2022 JRP Vol26 No5 Art

9/17/22, 11:01 AM Journal of Research in Pharmacy
Journal of Research in Pharmacy

An international open-access journal of pharmacy and pharmaceutical sciences
Formerly published as Marmara Pharmaceutical Journal
(https://jrespharm.com/)
ISSN : 1309-0801
Current Issue (https://jrespharm.com/content.php?id=78)
Articles in Press (https://jrespharm.com/content.php?id=46)
Archive (https://jrespharm.com/archive.php)
Search (https://jrespharm.com/search.php)
Author Benefits (https://jrespharm.com/static.php?id=11)
Editor-in-Chief
Hatice Kübra Elçioğlu
Vice Editors
Levent Kabasakal
Esra Tatar
Online ISSN
2630-6344
Publisher
Marmara University
Frequency
Bimonthly (Six issues / year)
Abbreviation
J.Res.Pharm.
Former Name
Marmara Pharmaceutical Journal
Editorial Board
Editor-in-Chief
Hatice Kübra
ELÇİOĞLU

Department of Pharmacology, Faculty of Pharmacy, Marmara University, Istanbul, Turkey

kubra.elcioglu@marmara.edu.tr (mailto:kubra.elcioglu@marmara.edu.tr)
https://jrespharm.com/static.php?id=2 1/8
Vice Editors
Levent KABASAKAL
lkabasakal@marmara.edu.tr (mailto:lkabasakal@marmara.edu.tr)
Esra TATAR
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
etatar@marmara.edu.tr (mailto:etatar@marmara.edu.tr)
Guest Editors for the Special Issue : The International Conference on Pharmaceutical Research and Practice
(ICPRP) 2022
Arba Pramundita RAMADANI
Department of Pharmacy, Universitas Islam Indonesia, Yogyakarta, Indonesia
arba.pramundita@uii.ac.id (mailto:arba.pramundita@uii.ac.id)
Asih TRIASTUTI
Department of Pharmacy, Universitas Islam Indonesia, Yogyakarta, Indonesia
asih.triastuti@uii.ac.id (mailto:asih.triastuti@uii.ac.id)
Analytical Chemistry & Therapeutic Drug Monitoring
Anisa ELHAMILI
Department of Medicinal & Pharmaceutical Chemistry, Faculty of Pharmacy, University of Tripoli, Tripoli, Libya
aaelhamili2000@gmail.com (mailto:aaelhamili2000@gmail.com)
Lorena MEMUSHAJ
Department of Pharmacy, Faculty of Medical Sciences, Aldent University, Tirana, Albania
lorena.memushaj@ual.edu.al (mailto:lorena.memushaj@ual.edu.al)
Mohd Younis RATHER

Multidisciplinary Research Unit, Government Medical College Srinagar, Srinagar, India
younis.rather78@gmail.com (mailto:younis.rather78@gmail.com)
Pablo MIRALLES IBARRA

1. Foundation for the Promotion of Health and Biomedical Research in the Valencian Region (FISABIO-Public
Health), Valencia, Spain
2. Department of Analytical Chemistry, Faculty of Chemistry, University of Valencia, Burjassot, Spain
miralles_pabiba@gva.es (mailto:miralles_pabiba@gva.es)
Pınar TALAY PINAR

Department of Analytical Chemistry, Faculty of Pharmacy, Yüzüncü Yıl University, Van, Turkey
ptalay@gmail.com (mailto:ptalay@gmail.com)
Biochemistry & Cancer Research
Derya ÖZSAVCI
Department of Biochemistry, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
derya.ozsavci@marmara.edu.tr (mailto:derya.ozsavci@marmara.edu.tr)
Gülberk UÇAR
Department of Biochemistry, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey
gulberk@hacettepe.edu.tr (mailto:gulberk@hacettepe.edu.tr)
Lokman AYAZ
Department of Biochemistry, Faculty of Pharmacy, Trakya University, Edirne, Turkey
lokmanayaz@yahoo.com (mailto:lokmanayaz@yahoo.com)
Biotechnology
Ali Demir SEZER

Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
adsezer@marmara.edu.tr (mailto:adsezer@marmara.edu.tr)
Ammad Ahmad FAROOQI

Department of Molecular Oncology, Institute of Biomedical and Genetic Engineering (IBGE), Islamabad,
Pakistan
farooqiammadahmad@gmail.com (mailto:farooqiammadahmad@gmail.com)
Murat DOĞAN
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Cumhuriyet University, Sivas, Turkey
mdogan@cumhuriyet.edu.tr (mailto:mdogan@cumhuriyet.edu.tr)
Uğur KARAGÖZ
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Trakya University, Edirne, Turkey
ugurkaragoz@trakya.edu.tr (mailto:ugurkaragoz@trakya.edu.tr)
Clinical Pharmacy & Social Pharmacy
Abdikarim Mohammed ABDI

