Faculty of Pharmacy - Mahidol University

Mahidol University Annual Research Abstracts, Vol. 37 315 

5-SUBSTITUTED PYRIDO[2,3-d]PYRIMIDINE, AN 

INHIBITOR AGAINST THREE RECEPTOR 

TYROSINE KINASES (NO. 0848) 

Naparat Kammasud 1 , Chantana Boonyarat 2 , Kingkan 

Sanpanya 1 , Maleeruk Utsintong 1 , Satoshi Tsunoda 3 , Hiroaki 

Sakurai 3 , Ikuo Saiki 3 , Isabelle Andre 4 , David S. Grierson 4 , Opa 

Vajragupta 1 

1 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, 

Mahidol Univerisyt, Bangkok 10400, Thailand; 2 Faculty of 

Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 

40002, Thailand, E-mail :pyovj@mahidol.ac.th; 3 Division of 

Pathogenic Biochemistry, Department of Bioscience, Institute of 

Natural Medicine, Japan; 4 UMR 176 CNRS, Institute Curie, 

Section Recherche, Bat. 110-112, 91405 Orsay cedex, France. 

Key words : 5-Substituted indolin-2-one, Anti-angiogenesis, Antiproliferation; 

Binding mode, Docking; FGFR-1, FGFR-1 inhibitor, 

SU6668, Virtual screening 

NP506, the 3-(2-4-dimethyl-5-(2-oxo-5-(N’-phenylhydrazinocarbonyl)-1,2-dihydro-indol-3-ylidenemethyl)-1H-pyrrol-3-yl)propionic 

acid, was designed as FGF receptor 1 inhibitor by 

computational study and found to be more active against endothelial 

proliferation of HUVEC after the rhFGF-2 stimulation than SU6668 

with minimum effective dose of 10 μM. NP506 inhibited the tyrosine 

phosphorylation in FGF, VEGF, and PDGF receptors and the 

activation of extracellular signal-regulated kinase (ERK), c-Jun-Nterminal-kinase 

(JNK) and AKT after the rhFGF-2 stimulation. The 

introduction of the phenyl hydrazide motif to the position 5 of the 

pyrido[2,3-d]pyrimidine scaffold led to the inhibitory effect in two 

signaling pathways : inhibition of AKT activation in the phosphatidyl 

inositol 3’-kinase (PI13K)/AKT signaling pathway and the inhibition 

of ERK and JNK activation in MAPK pathway 

(Bioorg Med Chem Lett, 2009; 19(3):745-750.) 

(Financial supported by the Thailand Research (TRF) through The 

Royal Golden Jubilee Project, The Commission of Higher Education, 

Ministry of Education, Thailand and the 21 st Century COE project 

from the Ministry of Education, Culture, Sports, Science and 

Technology, Japan.) 

VIRTUAL SCREENING AGAINS ALPHA-COBRA- 

TOXIN (NO. 0849) 

Maleeruk Utsintong 1 , Tood T. Talley 2 , Palmer W. Taylor 2 , Arthur 

J. Olson 3 , Opa Vajragupta 1 


Mahidol University, Bangkok, Thailand, E-mail : 

pyovj@mahidol.ac.th; 2 University of California, San Diego, 

Skaggs School fo Pharmacy & Pharmaceutical Sciences, San 

Diego, CA 92103 USA; 3 The Scripps Research Insitute, 

Department of Molecular Biology, Molecular Graphic 

Laboratory, La Jolla, CA 92037 USA. 

Key words : Virtual screening, Alpha-cobratoxin, Docking, 

Neurotoxin, Nicotinic Acetylcholine receptor 

Alpha-cobratoxin (Cbtx), the neurotoxin isolated from the venom of 

the Thai cobra, Naja kaouthia, causes paralysis by preventing 

acetylcholine (ACh) binding to nicotinic acetylcholine receptors 

(nAChRs). In the current study, the region of the Cbtx molecule that 

is directly involved in binding to nAChRs is used as the target for 

anticobratoxin drug design. The crystal structure (1Y15) of Cbtx in 

complex with the acetylcholine binding protein (AChBP), a soluble 

homolog of the extracellular binding domain of nAChRs, was selected 

to prepare an alpha-cobratoxin active binding site for docking. The 

amino acid residues (Ser182-Tyr192) of the AChBP structure, the 

binding site of Cbtx, were used as the positive control to validate the 

prepared Cbtx active binding site (root mean square deviation < 1.2A). 

