 10 ml of 70% acetone in 60 minutes extraction time. In conclusion,
high antioxidant activity was observed in C. nutans, which can be contributed as
natural antioxidant source for human consumption. More optimization options on C.
nutans can be conducted to cultivate more nutrients from the medicinal plant and thus
contributing for human health.
INTRODUCTION
214
promotes good health, prevents breast cancer, promote bowel movement, skin
care and goiter in Singapore (Lau et al. 2014). However, there is not much
study on its antioxidant activities. This study is focused on the effects of
solvents, extraction time and repeated extractions toward antioxidant
properties of C. nutans.
Freeze-dried stems were homogenized into 0.5 mm size before extraction. The
extraction procedure was conducted with (0.1 g) dried samples and 10 mL
extraction solvent for overnight. Solvents systems used were absolute
methanol, ethanol, acetone (Merck, Germany), and their aqueous solutions at
50%, 70%, and 100% concentrations (Musa et al. 2011). All extracted samples
were centrifuged using tabletop centrifuge (Kubota, Japan) for 10 min at
2,580×g. The supernatants were collected for further analysis. In the second
part of this study, extraction time (15, 30 and 60 min) and extraction sessions
(1st, 2nd and 3rd times) were studied using the best solvent selected in the first
part of the study. All tests such as Folin–Ciocalteu Index (Slinkard & Singleton
1977), Ferric Reducing/Antioxidant Power (Benzie & Strain 1996) and Radical-
Scavenging Activity (Musa et al. 2011) were performed at room temperature.
All the experiments were done in triplicates. The data obtained were
statistically analyzed using SPSS Version 22 (Chicago, Inc.) using one-way
ANOVA followed by Duncan’s Multiple Range Test.
Strong antioxidant activity was shown by solvent 70% acetone in TPC (177.69
mg of gallic acid equivalent per g of dried sample) and DPPH (231.87 mg of
trolox equivalent per g of dried sample), which was similar to the antioxidant
activity observed in pink-flesh guava of Musa et al. (2011), in a similar method.
For the effect of extraction time on antioxidant study, extraction time of 60
min exhibited the highest (p<0.05) antioxidant activities in TPC and FRAP
(221.64 mg gallic acid equivalent per g of dried sample in TPC, 3.26 mg of trolox
equivalent per g of dried sample in FRAP and 2.38 mg of trolox equivalent per g
of dried sample in DPPH). The TPC results of Lusia et al. (2015) stated 177.80 ±
19.10 mg TAE/ L for C. nutans tea, which is observed in 20 min infusion of
unfermented microwave-oven dried leaves. The results were much lower than
the current results of 15 min (886.84 mg gallic acid equivalent per g of dried
sample in TPC). There was an ascending trend on the antioxidant activities
when the extraction time was prolonged. The first extraction showed the
highest (p<0.05) antioxidant activity than the second and third extraction
215
(207.59 mg gallic acid equivalent per g of dried sample in TPC, 2.76 mg of trolox
equivalent per g of dried sample in FRAP and 2.69 mg of trolox equivalent per g
of dried sample in DPPH). More optimization options on C. nutans can be
conducted to cultivate more nutrients from the medicinal plant and thus
contributing towards human health. Future studies will focus on antimicrobial
activity, nutritive value and product development on C. nutans.
CONCLUSION
st
High antioxidant activity was shown on the 1 extraction session of 0.1 g of sample
immersed with 10 ml of 70% acetone in 60 min extraction time. Since high antioxidant
activity was observed in C. nutans, thus it can function as a good natural antioxidant
source for human consumption.
ACKNOWLEDGEMENT
The authors would like to thank Universiti Kebangsaan Malaysia for providing
the postgraduate scholarship (Zamalah), food antioxidant research lab facilities
and the following project funds: FRGS/1/2014/STWN03/UKM/02/1, GSP-2013-
019 and STGL-007-2008.
REFERENCES
Benzie, I. F. & Strain, J. J. (1996). The Ferric Reducing Ability of Plasma (FRAP) as a
Measure of Antioxidant Power the FRAP Assay. Analytic Biochemistry 239:
70─76.
Lau, K. W., Lee, S. K. & Chin, J. H. (2014). Effect of the Methanol Leaves Extract Of
Clinacanthus nutans on the Activity of Acetylcholinesterase In Male Mice.
Journal of Acute Disease: 3.
Lusia, B. M., Hasmadi, M., Zaleha, A. Z. & Mohd Fadzelly, A. B. (2015). Effect of
Different Drying Methods on Phytochemicals and Antioxidant Properties of
Unfermented and Fermented Teas From Sabah Snake Grass (Clinacanthus
nutans Lind.) Leaves. International Food Research Journal 22(2): 661─670.
Musa, K. H., Abdullah, A., Jusoh, K. & Subramaniam, V. (2011). Antioxidant Activity of
Pink-Flesh Guava (Psidium guajava L.): Effect of Extraction Techniques and
Solvents. Food Analysis Methods 4(2011): 100–107.
216
Siew, Y. Y., Zareisedehizadeh, S., Seetoh, W. G., Neo, S. Y., Tan, C. H. & Koh, H. L.
(2014). Ethnobotanical Survey of Usage of Fresh Medicinal Plants in
Singapore. Journal of Ethnopharmacology 155(3): 1450─1466.
Slinkard, K. & Singleton, V. (1977). Total Phenol Analysis; Automation and Comparison
with Manual Methods. American Journal of Enology and Vitivulture
28:49─55.
217
EFFECTS OF EQUAL DOSES OF SULFORAPHANE, CURCUMIN AND
QUERCETIN ON HEME OXYGENASE 1 GENE EXPRESSION IN MICE LIVER
ABSTRACT
INTRODUCTION
218
chemicals found in food and phenolic antioxidants are chemoprotective i.e.
they induce the expression of genes and proteins involved in cellular defense.
The increased expression of cellular defense proteins provided added
protection against oxidative/chemical stress. Some of these proteins belong to
phase II drug metabolizing enzymes, although other enzymes and antioxidant
proteins were also involved. These proteins are known as phase II proteins
(Wattenberg 1978, Prestera et al. 1993, Talalay & Fahey 2001). Phase II
proteins are mainly regulated by the nuclear factor E2-related factor 2/
antioxidant response element (Nrf2/ARE) system (Kitteringham et al. 2010,
Abdullah et al. 2012). Heme oxygenase-1 (HO-1) is an example of Phase II
proteins regulated by Nrf2 (He et al. 2001). HO-1 is a rate-limiting enzyme that
catalyzes the degradation of heme (a pro-oxidant) to carbon monoxide,
biliverdin and free iron (Keum et al. 2006). HO-1 induction is important in
terms of cellular defense mechanism due to the fact that HO-1 expression is
inducible in response to various forms of cellular insult. Moreover, the end
products of HO-1 catabolism exhibit anti-oxidative, anti-inflammatory and anti-
apoptotic properties (Keum et al. 2006). The objective of this study is to
determine the nature and potency of HO-1 expression induced by equal doses
of several chemicals commonly found in diet i.e. sulforaphane, curcumin and
quercetin.
Twelve ICR male white mice (25–30 g) were divided into 4 groups: (1)
sulforaphane treated group (n=3), (2) quercetin treated group (n=3), (3)
curcumin treated group (n=3) and (4) control (vehicle) group (n=3). The
chemicals were administered intraperitoneally at a dose of 50 mg/kg body
weight for 14 days. A vehicle (DMSO, tween 20 and normal saline in the ratio of
0.05:0.1:0.85) was similarly administered to the control group. At day 15,
animals were sacrificed and their livers isolated. Total RNA was extracted,
reverse transcribed and subjected to quantitative real‐time PCR to detect HO-1
expression. Agarose gel electrophoresis was also performed to verify the
specificity of the amplification.
219
Figure 1. Effects of intraperitoneal administration of 50mg/kg sulforaphane,
curcumin and quercetin for 14 days on HO-1 gene expression in the livers of
mice. Data is presented as mean ± SEM. VH1: vehicle control group, SUL:
sulforaphane group, CUR: curcumin group, QRC: quercetin group. Amplified
products were visualized by agarose gel electrophoresis and gene expression
was confirmed by identification of the appropriate bands. GAPDH served as a
reference gene. * P < 0.05 compared to controls.
220
CONCLUSION
Our findings indicate that at a dose of 50 mg/kg, sulforaphane has the greatest
effect in inducing HO-1 expression in mouse liver, followed by curcumin and
quercetin.
ACKNOWLEDGEMENTS
This research was funded by the Ministry of Education Malaysia & UKM, grant
code FRGS/1/2012/SKK03/UKM/02/2.
REFERENCES
Abdullah, A., Kitteringham, N.R., Jenkins, R.E, Goldring, C., Higgins, L.,
Yamamoto, M., Hayes, J. & Park, B.K. (2012). Analysis of the role of Nrf2
in the expression of liver proteins in mice using two-dimensional gel-
based proteomics. Pharmacological Reports: 680─697.
Bao, W., Li, K., Rong, S., Yao, P., Hao, L., Ying, C., Zhang, X., Nussler, A. & Liu, L.
(2010). Curcumin alleviates ethanol-induced hepatocytes oxidative
damage involving heme oxygenase-1 induction. Journal of
Ethnopharmacology: 549─553.
Cho, H.Y., Jedlicka, A.E., Reddy, S.P., Kensler, T.W., Yamamoto M., Zhang, L.Y. &
Kleeberger, S.R. (2002). Role of NRF2 in protection against hyperoxic
lung injury in mice. American Journal of Respiratory Cell and Molecular
Biology: 175─182.
He, C., Gong, P., Hu, B., Stewart, D., Choi, M., Choi, A. & Alam, J. (2001)
Identification of activating transcription factor 4 (ATF4) as an Nrf2-
interacting protein. Implication for heme oxygenase-1 gene regulation.
Journal of Biological Chemistry: 20858─20865.
Keum, Y.-S., Han, Y.-H., Liew, C., Kim, J.-H., Xu, C., Yuan, X., Shakarjian, M. P.,
Chong, S., & Kong, A.-N. (2006). Induction of heme oxygenase-1 (HO-1)
and NAD [P] H: quinone oxidoreductase 1 (NQO1) by a phenolic
antioxidant, butylated hydroxyanisole (BHA) and its metabolite, tert-
butylhydroquinone (tBHQ) in primary-cultured human and rat
hepatocytes. Pharmaceutical research: 2586─2594
221
Kitteringham, N.R., Abdullah, A., Walsh, J., Randle, L., Jenkins, R.E., Sison, R.,
Goldring, C.E., Powell, H., Sanderson, C., Williams, S., Higgins, L.,
Yamamoto, M., Hayes, J. & Park, B.K. (2010). Proteomic analysis of Nrf2
deficient transgenic mice reveals cellular defence and lipid metabolism
as primary Nrf2-dependent pathways in the liver. Journal of Proteomics.
Jun 16; 73(8): 1612–1631.
Noh, J.R., Kim, Y.H., Hwang, J.H., Choi, D.H., Kim, K.S., Oh, W.K. & Lee, C.H.
(2015). Sulforaphane protects against acetaminophen-induced
hepatotoxicity. Food and Chemical Toxicology: 193─200.
Prestera, T., Zhang, Y., Spencer, S., Wilczak, C. & Talalay, P. (1993) The
electrophile counterattack response: protection against neoplasia and
toxicity. Advances in Enzyme Regulation: 33: 281─296.
Steinmetz, K.A. & Potter, J.D. (1996). Vegetables, fruit, and cancer prevention:
a review. Journal of the American Dietetic Association: 1027─1039.
Yao, P., Nussler, A., Liu, L., Hao, L., Song, F., Schirmeier, A. & Nussler, N. (2007).
Quercetin protects human hepatocytes from ethanol-derived oxidative
stress by inducing heme oxygenase-1 via the MAPK/Nrf2 pathways.
Jounal of Hepatology: 253─261.
222
EVALUATION OF ANTIMICROBIAL AND ANTIOXIDANT PROPERTIES OF
ANETHUM GRAVEOLENS LEAF EXTRACTS
ABSTRACT
INTRODUCTION
223
and trans-anethole (11.0%). These phenolic compounds contribute to its
antioxidant and antimicrobial properties (Cao & Prior 1998; Koleva et al. 2001).
Also, a report by Singh et al. (2005) showed that dill extract retarded the
growth of Bacillus cereus, Bacillus subtilis and Staphylococcus aureus and
fungus Aspergillus niger, Fusarium gramenearum and Penicillium viridicatum.
It was also shown that dill possessed greater antioxidant activities compared to
synthetic antioxidant, BHA and BHT. The objective of this project is to study
and compare antioxidant and antimicrobial activities of various dill extracts.
Three different solvents were used to extract dill leaves, i.e. ethanol, methanol
and water.
Leaf Extraction
Dill leaves were washed and dried in oven at 50˚C overnight. Dried samples
were grinded and weighed, and soaked in ethanol (99.9%) at ratio 1:6
(sample:ethanol) (w:v) for 3 days. The extract was filtrated (Whatman No. 1)
using Buchner flask in vacuum condition. The extract was then concentrated
using rotary evaporator (Buchi R-200) at 55 ˚C. The obtained dried extract was
weighed and kept at 4˚C for testing. Extraction of dill leaves using methanol
was similar to ethanolic extraction. Methanol (99.7%) was used to replace
ethanol. Meanwhile, for water extraction, fresh dill leaves were soaked in
distilled water at ratio 1:3 (sample:water) (w:v) and grinded. Extract was
filtrated (Whatman No. 1) using Buchner flask in vacuum. Filtrate was then
concentrated using rotary evaporator (Buchi R-200) at 100 ˚C. All extracts were
then dissolved in distilled water to produce extract solutions with different
concentrations (200, 400, 600, 800, 1000 ppm).
Two methods were used to determine the antimicrobial activities of dill leaf
extracts, namely disc diffusion test (Gulcin 2003) and direct inhibition test
(Ramdas 1998) with slight modification. Meanwhile, to investigate the
antioxidant activities, three tests were used, i.e. ferric reducing antioxidant
power (Oyaizu 1986), ferric thiocyanate test (FTC) by Osawa and Namiki (1981)
and thiobarbituric acid test (TBA) by Kikuzaki and Nakatani (1993).
224
RESULTS AND DISCUSSION
Extraction Yield
Dill water extract showed the highest extraction yield (4.17%), followed by
methanol extract (2.53%) and ethanol extract (2.47%). All three extracts were
dark green in colour with strong aroma.
In the disc diffusion test, the dill water extract inhibited Bacillus cereus and
Staphylococcus aureus, while dill methanolic and ethanolic extracts were found
to be effective against Escherichia coli, Bacillus cereus, Staphylococcus aureus
and Saccharomyces cerevisiae. As the concentration of the extract increased,
the inhibition zone diameter increased. Singh et al. (2005) reported that in agar
well diffusion method, dill extract showed better results in comparison to
commercial bactericide (ampicillin). It was found effective towards Bacillus
cereus, Staphylococcus aureus and Bacillus subtilis, but less effective towards
Escherichia coli, Pseudomonas aeruginosa and Salmonella typhi. Carvone, the
major component of dill essential oil has been shown to inhibit the growth of
bacteria (Agrawal et al. 2002) and some fungus (Smid et al. 1995).
This method is also known as poison food technique, where the extract was
added into bacteria media growth. The dill ethanolic and methanolic extracts
inhibited E. coli, B. cereus and S. aureus whereas water extract inhibited B.
cereus and S. aureus. Reduction percentage of bacteria growth increased as the
concentration increased. At concentration 1,000 ppm, ethanolic extract
showed the highest percentage of growth inhibition (43.48% against E. coli,
88.23% against B. cereus, 91.81% against S. aureus) compared to other extracts
against these three bacteria (p<0.05). Using the same method, dill extract was
reported to be effective towards Penicillium citrinum, Aspergillus niger,
Fusarium gramenearum, Aspergillus ochraceus, Penicillium viridicatum (Singh
et al. 2005) and Colletotrichum lindemuthianum (Shridhar et al. 2003).
The reducing power of three dill extracts along with BHA and BHT increased
with concentration. Ethanolic extract showed the highest absorbance reading
compared to methanolic and water extracts, and positive control (p<0.05). At
1000 ppm, the sequence for reducing power was ethanolic extract >
methanolic extract > water extract > BHA > BHT. Singh et al. (2005) reported
225
that dill extract reduced ferum (III) better than synthetic antioxidant BHA, BHT
and propyl galate.
This method was used to evaluate the peroxide level during initial stage of
peroxidation. Low absorbance values would indicate high level of antioxidative
activity (Singh et al. 2005). After seven days, all extracts effectively inhibited
linoleic acid oxidation. Ethanolic and Methanolic extracts showed the best
results compared to water extract, BHA, BHT and control. Singh et al. (2005)
reported that Anethum graveolens extract effectively inhibited linoleic acid
oxidation compared to synthetic antioxidant.
This method was used to measure the secondary product of oxidation such as
aldehyde and ketone. Ethanolic and methanolic extracts showed the lowest
absorbance values at day 5 to 7 compared to water extract, BHA ,BHT and
control. This indicated that the amount of peroxidation was lower in ethanolic
and methanolic extracts until day 7, compared to other extracts.
CONCLUSION
It can be concluded that all three extracts of dill (A. graveolens) showed good
antimicrobial and antioxidant properties. However, dill extracted with ethanol
and methanol showed better antimicrobial and antioxidant activitie compared
to the water extract.
226
ACKNOWLEDGEMENT
The author would like to thank Food Science Department, Faculty of Sciences
and Technology, Universiti Kebangsaan Malaysia (UKM) for funding and
technical support.
REFERENCES
Agrawal, K.K., Khanuja, S.P.S., Ahmed, A., Santhakumar, T.R., Gupta, V.K. &
Kumar, S. (2002). Antimcrobial Activity Profiles of the Two Enantiomers
of Limonene and Carvone Isolated from Oils of Mentha spicata and
Anethum sowa. Flavour Fragrance Journal 17: 59─63.
Osawa, T. & Namaki, M. (1983). A Novel Type Antioxidant Isolated from Leaf
Wax of Eucalyptus Leaves. Agric Biol Chem. 45:735─739.
Shyu, Y.S., Lin, Y.S., Chang, Y.T., Chiang, C.J & Yang, D.J. (2009). Evaluation of
Antioxidant Ability Extract from Dill (Anethum graveolens L.) Flower.
Food Chemistry 115: 515─521.
Singh, G., Maurya S., de Lampasona, M.P & Catalan, C. (2005). Chemical
Constituents, Antimicrobial Investigations and Antioxidative Potentials
of Anethum graveolens L. Essential Oil and Acetone Extract: Part 52.
Journal of Food Sc. 70: 208─215.
Smid, E.J., de Witte, Y. & Gorris, L.G.M. (1995). Secondary Plant Metabolites as
Control Agents of Post Harvest Penicillium Rot on Tulip Bulbs.
Postharvest Biotechnology 6: 303─312.
