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Correlation of Bioactive Compound in Melicope Lunu and Cosmos Caudatus in Antibacterial Activity
Correlation of Bioactive Compound in Melicope Lunu and Cosmos Caudatus in Antibacterial Activity
C. 131
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2.3 Preparation of solvent fractionation 133
The methanol extract was diluted in 25 mL Methanol and mixed with 100 mL distilled 134
water then mixed with 125 mL of serial solvent( hexane, Dichloromethane, ethyl acetate, 135
butanol) in 1000 mL seperated funnel. The ratio of the mixer of Water: Methanol: Serial 136
solvent was 4:1:5. Each of the solvent fraction were removed by using Eyela rotary 137
evaporator (Tokyo Rikikai Co. Ltd, Japan) at 40
C. 138
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2.3 Microbial strains and media 140
Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 14579, Listeria 141
monocytogenes ATCC 19115, Campylobacter jejuni ATCC 29428, Salmonella typhimurium 142
ATCC 13311 and Escherichia coli ATCC 25922 were purchased from ALL EIGHTS (M) 143
SDN BHD (Subang, Selangor, Malaysia). Each bacterial strain was cultivated in Nutrient 144
Agar and incubated for 18-20 hours at 37
C for 18-20 hours in incubator (Memmert, Germany). Then three or five of the same 153
morphological type form on agar plate culture were took up and suspended in 10 ml sterile 154
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saline then vortex thoroughly. The bacterial suspension then is compared to 0.5 McFarland 155
standards. The inoculums should be used less than 15 minutes after prepare it. 156
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2.6 Determination of Minimum Inhibition Concentration by using Agar Dilution 159
method. 160
The minimum inhibition concentration (MIC) of the extracts were tested by using 161
Agar Dilution method according to method that proposed by Jennifer, (2001) with some 162
modification. The extracts were diluted using 10 % DMSO and adjusted to 8mg/mL as stock 163
solution then two-fold dilution were made from 4mg/mL up to 0.5mg/mL. The suitable 164
amount of extract was added on to the well of 24 well plate then added with suitable amount 165
of sterile Muller Hilton agar (MHA). The temperature of MHA was cold to 50 C before 166
poured to the well. Then was swirled up until thoroughly mixed. The plates were left for 10 167
minutes and allowed to solidified. Then 1 uL of suspensions from the prepared inoculums 168
were placed on the agar surface. Chloramphenicol was used as a positive control. While for 169
the blank was MHA only. The plates were incubated in an inverted position at 37
C in 170
incubator (Memmert, Germany) for 18-20 hours. The MIC was read as the first concentration 171
that inhibits the organism growth completely. In the present study, five replicates were 172
prepared for each sample. 173
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2.7 GCMS analysis 175
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2.8 Statistic analysis 177
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2.9 Multivariate analysis 179
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3. Result 3.2 Agar dilution methodTable 3.2: Minimum inhibition concentration (mg/mL) of 186
methanol extract of 17 salads plants. 187
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(-) no activity; BC= Bacillus cereus ATCC 14579; SA= Staphylococcus aureus ATCC 25923; 188
LM= Listeria monocytogenes ATCC 19115; CJ= Campylobacter jejuni ATCC 29428; EC = 189
Escherichia coli ATCC 25922; ST= Salmonella thyphii ATCC 13311 190
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Introduction 192
4. Discussion 193
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5. Conclusion 196
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the mode of action of antimicrobial agents also Conclusion 198
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4. Discussion. 225
5.Conclusion 226
Table 227
Table 1: Ethnomedicine uses of selected edible traditional medicinal plants salad in 228
Malaysia communtiy. 229
Species name Common name Part tested Ethnomedicinal uses
Pipper betle Sireh Leaves
Digestive, stimulative, carminative and
aphrodisiac (Sankaranarayanan et al.,
2010).