Department of Clinical Pharmacy, Faculty of Pharmacy, Yeditepe University, Istanbul, Turkey
abdikarim.abdi@yeditepe.edu.tr (mailto:abdikarim.abdi@yeditepe.edu.tr)
Betül OKUYAN
Department of Clinical Pharmacy, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
betul.okuyan@marmara.edu.tr (mailto:betul.okuyan@marmara.edu.tr)
Maja ORTNER HADŽİABDİĆ

Centre for Applied Pharmacy, Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia
mortner@pharma.hr (mailto:mortner@pharma.hr)
Mesut SANCAR
mesut.sancar@marmara.edu.tr (mailto:mesut.sancar@marmara.edu.tr)
Tarik CATİĆ
Department of Pharmacy, Sarajevo School of Science and Technology, Sarajevo, Bosnia and Herzegovina
tarik.catic@ssst.edu.ba (mailto:tarik.catic@ssst.edu.ba)
In Silico Studies
Gizem TATAR YILMAZ

Department of Biostatistics and Medical Informatics, Faculty of Medicine, Karadeniz Technical University,
Trabzon, Turkey
gizemtatar@gmail.com (mailto:gizemtatar@gmail.com)
Medicinal Chemistry
Bahadır BÜLBÜL
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Düzce University, Düzce, Turkey
bahadir.bulbul@yahoo.com.tr (mailto:bahadir.bulbul@yahoo.com.tr)
Entela HALOCI
Faculty of Pharmacy, University of Medicine, Tirana, Albania
entela.haloci@umed.edu.al (mailto:entela.haloci@umed.edu.al)
Hasan Erdinç SELLİTEPE

Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Karadeniz Technical University, Trabzon,
Turkey
esellitepe@ktu.edu.tr (mailto:esellitepe@ktu.edu.tr)
Somaieh SOLTANI
Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
soltanis@tbzmed.ac.ir (mailto:soltanis@tbzmed.ac.ir)
Microbiology & Immunology
Erkan RAYAMAN
Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
erayaman@marmara.edu.tr (mailto:erayaman@marmara.edu.tr)
Gülgün TINAZ
Department of Basic Pharmaceutical Sciences, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
gulgun.tinaz@marmara.edu.tr (mailto:gulgun.tinaz@marmara.edu.tr)
Zahraa AMER HASHIM

Department of Microbiology and Immunology, College of Pharmacy, Mosul University, Mosul, Iraq
hashimz@uomosul.edu.iq (mailto:hashimz@uomosul.edu.iq)
Pharmaceutical Botany & Pharmacognosy & Chemistry of Natural Products
Ahmet EMİR
Department of Pharmacognosy, Faculty of Pharmacy, Ege University, Izmir, Turkey
ahmet.emir@ege.edu.tr (mailto:ahmet.emir@ege.edu.tr)
Annalisa CHIAVAROLI
Department of Pharmacology, Faculty of Pharmacy, G. d′Annunzio University of Chieti-Pescara, Chieti, Italy
annalisa.chiavaroli@unich.it (mailto:annalisa.chiavaroli@unich.it)
Antoaneta TRENDAFILOVA
Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Sofia, Bulgaria
antoaneta.trendafilova@orgchm.bas.bg (mailto:antoaneta.trendafilova@orgchm.bas.bg)
Ayşe Esra KARADAĞ

Department of Pharmacognosy, Faculty of Pharmacy, Istanbul Medipol University, Istanbul, Turkey
ayseesraguler@gmail.com (mailto:ayseesraguler@gmail.com)
Ceren EMİR
Department of Pharmacognosy, Faculty of Pharmacy, Ege University, Izmir, Turkey
ceren.acir@ege.edu.tr (mailto:ceren.acir@ege.edu.tr)
Claudio FERRANTE
Department of Pharmacology, Faculty of Pharmacy, G. d′Annunzio University of Chieti-Pescara, Chieti, Italy
claudio.ferrante@unich.it (mailto:claudio.ferrante@unich.it)
İ. İrem TATLI ÇANKAYA

Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey
iremcankaya@gmail.com (mailto:iremcankaya@gmail.com)
Lejla KLEPO
Department of Chemistry, Faculty of Science, University of Sarajevo, Sarajevo, Bosnia and Herzegovina
klepolejla@gmail.com (mailto:klepolejla@gmail.com)
Nurettin YAYLI
Department of Pharmacognosy, Faculty of Pharmacy, Karadeniz Technical University, Trabzon, Turkey
yayli@ktu.edu.tr (mailto:yayli@ktu.edu.tr)
Pharmacognosy
Randolph ARROO
Leicester School of Pharmacy, De Montfort University, Leicester, UK
rrjarroo@dmu.ac.uk (mailto:rrjarroo@dmu.ac.uk)
Turgut TAŞKIN
Department of Pharmacognosy, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
turguttaskin@marmara.edu.tr (mailto:turguttaskin@marmara.edu.tr)
Vildan ÇELİKSOY
School of Optometry and Vision Sciences, Cardiff University, Cardiff, UK
celiksoyv@cardiff.ac.uk (mailto:celiksoyv@cardiff.ac.uk)
Zoran ZEKOVIĆ
Faculty of Technology, University of Novi Sad, Novi Sad, Serbia
zzekovic@tf.uns.ac.rs (mailto:zzekovic@tf.uns.ac.rs)
Pharmaceutics
Afife Büşra UĞUR KAPLAN