Virtual screening of the National Cancer Institute diversity set, a 

library of 1990 compounds with nonredundant pharmacophore 

profiles, using AutoDock against the Cbtx active site, revealed 39 

potential inhibitor candidates. The adapted in vitro radiolignand 

competition assays using 3 H-epibatidine and 125 I-bungarotoxin against 

the AChBPs from the marine species, Aplysia californica (Ac), and 

from the freshwater snails, Lymnaea stagnalis (Ls) and Bolinus 

truncates (Bt), revealed 4 compounds from the list of inhibitor 

candidates that had micromolar to nanomolar interferences for the 

toxin binding to AChBPs. Three hits (NSC42258, NSC 121865, and 

NSC134754) can prolong the survival time of the mice if administered 

30 min before injection with Cbtx, but only NSC121865 and 

NSC134754 can prolong the survival time if injected immediately 

after injection with Cbtx. These inhibitors serve as novel templates/ 

scaffolds for the development of more potent and specific 

anticobratoxin. 

(J Biomol Screen, 2009; 14(9):1109-18.) 

(Financial supported by the Thailand Research Fund (TRF) through 

The Royal Golden Jubilee Project, The Commission of Higher 

Education, Ministry of Education, Thailand and Mahidol University 

Research Fund.) 

REDIOCIDES A AND G AS POTENTIAL ANTITO- 

XINS AGAINST COBRA VENOM (NO. 0850) 

Maleeruk Utsintong 1 , Atchara Kaewnoi 2 , Wichet Leelamanit 3 , 

Arthur J. Olson 4 , Opa Vajragupta 1 


Mahidol University, Bangkok 10400, Thailand; 2 Department of 

Chemistry, Faculty of Science and Technology, Bansomdejchaopraya 

Rajabhat University, Bangkok 10600, Thailand; 

3 Department of Biochemistry, Faculty of Pharmacy, Mahidol 

University, Bangkok 10400, Thailand; 4 The Scripps Research 

Institute, Molecular Graphics Laboratory, Department of 

Molecular Biology, California 92037, USA. 

Key words : Rediocides A ang g, Trigonostemon reidioides, Naja 

Kaouthia, cobratoxin, antitoxins, Antivenoms, Detoxification 

Rediocides A and G, the principle components, of 

Trigonostemon reidioides (Kurz) Craib, which is known as 

Lotthanong in Thai, were investigated for detoxification mechanism 

against Naja kaouthia venom by in silico, in vitro, and in vivo 

methods. Molecular dockings of a-cobratoxin with rediocides A and 

G were performed, and the binding energies were found to be – 14.17 

and -14.14 kcal/mol, respectively. Rediocides bind to a-cobratoxin 

at the samelocation as a-cobratoxin binds to the nicotinic 

acetylcholine receptor (nAChR), i.e, at the Asp27, Phe29, Arg33,

316 

Gly34, Lys35, and Val37 residues. A-Cobratoxin cannot bind to 

nAChR, because some of its binding sites areoccupied with 

rediocides. From in vitro SDS-PAGE, it was found thatrediocides 

can diminish the bands of a-cobratoxin. In the presence of 

acetylcholine-binding protein (AcHBP), it was apparent that 

rediocides can bindboth a-cobratoxin and AChBP. From an in vivo 

test, it was found that injection of rediocides at 0.5 mg/kg immediately 

after a-cobratoxin dose of three timesLD50 cannot prolong the 

survival time of mice. However, rediocide can prolongthe survival 

time, if it is injected 30 min before the injection of a-cobratoxin. 

The in vitro SDS-PAGE and the in vivo results support the in silico 

detoxification mechanism of rediocides against cobra venom at a 

molecular level. 

(Chem Biodivers, 2009; 6(9):1404-14). 

(Financial supported by the Thailand Research Fund (TFR) through 

The royal Golden Jubilee Project, The Commission of Higher 

Education, Ministry of Education, Thailand.) 