227
ASAM GELUGUR POWDER RICH IN HCA (HYDROXYCITRIC ACID): A
POTENTIAL CROP FOR WEIGHT MANAGEMENT
ABSTRACT
INTRODUCTION
Garcinia atroviridis, also known as asam gelugur, asam gelugo, or asam keping
in Malay is a medium-sized fruit of a large rainforest tree native to Peninsular
Malaysia. This species grows wild throughout Peninsular Malaysia but is also
widely cultivated, especially in the northern states, owing to its economic and
medicinal value. Sun-dried slices of the fruits are commercially available and
popularly used as a seasoning in curries, sour relish and also for dressing fish
(Corner 1988). They also have been used to improve blood circulation, treat
coughs and as an expectorant and laxative (Amran et al. 2009). G. atroviridis is
228
one of the underutilised plants used frequently in medicine as fruit for
reducing weight and excess body fat by halting the glycogen process.
Optimization of Pre-Treatment
The HCA determination was carried out in three replicates. All determinations
were statistically analyzed by the analysis of variance (ANOVA. The Duncan
Multiple Range Test was used to detect the differences between samples
(Gomez & Gomez 1984).
229
RESULTS & DISCUSSION
Optimization of Pre-Treatment
From the statistical analysis, Table 1 shows that the second order on
percentage of enzyme has a significant effect (p<0.05) on HCA content. Same
goes to boiling duration that this pre-treatment also has a significant effect
(p<0.05) on HCA content.
Table 1. Regression coefficients (R2) and p (probability values) for HCA of asam
gelugur sample
HCA
Suggested model : quadratic
Regression coefficient
Constant, 2.96
* percentage of enzyme, b1 -3.193E-003
* boiling duration, b2 -0.13
* percentage of enzyme, b21 -0.35*
2
* boiling duration, b 2 -0.39*
* percentage of enzyme * boiling duration, b12 -0.11
R2 0.7603
P or probability 0.1133
Subscripts: 1 = ultrasound amplitude, 2 = sonication time
* Significant at 0.05 level, ** Significant at 0.01 level, *** Significant at 0.001 level
DESIGN-EXPERT Plot
HCA
X = A: Enzyme
Y = B: Blanched
2.97434
2.72452
2.4747
2.22488
HCA
1.97506
0.50
0.40
30.00
23.75 0.30
17.50 A: Enzyme
0.20
11.25
B: Blanched 5.00 0.10
230
Figure 1. HCA content in asam gelugur in comparison with commercial
products.
CONCLUSION
REFERENCES
Amran, A.A., Zaiton, Z., Faizah, O. & Morat, P. (2009). Effect of Garcinia
Atroviridis on Serum Profiles and Atherosclerotic Lesions in The Aorta of
Guinea Pigs Fed a High Cholesterol Diet. Singapore Med J. 50 (3):
295─299.
Bernad, W.D., Manashi, B., Gottumukkala, V.S., Micheal, A. S., Harry, G.P. &
Debasis, B. (2005). Bioefficacy of a Novel Calcium-Potassium Salt Of (-)-
Hydroxycitric Acid. Mutation Research. 579: 149─162.
Corner, E.J. (1988). Wayside Tress of Malaya. Malayan Nature Society, Kuala
Lumpur.
Gomez, K.A & Gomez, A.A. (1984). Statistical Procedures for Agricultural
Research 2nd ed., p.208─215. New York: John Wiley.
231
Jena, B.S., Jayaprakasha, G.K., Singh, R.P., & Sakariah, K.K. (2002). Chemistry
and Biochemistry of (-)-Hydroxycitric Acid from Garcinia. J.Agric. Food
Chem. 50:10─22
Ohia, S.E., Opere, C.A., Leday, M., Bagchi, D. & Bagchi S.J. (2002). Stohs, Safety
and Mechanism of Appetite Suppression by a Novel Hydroxycitric Acid
Extract (HCA-SX). Molecul Cell Biochem. 238:89─103
Preuss, H.G., Bagchi, D., Bagchi, M., Rao, C.V., Satyanayana, S. & Dey, D.K.
(2004). Efficacy of a Novel, Natural Extract Of (-)-Hydroxycitric Acid
(HCA-SX) and a Combination of HCA-SX, Niacin Bound Chromium and
Gymnema Sylvestre Extract in Weight Management in Human
Volunteers: a Pilot Study. Nutr. Res. 24: 45─58.
232
TOXICOLOGY STUDY OF HIBISCUS SABDARIFFA L. LEAVES EXTRACT ON
NORMAL SPRAGUE-DAWLEY RATS
ABSTRACT
INTRODUCTION
Hibiscus sabdariffa Linne, also known as roselle grows in many tropical and
sub-tropical countries and is one of highest volume specialty botanical
products in international commerce (Halimatul et al. 2007). The swollen
calyces are of commercial interest and the leaves can be used for animal
fodder and fiber. In addition, roselle leaves are rich in polyphenols (which act
as antioxidants that can remove harmful free radicals) have high potential to
be used as a health food product with pharmacological activities (Qi et al.
2005). In line with efforts to balance the conservation of biodiversity and
encouraging controlled exploitation of plant resources for economic gains
especially in biopharming, roselle leaves can be converted into high value
233
products (from agricultural waste). Therefore, it is important to maximize the
usage of this beneficial component for overall health and well being. However,
there is limited information on the safety of roselle leaves and never been
scientifically evaluated through standard in vivo toxicological studies.
Therefore, there is a pressing need to clarify the toxicological profile of the
leaves. In this study, a comprehensive safety evaluation on roselle leaves
extract (RLE) was conducted by performing a single dose acute and 28 days
repeated dose of sub-acute oral toxicity studies in Sprague Dawley (SD) rats.
Sample Preparation
The roselle leaves were dried at 40 °C (moisture content < 10%) and ground.
The extract of roselle leaves was prepared by infusing the dried leaves with
boiling water for 10 min. The extract were prepared at 3000 mg/kg of body
weight (BW) for acute study and three different concentrations (1000, 2000
and 5000 mg/kg BW) for sub-acute oral toxicity study.
Sprague Dawley rats were randomly divided into two groups (control group
and RLE treatment group) consisting of ten animals (5 males and 5 females
weighing 150-200 g per group) in each group. After overnight fasting (8–10 h),
a single RLE dose of 3000 mg/kg BW was given to the treatment group whereas
the control rats received only distilled water. All animals were observed for
clinical signs including mortality and any adverse reactions immediately after
dosing at 1, 2, 4 and 6 h, then once daily until day 14. The body weight was
measured once before the commencement of the dosing and then daily until
day 14 (Abdullah et al. 2009).
Twenty female SD rats weighing 200-250 g were randomly assigned into four
groups; a control and three treatment groups (n=5). During the oral
administration period, SD rats were respectively dosed with distilled water
(control group, C) or RLE at doses of 1000 mg/kg BW (LD, low dose), 2000
mg/kg BW (MD, medium dose) and 5000 mg/kg BW (HD, high dose) of extract
via drinking bottles as they can access ad libitum. On average, each rat will get
100 ml of sample per day. Any remaining sample left will be measured. General
appearance or behaviour of each rat was observed daily during the 28-days
study and the body weight was recorded every week (Ryu et al. 2004).
234
Haematology and Serum Biochemistry Analysis
The animals were fasted for approximately 12 h and blood samples for
haematological and biochemistry analyses were withdrawn under light ether
anesthesia from posterior vena cava. An aliquot of blood per animal
(approximately 20 µL) was treated in a 3 ml ethylen-diamino-tetracetic-acid
(K3-EDTA) tube to analyze haematological indexes. The blood sample was
analysed for complete blood profile by using Haematology Analyzer (Medonic
CA530, Italy). For serum biochemical blood analysis, one aliquot of blood per
animal was placed in a 5 ml Z-serum tube and centrifuged at 3,000 rpm for 20
min. The serum was analyzed by using Blood Clinical Analyzer (Vitalab Selectra
E, Italy).
There were neither significant toxicity symptoms nor death occurred to the
acute experimental rats during the first 6 hours and within 14 days in acute
toxicity studies. Figure 1 and Figure 2 showed no significant differences in
body weights increment between the RLE administrated group and the control
in both sexes. This result showed that the RLE is safe since there were neither
toxic signs nor mortality observed after administration of single high dose
(3000 mg/kg BW) extract.
235
Sub-Acute Oral Toxicity Study
Figure 5 showed the reduction of relative liver weight (p < 0.05) probably
due to the reduced final weight of the rats and cannot be concluded as a sign
of toxicity and the results also within the normal reference range of 2.43 to
3.46% (Han et al., 2010). The enzyme serum analysis (Figure 6) also showed no
signs of toxicity due to decreased levels of ALT (alanine aminotransferase) and
ALP (alkaline phosphatase) significantly. RLE is considered non-toxic due to the
lower level of these liver marker enzymes could indicate a degree of liver
protection (Zhang et al. 1996). There were also no significant changes observed
in serum protein profile (Figure 6) including total protein, albumin and globulin
level in the treated groups as compared to control. Figure 7 did not show any
kidney failure or malfunction since there were no significant difference noted
in urea (range from 5.48 to 6.27 mmol/l) and creatinine (range from 50.17 to
54.00 µmol/l) when dosage was increased from 1000 mg/kg to 5000 mg/kg as
compared to the control. Figure 8 showed that RLE reduce the glucose and TG
(trigliseride) level significantly (p < 0.05) in all dosages (1000 to 5000
mg/kg/day) as compared to control. However, the alterations were assumed to
be toxicologically irrelevant because they were within normal physiological
ranges (Petterino & Argentino-Storino 2006; Han et al. 2010), and were not
dose-related or reflected by any changes in other related parameters.
236
Figure 3. Normalized body weight Figure 4. Haematology data for
of rats for sub-acute study (n=5; RL female SD rats treated orally with
low dose, RM medium dose; RH roselle leaves extract (RLE) for sub-
high dose and C, control group) acute study (n-5)
237
CONCLUSION
In conclusion, the study showed RLE is considered non-toxic to the SD rats and
the NOAEL level for sub-acute test is 5000 mg/kg BW. These data provide
important reference of RLE for usage as a food supplement or in future clinical
trials as a medication.
ACKNOWLEDGEMENTS
The authors are thankful to the financial supports from MARDI and Malaysian
Ministry of Agricultural for the research development grant of P-161.
REFERENCES
Han, Z.Z., Xu, H.D., Kim, K.H., Ahn, T.H., Bae, J.S., Lee, J.Y., Gil, K.H., Lee, J.Y.,
Woo, S.J., Yoo, H.J., Lee, H.K., Kim, K.H., Park, C.K., Zhang, H.S. and
Song,S.W. (2010). Reference Data of the Main Physiological Parameters
in Control Sprague-Dawley Rats from Pre-clinical Toxicity Studies.
Laboratory Animal Research 26 (2): 153─164.
Halimatul, S.M.N., Amin, I., Mohd Esa, N. Nawalyah, A.G. and Siti Muskinah, M.
(2007). Protein Quality of Roselle Seeds. ASEAN Food Journal 14 (2):
131─140.
Qi, Y., Chin, K.L., Malekian, F., Berhane, M. and Gager, J. (2005). Biological
Characteristics, Nutritional and Medicinal Value of Roselle, Hibiscus
Sabdariffa. CIRCULAR – Urban Forestry Natural Resources and
Environment No. 604.
Ryu, S.D., Park, C.S., Baek, H.M., Baek, S.H., Hwang, S.Y. and Chung, W.G.
(2004). Anti-diarrheal and Spasmolytic Activities and Acute Toxicity
238
Study of Soonkijangquebo, a Herbal Anti-diarrheal Formula. Journal of
Ethnopharmacology 91: 75─80.
239
CLINACANTHUS NUTANS L.: SAFETY AND TOXICITY STUDY
ABSTRACT
This study was aimed to evaluate the effect of Clinacanthus nutans L. extract
on Sprague Dawley rats subjected to acute (single, high dose) and sub-acute
(28-day repeated doses) test. A single dose (3 g/kg of body weight) oral
administration did not show any acute oral toxicity in rats. In sub-acute study,
C. nutans L. extract was administered at 1 g/kg body weight (low dose, LD), 2
g/kg (medium dose, MD) and 5 g/kg (high dose, HD). Distilled water was given
to the rats as control. Administration of C. nutans L. extract did not cause
negative effect in blood haematology even though a statistically significant
(p<0.05) decreased in platelet level was noted. Result from serum biochemical
test showed that the consumption of the extract did not result in liver and
kidney failure since no significant changes was observed except for alanine
phosphatase (ALP), although the results were still within the normal range.
Based on these findings, the no-observed-adverse effect of C. nutans L. extract
is 5 g/kg of body weight per day.
INTRODUCTION
240
contain lupeol and β-sitosterol, which are pharmacological active components
that showed chemopreventive properties. There is still lack of scientific
research regarding its benefit and the content of bioactive compound in CNL
and the needs to determine and quantify its chemoprevention property.
Therefore, it is hoped that this study could provide scientific evidence to prove
its safety and beneficial to health.
The safety and toxicity of CNL extract was evaluated using acute and sub-acute
test. This study was approved by MARDI Animal Ethic Committee. All animals
were housed in a controlled environment with 12 hours light per day. They
were allowed to acclimatize for seven days before beginning treatment.
The acute study was conducted with a single high dose (3 g/kg body
weight (BW)) of the extract (Ryu et al. 2004) on five male and five female
Sprague-Dawley rats (150-200 g). Distilled water was used as control. The rats
were monitored within six hours after administration and for the next 14 days
if any toxicity symptoms occured. The body weight was measured during
experimental period.
241
RESULTS AND DISCUSSION
After a high, single administration of CNL extract (3.0 g/kg ), there was neither
significant toxicity symptoms nor death in the first 6 hours. The body weight
increments of rats are shown in Figure 1 and Figure 2. There was no significant
difference between groups.
Acute toxicity study shows that the CNL extract is safe since there was
neither toxic sign nor mortality observed after administration of a single high
dose (3.0 g/kg ).
There was no mortality or abnormalities in all treated rats with respect to hair
coat, eye colour, rashes and skin irritation of animal at any dose level tested
during the 28-days study of CNL extract.
242
Figure 3. Normalized body weight of rats in the sub-acute test (n=5)
Results for serum clinical test for liver parameters are listed in Table 2.
Administration of CNL extract did not show any significant difference in all
parameters except for globulin. Results on serum protein showed significant
difference in globulin level in rats given medium dose of extract as compared
to the control. However, the difference is not associated with any liver failure
or malfunction because the globulin level was still within the normal range of
30-36 g/l (Claudio & Alberta 2006).
243
Table 2. Blood chemistry parameter for liver function after 28-day repeated
dose
Analysis Control Dose Low Dose Medium Dose High Dose
a a a a
ALT (U/l) 80.78±10.85 88.60±10.53 73.02±39.85 90.80±22.80
a a a a
AST (U/l) 96.72±17.09 111.28±7.33 116.00±78.95 112.99±8.92
ab a b ab
ALP (U/l) 270.33±90.94 338.20±83.36 205.20±102.85 242.60±21.84
a a a a
Bilirubin 2.66±0.53 3.13±0.40 9.20±13.87 2.40±0.61
(µmol/l)
a a a a
Total Protein 72.93±3.17 73.86±5.25 55.89±31.48 70.67±4.77
(g/l)
a a a a
Albumin (g/l) 41.77±2.60 42.36±2.88 67.32±57.55 41.16±3.47
a a b ab
Globulin (g/l) 31.11±3.30 31.20±2.59 23.68±10.01 29.40±2.51
a a a a
A:G 1.36±0.20 1.35±0.05 1.85±0.78 1.40±0.16
Values are means ± standard deviation (SD) for five rats in each group. Means
with the same letter in the same row are not significantly different (p<0.05).
CONCLUSIONS
ACKNOWLEDGEMENT
The authors would like to thank the Ministry of Agriculture & Agro-based
Industry of Malaysia for the Development Grant and our contract staff Miss
Nur Hafiqa & Miss Siti Nur Aslina for a great help in the project.
244
REFFERENCES
Hilaly, J.E., Israili, Z.H. & Lyoussi, B. (2004). Acute and Chronic Toxicological
Studies of Ajugaiva in Experimental Animals. J. of Ethnopharmacology
91:43–50.
Ryu, S.D., Park, C.S., Baek, H.M., Baek, S.H., Hwang, S.Y. & Chung, W.G. (2004).
Anti-Diarrheal and Spasmolytic Activities and Acute Toxicity Study of
Soonkijangquebo, A Herbal Anti-Diarrheal Formula. Journal of
Ethnopharmacology 91: 75–80.
Sittiso, S., Ekalaksananan T., Pientong C., Sakdarat S., Charoensri N. &
Kongyingyoes B. (2010). Effects of Compounds from Clinacanthus nutans
on Dengue Virus Type 2 Infection. Srinagarind Medical Journal 25
(Suppl).
Teo, S., Stirling, D., Thomas, S., Hoberman, A. & Khetani, V. (2002). A 90-Day
Oral Gavage Toxicity Study of D-Methylphenidate and D,L-
Methylphenidate in Sprague-Dawley Rats. Toxicology 179: 183–196.
245
IN VITRO TOXICOLOGICAL EVALUATION OF 50 METHANOL EXTRACTS
FROM TRADITIONAL MEDICINAL PLANTS USED BY THE ORANG ASLI
MGH Khoo, S Rohana, M Nik Musaadah, J Fadzureena, MA Adiana, H Nuziah,
AL Tan, Z Nurul Husna, B Intan Nurulhani, HF Lim & H Norini
Forest Research Institute Malaysia, 52109 Kepong, Selangor
Tel: 03-6279 7342 Fax: 03-6272 9805 E-mail: mary@frim.gov.my
ABSTRACT
Plants are widely used in the health care regimen since ancient times. In recent
years, various studies have investigated the biological activity and potential
medicinal values of medicinal plants. In this study, 50 methanol extracts from
15 medicinal plants used by the Orang Asli were evaluated for their potential
toxicity. Toxicity was evaluated in vitro using Vero and WRL-68 cell lines. Out of
the 50 extracts tested, 6.0% of the extracts exhibited toxicity towards both
Vero and WRL-68 cell lines with median inhibition concentration (IC50) values
below 20 g/ml; 30.0% had IC50 values ranging between 20-100 g/ml; 24.0%
with IC50 values above 100 g/ml; and 12.0% did not negatively affect the
viability of cells more than 50% at the highest concentration (625 g/ml)
tested. The level of toxicity towards Vero and WRL-68 cell lines were different
for the remaining 28.0% of the extracts (i.e. 14 extracts). Out of these 28.0%
extracts tested towards Vero cell line, the IC50 values for 21.4% were below 20
g/ml; 42.9% had IC50 values ranging between 20-100 g/ml; 28.6%
demonstrated IC50 values in the category above 100 g/ml; and 7.1% did not
cause more than 50% of cell death at the concentrations tested. Meanwhile,
for the toxicity towards WRL-68 cells of these remaining 14 extracts, 14.3%
demonstrated IC50 values that fall under the category of less than 20 g/ml;
35.7% with IC50 values ranging between 20-100 g/ml; 42.9% with values
above 100 g/ml; and 7.1% with values of more than 625 g/ml. As the saying
“The dose makes the poison”, which means everything could lead to toxicity
when taken in too high a dose, these findings serve as a rough guideline for the
safe usage of the medicinal plants. The bigger the gap between the dose
required to exert a certain biological activity (effective concentration, EC50) and
IC50 value, the safer it is to be used.