Department of Pharmaceutical Technology, Faculty of Pharmacy, Atatürk University, Erzurum, Turkey
afife.busra.ugur@gmail.com (mailto:afife.busra.ugur@gmail.com)
Dinesh KUMAR
Department of Pharmaceutical Engineering & Technology, Indian Institute of Technology (BHU), Varanasi,
India
dinesh.phe@itbhu.ac.in (mailto:dinesh.phe@itbhu.ac.in)
Ebru ALTUNTAŞ
Department of Pharmaceutical Technology, Faculty of Pharmacy, Istanbul University, Istanbul, Turkey
ebru.altuntas@istanbul.edu.tr (mailto:ebru.altuntas@istanbul.edu.tr)
Enkelejda GOCI
Pharmacotherapeutic Research Center, Aldent University, Tirana, Albania
enkelejda.goci@ual.edu.al (mailto:enkelejda.goci@ual.edu.al)
Gülşah GEDİK
Department of Pharmaceutical Technology, Faculty of Pharmacy, Trakya University, Edirne, Turkey
gulsahgedik@trakya.edu.tr (mailto:gulsahgedik@trakya.edu.tr)
Ongun Mehmet SAKA

Department of Pharmaceutical Technology and Biotechnology, Faculty of Pharmacy, Ankara University,
Ankara, Turkey
omsaka@gmail.com (mailto:omsaka@gmail.com)
Oya KERİMOĞLU
Department of Pharmaceutical Technology, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
osipahigil@marmara.edu.tr (mailto:osipahigil@marmara.edu.tr)
Rezarta SHKRELI
Department of Pharmacy, Faculty of Medical Sciences, Aldent University, Tirana, Albania
rezarta.shkreli@ual.edu.al (mailto:rezarta.shkreli@ual.edu.al)
Saeideh SOLTANI
Novel Drug Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of
Medical Sciences, Isfahan, Iran
soltanisa@pharm.mui.ac.ir (mailto:soltanisa@pharm.mui.ac.ir)
Pharmacology & Toxicology
Ana V. PEJČİĆ
Department of Pharmacology and Toxicology, Faculty of Medical Sciences, University of Kragujevac,
Kragujevac, Serbia
anapejcic201502@yahoo.com (mailto:anapejcic201502@yahoo.com)
Ayfer BECEREN
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Marmara University, Istanbul, Turkey
ayfer.tozan@marmara.edu.tr (mailto:ayfer.tozan@marmara.edu.tr)
Büşra ERTAŞ
busra.ertas@marmara.edu.tr (mailto:ayfer.tozan@marmara.edu.tr)
Klodiola DHAMO
Faculty of Technical Medical Sciences, Aldent University, Tirana, Albania
klodiola.dhamo@ual.edu.al (mailto:klodiola.dhamo@ual.edu.al)
Nasrin MALEKI DIZAJI

Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
malekins2014@gmail.com (mailto:malekins2014@gmail.com)
Rümeysa KELEŞ KAYA

Department of Medical Pharmacology, Faculty of Medicine, Sakarya University, Sakarya, Turkey
rumeysakeles@sakarya.edu.tr (mailto:rumeysakeles@sakarya.edu.tr)
Ünzile YAMAN
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Katip Çelebi University, İzmir, Turkey
unzileyaman@gmail.com (mailto:unzileyaman@gmail.com)
Zeina ALTHANOON
Department of Pharmacology and Toxicology, College of Pharmacy, Mosul University, Mosul, Iraq
dr.zeina@uomosul.edu.iq (mailto:dr.zeina@uomosul.edu.iq)
Rümeysa Keleş Kaya

Department of Medical Pharmacology, Faculty of Medicine, Sakarya University, Sakarya, Turkey
rumeysakeles@sakarya.edu.tr (mailto:rumeysakeles@sakarya.edu.tr)
Copy Editor
Ayşe Nur HAZAR YAVUZ
ayse.hazar@marmara.edu.tr (mailto:ayse.hazar@marmara.edu.tr)
Büşra ERGEN
busra.ergen@marmara.edu.tr (mailto:busra.ergen@marmara.edu.tr)
Fatih Taha ÇİFTÇİ

fatih.ciftci@marmara.edu.tr (mailto:fatih.ciftci@marmara.edu.tr)
Müzeyyen AKSOY
muzeyyen.aksoy@marmara.edu.tr (mailto:muzeyyen.aksoy@marmara.edu.tr)
Ömer Faruk ÖZKANLI

oozkanli@marmara.edu.tr (mailto:oozkanli@marmara.edu.tr)
Şeyma GÖZELİZMİR
seyma.gozelizmir@marmara.edu.tr (mailto:seyma.gozelizmir@marmara.edu.tr)
Language Editor
Khadija ALJESRI
Department of Pharmacology, Institute of Health Sciences, Marmara University, Istanbul, Turkey
Biostatistics Editor
Gülnaz NURAL BEKİROĞLU
Department of Bioistatistics, Faculty of Medicine, Marmara University, Istanbul, Turkey
nural@marmara.edu.tr (mailto:nural@marmara.edu.tr)
Marmara University
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Journal of Research in Pharmacy