Lowsonia
inermis
Inai Leaves
Alleviating jaundice, skin diseases, venereal
diseases, smallpox and
spermatorrhoea (Chaudhary et al., 2010)
Melicipe lunu Tenggek Burung Leaves
Treatment of itches and wounds (Shaari et
al., 2006),
Borsenbergia
rotunda
Temu Kunci Rhizom
Ailment, illness and confinement. Rhizomes
are also taken as carminatives for relieving
flatulence.(Chan et al., 2008 )
Curcuma
xanthorhiza
Temu Lawak Rhizom
Treatment of stomach diseases, liver
disorders, constipation, bloody diarrhea,
dysentery, fever in children, hemorrhoids,
and skin eruptions (Yasni et al. 1994)
Cosmos
caudatus
Ulam Raja Leaves
Blood cleansing, induction of uterine
contractions and prevention or cure of
ailments such as diabetes, high blood
pressure, cardiovascular disease, arthritis,
fever and coughs (Abas et al., 2006)
Andrographis
paniculata
Hempedu Bumi Leaves
Treatment of upper GI tract and upper
respiratory infections, fever, herpes and
other chronic diseases. (Roy et al., 2010)
Muraya Kari Leaves Relieve nausea, indigestion, vomiting;
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koenigii treatment of diarrhea and dysentery
(Chowdhury et al., 2008).
Kaempferia
galangal
Cekur Leaves
Treatment of Tinea versicolor, and eye
diseases and seizures, rheumatism, asthma,
headaches, cough, toothaches, bruises and
wounds (Ridtitid et al., 2008)
Piper longum Kaduk Leaves
Treatment of respirotory tract, cough,
bronchitis, irritant, inflammation. (Manoj et
al., 2004)
Talinum
triangulare
Kerekot Belanda Leaves
Treatment of diuretic, gastro-intestinal
disorder.(Mensah Et al., 2008)
Sesbania
grandifolia
Turi Leaves
Aperient, diuretic, and tonic and
disinfect the mouth and throat (gowri1 and
Vasantha., 2010)
Psophocapus
tetragonolobus
Kacang Botol Pods
Treatment of skin sores such as boils and
ulcers (Perry, 1980).
Molinda
citrifolia
Mengkudu Leaves Relief joint pain (Rout et al., 2009)
Gynura
procumben
Sambung
Nyawa
Leaves
Treatment of malaria, general febrifuge, and
analgesic (Scott., 2006)
Justicia
gendarussa
Ganda Rusa Leaves
Treatment of fever, hemiplegia,
rheumatism, arthritis, muscle pain,
lumbago, headache and earache (Ahmad
and Holdworth 2003; Anonymous 1959)
Centella
asiatica
Pegaga Leaves
Against conjunctivitis and other eye injury,
wound healing but especially for the
treatment of skin diseases such as eczema,
leprosy and psoriasis. Treatment of burns,
itching and insect bites (Gupta et al., 1999;
Zainol et al., 2008)
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3.1 Disk diffusion method 256
Table : Results of weight collected from different fraction solvent. 257
Plants
Weight percentage collected %
Hexane Dichloromethane Ethyl acetate Butanol Water
Melicope lunu 11.8 1.42 74.912.65 6.260.3 26.080.51 5.880.42
Cosmos caudatus
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Table : minimum inhibition concentration of extract in different fraction layer. 261
Plants Extract of minimum concentration inhibition (mg/mL)
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fraction layer
Staphylococcus
aureus
Bacillus cereus
Listeria
monocytogenes
Melicope lunu
Hexane <0.5 <0.5 <0.5
Dichloromethane <0.5 <0.5 <0.5
Ethyl acetate <0.5 <0.5 <0.5
Butanol 4 2 2
water 1 <0.5 <0..5
Cosmos caudatus
Hexane 4 4 4
Dichloromethane 4 2 2
Ethyl acetate 2 1 2
Butanol 1 <0.5 <0.5
water 2 1 2
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3.2 Agar dilution m 275