It must however be noted that the data generated from this study is only
preliminary, further tests are required for extensive toxicological evaluation.
246
INTRODUCTION
Malaysia consists of Sabah and Sarawak (the East Malaysian States located at
the northern coast of Borneo) and West Malaysia, also known as Peninsular
Malaysia. Malaysia has a population of 28 million and 3.5 million are
indigenous people, which is about 12% of the total population. In Peninsular
Malaysia, the number of Orang Asli is around 150,000, representing 0.6% of
the national population. Orang Asli is the Malay term used to refer to
indigenous people in Peninsular Malaysia. Majority of the Orang Asli live in
rural and remote areas. They depend substantially on natural resources in the
ecosystem for subsistence, e.g. by hunting, fishing, gathering, swidden farming,
arboriculture, and trading forest products. The forest is the source of their
livelihood; it shapes their customs and culture, defines their identity, and
constitutes their local environment (Kardooni et al. 2014).
The Orang Asli also depends on the forest as a source of health care
remedies. Many plants in the forest contain medicinal values that are able to
cure various health problems. This traditional knowledge on medicinal plants is
inherited from generation to generation. As modernisation takes place, there is
a rapid change in the natural environment. Compounded with economic,
political, and cultural change on a global scale, many indigenous knowledge
systems are in danger of vanishing. Since the traditional knowledge of Orang
Asli might disappear without a trace, there is a need to identify and document
such knowledge as a basis for scientific research and enhance the
understanding. The Malaysian Ministry of Natural Resources and Environment
funded a study, Pangkalan data pengetahuan tradisi Orang Asli di
Semenanjung Malaysia, in an attempt to document the commonly used
traditional knowledge of the Orang Asli, specifically the traditional knowledge
on medicinal plants. Besides documentation, selected plant species were also
evaluated for various therapeutic and cosmeceutical potentials. The study
begins in 2007 and is still on-going up to this date. In this paper, the safety
aspect of 50 selected methanol extracts at cellular level will be discussed.
METHODOLOGY
A total of 15 medicinal plant species that are used by 5 communities of Orang
Asli – Jah Hut, Jakun, Kanaq, Semoq Beri, and Bateq were collected during
2011–2012. Voucher specimen for each species was prepared and stored at the
Traditional Knowledge (TK) Specimen Room, Forest Research Institute Malaysia
(FRIM).
247
The methanol extracts were prepared by separating the plants into
different parts, dried, and ground. Ground samples were then soaked in
methanol for 3 days before evaporation to produce dried extracts. The dried
extracts were stored at -20C until bioassay evaluation.
WRL-68 (liver-like) and Vero (kidney-like) cells were used for in vitro
toxicological evaluation. Cells (1 × 104 cells/well) were exposed to the extract
for 72 hours and viability was measured by the MTT (3-(4, 5-dimethylthiazolyl-
2)-2, 5-diphenyltetrazolium bromide) assay as described by (Mosmann 1983).
Dose-response curve for each extract was plotted and the median inhibitory
concentration (IC50) was determined by non-linear regression using a variable
slope model (GraphPad Prism 6, GraphPad Software Inc., California, U.S.A).
Upon closer inspection, out of the 50 extracts tested, only 72% exhibited
similar toxicity towards both Vero and WRL-68 cells (Figure 2a). The breakdown
is as follows: 6.0% of the extract had IC50 < 20 g/ml; 30.0% with IC50 20-100
g/ml; 24.0% with IC50 100-625 g/ml; 12.0% with
IC50 > 625 g/ml and 28.0% (14 extracts) had different toxicity level towards
Vero and WRL-68 cells. For this remaining 14 extracts, 21.4% was cytotoxic
towards Vero cells with IC50 value < 20 g/ml; 42.9% with IC50
20-100 g/ml; 28.6% with IC50 100-625 g/ml and 7.1% did not cause more
than 50% of cell death at the concentrations tested (Figure 2b). Meanwhile, for
the toxicity towards WRL-68 cells, 14.3% demonstrated IC50 values
< 20 g/ml; 35.7% with IC50 20-100 g/ml; 42.9% with values between
100-625 g/ml; and 7.1% with values > 625 g/ml (Figure 2b).
248
25
Vero
Number of extracts
WRL-68
20
15
10
0
< 20 g/ml 20-100 g/ml 100-625 g/ml > 625 g/ml
IC50 category
Figure 1. Number of extracts categorised according to their IC50 values.
< 20 g/ml
< 20 g/ml 14.3%
100-625 21.4%
g/ml
100-625
28.6% 20-100
g/ml
20-100 g/ml
42.9%
35.7%
g/ml
42.9%
Vero WRL-68
Figure 2. Percentage of extracts categorised according to their IC50
values, (a) extracts with similar toxicity towards Vero and WRL-68 cells,
(b) extracts with different toxicity levels towards Vero and WRL-68 cells.
249
In order to determine whether an extract is toxic or otherwise, it is best
to compare the IC50 value determined from a toxicological study to the median
effective concentration (EC50) for a desired therapeutic activity. This is because
toxicity is dose-dependent. Everything could become toxic when the dose
taken is too high. For example, water is an essential element for all living things
but if a person drinks too much water, this could lead to water intoxication.
Hence the saying “the dose makes the poison”. An extract has favourable
safety profile when the difference between IC50 and EC50 values is large.
CONCLUSIONS
Few medicinal plants commonly used by the Orang Asli appear to have very
low IC50 value (< 20 g/ml). These plants may cause toxicity should the dosage
required for desired therapeutic effect exceeds the mentioned IC50 value.
Contrary to popular belief that all natural are safe, the usage of plants with
very low IC50 value must be exercised with caution. The findings obtained from
this study are only preliminary and serve as a rough guideline for the safe
usage of the medicinal plants. Further tests are required for extensive
toxicological evaluation.
REFERENCES
Kardooni, R., Kari F., Yahaya, S.R. & Yusup, S.H. (2014). Traditional Knowledge
of Orang Asli on Forests in Peninsular Malaysia. Indian Journal of Traditional
Knowledge: 13: 282─291.
Mosmann, T. (1983). Rapid colourimetric assay for cellular growth and survival
application to proliferation and cytotoxicity assays. J. Immuno. Methods: 54:
55─63.
250
TOXICITY STUDY OF SARAWAK WILD PEPPER ROOT (PIPER
ARBORESCENS)
ABSTRACT
Sarawak wild pepper root (Piper arborescens) has been used in cooking as
health food for many generations and is generally considered as safe, mainly
due to its natural origin but without sufficient scientific studies. This study was
aimed to evaluate the effect of wild pepper root extract on Sprague-Dawley
rats subjected to sub-acute 28-day repeated doses. The rats were given wild
pepper root extract at the dose of 1 (low dose), 2 (medium dose) and 5 (high
dose) g/kg body weight. The control group was given distilled water. Based on
the body weight, organ relative weight and serum biochemistry parameters in
sub-acute toxicity results, wild pepper root showed no noticeable gross toxicity
in all treated Sprague-Dawley rats even though the white blood cell volume,
alanine aminotransferase (ALT) level and creatinine concentration were
significantly lower in rats treated with wild pepper root as compared to the
control group. This indicates that the no-observed-adversed-effect-level
(NOAEL) of wild pepper root extract is 5 g/kg body weight per day.
INTRODUCTION
The genus Piper belongs to Piperaceae family, comprises five genera and
approximately 1,400 species, distributed in the tropical and subtropical regions
(dos Santos et al. 2001). Various phytochemical investigations of Piper species
have led to the isolation of various classes of physiologically active compounds
(Parmar et al. 1997).
251
known in Sarawak, P. arborescens (lada hutan) is one of the most common
species widely distributed in the lowlands and hill rainforest. The perennial
roots and vines of this wild pepper have a peppery flavour, which is the reason
it is given the general name “Sarawak wild pepper root” or “akar lada liar
Sarawak”. The decoction of wild pepper root is used as household remedy and
reputed to help in treating rheumatism, gout, gastrointestinal diseases,
asthma, bronchitis, improvement of metabolism and provide body warmth
(Chai et al. 1989).
Wild pepper root has been consumed for many generations and is
generally considered as safe, mainly due to its natural origin but without
sufficient scientific studies. Since safety is regarded as one of the important
aspects in herbal product development, the objective of this study is to
evaluate the effect of wild pepper root extract on Sprague-Dawley rats
subjected to in vivo sub-acute (28-day) repeated doses.
Dried wild pepper roots were procured from the local market in Kuching,
Sarawak. The dried roots were cleaned thoroughly with fresh water and dried
at 45C, and then cut into small pieces using a blender. The extract was
prepared using traditional decoction method i.e. by boiling with water at the
ratio of 60 g/1.5 L for 30 minutes. Extracts were then filtered and stored at 4°C
in an airtight bottle until use.
252
Physical and Body Weight Assessment
Oral administration of wild pepper roots extract did not induce any mortality at
any dose level tested within the 28-day repeated feeding study. All the treated
rats also did not show any sign of gross toxicity, adverse pharmacological
effects or abnormalities with respect to hair coat, eye colour, rashes and skin
irritation. The gross examination of internal organs also did not show any
inflammation, changes in colour and no abnormal spots occurred as compared
to control group.
Serum Biochemistry
1. Liver Function
Administration of wild pepper root extract did not show any significant
difference in all parameters except for serum alanine aminotransferase (ALT)
(Table 1). Significant decreased in serum ALT level was observed in rats fed on
high dose wild pepper root extract – the higher the dose, the lower the ALT
level. However, the differences were not significant among all the three
treated groups. This implies that the extract might not cause any toxic effect on
the liver even at high dose. Reduction of ALT level is important for instilling
protection of the liver from oxidative damages (Jie et al. 2014).
2. Lipid Profile
There was also no significant difference observed in all the plasma lipid profiles
in all the three treated groups as compared to the control (Table 2). Even
though no significant difference was found compared to the control, an
increase in high density lipoprotein (HDL) level in the low- and medium-dose
groups, decrease of cholesterol level in medium- and high-dose group and
decrease of low density lipoprotein (LDL) level in low-, medium- and high-dose
groups were observed. These are indicators that the extract might have the
potential to reduce cardiovascular risk factors.
253
Table 1. Effects of wild pepper root extract intake on liver function in rats
(n = 5) after 28-day repeated dose
Parameter Control Low Dose Medium High Dose
Dose
ALT (U/l) 58.33 ± 53.83 ± 48.67 ± 45.67 ±
8.12a 12.51ab 3.83ab 6.44b
AST (U/l) 114.76 ± 109.10 ± 104.96 ± 99.43 ±
a a a
11.12 22.22 4.13 9.73a
ALP (U/l) 290.17 ± 215.00 ± 220.83 ± 243.83 ±
a a a
102.00 63.59 66.09 22.50a
Bilirubin 2.20 ± 1.31a 2.58 ± 0.49a 2.55 ± 0.63a 3.12 ± 0.45a
(µmol/l)
Total Protein 72.97 ± 71.98 ± 71.76 ± 73.23 ±
(g/l) 4.24a 4.52a 3.14a 3.92a
Albumin 45.42 ± 43.82 ± 44.32 ± 45.35 ±
a a a
(g/l) 1.55 1.18 1.04 1.87a
Globulin 27.67 ± 28.17 ± 27.17 ± 27.83 ±
(g/l) 3.83a 5.19a 2.71a 2.48a
a a a
Albumin/ 1.68 ± 0.27 1.61 ± 0.35 1.65 ± 0.17 1.63 ± 0.12a
Globulin
ratio
Glucose 4.80 ± 2.02a 5.50 ± 0.43a 5.75 ± 0.83a 5.53 ± 0.39a
(mmol/l)
*Values are means ± standard deviation (SD) for five rats in each group. Means with
the same letter in the same row are not significantly different (p<0.05).
Table 2. Effects of wild pepper root extract intake on lipid profile in rats
(n = 5) after 28-day repeated dose
Parameter Control Low Dose Medium High Dose
Dose
Cholesterol 1.53±0.68a 1.55±0.31a 1.23±0.45a 1.21±0.15a
(mmol/l)
HDL 0.44±0.15a 0.51±0.15a 0.45±0.08a 0.38±0.09a
(mmol/l)
LDL (mmol/l) 0.83±0.51a 0.80±0.52a 0.55±0.41a 0.50±0.16a
a a a
Trigliseride 0.56±0.15 0.57±0.05 0.54±0.07 0.60±0.17a
(mmol/l)
*Values are means ± standard deviation (SD) for five rats in each group. Means with
the same letter in the same row are not significantly different (p<0.05).
254
3. Kidney function
CONCLUSION
Based on the body and organ relative weight, blood hematology and serum
biochemistry parameters in this sub-acute toxicity study, wild pepper root
showed no noticeable gross toxicity in all treated Sprague-Dawley rats. The no-
observed-adversed-effect-level (NOAEL) of wild pepper root extract is
5 g/kg body weight per day. Even though no significant difference was found
between treated and the control group, there were indicators that wild pepper
root extract might have the potential to reduce cardiovascular risk factors.
Further studies are needed to identify the active phytochemicals present in
wild pepper root to elucidate their cardiovascular protection activities.
ACKNOWLEDGEMENTS
The authors thank the Ministry of Higher Education, Malaysia for their financial
support via Fundamental Research Grant Scheme (FRGS), research grant
number 03─03─03─064 EA 001.
255
REFERENCES
Chai, P.P.K., Lee, B.M.H. & Othman, I. (1989). Native Medicinal Plants of
Sarawak. Report No. FB 1. Sarawak: Forest Botany Unit, Forest
Department. Pp. 53–55.
dos Santos, P.R.D., Moreira, D.D., Guimaraes, E.F., & Kaplan, M.A.C. (2001).
Essential Oil Analysis of 10 Piperaceae Species from the Brazilian
Atlantic Forest. Phytochemistry 58: 547–551.
Jie, M., Cheung, W.M., Yu, V., Zhou, Y., Tong, P.H. & Ho, J.W.S. (2014). Anti-Proliferative
Activities of Sinigrin on Carcinogen-Induced Hepatotoxicity in Rats. PLoS ONE
9(10): e110145. doi:10.1371/journal.pone.0110145.
Parmar, V.S., Jain, S.C., Bisht, K.S., Jain, R., Taneja, P., Jha, A., Tyagi, O.D.,
Prasad, A.K., Wengel, J. Olsen, C.E. & Boll, P.M. (1997). Phytochemistry
of the Genus Piper. Phytochemistry 46(4): 597–673.
Ryu, S.D., Park, C.S., Baek, H.M., Baek, S.H., Hwang, S.Y. & Chung, W.G. (2004).
Anti-Diarrheal and Spasmolytic Activities and Acute Toxicity Study of
Soonkijangquebo, A Herbal Anti-Diarrheal Formula. Journal of
Ethnopharmacology 91: 75–80.
Tawan, C.S., Ipor, I.B., Fashihuddin, B.A. and Sani, H. (2002). A Brief Account on
the Wild Piper (Piperaceae) of the Crocker Range, Sabah. ASEAN
Review of Biodiversity and Environmental Conservation (ARBEC).
http://www.arbec.com.my/ pdf/art6julysep02.pdf.
WHO. (1993). Research Guidelines for the Evaluation of the Safety and Efficacy
of Herbal Medicines. Manila: WHO. Pp.35–40.
256
IN SILICO PREDICTION OF DRUG LIKENESS AND ADMET PROPERTIES OF
SOME CENTELLA COMPOUNDS
ABSTRACT
257
INTRODUCTION
a b c
d e f g
258
MATERIALS AND METHODS
ADMET Prediction
Toxicity prediction using OSIRIS property explorer revealed that all the
compounds were not mutagenic, tumorigenic, irritant and reproductive
effective. Prediction on toxicological hazard using OpenTox revealed that the
compounds shared common properties such as Verhaar scheme, start
biodegradability, skin irritation/corrosive and Kroes TTC which are class 5, class
259
2, not corrosive and class 1, respectively (Table 2). It is also observed that
asiaticoside and madecassoside shared the same profile for Cramers rule and
eye irritation, which are class III and not eye lesion R36 respectively. The other
compounds showed class I and not lesions R34, R35, R36 or R4. In general, all
compounds were predicted as substance that would not be expected to be a
safety concern and non inhibitor for all cytochrome P450 enzymes, except for
asiaticoside that could not be predicted for Cyp2C19.
CONCLUSION
ACKNOWLEDGEMENTS
REFERENCES
Sushko, I., Novotarskyi, S., Korner, R., Pandey, A.K., Rupp, M., Teetz, W., et al.
(2011). Online Chemical Modeling Environment (OCHEM): Web
Platform for Data Storage, Model Development and Publishing of
Chemical Information. Journal of Computer-Aided Molecular Design:
25; 533─554.
Bolton, E., Wang, Y., Thiessen, P.A., & Bryant, S.H. (2008). PubChem: Integrated
Platform of Small Molecules and Biological Activities. Chapter 12 Pp
217-241 in Wheeler R.A. & Spellmeyer D.C. (Eds.). Annual Reports in
Computational Chemistry Vol 4. Oxford, UK: Elsevier.
260
foodb.ca/compounds/
Hong, S.S., Kim, J.H. & Shim, C.K. (2005). Advanced Formulation and
Pharmacological Activity of Hydrogel of the Titrated Extract of C.
asiatica. Arch Pharm Res: 28(4); 502─508.
http://www.organic-chemistry.org/prog/peo/
http://www.molinspiration.com
http://toxtree.sourceforge.net
Xiong, Y., Ding, H., Xu, M. & Gao, J. (2009). Protective Effects of Asiatic Acid on
Rotenine- or H2)2-induced Injury in SH-SY5Y cells. Neurochem Res:
34(4); 746-754.
Mook-Jung, I., Shin, J.E., Yun, S.H., Huk, K., Koh, J.Y., Park, H.K., Jew, S.S. & Jung,
M.W. (1999). Protective Effects of Asiaticoside Derivatives Against
Beta-Amyloid Neurotoxicity. Journal of Neuroscience Research: 58(3);
417─425.
Park, B.C., Paek, S.H., Lee, Y.S., Kim, S.J., Lee, E.S., Choi, H.G., Yong, C.S.& Kim,
J.A. (2007). Inhibitory Effects of Asiatic Acid on 7, 12-
dimethylbenz[a]anthracene and 12-0-tetradecanoylphorbol 13-
acetate-induced Tumor Promotion in Mice. Biolo Pharm Bull: 30(1);
176─179.
261
Table 1. Drug likeness properties calculated by Molinspiration and OSIRIS property explorer.