An international open-access journal of pharmacy and pharmaceutical sciences
Formerly published as Marmara Pharmaceutical Journal
(https://jrespharm.com/)
ISSN : 2630-6344
Current Issue (https://jrespharm.com/content.php?id=78)
Articles in Press (https://jrespharm.com/content.php?id=46)
Archive (https://jrespharm.com/archive.php)
Search (https://jrespharm.com/search.php)
Author Benefits (https://jrespharm.com/static.php?id=11)
Editor-in-Chief
Hatice Kübra Elçioğlu
Vice Editors
Levent Kabasakal
Esra Tatar
Online ISSN
2630-6344
Publisher
Marmara University
Frequency
Bimonthly (Six issues / year)
Abbreviation
J.Res.Pharm.
Former Name
Marmara Pharmaceutical Journal
Other Issues :
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Biomedical application of microemulsion delivery systems: A review (abstract.php?id=1087)

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Pages 1052-1064
Nisha TIWARI,Amravati SIVAKUMAR
DOI : 10.29228/jrp.203
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Pages 1084-1101
Nada WALWEEL,Aybuke Ulku KUTLU,Ummugulsum YILMAZ,Zakarya AL-SHAEBI,Jalil CHARMI,Omer AYDIN
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Zarife Nigâr ÖZDEMİR KUMRAL,Alisina BULUT,Bahadır ÜZÜLMEZ,Mustafa VEZİRHÜYÜK,Zafer KÖK,Naziye ÖZKAN
YENAL,Berrak Ç. YEĞEN,Mehmet KOÇ
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Chemometrics-assisted content uniformity evaluation of extemporaneous preparation containing

ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl (abstract.php?id=1115)
Research Article
Pages 1403- 1410
Sri Hartati YULIANI,Clara Angelika SINULINGGA,Dina Christin Ayuning PUTRI,Michael Raharja GANI,Florentinus Dika Octa
RISWANTO,Dita Maria VIRGINIA
DOI : 10.29228/jrp.233
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Mohammed KHALIL,Akram AL-SALEH,Hana MAKHAMREH,Rabab TAYEM,Mohammad ABUFARAH,Abdullah ZUBI,Omaimah
NAJIB,Mohammad BADER,Naji NAJIB,Suliman ALFAYOUMI,Nasir IDKAIDEK
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Viktor STAVYTSKYI,Oleksii ANTYPENKO,Oleg DEVINYAK,Oleksii VOSKOBOİNİK,Sergiy KOVALENKO
DOI : 10.29228/jrp.235
Abstract
Journal of Research Article
Research in Pharmacy www.jrespharm.com
Chemometrics-assisted content uniformity evaluation of