TLC DENSITOMETRIC METHOD FOR THE 

QUANTIFICATION OF CREBANINE IN 

Stephania venosa (NO. 0851) 

Bongkod Wongpan 1 , Omboon Luanratana 1 , Nongluck 

Ruangwises 2 , and Ampol Mitrevej 3 

1 Department of Pharmacognosy; 2 Department of Pharmaceutical 

Chemistry; 3 Department of Manufacturing Pharmacy, Faculty 

of Pharmacy Mahidol University, Bangkok, Thailand. 

Key words : Stephania venosa, Crebanine, TLC-Densitometry 

Crebanine (aporphine alkaloid) was found in the tubers 

of Stephania venosa (BI) Spreng., known in Thai as Sabu-leud, 

belongs to the Menispermaceae family. This compound has been 

shown to give interesting biological activities, therefore it could be 

used as biomarker. The validate analytical methods were developed 

for the effective quantification of this compound by TLC- 

Densitometry with a good correlation coefficient value of 0.9991. 

The method was found to be precise with %RSD of 0.77 (intra-day) 

and 0.83 (inter-day). Instrumental precision was 0.49 (%RSD). 

Accuracy of the method was evaluated by managing recovery study 

at two different levels concentration of crebanine and the average 

percentage recovery was found to be 100.06%. The content of 

crebanine in various extracts of S. venosa were estimated and found 

to be in the range of 0.10-0.52%, the highest content was found in 

the ethyl acetate extract obtained by soxhlet extraction (0.52%). This 

developed method is simple, accurate and precise, and can be suitably 

applied for the quantitative analysis of crebanine content in crude 

extract of S. venosa tubers. 

(This work is a part of Master Thesis in Pharmaceutical Chemistry 

and Phytochemistry Programme, Department of Pharmacognosy, 

Faculty of Pharmacy, Mahidol University, Bangkok, Thailand, 2008) 

(The paper was submitted for publication in Journal of Planar 

Chromatography on December 5, 2009) 

THE OBSTRACLES OF ASIAN MEDICINE UTILI- 

SATION AND SOLUTIONS (NO. 0852) 

Omboon Luanratana 

Department of Pharmacognosy, Faculty of Pharmacy, Mahidol 

University, Bangkok 10400, Thailand; pylon@mahidol.ac.th 

Key words : Asian medicine,holistic healing IASTAM 

Asian medicine are practice throughout the world but it 

is not officially recognized by most governments since it cannot be 

reimbursed. The obstacles which hold back this art of healing and 

its consequence to cancer patients will be presentedas an example. 

The imposition of western biomedical sciences and its paradigm on 

traditional medicine reflected in laws and regulations against 

traditional medicine and its practitioners. Simple analogy i.e. people 

try to use rugby rules with soccer game which is impossible, however, 

some soccer players and the audiences keep trying to apply rugby 

rules with the soccer game. Another obstacle is the commercial 

influence from western medicine on modern medicine schools. These 

medical students were framed their thought with the paradigm of 

scientific based medicine only and believe that it is the only reliable 

method of healing and subsequently refuse to look at other methods. 

Asian medicine is a philosophy, the art of holistic healing derived 

from real practices on man throught history of mankind which can 

be described well with ‘the black box theory’ rather than the 

mechanistic biomedical sciences which believe in reductionism. The 

solutions can be found it the government and healthcare personels 

regard patients as the center of curing not their belief. Both western 

medicine and eastern medicine should be used to complement each 

other and clinical case reports should be sufficient evidences for its 

effectiveness. Any stringent rules and laws should be uplifted and 

special appropriate laws and rule should be designed for traditional 

medicine. International voice through IASTAM could be better heard 

than a single national voice. 

(Oral Presentation in Asian Medicine, Cultivating, Traditions and 

the Challenges of Globalisation, Thimphu, Bhutan, 6-13, July 2009.) 