262
Table 2. Toxicological hazard classification and ADME prediction results by OpenTox and OCHEM
Asiaticoside Asiatic acid Madecassic acid Madecassoside Brahminoside Madasiatic acid Brahmoside
OpenTox
Cramers rule High (Class III) Low (Class I) Low (Class I) High (Class III) Low (Class I) Low (Class I) Low (Class I)
Verhaar scheme Class 5 (Not Class 5 (Not Class 5 (Not Class 5 (Not Class 5 (Not Class 5 (Not Class 5 (Not
possible to possible to possible to possible to possible to possible to possible to
classify according classify according classify according classify according classify according classify according classify according
to these rules) to these rules) to these rules) to these rules) to these rules) to these rules) to these rules)
Start biodegradability Class 2 Class 2 Class 2 Class 2 (persistent Class 2 Class 2 Class 2
(persistent (persistent (persistent chemical) (persistent (persistent (persistent
chemical) chemical) chemical) chemical) chemical) chemical)
Skin Not corrosive Not corrosive Not corrosive Not corrosive Not corrosive Not corrosive Not corrosive
irritation/corrosion
Eye Not eye irritation Not lesions R34, Not lesions R34, Not eye irritation Not lesions R34, Not lesions R34, Not lesions R34,
irritation/corrosion R36 R35, R36 or R41 R35, R36 or R41 R36 R35, R36 or R41 R35, R36 or R41 R35, R36 or R41
Kroes TTC (Threshold 1 (Substance 1 (Substance 1 (Substance 1 (Substance 1 (Substance 1 (Substance 1 (Substance
Toxicological Concern) would not be would not be would not be would not be would not be would not be would not be
expected to be a expected to be a expected to be a expected to be a expected to be a expected to be a expected to be a
safety concern) safety concern) safety concern) safety concern) safety concern) safety concern) safety concern)
OCHEM
CYP450 modulation Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor
(CYP3A4) (90%) (94%) (94%) (86%) (94%) (94%) (94%)
CYP450 modulation Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor
(CYP2D6) (93%) (95%) (96%) (93%) (96%) (95%) (96%)
CYP450 modulation error Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor
(CYP2C19) (94%) (94%) (89%) (94%) (94%) (94%)
CYP450 modulation Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor
(CYP2C9) (95%) (94%) (93%) (97%) (94%) (94%) (94%)
CYP450 modulation Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor Noninhibitor
(CYP1A2) (96%) (96%) (94%) (96%) (94%) (94%) (94%)
263
NUTRIGENOMIC EFFECTS OF CURCULIGO LATIFOLIA ON TYPE 2
DIABETIC MODEL
ABSTRACT
Key words: Curculigo latifolia, gene expression, high fat fed diet, insulin, type 2
diabetes mellitus
INTRODUCTION
Diabetes mellitus (DM) is a metabolic disorder which has become one of the
major worldwide health issues. It has been defined as a continual metabolic
264
disorder which leads to the deficiency in the production of insulin by the
pancreas along with insulin resistance. It has been reported that 371 million
peoples have been diagnosed with diabetes in 2012 and this number is
expected to increase by 2030. This scenario leads to a prediction that by 2030,
diabetes will become the seventh leading cause of death in the world (IDF
Diabetes Atlas Group 2015). In Malaysia, the National Health and Morbidity
Survey (NHMS) 2011 has shown that the prevalence of diabetes has increased
by 31.0% in the space of just 5 years, from 11.6% in 2006 to 15.2% in 2011
(National Diabetes Registry Report 2012).
265
concentration of C. latifolia fruit:root extracts used in this study were 50, 100
and 200 mg/kg body weight (b.w).
The normal pellet diet (NPD) from Miba Mansura (Malaysia) are consists of
46% of cornstarch, 26% of palm kernel meal, 4% of soybean oil, 3.5% of
minerals mixture, 1% of vitamins mixture, 0.25% of choline bitartrate and
0.18% of L-cystine. Meanwhile, the high fat diet was formulated based on the
composition provided by Levin et al. (1989). It has been prepared from a
mixture of 50% normal rat chow pellet, 24% of corn oil (Mazola brand), 20% of
full-cream milk powder (Nespray brand from Nestlé) and 6% sugar.
Animal Study
266
Treatment was carried for 30 days. At the end of experimental
period, all rats were fasted for 15 hr prior to sacrifice. Blood samples were
collected by cardiac puncture in order to measure glucose, lipid and insulin
levels. Meanwhile, adipose tissues were excised and stored at -80oC prior
used.
Biological Assays
Blood samples were collected using K2EDTA blood collection tube (BD
Diagnostics, New Jersey). Plasma was collected after blood was centrifuged at
3,000 rpm for 10 min. Biological assay such as glucose, total cholesterol (TC),
triglycerides (TG), low density lipoprotein (LDL), high density lipoprotein (HDL)
were measured using Selectra XL clinical chemistry analyzer (Vital Scientific,
Netherlands). Insulin level was measured using rat insulin ELISA kit (Mercodia
AB, Uppsala, Sweeden) with rat insulin as a standard.
Total RNA was extracted from adipose tissues using RiboPureTM Isolation of
High Quality Total RNA (Ambion, USA). Frozen tissues were thawed before
starting the disruption procedure. The RNA extraction was carried out as
described in RiboPureTM protocol. The quantification of different express
gene between group treatments was using GenomeLab GeXP Start Kit.
Multiplex primers for GLUT4, Lep and IRS1 were design designed by using
eXpress designer module of the GenomeLab eXpress Profiler software.
Samples were prepared and were added to the appropriate wells of 96-well
sample microplate. All the samples were run triplicates. Besides, all the data
were analyzed using eXpress Analysis software where fragment data is easily
identified. Multiplex genes were normalized with 18S by dividing the peak
area of each gene by peak area of 18S gene. The expression level was
calculated according to this formula:
Fold change = Normalized data of the gene from treated samples
Normalized data of the gene from untreated samples
Statistical Analysis
All data were expressed as mean ± standard deviation and each value
represents a minimum of three (n=3-6) replicate experiments and all assay
conditions were performed in triplicate. Data were analyzed using one-way
analyses of variance (ANOVA) followed by Turkey’s post hoc test. Level of
significance was set at p<0.05.
267
RESULTS AND DISCUSSION
268
Hyperlipidemia has developed in diabetic rats after 30 days induced with high
fat diet and low dose STZ. Meanwhile, lipid profiles in normal rats were in
normal range. Plasma TC, LDL and TG levels in obese rats (Group 2) were
significantly higher (p<0.05) while HDL level was lower than normal rats (Group
1) after 30 days of study (Table 1). Diabetic control rats in Group 3 also showed
a similar pattern as obese rats where plasma TC, TG and LDL levels increased
56.2%, 52.6% and 75.4% respectively as compared to normal rats. It has been
shown that rats administrated with HFD caused dyslipidemia and other
syndromes in diabetic (Rossi et al. 2010). Besides, defect in insulin secretion
due to STZ also caused defect in lipogenic activity. Insulin plays an important
role in stimulating lipogenesis in mammals, by low secretions of insulin it
implicates of high level of lipid in plasma (Rossi et al. 2010). However, plasma
HDL level in diabetic control rats decreased by 51%. C. latifolia has significantly
reduced (p<0.05) plasma TC, TG and LDL levels. Besides, HDL level also
increased after 30 days treatment in diabetic rats in Group 4, 5 and 6.
269
Figure 2. Fasting plasma insulin
Data are means SD for fasting plasma insulin level at day 0 and day 30 treatment. * are
significant different at p<0.05 compared with diabetic control i.e. group 3.
Indicator: ( ) before treatment, ( ) after 30 days of treatment
This study showed that plasma insulin level decreased in obese rats. Disruption
of pancreatic β-cells by STZ in diabetic rats has caused insufficient insulin
secretion in blood. This STZ is being uptake into pancreatic cells by GLUT2 and
it causes DNA damage via reactive oxygen species generation (Shingo et al.
2012). However, C. latifolia fruit:root extracts in all concentration has
prevented further disruption of cells. There was 16% of insulin increasing in
rats in Group 4 and followed by 13% in Group 5, 12% in Group 6 and 11% in
Group 7. Previous study also indicated that C. latifolia extracts elicited
insulinotropic properties by stimulating insulin secretion either in basal and
glucose.
Present results showed that the expression of GLUT4, IRS1 and leptin genes in
obese (Group 2) and diabetic rats (Group 3) were downregulated in adipose
tissues compared to normal group (Group 1) while leptin gene was
upregulated. However, GLUT4 and IRS1 genes were significantly upregulated
higher in treated diabetic rats in Group 6 followed by group 5, 4 and 7. Those
genes are responsible in insulin signalling, glucose transportation and also in
metabolism of glucose and lipid. According to Rakhshandehroo et al.(2010)
high fat diet may down regulates several transcription factors such as nuclear
receptor (PPAR) and sterol regulatory binding proteins (GLUT). This study is
parallel to finding of Rakhshandehroo et al. (2010) because our diabetic model
270
that has been developed through HFD with combination of STZ also shows
similar results. Downregulation of IRS-1 and GLUT4 genes may defect insulin
signaling and glucose uptake activity. Meanwhile, downregulation of leptin
gene may affect glucose and lipid metabolisms in diabetic rats (Wang et al.
2009).
Table 2. Expression of of GLUT4, Lep and IRS1 genes in adipose tissues after 30
days treatment with C. latifolia fruit:root
CONCLUSION
ACKNOWLEDGEMENTS
This work was supported by the Fundamental Research Grant Scheme (FRGS)
from the Ministry of Higher Education, Malaysia.
REFERENCES
Bansal, P., Paul, P., Mudgal, J., Nayak, G., Thomas, P.S., Priyadarsini, K.I, et al.
(2011). Antidiabetic, Antihyperlipidemic and Antioxidant Effects of the
Flavonoid Rich Fraction of Pilea microphylla L. in High Fat
Diet/Streptozotocin-induced Diabetes in Mice. Experiment Toxicology
and Pathology. doi:10.1016/j.etp.2010.12.009.
271
Chooi, O. H. (2006). Tumbuhan liar: khasiat ubatan dan kegunaan lain. Utusan
Publications and Distributors Sendirian Berhad, Kuala Lumpur, p. 116.
Ibuka, A.S., Morita, Y., Terada, T., Asakura, T., Nakajima, K., Iwata, S., Misaka,
T., Sorimachi, H., Arai, S., and Abe, K. (2006). Crystal Structure of
Neoculin: Insights into its Sweetness and Taste-modifying Activity.
Journal Molecular Biology. 359:148─158.
Nur Akmal, I., Maznah, I., Muhajir, H., Zalinah, A. and Siti Aisyah, A. G. (2013).
Antidiabetic and Hypolipidemic Activities of Curculigo latifolia fruit:root
Extract in High Fat Fed Diet and Low Dose STZ induced Diabetic Rats.
Evidence-based Complementary and Alternative Medicine.
http://dx.doi.org/10.1155/2013/601838.
Oribe, J., Kakuma, T., Haranaka, M., Okamoto, K., Seike, M. and Yoshimatsu, H.
(2012). Intraperitoneal Administration Attenuates Thiazolidinedione-
induced Hepatic Steatosis in KKAy Mice with Increased Hepatic
Peroxisome Proliferator-activated Receptor (PPAR)γ mRNA Expression.
Obesity Research & Clinical Practice. 6(3): 249─261.
272
Shingo, A. S., Kanabayashi, T., Murasea, T. and Kito, S. (2012). Cognitive Decline
in STZ-3V Rats is Largely due to Dysfunctional Insulin Signaling through
the Dentate gyrus. Behavioural Brain Research. 229:378─383.
Wang, Y., Zhang, Di., Liu, Y., Yang, Y., Zhao, T., Xu, J. et al. (2009). Association
Study of the Single Nucleotide Polymorphisms in Adiponectin-
associated Genes with Type 2 diabetes in Han Chinese. Journal of
Genetics and Genomics. 36(7):417─423.
Wheeler, S., Moore, K., Forsberg, C. W. et al. (2013). Mortality among Veterans
with Type 2 Diabetes Initiating Metformin, Sulfonylurea or
Rosiglitazone Monotherapy. Diabetologia. 56: 1934–1943.
Yokoyama, H., Araki, S., Kawai, K., Hirao, K., Oishi, M., Sugimoto, K., Sone, H.,
Maegawa, H., Kashiwagi, K. (2015). Pioglitazone Treatment and
Cardiovascular Event and Death in Subjects with Type 2 Diabetes
without Established Cardiovascular Disease (JDDM 36). Diabetes
Research and Clinical Practice. 109 (3): 485─492.
Yu, O. H. Y., Yin, H. & Azoulay, L. (2015). The Combination of DPP-4 Inhibitors
Versus Sulfonylureas with Metformin After Failure of First-line
Treatment in the Risk for Major Cardiovascular Events and Death.
Canadian Journal of Diabetes. 39(5):383–389.
Zheng, X.Y., Li, Y.J., Zhang, L., Feng. W.S., & Zhang, X. (2011). Antihyperglycemic
Activity of Selaginella tamariscina (Beauv). Spring. Journal of
Ethnopharmacology 133:531─53.
273
PROTEIN EXPRESSION PATTERN IN SWIETENIA MACROPHYLLA SEED
ABSTRACT
INTRODUCTION
274
system lead to the recognition of proteome technology as an important
research that complements genome studies.
Plant Materials
Plant species used in this research were collected from Yan, Kedah. Swietenia
macrophylla fruits were stored in room temperature for at least 7 days for the
fruits to crack. The fruits were processed to extract the seeds.
Seed Storage
The seeds were randomly selected and then stored in an airtight aluminum bag
for subsequent studies. The seeds were stored at 10°C and 4°C for a period of 6
months to determine seed germination rate after storage.
Protein Extraction
S. macrophylla embryo from the control and 6-month stored seeds were frozen
in liquid nitrogen and ground into fine powder. Protein was extracted using 9
M urea, 4% CHAPS, 0.5% triton X-100 and 100 mM DTT, and incubated for 2 hr
at 4°C. The extract was centrifuged at
12,000 rpm for 10 min. The supernatant was collected to determine the
protein concentration.
275
and finally 72 mA per gel. Gels were stained with silver nitrate and scanned
using ImageScanner (GE Healthcare). The IMAGE MASTER 2D PLATINUM
software by GE Healthcare was used to capture gel images and analysis of all
the 2D gels was performed according to the manufacturer’s instructions.
Vertical and horizontal streakings were also removed and background intensity
was substracted during spot detection. The gels were then matched by
landmarking common spots found in different gels. Normalization was
performed to correct variations in spot size and intensity between gels.
MALDI-TOF/TOF
Protein spots were chosen from the 2-DE gels, excised and identified by mass
spectrometry. Peptides were extracted from the gel plugs by three cycles of
sonication in water, followed by acetonitrile for 15 min. The extracted peptides
were concentrated using vacuum centrifugation and mixed with saturated
matrix. The mixture was spotted onto slides for matrix assisted laser
desorption/ionizing-time of flight (MALDI-TOF) mass spectrometry analysis.
The generated peptide mass values were searched against the Swiss Prot and
NCBInr protein databases.
The present study demonstrated the optimized protein extraction protocol for
S. macrophylla embryo and subsequently the proteomes of two different
sources of seeds i.e. control and stored seeds. This study was an early effort to
establish a platform to identify characteristic proteins involved in the
biochemical and molecular process under cold-stress conditions.
276
assigned sequences in protein sequence databases. However, we did not
proceed to predict the function of hypothetical protein detected in the present
study.
277
CONCLUSION
The proteins identified in the present study may have roles in the regulation of
cell cycle, metabolic pathway and cell defense mechanism or seed maturation.
Changes in their composition could lead to the possibility of protein
deterioration. Information generated from this study will give an impact on the
future of forestry practices and management of seed storage. Further research
can also be conducted by monitoring the possibility of protein-protein
interaction on secondary metabolites changes after seed storage related to the
medicinal value of this species.
REFERENCES
Sheng, W., Hua, X.Y., Sheng, W.W. & Li, X.B. (2003). Protein Phosphatase 2A: Its
Structure, Function and Activity Regulation. Acta Biochimica et
Biophysica Sinica 35(2): 105–112.
Cairns, B.R., Kim, Y.J., Sayre, M.H., Laurent, B.C. & Kornberg, R.D. (1994). A
Multisubunit Complex Containing the SWI/ADR6, SWI2/SNF2, SWI3,
SNF5, and SNF6 Gene Products Isolated from Yeast. Proc. Natl. Acad.
Sci. USA 91: 1950–1954.
Cho, M.H., Shears, S.B., & Boss, W.F. (1993). Changes in Phosphatidylinositol
Metabolism in Response to Hyperosmotic Stress in Daucus carota L.
Cells Grown in Suspension Culture. Plant Physiology 103(2): 637─647.
278
PROTEOME PROFILES OF SEEDS FROM SWIETENIA MACROPHYLLA
AFTER COLD STORAGE
ABSTRACT
INTRODUCTION
279
With the aim of profiling proteins in seed samples, the focus of our
study is to develop a method for the 2-D PAGE analysis of Swietenia
macrophylla seed protein which allows the visualization of a maximum of
proteins from the sample. Preparation of high-quality protein from plant has
been problematic using standard protocols due to high salts, polysaccharides,
lipids and proteases. The improved techniques in protein extraction and 2-D
PAGE enable comprehensive protein visualization on 2-D PAGE gels.
The seed embryos were excised and the protein was homogenised in lysis
buffer containing 9 M urea, 100 mM DTT, 4% CHAPS dan 0.5% triton X-100.
Protein concentration was determined using the 2-D Quant Kit (Amersham
Biosciences, Uppsala, Sweden). The quantification protocol was performed as
described by the manufacturer using Bovine Serum Albumin (BSA) as protein
standard. A standard calibration curve of absorbance values over several
protein concentrations was constructed.
Protein sample (50 μg) was diluted in rehydrating buffer containing urea (6 M),
SDS (2%), Tris/HCL pH 8.8 (0.375 M), gliserol (25%) dan DTT (2%) and applied to
7 cm IPG strips of pH 3–10 overnight. The strips were subjected to two-step
equilibration in buffers containing urea (6 M), SDS (2%), Tris/HCL pH 8.8 (0.375
M), gliserol (25%) and DTT (2%) and followed by 250 mM iodoacetamide and
applied to the 12 % SDS-PAGE gels. The second dimension electrophoresis was
performed by SDS-PAGE with an SE 600 Ryby (GE Healthcare, Sweden)
electrophoresis unit. Gels were stained with silver stain according to
Berkelman and Stenstedt (1998). Digital images of the analytical gels were
acquired and analysed quantitatively for differentially expressed proteins using
ImageMaster 2D Platinium 7.0 analysis software (GE Healthcare, Sweden).
From the protein extraction, seeds that underwent 3 months storage at 4oC
yielded a total protein of 5.2 ± 0.14 μg/μL. The proteins profiles obtained from
2-Dimensional Gel Electrophoresis (2-DE) were established using immobilized
pH-gradient (IPG) strips pH 3–10, 7 cm.