extemporaneous preparation containing ambroxol HCl,
pseudoephedrine HCl, and triprolidine HCl
Sri Hartati YULIANI1 * İD , Clara Angelika SINULINGGA 1 İD , Dina Christin Ayuning PUTRI1 İD ,
Michael Raharja GANI 2 İD , Florentinus Dika Octa RISWANTO 2 İD , Dita Maria VIRGINIA 3 İD
1 Division of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Universitas Sanata Dharma,
Yogyakarta, Indonesia
2 Division of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Sanata Dharma,
Yogyakarta, Indonesia
3 Division of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Universitas Sanata Dharma, Yogyakarta,
Indonesia
* Corresponding Author. E-mail: srihartatiyuliani@usd.ac.id (S.H.Y.); Tel. +62-274-883037.
Received: 3 March 2022 / Revised: 5 April 2022 / Accepted: 5 April 2022
ABSTRACT: Split powder preparations containing ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl were
frequently dispensed at a private hospital in Semarang, Central Java, Indonesia. Triprolidine HCl was reported to have a
low composition in the divided powder. Content uniformity is one of the most important characteristics of
extemporaneous formulations, especially for low-dose active ingredients. The aim of this study was to evaluate the dose
uniformity of the divided powder preparation containing ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl
prepared by a pharmacist in a private hospital in Semarang, Central Java, Indonesia. In this study, the
spectrophotometric method was used in combination with the chemometric analysis to determine the drug
concentration in the divided powder. The accuracy of the models was evaluated by the highest value of the coefficient of
determination of calibration, cross-validation and validation, while the precision of the models was evaluated by the
lowest value of the mean square error of calibration, cross-validation and prediction. The selected model for each
compound was used to build predictive models for quantitative analysis. It was found that the content of ambroxol HCl,
pseudoephedrine HCl, and triprolidine HCl in the subdivided powder samples were 8.120 ± 1.167 mg, 30.142 ± 3.965 mg,
and 1.193 ± 0.221 mg, respectively. The results on the uniformity of the content did not meet the requirements, as it was
found that the coefficient of variance of these determinations was more than 5%. Surprisingly, the divided powder can
be considered as a safe and acceptable dosage form due to its wide therapeutic window.
KEYWORDS: Ambroxol HCl; pseudoephedrine HCl; triprolidine HCl; divided powder; quality.
1. INTRODUCTION
Extemporaneous preparation was still needed especially for pediatric patient since the license drug for
pediatric was limited [1]. Many problems have been identified in the formulation of pediatric medicines [2].
Extemporaneous preparation for children were unique due to their physical and chemical properties.
Pharmacists must ensure that the quality of the drug, especially its stability and tolerability, is met. [3]. The
lack of information on stability, bioavailability, pharmacodynamics, pharmacokinetics, effectiveness and
safety data makes it important to develop an appropriate pediatric therapeutic formulation [4]. Hence, it is
important to guarantee the safety, efficacy and quality of extemporaneous preparation in the community
service.
The quality of extemporaneous preparation can be recognized by its appearance, uniformity of active
ingredient, stability of active ingredient, and compatibility between active ingredient and excipient [5,6]. The
manipulation to the license drug during extemporaneous preparation will change the specification of the
drug including the stability of the active ingredient in the dosage forms [7]. The incompatibility of the other
active ingredient was also unknown [8,9]. The compatibility between active ingredient and active ingredient-
excipient will affect the dose of the drug in the preparation and potentially impact the safety and
effectiveness of the pharmaceutical dosage form [10,11].
How to cite this article: Yuliani SH, Sinulingga CA, Putri DCA, Gani MR, Riswanto FDO, Virginia DM. Chemometrics-assisted content uniformity
evaluation of extemporaneous preparation containing ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl. J Res Pharm. 2022; 26(5): 1403-
1410.
© 2022 Marmara University Press http://dx.doi.org/10.29228/jrp.233