A POTENTIAL HERBAL EXTRACT FOR THE 

CURE OF MELANOMA (NO. 0853) 

Omboon Luanratana 1 , and Pongpun Siripong 2 

Faculty of Pharmacy 

1 Department of Pharmacognosy, Faculty of Pharmacy, Mahidol 

University, Bangkok 10400, Thailand, pylon@mahidol.ac.th; 

2 Medicinal Plant Research Unit, National Cancer Institute, 

Department of Medicine, Ministry of Public Health, Bangkok 

10400, Thailand. 

Key words : Morus alba NM-60, antimelanoma, antioxidant 

At present we are facing with Global warming and the 

increase intensity of UV-A and UV-B in the sun light. Skin cancer 

patients are increasing in number especially those with melanoma 

which is fatal due to its ability in metastasis. WHO reported 160,000 

patients suffered from melanoma per year and 48,000 patients died 

each year. At present, there is no effective drug for this type of cancer, 

moreover the chemical drugs often have adverse effects to the normal 

cell and can also induce resistance in the cancer cells. It is the aim of


this study to find a safe and effective drug for the treatment of 

melanoma. Antiproliferative property and antiangiogenesis are 

considered to be the crucial properties of anticancer drugs. To be 

safe and effective the drug should be able to quench the toxic effects 

that cancer cells exert on normal cells and due to the over production 

of melanin pigment the drug should be able to reduce this overactive 

biosynthesis. Preliminary study showed that various extracts of Morus 

alba sericulture NM-60 contained inhibitory activities against 

melanoma cell line. Moreover, reduction of skin tags was also 

reported by those using Mulberry containing consmetics. Methods : 

The twigs and roots of Mulberry were extracted and tested against 

B-16 melanoma cell line. The antioxidant property and the 

antityrosinase property were also studied. The IC 50 values were 

compared with those of the markers; oxyresveratrol and betulinic 

acid. Results : The stem heartwood and the root bark of Mulberry 

were active against melanoma. Both dichloromethane extract and 

methanolic extract were active. One of the active component in the 

stem was oxyresveratrol while betulinic acid was among the active 

components in the root bark. The IC 50 values of antioxidant and 

antimelanoma properties were in the level of micrograms while that 

of antityrosinase was in the level of milligrams. Antiangiogenesis is 

under investigation. 

(Poster presentation in The 2009 DMS Annual Scientific Conference 

and the 10 th National Cancer Conference : A joint Meeting, Miracle 

Grand Convention Hotel, Bangkok, November 2-4, 2009.) 

HIGH-PERFORMANCE LIQUID CHROMATO- 

GRAPHIC ANALYSIS OF SELECTED BIOACTIVE 

COMPONENTS IN Zingiber cassumunar Roxb. 

(NO. 0854) 

Sompol Paramapojn 1 , Santi Tip-pyang 2 , Wandee Gritsanapan 1 , 


University; 2 Department of Chemistry, Faculty of Science, 

Chulalognkorn University. 

Key words : Zingiver cassumunar, anti-inflammatory, (E)-4-(3’,4’ 

–dimethoxyphenyl) but-3-en-1-o1 

The rhizomes of Zingiber cassumunar Roxb. have long 

been used as a traditional medicine for treatment of asthma, and 

muscle and joint pain. Major constitutents, (E)-4-(3’, 4’dimethoxyphenyl) 

but-3-en-1-o1 (A), curcumin (C), and cis-3- 

(2’,4’,5’-trimethoxyphenyl)-4-[(E)-2", 4",5"-trimethoxystyryl] 

cyclohex-1-ene (G) have been reported to be the active antiinflammatory 

constituents in the rhizomes of this plant. A photodiode 

array HPLC method has been developed and validate for quantitation 

of the major bioactive compounds in the methanolic extracts of the 

rhizomes of Zingiber cassumunar collected from fourteen different 

locations in the North, North-East, East, Central and South of 

Thailand. The content of (A) in the dry powdered rhizomes was 

found in the range of 0.400 + 0.001 to 1.642 + 0.005% w/w while 

those of (C) and (G) were 0.066 + 0.001 to 0.265 + 0.002, and 0.027 

+ 0.001 to 0.287 + 0.004 % w/w, respectively. The proposed HPLC 

method was found to be accurate, precise, specific and suitable for 

quantitative analysis of these bioactive constituents. The method 

can be used for quality control of raw material of Zingiber 

cassumunar, extract and its pharmaceutical products. It can be also 

applied in quantifying these marker compounds in other drugs. The 

information of ranges of the contents of bioactive compounds will 

be useful as a guidance for standardization of Z.cassumunar powder 

its extract, and finding sources of good quality of Z. cassumunar. 