280
Gel image analysis was performed by comparing the occurrence of
every spot among the sets of protein profiles from fresh sample and cold
storage treatment. Proteome profiling experiment via 2-DE detected at least
240 protein spots expressed in fresh samples and 12 proteins were
differentially expressed after storage. From the analysis of 12 differentially
expressed protein spots (parameters: >1.5 folding value & p<0.05); seven
proteins were down-regulated and five proteins were up-regulated. Figure 1
shows the representative gel images of fresh (b) and seeds underwent cold
storage. While Figure 2 shows the representatives’ gel spots of selected protein
of interest.
+ +
+ +
+
+ + +
66 + 7
+ +
3 6
3 6
+
+ + +
+ of interest.
+ (a) down-
Figure 2. Representatives
+ gel spots of selected
8
protein
77 8 +
4 and +
regulated (b) up-regulated protein spots. +
7
4 7 +
+
+
+ ++ 281 9
9
88 + + + +
5 8 + +
+ + +
810 +
5 +
+ 10
99 +
CONCLUSION
ACKNOWLEDGEMENTS
REFERENCES
Berkelman, T., & Stenstedt, T., (1998). 2-D Electrophoresis- Principle and
Methods. Sweden: Amersham Bioscience.
282
EFFECT OF pH ON ADSORPTION OF ORGANIC ACID AND PHENOLIC
COMPOUNDS IN NONI (MORINDA CITRIFOLIA L.)
ABSTRACT
INTRODUCTION
Morinda citrifolia L. or noni has been used traditionally to treat a broad range
of diseases for over 2000 years (Dixon et al. 1999). About 160 phytochemical
compounds have already been identified in noni plant, with the major
micronutrients being phenolic compounds, organic acids and alkaloids (Wang &
Su 2001). The unpleasant odour of noni extract is contributed by medium chain
fatty acids such as decanoic, hexanoic and octanoic acids (Norma et al. 2004).
Recently, more attention has been focused on the role of natural antioxidants,
in particular, phenolic compounds. Scopoletin is a characteristic phytochemical
in noni, while rutin and quercetin are bioactive flavonoids (Deng et al. 2010).
Due to the beneficial role of antioxidants, it is important that deacidification
did not reduce the antioxidant activity.
283
This study will focus on the adsorption behaviour of organic acids
(octanoic acid and hexanoic acid) and phenolic compounds (rutin, scopoletin
and quercetin) onto Amberlite IRA 67 resin in a model system. Apart from that,
this research is also important to obtain more information on the interaction of
organic acids constituents and phenolic compounds with the resin using multi-
compounds system.
Materials
Weak base anion exchanger Amberlite IRA 67 (food grade) and standard
compound of octanoic acid, hexanoic acid, rutin, scopoletin and quercetin were
purchased from Sigma Aldrich Corporation. Methanol (purity > 99.9%), HPLC
grade was purchased from Fisher Scientific.
The adsorption studies were carried out in Erlenmeyer flasks where 0.1 g of the
resin and 0.02 l of each samples which were octanoic acid (250 mg/l), hexanoic
acid (100 mg/l), rutin (25 mg/l), scopoletin (25 mg/l), and quercetin (25 mg/l),
were added without pH adjustment. The flasks were agitated in the orbital
shaker at 120 rpm for 180 min to achieve equilibrium. After which the
adsorption of each of the organic acids was measured using a Gas
Chromatography. While adsorption of each of the phenolic compounds was
measured using HPLC. The amount of organic acids and phenolic compounds
adsorbed at equilibrium, qe(mg/g), were calculated based on the equation
below:
Where Co and Ce are the concentrations of the each organic acids and phenolic
compounds at the beginning and in the equilibrium, respectively (mg/L); V is
the volume of the solution (L); w is the mass of the dry anion exchanger (g).
pH Determination
pH determination was carried out on all samples using a pH meter (Model PB-
10, Sartorius Basic Meter, Germany).
284
Determination of Ferric Reducing Assay (FRAP)
FRAP assay was conducted according to the methods of Benzie and Strain
(1996). The results were expressed as μmol/ g fresh weight (FW) sample.
The antioxidant activity of all juices were evaluated based on free radical
scavenging effect according to the methods of Akowuah et al. (2005).
Percentage of free radical scavenging activity was calculated based on the
formula below:
285
Figure 1. The effect of organic acids of multi-compounds (A0 = 25 ppm; OA =
250 ppm; HA = 100 ppm) using Amberlite IRA 67 resin at different pH (Resin
amount 100 mg; agitation rate: 120 rpm)
286
Figure 2. HPLC determination of multi-compounds (A0 = 25 ppm; OA = 250
ppm; HA = 100 ppm) using Amberlite IRA 67 resin at different pH (Resin
amount 100 mg; agitation rate: 120 rpm)
CONCLUSION
ACKNOWLEDGEMENTS
The authors would like to thank The Ministry of Education (MOE) for the
scholarship, The Ministry of Science, Technology and Innovation (MOSTI) for
financing the project under the ERGS/1/2013/STWN03/UKM/02/1 grant and
Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia.
REFERENCES
Akowuah, G. A., Ismail, Z., Norhayati, I. & Sadikun, A. (2005). The Effects of
Different Extraction Solvents of Varying Polarities on Polyphenols of
Orthosiphon stamineus and Evaluation of The Free Radical-Scavenging
Activity. Food Chemistry 93(2): 311–317.
287
Benzie, I.F.F. & Strain, J. J. (1996). Ferric Reducing Ability of Plasma (FRAP) as a
Measure of Antioxidant Power: The FRAP Assay. Analytical
Biochemistry 239: 70-76.
Deng, S., West, B.J. & Jensen, C.J., 2010. A Quantitative Comparison of
Phytochemical Components in Global Noni Fruits and Their Commercial
Products. Food Chemistry, 122(1), 267–270.
Duh, P. D., Tu, Y. Y., & Yen, G. C. (1999). Antioxidant Activity of Water Extract of
Harn Jyur (Chyrsanthemum morifolium Ramat).
LebensmittelWissenschaft und-Technologie-Food Science and
Technology 32: 269–277.
Norma, H., Normah, A., Ahmad, A.W., Rohani, M.Y., Muhammad Gawas, M. &
Sharizan, A. (2004). Reducing The Smelly Compounds (Caproic, Caprylic
and Capric acids) in Noni by Treating The Juice with Activated Charcoal
Powder. Proceeding of the National Food Technology Seminar.
Wang, M.Y. & Su, C. 2001. Cancer Preventive Effect on Morinda citrifolia
(Noni). Annals of the New York Academy of Sciences 952:161─168.
288
EFFECT OF ORGANIC-BASED FERTILIZER RATE AND PLANTING DISTANCE
ON BIOMASS YIELD OF BELALAI GAJAH (CLINACANTHUS NUTANS)
ABSTRACT
289
INTRODUCTION
Belalai gajah (Clinacanthus nutans is one of the highly valued herbs that has
been identified for developement under Entry Point Project (EPP1). Belalai
gajah (BG) can be traditionally used for diabetes mellitus, fever, diarrhoea and
dysuria (Uawonggul et al. 2011). In recent years, various pharmacological
properties such as immunomodulatory (Sriwanthana et al. 1996), cytotoxic
(Thongrakard & Tencommao 2010), anti-oxidant (Yong et al. 2013) and anti-
viral (Sakdarat et al. 2009) have been previously reported. BG is a small shrub
that native to tropical Asia countries and a member of the Acanthaceae family.
The plant grows well under the scattered forests and bushes on the
low altitude area up to 620 meters. In recent years, the plant has gained
popularity among Malaysian especially for treating cancers. The leaf is the
most useful plant part due to the various phytochemicals and flavanoids
content. Thus, a study was conducted to determine optimum fertilizer rate
and planting distance to increase the biomass leaf yield of BG.
The field experiment was conducted in MARDI Serdang, Selangor. The plants
were propagated using stem cuttings. Stem was cut into 2 – 3 nodes and was
placed in germination trays (104 plugs) containing mixture of subsoil and
Holland peat at the ratio of 3:1. The germination tray were exposed under 25
% shade for better germination rate and the seedlings were ready to be
transplanted after 30 days. Land preparation for field planting was done by
one round of disc-ploughing about 25 – 30 cm deep and after 1-2 week, land
was rotovated for another round. The soil beds were raised about 30 cm
(height), 1.2 m (width) and 4.5 m (length). The beds were later covered by
plastic mulching (UV silver shine) to control weeds. Planting holes were set up
a day before transplanting and one seedling was transplanted into each hole.
The seedlings were planted at a spacing of 40 cm between rows and different
interplant spacing at 30, 40, 50 and 60 cm.
290
to avoid further deterioration.
The interaction between fertilizer rate and planting distance was observed only
on biomass yield at first harvest but no interaction for the second, third and
total harvest. Fresh yield of BG at first harvest had increased with the
increasing rate of fertilizer for 40 cm x 40 cm and 40 cm x 50 cm. After the
application rate of 220 kg fertilizer/ha at 40 cm x 40 cm and after 200 kg
fertilizer/ha for 40 cm x 50 cm, the fresh yield had decreased. However, the
highest yield was at 40 cm x 30 cm and 40 cm x 40 cm planting distance. The
interaction showed that the optimum fertilizer rate was 220 kg/ha with
planting distance at 40 cm x 40 cm. At these rates and distances, fresh yield
and dry yield were at 300 kg/ha (Figure 1) and 80 kg/ha (Figure 2) respectively.
2 2 2
*S1, y=219.26 + 0.4776x (R = 0.95), S2, y=186.07 + 1.992x – 0.0062x (R =0.83),
2 2 2
S3, y=106.92 + 0.407x – 0.0009x (R =0.95), S4, y=90.19 + 0.3009x (R =0.96).
Figure 1. Interaction between fertilizer rate and plant spacing on fresh yield of
belalai gajah at first harvest
291
2 2 2
*S1, y=58.84 + 0.1123x (R = 0.93), S2, y=45.24 + 0.517x – 0.0016x (R =0.85),
2 2 2
S3, y=27.48 + 0.1117x – 0.0003x (R =0.87), S4, y=22.93 + 0.0745x (R =0.96).
Figure 2. Interaction between fertilizer rate and plant spacing on dry yield of
belalai gajah at first harvest
At the third harvest, the highest biomass yield (888 kg/ha of fresh
yield and 224 kg/ha of dry yield) was at fertilizer rate of 300 kg/ha. Similar
trend was observed with second harvest on planting distance of 40 cm x 30 cm
and 40 cm x 40 cm (Table 2).Total yield from all harvests showed that biomass
yield was highest at fertilizer of 300 kg/ha for both fresh and dry yield. The
results also showed that planting distance of 40 cm x 30 cm and 40 cm x 40 cm
was better than 40 cm x 50 cm and 40 cm x 60 cm.
292
Based on research done by Mengel (1982), the highest yield increment
occurred at the lower rate of nitrogen fertilizer applied but with successive
applications of nitrogen, the yield increments become progressively smaller.
On the other hand, according to the Acquaah (2001), when the nutrient are
present in non-limiting amounts, plants have a tendency to use more than they
need. Thus, up to a certain point, uptake of nutrients does not translate into
increased biomass yield or plant productivity. While a research done by
Maheshwar (2005) on stevia in Karnataka, India showed that application of 105
kg N ha-1, 30 kg P ha-1 and 45 kg K ha-1 recorded significantly higher dry leaf
yield. It was due to maximum number of leaves per plant and branches per
plant as compared to lower doses of N under loamy soil. The interaction
between fertilizer rate and planting distance occurred in this study is probably
due to the difference of fertilizer amount received by the plants and the
number of plant population at different plant spacing. There were increments
on both fresh and dry yields with later harvests probably due to increase in
plant maturity and increasing in number of new branches and leaves after
several ratoon were done.
Table 1. Fresh and dry yield of BG leaves at different fertilizer rate for harvest
2, 3 and total harvest
Fertilizer Fresh Yield (kg/ha) Dry Yield (kg/ha)
Rate Total Total
(kg/ha) Harvest 2 Harvest 3 Yield Harvest 2 Harvest 3 Yield
0 301c 442d 899d 76c 111d 558d
100 377b 662c 1233c 94b 167c 806c
200 473a 754b 1481b 118a 189b 937b
300 527a 888a 1637a 131a 224a 1075a
Means with the same letter for each harvest are not significantly different by DMRT at p< 0.01.
Table 2. Fresh and dry yield of BG leaves at different plant spacing for every
harvest and total harvest
Plant Fresh Yield (kg/ha) Dry Yield (kg/ha)
Spacing
Total Total
Harvest 2 Harvest 3 Yield Harvest 2 Harvest 3 Yield
40cm x30cm 472a 777a 1540a 117.27a 197.32a 1540.11a
40cm x 40cm 449ab 712ab 1427a 113.2a 178.39ab 1426.91a
40cm x 50cm 399bc 640bc 1175b 99.29b 159.92bc 1175.03b
40cm x 60cm 358c 617c 1110b 89.37b 155.4c 1110.01b
Means with the same letter for each harvest are not significantly different by DMRT at p< 0.01.
293
CONCLUSION
The study concluded that optimum fertilizer rate (NPK 10:5:5) for BG is 210 ±
10 kg/ha for first and second harvest and 300 kg/ha for third harvest based on
biomass yield. It also showed that BG was best cultivated using planting
distance of 40 cm between row and 35 ± 5 cm within row.
ACKNOWLEDGEMENTS
The authors wish to thank Mr. Zakry Al-Asyraf Abd Latiff, Norsyamiza Mat
Yusof and Noor Kamilah Omar for their assistance in implementing the
experiment. This project was funded by MARDI under MEGA Project (P161).
REFERENCES
Uawonggul, N., Thammasirirak, S., Chaveerach, A., Chuachan, C., Daduang, J &
Daduang, S.(2011). Plant Extract Activities Against The Fibroblast Cell
294
Lysis by Honey Bee Venom. Journal of Medicinal Plant Research
5(10):1978─1986.
Yong, Y. K., Tan, J.J., Teh, S. S., Mah, S. H., Ee, G. C., Chiong, H. S., & Ahmad,
Z.(2013). Clinacanthus nutans Extracts are Antioxidant with
Antiproliferative Effect on Cultured Human Cancer Cell Lines. Evidence-
Based Complementary and Alternative Medicine. p:462─751.
295
THE EFFECT OF PACKAGING TECHNIQUE ON QUALITY OF DRIED MAS
COTEK (FICUS DELTOIDEA)
ABSTRACT
INTRODUCTION
Malaysian rainforest has been reported to host more than 2,000 plants with
medicinal value, and in most village around the country, the older generation
still use myriad brews, poultices and pastes derived from herbal plants for
everything from physical ailments to beauty. The use of herbal plants and their
296
related products have become increasingly important worldwide over the past
two decades. Global trade of natural plant products is projected to triple by
2020, with the herbal medicine market expected to grow by 10 and 20 percent.
On the domestic front, herbal industry in Malaysia is estimated to grow at the
rate of 15% per annum, with the market value rising from 7 billion ringgit in
2010 to 29 billion ringgit in 2020 (Nor Mohamad 2011).
297
In the case of prolonging the storage life of the fresh produce, suitable
packaging materials and temperature need to be established. Shelf life of fresh
produce can be further extended under low temperature storage with proper
packaging materials such as perforated LDPE bag. Mariana et al (2011)
reported that proper packaging material of thyme and rosemary had shown
negligible quality loss up to one year of storage. Similar results also found in
coneflower where freeze-dried sample packed in polyethylene bag, aluminium
foil and nylon polyethylene bag have shown highest retention of bioactive
compound (Lin et al. 2011) Thus, there is a need to evaluate the stability of the
phytochemicals and shelf life under storage conditions in order to meet fresh
and dried market demand for leafy herbs.
The plant (mas cotek) was harvested at Farmer’s farm, Pedas, at optimum
maturity of 12 months after planting. The herb was washed and dried in solar
dryer until the moisture content dropped to 10%. Then, the plant was
transported to Post Harvest Laboratory at MARDI, Serdang and packed in
various types of packaging materials, namely low density polyethylene bag,
Nylon-PE bag and laminated aluminium bag. The thickness of the bag is 0.08
mm. The packed herb was stored for 6 months under ambient condition (room
temperature of 25°C). Each packaging materials contain 100 g (0.1 kg) of dried
leaves. The physical and chemical analyses were done at intervals of 45, 90,
135 and 180 days of storage.
Moisture content was analysed using moisture analyser model AND 300. Total
phenolic content and total flavonoids content were determined by method
described by Marinova et al. (2005). The total antioxidant activity was
determined by the method described by Chan et al. (2008). All analyses were
done in 4 replicates.
Statistical Analyses
298
RESULTS AND DISCUSSION
12
Moisture content (% w.b)
10
4 Nylon PE
2 LDPE
Aluminium
0
0 45 90 135 180
Days of storage
299
Table 1. The effect of different packaging technique on antioxidant activity of
mas cotek
Storage period 0 Days 45 Days 90 Days 135 Days 180
Days
Packaging
technique
Nylon PE 86.61 87.59 84.13 86.36 86.5
LDPE 86.61 87.27 82.54 86.00 86.23
Aluminium 86.61 88.32 82.71 88.11 87.06
Packaging
technique
Nylon PE 377.78 447.78 508.7 329.26 371.48
LDPE 377.78 386.85 496.48 392.78 396.00
Aluminium 377.78 387.22 540.00 316.85 362.78
Packaging
technique
Nylon PE 377.78 447.78 508.7 329.26 371.48
LDPE 377.78 386.85 496.48 392.78 396.00
Aluminium 377.78 387.22 540.00 316.85 362.78
300
Table 4. Statistical analysis of mas cotek in different packaging technique
Response Total Total Total Moisture
flavonoids phenolics antioxidant content (%
(mg/100g CE) (ug/g GAE) activity (%) w.b)
Storage time n.s ** ** **
(days)
0 222.57 a 377.78 c 86.59 b 7.58 d
45 206.70 b 407.28 b 87.73 a 8.23 c
90 235.76 a 515.06 a 83.14 c 8.56 b
145 215.05 b 346.30 d 86.85 b 9.72 a
180 217.01 b 376.73 d 86.60 b 8.78 b
Packaging n.s n.s n.s **
material
Nylon PE 226.92 a 409.96 a 85.74 b 9.34 a
LDPE 209.18 b 407.00 a 86.24 ab 9.12 b
Laminated 222.16 a 396.93 a 86.56 a 6.96 c
aluminium
Storage time n.s * n.s **
X packaging
material
CONCLUSION
Improper packaging technique could promote bacteria and fungi growth during
storage. So, selecting the best packaging material is crucial to ensure the safety
of herbal product. Based on this study, it is recommended to use LDPE bag for
bulk packaging because it is cheap, reliable and readily available. The cost of
LDPE bag is comparable with Nylon PE bag but LDPE bag is much stronger with
high tensile strength. Nylon PE is not suitable for bulk packaging because the
material is easily broken and could not withstand heavy load. The cost for
aluminium bag is more expensive compared to others but this material provides
good moisture barrier and it is suitable for retail packaging.