ISSN: 2630-6344
1403
Yuliani et al. Journal of Research in Pharmacy
Chemometrics-assisted content uniformity evaluation Research Article
Extemporaneous preparation of divided powder is still widely used in Indonesia, especially for
pediatric patient. The mixture of ambroxol hydrochloride (HCl), pseudoephedrine hydrochloride (HCl), and
triprolidine hydrochloride (HCl) is high frequency drug combination to treat cough in the pediatric patient.
Ambroxol is found in numerous over-the-counter dosage forms as a mucolytic agent [12]. A study engaging
1300 pediatric patients indicated that ambroxol is well tolerated, even in the early infant for acute and
chronic respiratory disease [13]. On the other hand, pseudoephedrine and triprolidine are frequently
formulated as a combination dosage form. Pseudoephedrine is considerably safe as a nasal decongestant
among children aged 6 to <12 years with the common cold [14,15]. Triprolidine HCl is the first generation of
antihistamine indicated for allergic rhinitis [16]. The previous study observing the pharmacokinetic of
triprolidine in the combination of triprolidine and pseudoephedrine reported that the level of triprolidine
was higher in combination than in single agents [17]. However, the dosage variant increases the risk of
under- or overdosing of the individual components. Remarkably, the combination of antihistamine and
decongestant in children increase dystonic reaction and sympathomimetic toxicity [18–20].
Children are vulnerable patients. The limited communication with patient may lead to the adverse
effect of the treatment they receive [7]. The problems that are often encountered in the preparation of
compound pediatric preparations are achieving dose uniformity [21]. Hence, it is necessary to study the
quality of the extemporaneous preparation of divided powder containing ambroxol HCl, Pseudoephedrine
HCl and Triprolidine HCl. The quality in this study was defined as the uniformity of active ingredient
during administration of the preparation.
Chemometric, a combination of mathematical and statistical techniques to solve several problems in
chemistry, was stated as a powerful tool to predict and classify drug concentration in pharmaceutical
formulas [22,23]. Previous study proved that chemometrics techniques were successfully applied for
determining the content of paracetamol and tramadol concentration divided powder dosage form [24]. The
other study revealed that the combination of UV spectroscopic techniques and chemometrics was a
favorable tool to predict the concentration of acetaminophen, caffeine, and propyphenazone in tablet
samples [25]. Ultraviolet (UV) spectroscopic method and chemometrics techniques allowed a rapid and
efficient analytical stage to be developed, compared to the other analytical technique such chromatographic
method which requires more stages to separate one analyte from the others with the similar sample matrices
[26-28]. A fast, simple and effective method was developed for pharmaceutical analysis by combining UV
spectroscopic technique and chemometrics of multivariate calibration in order to determine the content of
ambroxol HCl, pseudoephedrine HCl and triprolidine HCl in extemporaneous preparation of divided
powder.
2. RESULTS
Drug combination composed by ambroxol HCl, pseudoephedrine HCl and triprolidine HCl were
commonly prescribed for pediatrics in a private hospital in Semarang, Central Java, Indonesia. Since it was
important to maintain patient obedience, divided powders containing ambroxol HCl, pseudoephedrine HCl,
and triprolidine HCl were prescribed for children. Tablets of ambroxol HCl were compounded with
commercial tablets containing pseudoefedrin HCl of 60 mg and triprolidine HCl of 2.5 mg to produce
divided powders for pediatric patients. The potential interaction between drug and excipient may have
occurred during the preparation of the dosage form. Uniformity of drug content plays an important role for
good quality of therapy.
The multivariate calibration techniques were implemented in this study. Two multivariate calibration
models namely principal component regression (PCR) and partial least square (PLS) were applied as
chemometrics techniques to build predictive models for amboxol HCl, pseudoephedrine HCl, and
triprolidine HCl. Spectral preprocessing of the UV spectroscopic data was carried out to achieve various
types of spectra including original, first derivative, second derivative, standard normal variate (SNV), and
Savitzky-Golay(SG). The accuracy of the models was assessed by evaluating the highest value of coefficient
of determination (R2) including coefficient of determination of calibration (Rcal2), coefficient of
determination of cross validation (RCV2), and coefficient of determination of validation (Rval2), whereas the
precision of the models were assessed by evaluating the lowest value of root mean square error of calibration
(RMSEC), root mean square error of cross validation (RMSECV), and root mean square error of
prediction(RMSEP). Performance of the multivariate calibration models are presented in Table 1.
http://dx.doi.org/10.29228/jrp.233
J Res Pharm 2022; 26(5): 1403-1410
1404
Table 1. The performance of PCR and PLS for predicting the content of ambroxol HCl, pseudoephedrine HCl, and
triprolidine HCl
Multivariate Number of
Analytes Types of spectra Rcal2 RMSEC RCV2 RMSECV Rval2 RMSEP
calibration components
Original 2 0.989 0.500 0.986 0.574 0.992 0.578
First derivative 2 0.991 0.463 0.987 0.549 0.993 0.512
PCR Second derivative 8 0.994 0.366 0.988 0.535 0.993 0.543
SNV 9 0.970 0.837 0.938 1.202 0.969 1.110
SG 2 0.989 0.502 0.986 0.576 0.992 0.581
AMB
Original 2 0.989 0.500 0.986 0.574 0.992 0.577
PLS Second derivative 2 0.992 0.451 0.986 0.566 0.994 0.504
SNV 4 0.958 0.997 0.932 1.262 0.952 1.383
SG 2 0.989 0.502 0.986 0.576 0.992 0.581
Original 11 0.955 1.415 0.891 2.197 0.965 1.106
SNV 7 0.868 2.425 0.760 3.265 0.805 2.595
SG 8 0.952 1.467 0.902 2.089 0.919 1.669
PSE
Original 5 0.951 1.471 0.892 2.188 0.974 0.940
SNV 5 0.883 2.277 0.720 3.525 0.802 2.613
SG 7 0.957 1.388 0.897 2.142 0.901 1.849
Original 2 0.992 0.263 0.990 0.289 0.938 0.608
SNV 4 0.943 0.693 0.914 0.850 0.882 0.844
SG 2 0.992 0.263 0.990 0.289 0.939 0.608
TRP
Original 2 0.992 0.263 0.990 0.289 0.938 0.608
SNV 4 0.944 0.682 0.913 0.852 0.884 0.834
SG 2 0.992 0.263 0.990 0.289 0.939 0.608
Note: Selected model of calibration for each compound were marked with bold. AMB: ambroxol HCl; PSE: pseudoephedrine HCl; TRP:
triprolidine HCl; PCR: Principal Component Regression; PLS: Partial Least Squares; SNV: Standard Normal Variate; SG: Savitzky-Golay
smoothing with polynomial order of 3 and window width of 11 points.
Figure 1. Variable selection plot (a), regression coefficient plot (b), and multivariate calibration plot (c) of ambroxol HCl
J Res Pharm 2022; 26(5): 1403-1410
1405
According to the data presented in Table 1, it was obtained that the selected model for ambroxol HCl
was PCR model on second derivative spectra with the Rcal2, RCV2 and Rval2 were 0.994, 0.988, and 0.993,
respectively. The error of the model was expressed from the RMSEC, RMSECV, and RMSEP with the value
of 0.366, 0.535, and 0.543, respectively. Multivariate predictive equation for ambroxol HCl was y = 1.002x -
0.386. The variable selection plot, regression coefficient plot, and multivariate calibration plot of ambroxol
HCl were depicted in Figure 1.
The selected model for pseudoephedrine HCl was PCR model on SG spectra with the Rcal2, RCV2 and
Rval were 0.952, 0.902, and 0.919, respectively. The error of the model was expressed from the RMSEC,
2
RMSECV, and RMSEP with the value of 1.467, 2.089, and 1.669, respectively. Multivariate predictive
equation for pseudoephedrine HCl was y = 1.078x - 3.832. The variable selection plot, regression coefficient
plot, and multivariate calibration plot of pseudoephedrine HCl were depicted in Figure 2.
Figure 2. Variable selection plot (a), regression coefficient plot (b), and multivariate calibration plot (c) of pseudoephedrine HCl
Figure 3. Variable selection plot (a), regression coefficient plot (b), and multivariate calibration plot (c) of triprolidine HCl
The selected model for triprolidine HCl was PLS model on SG spectra with the Rcal2, RCV2 and Rval2
were 0.992, 0.990, and 0.939, respectively. The error of the model was expressed from the RMSEC, RMSECV,
and RMSEP with the value of 0.263, 0.289, and 0.608, respectively. Multivariate predictive equation for
triprolidine HCl was y = 0.8906x + 0.636. The variable selection plot, regression coefficient plot, and
multivariate calibration plot of tripolidine HCl were depicted in Figure 3.
J Res Pharm 2022; 26(5): 1403-1410
1406
3. DISCUSSION
Aided by the selected models for ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl, the
content uniformity of the samples was evaluated. The sample used in this study were compounded to the
target content of 10 mg, 30 mg, and 1.25 mg for ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl,
respectively. Content of ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl found in divided
powder samples were 8.120±1.167 mg, 30.142±3.965 mg, and 1.193±0.221 mg, respectively. Graphical results
of content uniformity were presented in Figure 4.
Figure 4. Content uniformity evaluation for ambroxol HCl (a), pseudoephedrine HCl (b), and triprolidine HCl (c)
The coefficient of variance for percentage content uniformity of ambroxol HCl, pseudoephedrine HCl,
and triprolidine HCl, were 14.002%, 12.822%, and 18.094%, respectively. Pharmaceutical dosage form was
stated to have dose uniformity only if the variance of the content was less than 5 % [29]. The coefficient of
variance for triprolidine HCl was found in the highest percentage compared to ambroxol HCl and
pseudoephedrine HCl. The small amount of triprolidine HCl in the combination of dose may increase the
random error of compounding due to the high variance of the samples. Both underdose and overdose
therapies are possible if the dose of the active ingredient is present in the sample at the different
concentration. However, ambroxol HCl was reported as a well-tolerated drug with low risk of adverse
effects even when it was received in higher therapeutic dose [30]. The highest amount of pseudoephedrine
HCl found in the sample was 38.90 mg (129.678%). It can be assumed that the maximum daily dose of
pseudoephedrine HCl was 116.70 mg since the divided powder has been prescribed for three times a day.
The maximum permissible daily dose of pseudoephedrine HCl was 120 mg for children aged 6–12 years [15].
Thus, it can be stated that in the overdose content obtained from the sample, the potential toxicity can be
avoided with the consideration of daily dose administration.
4. CONCLUSION
Drug combination of mucolytic agent, nasal decongestant, and antihistamine were commonly
prescribed for pediatric patients. In a private hospital in Semarang, Central Java, Indonesia, combination of
drugs including ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl were frequently prescribed as
extemporaneous preparation in the form of divided powder. It was important to ensure the content
uniformity of the divided powder in order to enhance the quality of the therapy given. UV spectroscopy
method combined with chemometrics techniques were applied to determine content uniformity of the
samples. The best predictive models for each compound were selected along with the evaluation of accuracy
and precision of the models.
Content uniformity was evaluated from the sample of divided powders. It was found that the
coefficient of variance (CV) for percentage content uniformity of ambroxol HCl, pseudoephedrine HCl, and
triprolidine HCl, were 14.002%, 12.822%, and 18.094%, respectively. It can be assumed that the sample of
divided powders compounded by pharmacist in private hospital did not meet the requirements of content
uniformity since the percentage of coefficient of variance were more than 5%. However, the daily doses of
therapy of ambroxol HCl, pseudoephedrine HCl, and triprolidine HCl were still in the safety therapeutic
range. Pharmacists are strongly encouraged to improve their knowledge and skills in the practice of
compounding in order to provide patient-centered health care services.
5. MATERIALS AND METHODS