(Published in International Journal of Biomedical and 

Pharmaceutical Science (IJBPS) 3(1), 34-38, 2009.) 

(This Research work was granted by the Thailand Research Fund 

(TRF)-RGJ-PhD. Program) 

DETERMINATION OF ANTHRAQUINONE GLYCO- 

SIDE CONTENT IN Cassia fistula LEAF EXTRACTS 

FOR ALTERNATIVE SOURCE OF LAXATIVE 

DRUG (NO. 0855) 

Aurapa Sakulpanich, Wandee Gritsanapan 


University, Bangkok 10400, Thailand. E-mail : 

pywgs@mahidol.ac.th 

Key words : Indian Laburnum, Golden shower, UV-vis 

spectrophotometry 

The pod of Cassia fistula Linn. has been used as a laxative 

drug in Thai traditional medicine for a long time. The pods and 

leaves contain anthraquinone aglycones and anthraquinone 

glycosides which are the active laxative form, while rhein is a major 

component. The degree of laxative potency is depended on the content 

of anthraquinone glycosides. This study determined the content of 

tatal anthraquinone glycosides in leaves of C. fistula collected in 

early summer (February-March, 2008) from 10 provinces in the North, 

North-East, Central, and South of Thailand. The leaves were extracted 

by decoction which was found to be a suitable method for extraction 

of anthraquinone glycosides from C. fistula. All extracts were 

analyzed using a UV-visible spectrophotometric method. The 

contents of total anthraquinone glycosides in the decoction leaf 

extracts were 0.62-2.01% dry weight (average 1.52% dry weight) 

while in the dried leaves were 0.09-0.63% w/w(average0.36% w/w) 

calculated as rhein. The leaves collected from the Central and the 

North-East area, where the weather is warm in summer, contained 

high amounts (average 0.46 and 0.45% dry weight of total 

anthraquinones glycosides, respectively) while the samples from the 

South, where the weather is cool and raining throughout the year, 

contained a lower amount (average 0.13% dry weight). According 

to the standard of ASEAN Herbal Medicine, the central and northeastern 

leaf samples of C. fistula, which contained about 0.5% of 

total anthraquinone glycosides, might be used a source of laxatives 

just as the ripe pods. The decoction extract of the leaves containing 

and average total anthraquinone glycosides 1.52% w/w might be used 

as an alternative source of raw material for various laxative 

preparations. 

(International Journal of Biomedical and Pharmaceutical Sciences 

(IJBPS) 3(1), 42-45, 2009.) 

(This research was granted by the Thailand Research Fund (TRF) 

with the Office of Small and Medium Enterprises Promotion 

(OSMEP).

318 

RAPID CAPILLARY ELECTROPHORETIC 

ANALYSIS OF BERBERINE IN THE STEM 

EXTRACTS OF Coscinium fenestratum (Gaertn.) 

Colebr. (NO. 0856) 

Piyanuch Rojsanga 1 , Leena Suntornsuk 1 , Wandee Gritsanapan 2 , 

1 Department of Pharmaceutical Chemistry, 2 Department of 

Pharmacognosy, Faculty of Pharmacy, Mahidol University, E.mail 

: pywgs@mahidol.ac.th 

Key words : capillary electrophoresis, standardization, isoquinoline 

alkaloid 

Coscinium fenestratum (Gaertn.) Colebr. 