REFERENCES
Chan, E.W.C., Lim, Y.Y., Wong, L.F., Lianto, F.S., Wong, S.K., Lim, K.K., Joe, C.E. &
Lim, T.Y. (2008). Antioxidant and Tyrosinase Inhibition Properties of
Leaves and Rhizomes of Ginger Species. Food chemistry 109: 477–483.
301
Lin, S., Sung, J.M., & Chen, C. (2011). Effect of Drying and Storage Conditions on
Caffeic Acid Derivatives and Total Phenolics of Echinacea Purpurea
Grown in Taiwan. Food Chemistry 125: 226–231.
Jones, R., Premier, R. & Tomkins, B. (2006). The Effect of Postharvest Handling
Conditions on Phytochemicals Important for Human Health Contained
in Fruits and Vegetables. In Advances in postharvest technologies for
horticultural Crops. CABI publication. Pp. 1–19.
Lachman, J., Proněk, D., Hejtmánková, A., Dudjak, J. & Pivec, K. (2003). Total
Polyphenol and Main Flavonoid Antioxidants in Different Onion (Allium
cepa L.) Varieties. Hort. Sci. (Prague) 30(4): 142–147.
Marianna, U., Mauro, M., Marzia, F., Alessandra, D.C. & Roberta, D. (2011).
Influence of Different Stabilizing Operations and Storage Time on the
Composition of Essential Oil of Thyme (Thymus officinalis L.) and
rosemary (Rosmarinus officinalis L.). LWT – Food Science and
Technology 44: 244–249.
Marinova, D., Ribarova, F. & Atanassova, M. (2005). Total Phenolics and Total
Flavonoids in Bulgarian Fruits and Vegetables. Journal of the University
of Chemical Technology and Metallurgy 40(3): 255–260.
Matthews, R.F., Locasio, S.J. & Ozaki, H.Y. (1975). Ascorbic Acid and Carotene
Contents of Pepper. Proc. Florida State of Horticultural Science Society
88: 263–265.
302
POTENTIAL OF MEDICINAL PLANTS USED BY THE JAKUN PEOPLE AS
ANTITUBERCULOSIS AGENTS
ABSTRACT
Tuberculosis (TB) is a global health problem that causes high rate of mortality
by a single pathogen called Mycobacterium tuberculosis. The effective yet
problematic antituberculosis drugs and the emerging resistant strains of TB
persist to bring challenges in TB medication. Therefore, the investigation of
traditionally used plants by the Jakun people to treat TB and its related
symptoms could be an ideal approach in searching for novel antituberculosis
agents to improve the circumstances. This study was carried out to screen
antimycobacterial activities of eight selected medicinal plants, namely
pengesep (Rourea mimosoides), keruing air (Dipterocarpus sublamellatus),
perut keletung (Thottea praetermissa), hempedu beruang (Thottea
grandiflora), akar ipoh (Stryhnos ignatii), akar empelas (Tetracera macrophylla)
and segindu (Rennellia elliptica), extracted in different solvents (aqueous, 80%
MeOH, hexane, ethyl acetate and 100% MeOH). Antimycobacterial activities
were assessed through zone of inhibition (ZOI) by agar disk diffusion, minimum
inhibitory concentration (MIC) by microbtroth assay and minimum
mycobactericidal concentration (MMC) by conventional plating. In this study, a
model organism of TB strain, Mycobacterium smegmatis mc2 155 (ATCC
700084), was shown to be susceptible against the plant crude extracts by
showing different ranges of antimycobacterial activities. Ethyl acetate root
extract of T. grandiflora exhibited the largest ZOI value of 15.08 ± 0.86 mm
while both hexane and ethyl acetate extracts of D. sublamellatus showed the
lowest MIC and MMC values of 0.78 mg/ml and 3.13 mg/ml respectively. The
results suggest that majority of traditionally used plants by the Jakun people
possess antimycobacterial properties and their active crude extracts have a
great potential in the development of future antituberculosis agents.
303
INTRODUCTION
Prior Informed Consent (PIC) was obtained from the Jakun’s traditional herbal
practitioners (THM) to document their knowledge about medicinal plants used
to treat TB and its related symptoms. Plant samples were collected under a
permit granted by Perbadanan Taman Negara Johor (PTNJ). The samples were
collected from areas mentioned by the THM around Kampung Peta and Taman
304
Negara Johor Endau Rompin in March, 2014. Voucher specimens were
prepared and identification of plant species were verified by Mr. Kamarudin
Salleh (Forest Research Institute Malaysia, FRIM). The species of plants and
parts used were detailed in Table 1.
All samples were allowed to air-dry (AD) for four weeks and pulverized into fine
powder. The powdered samples were extracted in five solvents by:
1) decoction in distilled water for 15 min; 2) maceration in 80% MeoH for 2
hours with constant agitation and repeated three times; and
3) consecutive maceration in hexane, ethyl acetate and 100% MeOH for 24
hours without agitation and repeated three times. Crude extracts were filtered
through Whatman no. 1 filter paper. Aqueous extracts were lyophilized using
freeze-drier while organic extracts were concentrated using rotary evaporator.
305
incubated overnight at 37°C and assessed for color development. A change
from blue to pink indicates reduction of resazurin due to bacterial growth. The
MIC was defined as the lowest concentration of a sample that prevented this
colour change. Prior to determination of MIC method,
100 l of broth suspension from the microplate wells that showed no colour
change were inoculated onto fresh agar plates. The agar plates were incubated
for 3 days at 37oC. The MMC was defined as the lowest concentration that did
not harbour any bacterial growth on agar plates. Tests were performed twice
in triplicates.
The results suggest that majority of traditionally used plants by the Jakun
people possess antimycobacterial properties against M. smegmatis and their
active crude extracts have great potential in the development of future
antituberculosis agents. As shown in Table 1, ethyl acetate extract of T.
grandiflora exhibited the largest ZOI value of 15.08 ± 0.86 mm, followed by
ethyl acetate extract of T. praetermissa (12.08 ± 2.15 mm) and aqueous extract
of T. grandiflora (11.33 ± 0.52 mm) while all crude extracts of R. elliptica
showed no ZOI value at all.
Table 1. ZOI when M. smegmatis was treated with plant extracts in different
solvents at 100 mg/ml
Samples/ Part ZOI (mm)
used
Aq 80% Hex EA 100%
MeoH MeOH
r
R. mimosoides 9.58±0.49 8.92±0.20 7.75±0.69 8.08±0.74 7.00±0.32
s
H. macrocarpa NI NI 7.08±0.38 7.75±0.88 6.92±0.38
r
R. elliptica NI NI NI NI NI
sb
D. sublamellatus 9.67±0.41 8.00±0.63 9.17±0.26 8.75±0.94 7.58±0.92
r
T. praetermissa 8.83±0.26 NI 7.42±0.49 12.08±2.15 8.42±0.92
r
T. grandiflora 11.33±0.52 NI 7.25±0.27 15.08±0.86 9.75±0.69
s
S. ignatii 7.42±1.02 NI 7.92±0.86 8.58±0.49 NI
s
T. macrophylla 8.00±0.32 8.00±1.00 9.42±1.20 8.42±0.38 7.58±0.38
Aq- Aqueous, Hex- Hexane, EA-Ethyl acetate, MeOH- Methanol, NI- No inhibition, r- root, s-
stem, sb- stem bark
306
macrophylla, and both hexane extracts of D. sublamellatus and T. macrophylla
exhibited the lowest MMC value (3.13 mg/ml).
Table 2. MIC and MMC when M. smegmatis was treated with plant extracts in
different solvents
MIC/MMC (mg/ml)
Samples/ Part used Aq 80% MeoH Hex EA 100%
MeOH
r
R. mimosoides 3.13/25.0 25.0/NA 12.5/25.0 6.25/6.25 12.5/25.0
s
H. macrocarpa NA/NA 25.0/NA 6.25/25.0 6.25/12.5 25.0/25.0
r
R. elliptica NA/NA 25.0/NA 25.0/NA 25.0/NA 6.25/6.25
sb
D. sublamellatus 6.25/NA 12.5/12.5 0.78/3.13 3.13/6.25 6.25/25.0
r
T. praetermissa NA/NA 6.25/6.25 6.25/12.5 6.25/6.25 6.25/25.0
r
T. grandiflora NA/NA 3.13/3.13 6.25/12.5 3.13/12.5 12.5/12.5
s
S. ignatii NA/NA 25.0/25.0 3.13/6.25 3.13/6.25 25.0/25.0
s
T. macrophylla 0.78/NA 12.50/12.5 0.78/3.13 1.56/3.13 12.5/25.0
NA- No activity
CONCLUSION
ACKNOWLEDGEMENTS
307
Research Institute Malaysia (FRIM), Perbadanan Taman Negara Johor (PTNJ),
Perbadanan Bioteknologi dan Biodiversiti Negeri Johor (J-Biotech), and the
Jakun people who unreservedly shared their precious knowledge.
REFERENCES
Chee, B.J. (2005). Medicinal Properties and Common Usages of Some Palm
Species in the Kampung Peta Community of Endau-Rompin National
Park, Johor. J. Trop. Med. Plant 6(1): 79–83.
Cox, P.A. (2000). Will Tribal Knowledge Survive the Millennium? Science
287(5450): 44–45. doi:10.1126/science.287.5450.44.
Seow, T.W., Mohamed, M., Nur, M., Bin, S., Asli, O. & Ehwal, J.H. (2013).
Pembangunan Sosioekonomi Komuniti Orang Asli di Malaysia.
Persidangan Kebangsaan Geografi & Alam Sekitar Kali Ke-4 Anjuran
Jabatan Geografi dan Alam Sekitar, Fakulti Sains Kemanusiaan, hlm.
755–761. Universiti Pendidikan Sultan Idris.
Zumla, A.I., Gillespie, S.H., Hoelscher, M., Philips, P.P.J., Cole, S.T., Abubakar, I.,
McHugh, T.D., Schito, M., Maeurer, M. & Nunn, A.J. (2014). New
Antituberculosis Drugs, Regimens, and Adjunct Therapies: Needs,
Advances, and Future Prospects. The Lancet. Infectious diseases 14(4):
327–40. doi:10.1016/S1473-3099(13)70328─1.
308
MEDICINAL PLANTS USED FOR WOMEN’S HEALTHCARE AMONG THE
JAKUN COMMUNITY IN KG. PETA: A PRELIMINARY STUDY
ABSTRACT
Women in Jakun community have been using medicinal plants to cure their
health problems. It is interesting to note their knowledge in this field. This
preliminary study aims to study the medicinal plants used for Jakun women
healthcare. This research was conducted from January 2015 to June 2015 in
Kg. Peta, Endau Rompin Forest. Four key informants from Jakun community
were chosen to participate in this study. Snowball sampling method was used
to determine the highly experienced practitioner. Semi-structured
questionnaire and informal discussion were conducted. The total number of
medicinal plants that were used for Jakun women’s healthcare in Kg. Peta was
12 species from nine families. Seven species of plants were used by women is
for post-partum recovery. Future investigation and proper documentation
need to be done in order to preserve their knowledge from erosion.
INTRODUCTION
309
of male traditional healers and scholars, without taking into account the
knowledge that is adopted by women (Pfeiffer and Butz 2005; de Boer &
Cotingting 2014). As women are also the main user of the herbs, it is
interesting to note their knowledge on this field. This preliminary study aims to
study the medicinal plants used for Jakun women healthcare.
METHODOLOGY
This preliminary study was conducted from January 2015 to June 2015 in Kg.
Peta, Endau Rompin Forest. Four key informants from Jakun community were
chosen to participate in this study. Snowball sampling method was used to
determine the highly experienced practitioner (d’Avigador et al. 2014). Two
criteria that were taking into account when choosing informants are i) the
recognition that they are local practitioner by the Jakun community, and ii) the
ability to identify plants and explain their uses. Semi-structured questionnaire
was used during the interview session followed by informal discussion during
fieldwork. The available plant specimens were photographed, collected and
processed according to the plant taxonomic method.
From the preliminary study the total numbers of medicinal plants that were
used for Jakun women’s healthcare in Kg. Peta are 12 species from 9 families
(Table 1). Seven species of plants were used by women is for post-partum
recovery. From the result, Rourea mimosoides and Cnestis palala had the same
local name (i.e. pengesep) among Jakun people. These two species belongs to
the same family which is Connaraceae. Most of the mode of administration of
herbal medicine is taken orally in the form of decoction.
The plant part that was mostly used is root. Based on the interview, the
root is the main plant parts used in the Jakun traditional medicine practice.
Among Jakun women, kacip fatimah (Labisia pumila) was used for post-partum
treatment. It could induce uterus shrinking. This is in agreement with the data
collected by Taylor and Wong (1987). The mixture of medicinal plants was also
used for the post-partum treatment. According to the informant, after
childbirth, Jakun women usually drink the mixture of L. pumila root, R.
mimosoides or C. palala root and Microporus xanthopus decoction. This is to
ensure full recovery and provide energy to the mother. Interestingly, Jakun
women also used one species of fungi which is M. xanthopus.
310
Table 1. Medicinal plants used for women healthcare in Jakun community.
Botanical Parts Methods of Application Ways of
information used preparation administrati-
on
1. L. pumila Benth. Root Decoction in Post-partum recovery Oral : drink
Kacip fatimah water (inducing uterus
Myrsinaceae shrinking); Fertility &
health tonic
2. C. palala Root Decoction in Post-partum recovery Oral : drink
Pengesep water (inducing uterus
Connaraceae shrinking); Health tonic
3. R. mimosoides Root Decoction in Post-partum recovery; Oral : drink
(Vahl.) Planch water Health tonic
Pengesep
Connaraceae
4. M. xanthopus Fruiting Decoction in Post-partum recovery Oral : drink
Kulat kelentit kering body water (inducing uterus
(3K) (decoction shrinking)
Polyporaceae with L. pumila
and C. palala)
5. Helminthostachys Root Decoction in Post-partum recovery; Oral : drink
zeylanica (L.) Hook water For beauty purposes Topical :
Paku tunjuk langit Flower Raw applied to
Ophioglossaceae skin
6. Rennellia elliptica Root Decoction in Facilitate childbirth Oral : drink
Korth. water
Segindu
Rubiaceae
7. Mapania sp. Root Decoction in Post-partum recovery Oral : drink
Pandan serapat water (inducing uterus
Cyperaceae shrinking)
8. Penggugur Root Decoction in Abortifacient Oral : drink
water
9. Ixonanthes icosandra Root Decoction in Induce fertility Oral : drink
Jack water
Pepagar
Ixonanthaceae
10. Cinnamomum sp. Root Decoction in Post-partum recovery Oral : drink
Teja lawang water
11. Melastoma sp. Leaves Decoction in Post-partum recovery; Oral : drink
Sekenduduk putih water Internal body pain Eaten raw
Melastomataceae Flower Raw
311
CONCLUSION
The results of our preliminary study shows that the women in Jakun
community still rely on the medicinal plants to treat women’s disorder. Future
investigation and proper documentation need to be done in order to preserve
their knowledge from erosion.
ACKNOWLEDGEMENT
The authors wish to thank the Johor National Park Corporation for their
assistance during fieldwork and Jakun community in Kampung Peta, Endau-
Rompin for their time, effort and willingness to share their knowledge with us.
REFERENCES
D’Avigdor, Elizabeth, Hans Wohlmuth, Zemede Asfaw, & Tesfaye Awas. 2014.
The Current Status of Knowledge of Herbal Medicine and Medicinal
Plants in Fiche , Ethiopia. Journal of Ethnobiology and Ethnomedicine
10 (38): 1–32.
Pfeiffer, J.M. & Butz, R.J. 2005. Assessing Cultural and Ecological Variation in
Ethnobiological Research : the Importance Of Gender. Journal of
Ethnobiology 25: 240–278.
Taylor, C. E. & Wong, K. M. 1987. Some Aspects of Herbal Medicine Among the
Orang Hulu Community of Kampung Peta, Johore, Malaysia. Malaysian
Heritage & Scientific Expedition: Endau Rompin. The Malayan Nature
Journal, 41 (2 &3): 317─328.
312
KAEMPFERIA L.: HERBS AND ORNAMENTAL POTENTIAL OF
ZINGIBERACEAE SPECIES
R Izlamira1, MA Zulkhairi2, MZ Nurin Izzati2 & B Suryanti2
1
Programme of Genetic Resources and Germplasm Conservation Management
(GB1), 2Genebank and Seed Centre (GB), MARDI Jerangau, 21820 Ajil,
Terengganu
Tel: 013-2752142 Fax: 09-838 4208 E-mail: izlamira@mardi.gov.my
ABSTRACT
Kaempferia is a medium-sized genus of small herbaceous in the Zingiberaceae
family. This genus is known for its outstanding beautiful foliage and flower.
Seven species were collected during the study. There are K. galanga L., K.
pulchra Ridl., K. angustifolia Roscoe, K. rotunda L., K. parviflora Wall. ex Baker,
K. gilbertii W. Bull and K. elegens (Wall.) Baker. Most of the Kaempferia species
are generally used as medicinal properties and ornamental except for K.
gilbertii and K. elegens. These two species is less known for its medicinal value,
however showing a great potential as ornamental. This is based on the
beautiful morphological characteristics of their foliage which usually variegated
on the upper surface. All the accessions of collected species were conserved in
MARDI Jerangau germplasm. The genetic diversity serves as the gene pool that
will provide important genes for the future study in the broader scope.
INTRODUCTION
The genus Kaempferia L. comprises about 70 species geographically distributed
from tropical Africa to India and throughout Southeast Asia (Kumar et al.
2013). In Peninsular Malaysia, several taxa such as Kaempferia. galanga, K.
pulchra, K. parviflora and K. rotunda are known locally for their medicinal
properties. The rhizomes of K. galanga are carmative and useful for skin
problems, sore eyes, tonics, inflammation, childbirth, cough, sore throat and
fever (Idha & Helmy 2011). The leaves are also being used in traditional
medicine to treat swelling, headache and rheumatism. K. parviflora is
traditionally used for health promoting, stimulating and vitalizing. It is also
used in several treatments such as dysentery, impotence, constriction and colic
disorders. K. rotunda is widely used in traditional medicine as a wound healing
agent (Syed et al. 2013).
313
Other than its medicinal values, Kaempferia species also have a great
potential as ornamental or landscape plant. It is reported to be one of the most
common genera used as an ornamental other than Curcuma and Globba
(Kuehny 2001, Kuehny et al. 2002). This genus is known for its outstanding
beautiful foliage and flower with most of them having a variegated silver to
purple feather pattern on the upper surface of the leaf. K. pulchra, a wild
species which discovered in the limestone hills of Langkawi Island has been
successfully brought into cultivation, growing in pots, on the ground in shaded
areas and gardens (Larsen et al. 1999). Several other species, including K.
galanga and K. elegens are also often used as a pot plant and decorative
garden plant (Picheansoonthon & Koonterm 2008). The morphological
characteristics of Kaempferia such as size, colour of flower and unique foliage
make this genus gaining increase recognition as an attractive ornamental
species.