5.1. Materials
Materials used in this study were working standard of ambroxol HCl obtained from PT. Ifars
Pharmaceutical Laboratories (Surakarta, Central Java, Indonesia), pseudoephedrine HCl and triprolidine
J Res Pharm 2022; 26(5): 1403-1410
1407
HCl working standards obtained from PT. Dexa Medica (Palembang, South Sumatra, Indonesia). Samples of
divided powders were compounded by pharmacist in a private hospital in Semarang, Central Java,
Indonesia. These samples were compounded from the ambroxol HCl tablet mixed with Alerfed® tablets
containing pseudoephedrine HCl and triprolidine HCl.
5.2. Instrumentation and Software
The UV-Vis Spectrophotometer type UV 1800 (Shimadzu, Japan) with a set of quartz cuvette 1 cm
(Hellma, USA), analytical balance of OHAUS® (China) series of PA224C, and a set of Socorex®(Switzerland)
micropipettes were used in this study. The UV spectral data were exported to Excel (Microsoft, USA) and
converted into .csv formatted files for further data processing. Statistical analysis and multivariate
calibrations were performed using R software version 1.4.1717 with “pls” and “prospectr” packages.
5.3. Methods
5.3.1. Sample Preparation
Samples used in this study were divided powder containing ambroxol HCl and commercial tablet
(containing pseudoefedrin HCl of 60 mg and triprolidine HCl of 2.5 mg) that prepared by pharmacist in a
private hospital in Semarang, Indonesia. This study was approved by the Health Research Ethics Committee
of Faculty of Health Science, Universitas Respati Yogyakarta (220.3/FIKES/PL/X/2021). The formula of the
divided powder was prescribed as follow:
R/ Ambroxol HCl 30 mg 1/3 tab
Alerfed® ½ tab
Mf. Pulv. dtd no.X
The preparation of divided powder was triplicated, hence a total of 30 dose units were obtained.
5.3.2. Calibration and Validation Solutions
An accurate weight of 10.0 mg of ambroxol HCl, pseudoephedrine HCl and triprolidine HCl were
transferred into three separated 10 mL volumetric flasks for each compound followed by dilution with
methanol into the volume. These solutions were labeled as ambroxol HCl, pseudoephedrine HCl and
triprolidine HCl stock solution.
Calibration and validation set solutions were prepared by mixing synthetic combination of ambroxol
HCl, pseudoephedrine HCl and triprolidine HCl from the stock solutions to obtain 45 compositions of
standards solution mixture as presented in Table 2. Each composition was scanned using spectroscopic
method at the range of 240-300 nm. The absorbance for each composition were recorded with the interval of
2 nm. The obtained absorbance data for each wavelength were assigned randomly and used for building
calibration and validation models.
Table 2. Calibration and validation data sets information for developing multivariate calibration models of ambroxol
HCl, pseudoephedrine HCl and triprolidine HCl
Items Data sets
Calibration Validation
Number of mixture standards 30 15
Ambroxol HCl Concentration (µg/mL)
Mean 12.953 14.312
Range 5.96 – 23.85 4.97 – 24.85
Pseudoephedrine HCl Concentration (µg/mL)
Mean 30.750 28.101
Range 20.12 – 40.24 20.12 – 40.24
Triprolidine HCl Concentration (µg/mL)
Mean 5.672 4.766
Range 1.01 – 10.07 1.01 – 10.07
Multivariate calibration models PCR PCR
PLS PLS
Evaluated parameters for model selection R2 R2
RMSEC RMSEP
RMSECV*
Note: * cross validation was performed using leave one out technique
5.3.3. Multivariate Calibration Analysis