(Menispermaceae) is a medicinal plant widely used in the Indochina 

region. The sterm has been claimed for the treatment of hypertension, 

hypercholesterolemia, cancer and diabetes mellitus. The major and 

active constituent is known to be berberine alkaloid. In the present 

study, a simple and rapid capillary electrophoresis (CE) was developed 

and validated to determine berberine content in the stem extracts of 

C. fenestratum. The background electrolyte system was composed 

of 0.1 M phosphate buffer pH 7.0: methanol (65:35). Linear 

calibration range for berberine was 74.9-374.9 μg/mL (r 2 = 0.999, n 

= 3) with relative standard deviation from intra- and inter-day 

precisions of less than 4.7%. The recovery of berberine was found to 

be 95.8-99.6% with a limit of detection (LOD) and limit of 

quantitation (LOQ) of 4.5 and 15.0 μg/mL, respectively. Capillary 

electrophoresis is a satisfactory system for the standardization of C. 

fenestratum stem extract. 

(Published in International Journal of Biomedical and 

Pharmaceutical Sciences (IJBPS) 3(1), 39-41, 2009.) 

BIOLOGICAL AND CHEMICLA STABILITY OF 

MANGOSTEEN FRUIT RIND EXTRACT (NO. 0857) 

Werayut Pothitirat 1 , Pimolpan Pithayanukul 2 , Mullika Traidej 

Chomnawang 3 , Roongtawan Supabphol 4 , Wandee Gritsanapan 1 


University; 2 Department of Pharmacy, Faculty of Pharmacy, 

Mahidol University; 3 Department of Microbiology, Faculty of 

Pharmacy, Mahidol University; 4 Department of Physiology, 

Faculty of Medicine, Srinakarintarawirot University. 

Key words : Acne vulgaris, antioxidant, Propionibacterium acnes 

The stability of α-mangostin, antiradical activity against 

DPPH radical and antibacterial activity against bacteria-inducing 

acne. i.e. Propionibacterium acnes and Staphylococcus epidermidis 

of Garcinia mangostana fruit rind extract were studied. The extract 

was kept in an amber glass vial and in an aluminium foil bag at 

different temperatures (4-8, 25-28 and 45 o C) for 120 days. The 

content of α-mangostin in the extract stored at each condition was 

analyzed bythe validated RP-HPLC method, while free radical 

scavenging and antibacterial activity were performed using the DPPH 

scavenging assay and broth microdilution method, respectively. The 

results revealed that packaging material and storage temperature had 

no effect on the content of α-mangostin, antiradical and anti-acne 

activities of the G. mangostana fruit rind extract. However, a small 

change in color and characteristic of the extract was observed when 

the extract was kept at 25 and 45 o C. These data indicate good 

chemical biological stabilities of G. mangostana fruit rind extract 

for appropriate antioxidant and anti-acne raw materials in 

pharmaceutical preparations. 

(Published in Functional Plant Science and Biotechnology (FPSB) 

3(1), 16-18, 2009) 

TOTAL PHENOLIC AND FLAVONOID CONTENTS 

AND FREE RADICAL-SCAVENGING CAPACITY 

OF SIAMES NEEM FLOWER EXTRACTS (NO. 0858) 

Worarat Chaisawangwogn and Wandee Gritsanapan 


University, Bangkok 10400, Thailand. E-mail : pywgs@mahidol. 

ac.th 

Key words : Azadirachta indica, scavenging activity, Siames neem 

The free radical scavenging activity and the contents of 

total phenolic compounds and total flavonoids in the aqueous extracts 

of Siamese neem young flowers collected from 14 different locations 

in Thailand were determined. The aqueous extracts were purified by 

partition with butanol. The butanol extract containe total phenolic 

compounds and total flavonoids ranging from 59.86 – 120.61 mg 

gallic acid equivalent (GAE) and 28.03-74.18mg rutin equivalent 

(RE) per gram butanol extract, respectively. EC50 values determined 

by DPPH scavenging assay of the butanol extracts were within the 

range of 9.32-46.97 μg/mL. Extracts from samples from the central 

area of Thailand had the strongest scavenging activity (average EC50 

= 18.33 μg/mL), the highest total phenolic content (94.08 + 19.09 

mg GAE/g butanol extract) and total flavonoid conten (60.08 + 14.86 

mg RE/g butanol extract). The results suggested correlation between 

the contents of total flavonoids and total phenolic compounds, and 

the scavenging activity of most extracts. The high free radical 

scavenging activity of Siamese neem young flower extracts supports 

the ethnomedical use of this plant. 