314
a b c
d e f g
Figure 1. (a) K. galanga, (b) K. pulchra, (c) K. angustifolia, (d) K. rotunda, (e) K.
parviflora, (f) K. gilbertii and (g) K. elegens
K. galanga L.
Local name: Cekur, cekur Jawa, cengkur or kencur
Distinguished characters: Rhizome strongly aromatic; leaves green, often
horizontal and flat on the ground, lamina elliptic to broad elliptic; inflorescence
enclosed in the two leaf-sheaths; flower white with violet bands in the basal
half.
Uses: ‘Ulam’, medicinal and ornamental; growing as pot plants, the leaves and
rhizomes are used for food and medicine, the rhizomes are an ingredient of
post partum medicine and to treat common cold.
K. pulchra Ridl.
Local name: Cekur batik, cekur hitam or cekur mas
Distinguished characters: Leaves broad elliptic, dark purple to dark green with
or without greyish spot; sheath closely imbricating and enclosing the
inflorescence; flower lilac except for a small white at the base.
Uses: ‘Ulam', medicinal and ornamental; planted as garden ground cover and
pot herbs.
K. angustifolia Roscoe
Local name: Cekur rumput or kunci pepet
Distinguished characters: Leaves erect, green, elliptic-oblong to lanceolate;
inflorescence borne in the two innermost leaf-sheath; flower lilac with a purple
blotch at the centre.
315
Uses: Spiritual and medicinal purposes, the small roots and tubers have
astringent properties and are used in the treatment of cough, dysentery and
diarrhoea.
K. rotunda L.
Local name: Kunyit putih or temu putih
Distinguished characters: Leaves oblong lanceolate, mottled green above,
purple beneath; Inflorescence appearing before the leaves. This genus easily
recognize as the flower consist of four lobes.
Uses: ‘Ulam', medicinal and ornamental; good for healing fresh wounds, leaves
and rhizomes are eaten fresh or cooked and used in cosmetic powder and as a
food flavouring agent.
K. gilbertii W. Bull
Local name: Unknown
Distinguished characters: Leaves are striped at the margins with white
variegation.
Uses: Growing for its beautiful foliage as potting ornamental plant.
316
CONCLUSION
Other than its medicinal value Kaempferia species also have a great potential
as ornamental or landscape plant.
REFERENCES
Kumar, K.M., Asish, G.R., Sabu, M. & Balachandran, I. (2013). Significance of
gingers (Zingiberaceae) in Indian System of Medicine ─ Ayurveda: An
overview. Anc Sci Life: 32(4): 253–61.
Syed, A.I., Saroj, K.R., Niranjan, S., Uma, S.S. & Ranju, S. (2013). Wound
healing activity of Kaempferia rotunda Linn leaf extract. International
Journal of current microbiology and Applied Sciences: 2(12): 7478.
Kuehny, J.S., Sarmiento, M.J. & Branch, P.C. (2002). Cultural studies in
ornamental ginger. Trends in New Crops and New Uses: 477–479.
Larsen, K., Ibrahim, H., Khaw, S.H. & Saw, L.G. (1999). Gingers of Peninsular
Malaysia and Singapore. Natural History Publications (Borneo), Kota
Kinabalu.
317
DISTRIBUTION OF HERBS CULTIVATION IN PENINSULAR MALAYSIA
A Nur Syazni, AR Rohana, AB Ariff Fahmi, Z Nur Fazreen, S Siti Zubaidah & M
Marzalina
Forest Research Institute Malaysia (FRIM), 52109 Kepong, Selangor Darul
Ehsan, Malaysia
E-mail: nursyazni@frim.gov.my
ABSTRACT
In general, herbs are plants that are used for medicinal, food, flavoring and
perfume purposes. Recently, there is an increse on the interest of herbal
products in Malaysia. Under the National Key Economic Areas (NKEA), high
value herbal products is listed under Entry Point Project 1 (EPP 1) focuses in
improving quality and marketing effort. This includes in ensuring sufficient
supply of raw material to fulfil market needs. However, there are issues on lack
of information on location of the herbs cultivation in this country. This paper
highlights distribution of herbs cultivation in Peninsular Malaysia. A census was
carried out from September 2014 to February 2015 to identify location of
herbs cultivators along the roads in Peninsular Malaysia. The information was
gathered based on questionnaire and through face-to-face interview. A total of
462 cultivators were identified in Peninsular Malaysia. Most of the herbs were
cultivated in Pahang, Selangor, Johor and Perak with a total number of 110, 83,
69 and 67 cultivators respectively. In Pahang the most cultivated herb was
Hibiscus sabdariffa (roselle), meanwhile in Selangor, Johor and Perak the
famous herb cultivated were Piper betle (sireh), Aquilaria malaccensis (karas)
and Citrus aurantifolia (limau nipis), respectively. The findings will be useful as
baseline information to further strengthen the industry by identifying the
stakeholders and market structure.
INTRODUCTION
Herbs are known as plants, seeds, or any plant parts that are used as medicine,
flavoring and perfume purposes and even some are taken raw. According to
Sahri (2012), herbs also have been recognized as an alternative medicine and
economical resources. Recently there has been a shift in universal trend from
synthetic to herbal medicine, which can be said as ‘Return to Nature’ (Sharma
et al. 2008). The herbs became famous when there are perceptions that herbal
318
products have fewer or less harmful side effects compared to synthetic or non-
herbal products. Most of the information on usage as well as benefits of herbs
are only based on traditional knowledge from various ethnics inherited from
the ancestors and only few are documented. Malaysia has been put as having a
vast potential size in terms of herbal based market with the diversity of genetic
resources, excellent tropical climate, increasing research and development
interest, increasing demand for natural products and indigenous knowledge
(Ibrahim 2006).
Study Area
This study was conducted across the Peninsular Malaysia which included 12
states. The target respondents for this census study were all herbal cultivators
found in the study area.
Data collection
Data Analysis
The data was analyzed according to number of cultivators by states and herbal
species that were cultivated by states. Further analysis was done using Chi-
319
square test to understand the correlation between the five most cultivated
herbal species and ethnicity of the herbal cultivators.
320
Table 2. Types of herbs cultivated in states across Peninsular Malaysia
Species/ States JHR KDH KEL MEL N.S PHG PRK PER P.P SEL TGG Total
Hibiscus sabdariffa (Roselle) 15 2 1 2 0 52 6 2 6 0 2 88
Orthosiphon stamineus (Misai kucing) 4 1 6 2 5 2 8 1 7 2 0 38
Eurycoma longifolia (Tongkat ali) 4 0 8 0 3 9 1 0 1 0 1 27
Clinacanthus nutans (Belalai gajah) 5 0 4 0 3 5 5 1 5 1 0 29
Ficus deltoidea (Mas cotek) 7 1 1 0 4 2 1 0 1 6 0 23
Labisia pumila (Kacip fatimah) 2 1 1 0 0 0 0 0 1 0 0 5
Momordica charantia (Peria katak) 1 1 0 1 0 0 2 0 1 1 0 7
Piper betle (Sireh) 5 0 1 2 1 1 3 0 1 65 1 80
Cymbopogon nardus (Serai wangi) 4 1 3 2 0 2 1 0 1 0 0 14
Morinda citrifolia (Mengkudu) 2 0 0 0 0 1 1 0 0 0 0 4
Phaleria macrocarpa (Mahkota dewa) 4 1 5 0 3 2 5 0 0 0 1 21
Senna alata (Gelenggang) 1 3 1 0 1 2 1 0 6 0 0 15
Centella asiatica (Pegaga) 2 0 0 1 0 0 1 2 1 2 0 9
Aquilaria malaccensis (Karas) 17 1 4 5 15 12 13 0 0 3 0 70
Citrofortunella japonica (Limau kasturi) 0 0 0 6 0 5 5 1 0 0 0 17
Citrus aurantifolia (Limau nipis) 1 1 0 4 0 14 28 1 0 1 1 51
Andrographis paniculata (Hempedu bumi) 1 0 0 0 2 1 0 0 2 0 1 7
Zingiber officinale (Halia) 0 0 1 0 0 5 0 0 2 0 0 8
Melastoma malabathricum (Senduduk) 1 1 1 1 0 0 0 0 0 0 0 4
Melastoma candidum (Senduduk putih) 1 1 0 0 0 0 0 0 1 0 0 3
Moringa oleifera (Merunggai) 0 0 0 0 1 0 0 0 2 0 0 3
Phyllantus niruri (Dukung anak) 0 0 0 0 2 0 0 0 1 0 0 3
Others 18 8 7 4 4 14 13 0 5 7 1 81
Source: FRIM’s survey, 2014-2015.
321
Table 3 shows the correlation between the number of respondents
based on herbal species and ethnicity. Based on the table, majority of the
cultivators were Malay. According to reports on socio-economic status of
farmers in North West Selangor area by Alam et al. (2010), variations in factors
such as education, politic, wealth, health status, accession to technology,
formal and informal capital were responsible for the variations in socio-
economic characteristics of farmers.
It was noted that P. betle was mostly cultivated or planted herb by the
Indian community and used as medicinal plants. It was also believed that the
species is related to symbol of etiquette and civility among the community
(Kumar 1999).
CONCLUSION
ACKNOWLEDGEMENT
We would like to acknowledge FRIM’s Director General, Y.Bhg. Dato’ Dr. Abd
Latif Mohmod for his support, motivation and assistance. We also thank MOA
for providing the financial support through NRGS fund. Our deepest
appreciation to Herbal Development Division for their support, collaborating
agencies, DoA, MARDI, IMR, FAMA, FELCRA, FELDA, LTKN, KESEDAR, KETENGAH
322
and RISDA for their cooperation, and to the research team members and all
herbal cultivators who were involved in this study.
REFERENCES
Alam, M.M., Siwar, C., Murad, M.W., Molla, R.I., and Mohd Ekhwan. T. (2010).
Socioeconomic Profile of Farmer in Malaysia: Study on Integrated
Agricultural Development Area in North-West Selangor, Agricultural
Economics and Rural Development, Vol. 7(2), pp. 249-265. Available at
<http://www.ipe.ro/RePEc/iag/iag_pdf/AERD1013_249-265.pdf > (ISSN
1841-0375; Publisher- Institute for Agricultural Economics, National
Institute of Economic Research, Romania; India; Indexed in EconLit,
RePEc)
Kumar, N. (1999). Betelvine (Piper Betle L.) cultivation: a unique case of plant
establishment under anthropogenically regulated microclimatic
conditions. Indian Journal of History of Science, 34(1)
Rohana, A.R, Nur Fazreen Z, Ariff Fahmi Ab, Nur Syazni A, Siti Zubaidah S, Lim
Hf & Marzalina M. 2015. Value chain study for holistic development of
herbal industry. Pp. 211–219 in Proceeding 3rd International
conference on Rural Development and Entrepreneurship (ICORE) 2015,
9–11 May, Hebei Province, China.
Rohana, A.R., Nur Fazreen, Z., Ariff Fahmi, A.B., Nur Syazni, A., Siti Zubaidah, S.,
Lim, H.F., Mohd Shahidan, M.A., Rosniza, R., Marzalina, M. & Abd Latif,
M. 2015. Directory of Herbal Cultivators in Peninsular Malaysia, Forest
Research Insitute Malaysia.
Sharma, A., Shanker, C., Tyagi, L. K., Singh, M., & Rao, C. V. (2008). Herbal
medicine for market potential in India: an overview. Acad J Plant
Sci, 1(2), 26─36.
323
HALAL HERBAL PRODUCTS INTEGRITY RISK THROUGH SUPPLY CHAIN: A
CONCEPTUAL STUDY
ABSTRACT
Keywords: herbal products, risks, halal supply chain, integrity, halalan toyyiban
324
INTRODUCTION
Malaysia is well known for its rich natural resources and dense forests.
According to MARDI (2006), the rich flora of Malaysia includes 15,000 known
plant species, of which 2,000 species are used for its medicinal value. This
plentiful supply has contributed to the development of natural products, as
ingredients for food and drinks, cosmetics, and natural health & dietary
supplements. The most popular herb species used by the Malaysian herbal
industry for development of natural products are belalai gajah, dukung anak,
gelenggang, halia, hempedu bumi, kacip fatimah, karas, makhota dewa, mas
cotek, mengkudu, merunggai, misai kucing, pegaga, peria katak, rozel,
sambung nyawa, senduduk putih, serai wangi, sirih, and tongkat ali (FRIM
2015). The increasing health consciousness among Malaysians, coupled with an
increase in expendable income, higher levels of education, and population
growth has increased the demand for natural foods and drinks, natural
cosmetics as well as health supplements. Recently, collagen has gained the
attention of Malaysians, as an ingredient in food and drink products, cosmetics,
medical, and health supplements. However, it is not blindly accepted by the
Muslim consumers. Collagen extracted from ungulates (cattle, pig, and sheep)
are not easily marketable in Malaysia, due to religious obligations of halalan
toyyiban (Mohd. Rizal & Adham 2010). Even though medicines are exempted
from halal food regulations, a prescription medicine encapsulated in a
prohibited gelatine capsule made of porcine gelatine is still considered haram
and unacceptable by Muslims. Due to this, efforts are ongoing in research into
possibilities in halal pharmaceuticals as seen in the case of halal vaccines, as
well as research on nutraceuticals focusing on high-value herbal products like
tongkat ali, misai kucing, hempedu bumi, dukung anak, and kacip fatimah.
HALAL DEFINITION
Halal originates from an Arabic term which means allowed, permissible, lawful
or legal. This term specified that goods or services deemed not harmful and
emphasised as safe to be consumed, by the Syariah law, are permissible or
allowable. The opposite is haram or non-halal which means forbidden,
unlawful or illegal (Tieman 2011). Thereby, according to the Islamic principles
some things must be avoided (Ibn Abd al-Barr 2000). In general, halal and
haram has become important elements for consideration in a Muslim’s life.
The Muslims want assurance that the products they consume, not only follow
the Islamic law , but must be toyyib, which means pure and good (World Halal
Forum 2009; Anas et al. 2010).
As pointed out in the Syariah guidelines, any food, drink, cosmetics and
health supplements must not be najs; or contain things that are najs in or for
325
their processing; or processed using equipment or utensils that are najs; or
come into contact at any time during their preparation, processing and storing.
Najs includes meat from inappropriate slaughtered animals, refuse, faeces,
alcoholic beverages, carrion, as well as pork and all of its by products (Sabiq
2008). Furthermore, according to JAKIM (2009), activities of supply chain such
as processes, manufacturing and packaging, receiving, handling, storing and
delivery of products must be verified and certified as halalan toyyiban by
authorized persons.
HALAL INTEGRITY
Nowadays, consumers are concern about food safety, quality, hygiene and
humane approach along the entire production supply chain. Therefore,
integrity of halal food supply chains has become an important subject matter
(Zailani et al. 2010; Lam & Alhashmi 2008). A number of reasons influence the
integrity of halal food chains. One of the main causes is the length and
complexity of food supply chains that involve lots of handling as it is moved
across great distances before reaching the end consumers. The task of ensuring
halal integrity along such chains is a humongous task to manage and monitor.
The reality of this situation has raised questions on the credibility of halal
status of food products. According to Lodhi (2009); Khan (2009); Tieman (2011)
halal integrity is one of the main contributors to developing a well trusted halal
food supply chain. The halal integrity of a product, particularly herbal-based
products will not be compromised as it moves from upstream to downstream
along the herbal supply chain, which starts from the procurement of herbal
raw materials till the finished products reach the consumers. Previous
literature has revealed that all players in the supply chain should take the
responsibility without leaving the huge tasks to a single player (Mohd Hafiz et
al. 2013; Melatu Samsi et al. 2011). Past studies on halal indicated that the
halal integrity of a product is only intact when the product is still in the custody
of an organization (Jaafar et al. 2011). Once a product is moved along the
supply chain, the quality assurance depends on the next player who handles
that particular product. Thus, the integrity of a product cannot be guaranteed if
the other players in the supply chain do not practice similar halal concepts. The
halal herbal products supply chain has similar fundamental principles as other
food supply chains that is, “from farm to fork” concept. The long and complex
herbal-based product supply chains still remain an obstacle to ensure halal
integrity, due to the involvement of many players including suppliers and
producers/manufacturers. Therefore, there is also a risk to food integrity along
the supply chain. Past literatures revealed six risks associated with halal
integrity along supply chains; namely production risks, raw material risks, food
security risks, outsourcing practices, service risk, and logistics risk. Table 1
shows the six types of risks correlated with halal integrity.
326
Table 1. Halal Integrity Risks
Types of Explanation
Risks
Production Production is known as the most important stage in determining the
quality of the food for the customer. Along the long and complex halal
food supply chain, the most significant risk at the production stage is
the contamination from non-halal elements used in halal certified
ingredients. It is a must for manufacturers to obtain halal certification
which covers the ingredients used for production. Besides, Talib and
Johan (2012) mentioned that contamination from the equipment may
affect halal integrity.
Raw Very few countries aside from Malaysia implement halal food
Materials certification and standards through the government’s collaboration.
Due to this, the integrity of imported raw materials cannot be assured.
Tse and Tan (2011) supported that there is high risk to halal integrity
especially when there is low traceability of material origins. It is
understandable that the confirmation of halal certification practices by
suppliers is only set up at the very early stage of appointment.
Food As the process of supplier selection becomes inflexible and strict, firms
Security have limited options to leverage the potential source available in the
market. As a result of this, the local halal food industry directs the firm
to source from the international market. Other than that, halal food
security can be breached due to unexpected situations such as unusual
and/or seasonal demands, product shortages, poor management, and
price fluctuations of certain raw materials in the small local market. This
often leads to high risks for halal integrity (Mohd Helmi et al. 2013).
Services According to Wildes (2005), the most critical issue in an effective service
delivery system is the chance of human errors. From the perspective of
halal food production, a specialized and dedicated group of workers
should handle the production in order to minimize the probabilities of
human error. Generally, halal integrity concentrates on the risk of
human error and cross contamination particularly in a restaurant
setting (Talib & Johan 2012).
Outsourcing Current trends show that firms intend to outsource various processes
such as logistics, warehousing, packaging, marketing and other related
activities in order to lower their production costs. Their main goal is to
focus on core business within the firms and outsource other activities to
other service providers. The outsourcing makes the supply chain longer
and difficult to keep track of along the chain. As a result, firms often
lose control and lead to high integrity risks (Lyless et al. 2008).
Logistics The halal guidelines clearly state that halal products should not be
contaminated with non-halal products. Jaafar et al. (2011) revealed in
his study that transportation vehicles need to be cleaned and
consignments properly segregated in order to ensure that the
requirements are observed and the logistics dedicated to the halal
product shipment. This has become a major challenge for the industry
as there are very few halal logistics providers to cater to this demand.
327
It is inarguable that ensuring the halal integrity of herbal-based
products remains the biggest challenge in the halal herbal industry. The halal
food market including herbal-based market is a huge and lucrative business. It
is estimated to grow significantly in the next decade. Thus, the herbal-based
products supply chain players must ensure that there is no contamination,
intentionally or unintentionally in producing the products. More importantly,
halal integrity has become one of the key successes for the entire halal
industry, and without halal integrity, we as consumers would be living in a
maze of 'false integrity'. Thus, this concept paper aims to study on the
relationship between risks and halal integrity of herbs and herbal products
along the herbal supply chain.