Absorbance data of calibration and validation set solutions were preprocessed into five types of
spectra namely normal/original, first derivative, second derivative, SNV, and SG smoothing (window width
J Res Pharm 2022; 26(5): 1403-1410
1408
of 11 points, polynomial order of 3). Multivariate calibration models namely PCR and PLS were built for
ambroxol HCl, pseudoephedrine HCl and triprolidine HCl. The generated multivariate calibration models
were statistically evaluated by assessing several performances such as R2, RMSEC, RMSECV, and RMSEP.
The selected model for each compound was identified by evaluating the highest value of R2 (near 1) and
lowest value of RMSEC, RMSECV, and RMSEP [28].
5.3.4. Content Uniformity Evaluation
Selected multivariate calibration model for each compound were used to evaluate the content
uniformity of the active pharmaceutical ingredients namely ambroxol HCl, pseudoephedrine HCl, and
triprolidine HCl. The actual value of the content was calculated by plotting the predictive value obtained
from the multivariate calibration model. Content of ambroxol HCl, pseudoephedrine HCl and triprolidine
HCl from twenty samples of divided powders were determined. The mass of each compound was
calculated as well as their standard deviation. The percentage of the content was also determined and
presented as graphical chart to visually observe the variance occurred in each dose.
Acknowledgements: We thank to Institute of Research and Community Services, Sanata Dharma University for
financially fund with Centre Research Grant (“Penelitian Pusat Studi”) of 2021, with contract number ofNo.
013/Penel/LPPM-USD/II/2021. We thank to Faculty of Pharmacy Universitas Sanata Dharma, Yogyakarta,
Indonesia for providing laboratory facilities. We also thank to PT. Ifars Pharmaceutical Laboratories and PT. Dexa
Medica for providing working standards used in this study.
Author contributions: Concept – S.H.Y; Design – F.D.O.R., D.M.V.; Supervision – S.H.Y.; Resources – M.R.G.,
D.C.A.P.; Materials – M.R.G., D.C.A.P.; Data Collection and/or Processing – C.A.S; Analysis and/or Interpretation –
C.A.S., F.D.O.R.; Literature Search – D.M.V., D.C.A.P.; Writing – S.H.Y., M.R.G.; Critical Reviews – S.H.Y., F.D.O.R.,
M.R.G., D.C.A.P., D.M.V.
Conflict of interest statement: The authors declared no conflict of interest.
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