METHODOLOGY
CONCLUSION
328
future, the risks must be given serious attention in order to ensure the needs
and well-being of the halal food consumers. Based on the findings, it is hoped
that the major players in the halal industry especially the suppliers, producers,
logistic service providers, wholesalers, traders, and government will give higher
consideration to the factors that influence the integrity of halal herbal food
supply chain. Besides that, it is also hoped that the outcome from this study
could improve and enhance competitiveness of the Malaysian halal herbal
industry.
REFERENCES
Jaafar, H.S., Endut, I.R., Faisol, N. & Omar,E.N. (2011). Innovation in Logistics
Services - Halal Logistic. In Proceedings of the 16th International
Symposium on Logistics (ISL), Berlin, Germany, no. 34665, 844–851.
Lam, Y. & Alhashmi, S.M. (2008). Simulation of Halal Food Supply Chain with
Certification System: A Multi-agent System Approach. PRIMA ’08:
Proceedings of the 11th Pacific Rim International Conference on Multi-
agents: Intelligent Agents and Multi-agent System
Lodhi, A.-u.-H. (2009). Understanding Halal Food Supply Chain, London: HFRC
UK Ltd
Lyles, M.A., & Flynn, B.B. & Frohlich, M.T. (2008). All Supply Chains Don’t Flow
Through: Understanding Supply Chain Issues in Product Recalls.
Management and Organization Review, 4(2), 167–182
Melatu Samsi, S.Z., Tasnim, R. & Ibrahim, O. (2011). Stakeholders' Role for an
Efficient Traceability System in Halal Industry Supply Chain, in Annual
International Conference on Enterprise Resource Planning + Supply
Chain Management (ERP + SCM 2011): Penang, Malaysia.
329
Mohd Hafiz, Z., Marhani, M.A. & Mohamed Syazwan, Ab.T. (2014). Conceptual
Framework on Halal Food Supply Chain Integrity Enhancement.
Procedia - Social and Behavioral Sciences, (121), 58 – 67.
Mohd Helmi, A., Kim, H.T. & Zafir, M.M. (2013). Mitigating Halal Food Integrity
Risk through Supply Chain Integration. Asia Pacific Industrial
Engineering and Management System.
Sabiq, S. (2008). Fiqih Sunnah. Jakarta: Pena Pundi Aksara, Page: 25–65. [in
Malay].
Tse, Y.K & Tan, K.H. (2011). Managing Product Quality Risk in a Multi-tier Global
Supply Chain. International Journal of Production Research, 49(1),139–
158
Wildes, V.J. (2005). Stigma in Food Service Work: How it Affects Restaurant
Servers’ Intention to Stay in the Business or Recommend a Job to
Another, Tourism and Hospitality Research, 5(3), 213–233
Zailani, S., Arrifin, Z., Abd Wahid, N., Othman, R. & Fernando, Y. (2010). Halal
Traceability and Halal Tracking Systems in Strengthening Halal Food
Supply Chains for Food Industry in Malaysia (a review). Journal of Food
Technology, 8(3), 74─81.
330
CONSUMER PREFERENCES IN SELECTING HERBAL PRODUCTS
ABSTRACT
Malaysia has identified that herbal industry could help in generating gross
national income (GNI). Under National Key Economic Area (NKEA), in
transitioning from agriculture to agribusiness, the first entry point project is
unlocking value from Malaysia’s biodiversity through high-value herbal
product. One of the efforts to produce high-value herbal products is the
industry need to properly strategize their production based on actual
consumer preference. In order to understand consumers’ behavior on herbs
and herbal based products, a study on Consumer Preference and Demand
towards Herbal Based Product in Peninsular Malaysia was conducted in 2013.
The survey was conducted by using stratified random sampling based on
population in each district of Peninsular Malaysia. A total of 4,452 respondents
were interviewed using structured personal interview. The result from this
study highlights status on the use of herbal based products among the
respondents and their criteria in selecting herbal products. The findings
showed that 73% of respondents were currently using herbal based products.
A total of 2,499 respondents (76%) agreed that products with certificate from
Ministry of Health (MoH) are the most important aspect in herbal based
product selection. Aside from MoH certificate, Good Manufacturing Practice
(GMP) certificate was also an important concern of 72% of the respondents
concern. This study showed that the industry, especially manufacturers should
produce herbal based product with good quality and safety. Related agencies
should also help the manufacturer acknowledged to better understand the
procedure to get certificates.
INTRODUCTION
Import value for herbal based products such as plant based, ginseng root,
additional food, skincare products and pharmaceutical in 2012 was USD 402
million (Rohana et al. 2013). This indicated that demand for herbal based
products among Malaysians is quite high. Rohana et al. (2014) reported that
45% of the herbs and herbal products consumers in Malaysia used herb in
traditional way while 46% of them used contemporary value-added products.
331
In order to produce high-value herbal products that meet consumer demand,
production based on the actual consumer preferences are needed.
The respondents for this study were indentified by stratified random sampling
based on population in each Majlis Perbandaran with consideration on the
gender and age to represent the whole Peninsular Malaysia. A total of 4,452
respondents were interviewed via personal interview using structured
questionnaire.
The objective of this study was to assess the pattern of consumer behavior on
herbs and herbal based products. The Theory of Planned Behavior was used as
a guideline to develop the question in the Section B. In the questionnaire, there
were twelve criteria with five likert scale used to measure the consumer’s
preference in selecting herbal based products.
The information gathered from the questionnaire was analyzed using reliability
test, correlation test and descriptive statistic.
The status on the use of herbs and herbal based products was shown in Table
1. A total of 3270 (73.5%) out of 4452 respondents used herbs and herbal
based products in 2013. Further discussion is to focus on the 3270
respondents.
332
Mallery (2003) the twelve criteria were acceptable to be used in measuring the
consumer’s preference in selecting herbal based products as the α = 0.7.
333
When the level of knowledge on herbal products was high, there was more
tendency of respondents to choose products with GMP compliance as the most
important criterion.
334
Table 5. ANOVA test for consumer preferences based on importance of herbal based
products with low price from the aspect of yearly expenditure.
Likert scale N Mean Std. Std. 95% Confidence Min Max
score Deviation Error Interval for Mean
Lower Upper
Bound Bound
Very
73.00 763.83 1655.79 193.80 377.51 1150.16 0.00 11760.00
Unimportant
Unimportant 510.00 594.35 1069.59 47.36 501.30 687.40 0.00 8492.00
Moderate 1048.00 507.92 796.59 24.61 459.64 556.20 0.00 11544.00
Important 895.00 475.99 834.97 27.91 421.21 530.77 0.00 12000.00
Very
744.00 442.55 547.74 20.08 403.13 481.97 0.00 6000.00
Important
Total 3270.00 503.50 839.32 14.68 474.72 532.28 0.00 12000.00
Note: F= 4.497, p= 0.001
Source: Survey, 2013
CONCLUSION
REFFERENCES
Rohana, A.R., Intan Nurulhani, B., Mohd Asraf, I. & Lim, H.F. 2014. Penggunaan
Herba di Semenanjung Malaysia: Tradisi atau Kontemporari?. Pp. 32-35
in M. Mastura, B.J. Chee, A.L. Tan, M.A. Nor Azah, A.R. Rohana & M.
Noorhazmira (Eds.). Prosiding Seminar Pemuliharaan & Pemerkasaan
Pengetahuan Tradisi, 23 to 24 May 2014 at Putra World Trade Center,
Kuala Lumpur.
Rohana, A.R., Lim, H.F. & Ismariah, A. (2013). Socio-economic Backgrounds and
Income Generation of Herbal Cultivators in Peninsular Malaysia. Paper
Presented at the Conference on Forestry and Forestry Products 2013
(CFFPR 2013), 11─12 November 2013, Sunway Putra Hotel, Kuala
Lumpur.
335
CONSUMERS’ PERCEPTION TOWARDS LOCAL HERBAL SUPPLEMENT
PRODUCTS
ABSTRACT
Tongkat ali (Eurycoma longifolia), kacip fatimah (Labisia pumila), dukung anak
(Phyllanthus niruri), misai kucing (Orthosiphon stamineus), and hempedu bumi
(Andrographis paniculata) are some of the well-known local herbal food
supplements. However, one of the biggest challenges for local herbal products
is the lack of consumers’ interest in purchasing local herbs as their
supplements. Although several wellness lifestyle campaigns emphasized on the
use of local herbs in daily diet or health supplement, consumers’ awareness on
its benefits were found still lacking. Thus, the purpose of the study was to
determine consumers’ perception towards local herbal supplements. A face-to-
face interview with 500 respondents in Klang Valley was carried out using a
structured questionnaire. The data collected was analyzed using descriptive
analysis, chi-square analysis, and factor analysis methods. The results indicated
that majority of the respondents consumed local herbal products as
supplement. Local herbs namely pegaga, tongkat ali, and kacip fatimah were
the most preferred herbs consumed by majority of the respondents. Potential,
awareness of negative effect, effectiveness, and suitability were the factors
revealed to influence consumers’ perception towards local herbal supplement
products. This study concluded that if consumers have a well-informed
information on the potential and effectiveness of local herbal supplement
products towards their health, it will improve their acceptance of herbal
products.
INTRODUCTION
The use of herbal medicine has been growing steadily worldwide. Considering
that Malaysia is blessed with abundant flora including medicinal plants, there is
immense opportunity locally and internationally for the Malaysian herbal
336
industry. Some of the popular local herbal plants are kacip fatimah (Labisia
pumila), pegaga (Centella asiatica), peria (Momordica charantia), misai kucing
(Orthosiphon stamineus), mas cotek (Ficus deltiodea), and limau purut (Citrus
hystrix). The use of herbs as medicine has been growing steadily worldwide.
There is a great potential for the local herbal industry to expand, as the
popularity of Malaysian herbal products is growing locally and internationally.
Under the herb Entry Point Project 1 (EPP1), the herbal industry is expected to
produce high-value products and generates income totaling RM2.2 billion of
the Gross National Income (GNI), focused on five herbs i.e. as tongkat Ali, kacip
fatimah, misai kucing, hempedu bumi, and dukung anak. Furthermore, the
trade value of the herbal industry was expected to soar over RM2 trillion by
2020. However, the herbal industry in Malaysia is still lagging behind other
countries such as China and other Association of South-East Nations (ASEAN)
countries (Ahmad & Othman, 2013). Most of the raw materials used locally
were imported from China, Indonesia, and Thailand. The local herbal industries
do not have the full support of the corporate sector, to support its
development and growth for mass production. Due to this, Malaysia is far
behind on technological advancement in herbal medicine. One of the biggest
challenges for the local herbal industry is the lack of consumer interest in
purchasing local herbs as a supplement. The consumers’ interest in herbal
products is prompted by their health problems. Consumers’ perception and
purchasing pattern are important for the development and commercialization
of herbal products. However, the true potential of local herbs is yet to be
explored. Thus, the objective of this study is to determine consumers’
perception towards local herbal supplements.
337
RESULTS AND DISCUSSION
Socio-demographic Profiles
338
Table 1. Socio-demographic profile of respondents
Variables Frequency (n) Percentage (%)
Gender Male 250 50.0
Female 250 50.0
Age (year) 18-25 135 27.0
26-35 125 25.0
36-45 90 18.0
>45 150 30.0
Education Level Never been to school 5 1.0
Primary 20 4.0
SRP/PMR 65 13.0
SPM 185 37.0
STPM/Matriculation 50 10.0
University 145 29.0
Others 30 6.0
Marital Status Married 280 56.0
Single 185 37.0
Divorced 35 7.0
Race Malay 470 94.0
Chinese 15 3.0
Indian 10 2.0
Others 5 1.0
Income level < 1000 145 29.0
1001- 2000 130 26.0
2001-3000 90 18.0
3001-4000 30 6.0
>4000 105 21.0
Household Size Staying alone 25 5.0
2-4 280 56.0
>5 195 39.0
Occupation Government sector 185 37.0
Private sector 100 20.0
Self-employed 55 11.0
Housewife 45 9.0
Student 90 18.0
Unemployed 15 3.0
Others 10 2.0
Note: n=500 respondents
339
Table 2: Relationship between Respondents’ Socio-demographic Profiles and
Consumption of Herbs Product
Chi-Square
Variable df Significant Decision
Test
Gender 3.793 1 0.05** Reject H0
Age 10.646 3 0.014** Reject H0
Race 1.162 3 0.762 Failed to Reject H0
Education Level 3.598 6 0.731 Failed to Reject H0
Occupation 13.466 6 0.036** Reject H0
Income Level 7.969 4 0.093 Failed to Reject H0
Marital Status 7.272 3 0.026** Reject H0
Household Size 2.424 3 0.298 Failed to Reject H0
Note: *Statistically significant at 0.01, **Statistically significant at 0.05,
***Statistically significant at 0.10
Factor analysis was carried out to investigate various factors that influenced
consumers on purchasing local herbal products. Kaiser-Meyer-Olkin Measure
of Sampling Adequancy (KMO) and Barlett’s test of Sphericity were used to
measure sampling adequacy. Results indicated 0.719 and p=0.0000
respectively, and thus factor analysis is appropriate in this study. Based on
Table 3, five factors extracted from factor analysis. The five factors were
‘potential of herbal supplement’, ‘awareness of the negative side of herbal
supplement’, ‘consumer knowledge on local herbal supplement’, ‘effectiveness
of herbal supplement’ and ‘suitability of herbal supplement’. The first factors
indicated 22.451% of variance, the second factor was accounted for 12.586%,
third factor was 8.396%, forth factor accounted for 7.496% and the fifth factor
was 6.339%.
340
Table 3. Factor analysis result
Sub- % of Eigen Cronbach’
Factors variables variance values s Alpha
explained
Factor 1 – Potential of herbal supplement 22.451 3.817 0.815
i. Herbs is effective as nutritional supplement for 0.542
body
ii. You know about the herbs as supplement 0.643
iii. You believe that herbs is effective as 0.145
supplement
iv. The more often you consume herbal 0.589
supplement, the more you are feeling well
v. Herbal supplements are readily available in the 0.618
market
vi. Herbal supplements are sold at a reasonable 0.741
price
vii. Herbal supplements sold in stores or drug 0.522
stores is more reliable than direct sales
viii. More information about supplements from the 0.693
mass media
ix. Herbal supplements should be more campaigns 0.470
to introduce it to consumers
Factor 2 - Awareness of the negative side of herbal 12.586 2.140 0.764
supplement
i. Herbal supplements effects to the development 0.611
of the mind
ii. Herbal supplements only for those who can 0.736
afford
Factor 3 – Consumer knowledge on local herbal 8.396 1.427 0.701
supplement
i. Each herb has different nutritional value 0.433
ii. Most people do not use herbs for lack of 0.673
knowledge about herbs
iii. Herbs is good for ill treatment 0.732
Factor 4 – Effectiveness of herbal supplement 7.496 1.274 0.621
i. Most people not consume herbs because it is 0.726
not effective
ii. Most people know the benefits of herbs, but do 0.755
not believe in the effectiveness
Factor 5 – Suitability of herbal supplement 6.339 1.078 0.578
i. Herbal supplement only suitable for old people 0.886
341
CONCLUSION
ACKNOWLEDGEMENTS
The authors would like to express their gratitude to the Faculty of Agriculture,
Universiti Putra Malaysia for supporting the research.
REFERENCES
Ahmad, S. & Othman, N. (2013). Strategic Planning, Issues, Prospects and the
Future of the Malaysian Herbal Industry. International Journal of
Academic Research in Accounting, Finance and Management
Sciences, 3(4): 91─102.
342
THE IMPORTANCE OF LONG-TERM RELATIONSHIP FOR THE
SUSTAINABILITY OF MALAYSIAN HERBAL INDUSTRY
ABSTRACT
Herbal industry is one of the most promising industries in the future. The rapid
development of the herbal industry in Malaysia is reflected by its trade. Herbal
industry is one of the EPP in NKEA and government wants to improve the
product quality and marketing efforts of the herbal industry. The government
also targeted both upstream and downstream segments of the industry to help
in developing the industry. Long-term relationship has been acknowledged as
capable in increasing the competitiveness of an industry. It has been found that
retailers or buyers with long-term relationships can achieve a competitive
advantage by receiving merchandise in short supply, information on new and
best-selling products and competitive activity, best allowable prices, and
advertising and markdown allowances. Furthermore, buyers and sellers are
interdependence in the industry thus, to increase the efficiency and probability
to success, it is necessary to identify the factors that affecting the development
of long-term relationship among supply chain players in the Malaysian herbal
industry. Based on previous studies, there are a number of variables that can
contribute to the maintenance of collaborative long-term relationship such as
commitment, satisfaction, trust, power-dependence, communication,
reputation and others. It has been identified that the relationship building
efforts are theorized to enhance their membership’s commitment to the
relationship as well as the membership’s relationship behaviors. Satisfaction
has also been acknowledged as the buyer’s cognitive state of being adequately
rewarded for the sacrifices undergone in facilitating the exchange which can
help in sustaining the industry. Satisfaction also encourages greater loyalty and
a long-term working relationship. Trust is also one of the important factors that
can increase the sustainability of the industry. Trust is the willingness to rely
upon an exchange partner in whom who has confidence and facing the risk
together. Many benefits can be obtained by buyers and sellers through
development of long term relationships. Through a long-term relationship,
supply chain players in the Malaysian herbal industry are able to gain
advantages that may be not realized under a traditional relationship.
343
INTRODUCTION
The objective of this study is to develop the conceptual framework that can be
used to analyze the importance of long-term relationship for the sustainability
of Malaysian Herbal Industry using the following methodology framework.
344
RESULTS AND DISCUSSION
345
Relationship
Commitment Trust Cooperation Satisfaction Specific Technology
Investment
CONCLUSION
REFERENCES
Anderson, E. & Weitz, B. A. (1992). The Use of Pledges To Build And Sustain
Commitment in Distribution Channels. Journal of Marketing Research:
Vol 29 (1): 18─34.
346
Ford, D. (1980), The Development Of Long-Term Orientation in Buyer-Seller
Relationships. Journal of Marketing: Vol. 58 (April): 1─19.
Han, S. L., Wilson, D.T. and Dant, S. P. (1993) Buyer-Seller Relationships Today,
Industrial Marketing Management. Vol. 22: 331─338.
Cannon, J. P., Doney, P. M., Mullen, M. R., & Petersen, K. J. (2010). Building
Long-Term Orientation in Buyer-Supplier Relationships: The
Moderating Role of Culture. Journal of Operations Management: 28(6),
506─521.
Louis, D., & Lombart, C. (2010). Impact of Brand Personality on Three Major
Relational Consequences (Trust, Attachment, And Commitment to The
Brand). Journal of Product & Brand Management: 19(2), 114─130.
Jayachandran, S., Sharma, S., Kaufman, P., & Raman, P. (2005). The Role of
Relational Information Processes and Technology Use in Customer
Relationship Management. Journal of marketing: 69(4), 177─192.
347