(Published in The Philippine Agricultural Scientist 92(4), 397-402, 

December 2009.) 

FREE RADICAL SCAVENGING ACTIVITY DETER- 

MINATION AND QUANTITATIVE ANALYSIS OF 

CURCUMINOIDS IN Curcuma zedoaria RHIZOME 

EXTRACTS BY HPLC METHOD (NO. 0859) 

Somphol Paramapojn and Wandee Gritsanapan 



University, Bangkok, Thailand. E-mail : pywgs@mahidol.ac.th 

Key words : Bisdemethoxycurcumin, Curcuma zedoaria, free radical 

scavenging activity 

Curcuma zedoaria (Zingiberaceae) is a medicinal plant 

containing antioxidant and anti-inflammatory curcuminoids similar 

to the well-known Curcuma longa. Sofar, the content of each 

curcuminoid in the rhizome extract of C. zedoaria in Thailand has 

not been reported. A high-performance liquid chromatography


(HPLC) method was developed and validated for quantitative analysis 

of curcuminoids (curcumin, demethoxycurcumin and 

bisdemethoxycurcumin) content in 70% ethanolic extracts of the 

rhizomes of C. zedoaria collected from 10 different locations in 

Thailand. The method demonstrated good linearity, precision and 

accuracy. The content of curcumin in all crude extracts was found in 

the range of 1.46 + 0.45 to 5.73 + 0.11 % w/w (average 2.73 + 1.24 

%w/w) whereas the contents of demethoxycurcumin, which is a major 

component, and of bisdemethoxycurcumin were in the ranges of 3.15 

+ 0.15 to 10.98 + 0.28% w/w (average 7.37 + 2.71 %w/w) and 0.49 

+ 0.02 – 2.99 + 0.02%w/w (averge 1.40 + 0.82% w/w) respectively. 

The highest average total curcuminoids content in the crude extracts 

was 16.83 + 0.62% w/w whereas the lowest content was 6.09 + 1.79% 

w/w. This information will be useful as a guidance of further 

standardization of C. zedoaria raw materiala and extracts. Free radical 

scavenging activity of all extracts was determined using 1,1 – 

diphenyl-2-picrylhydrazyl scavenging assay. Values of EC 50 of all 

extracts were found in the range of 18.29 + 0.05 to 40.33 + 2.24 μg/ 

mL (average 25.71 + 7.54 μg/mL). Free radical scavenging activity 

of the separated pure compounds was found in descending order of 

curcumin > demethoxycurcumin > bisdemethoxycurcumin. 

(Published in Current Science 2009, 97(7): 1069-1073) 

CELLULAR TRANSPORT OF ANTI-INFLAMMA- 

TORY PRODRUGS ORIGINATED FROM A 

HERBAL FORMULATION OF Zingiber cassumunar 

AND Nigella sativa (NO. 0860) 

Prasan Tangyuengyongwatana 1 , Jariya Kowapradit 2 , Praneet 

Opanasopit 2 , and Wandee Gritsanapan 1 


University; 2 Department of Pharmaceutical Technology, Faculty 

of Pharmacy, Silpakorn University, Nakhon Pathom 73000, 

Thailand. E-mail : pywgs@mahidol.ac.th 

Key words : Zingiver cassumunar, cellular transport, Nigella sativa 

The rhizome of Zingiber cassumunar and the seed of 

Nigella sativa are two ingredients in Thai traditional medicine to 

relieve dysmenorrheal and adjust the menstrual cycle. Mixture of 

these two herbs produces three esters, namely (E)-4-(3,4dimethoxypheynl) 

but -3-en-1-yl linoleate (1), (E)-4-(3, 4dimethoxyphenyl) 

but -3-en-1-yl oleate (2) and (E)-4-(3,4dimethoxyphenyl) 

but-3-en-1-yl palmitate (3). The aim of this study 

is to examine in vitro absorption of these esters and evaluate their 

transport across the membrane. In vitro transport of these three esters 

was observed in Caco-2 cell monolayers. The ester compounds 1,2 

and 3 at a concentration of 10 μM were hydrolyzed by porcine liver 

esterase. All esters transported across the Caco-2 cell without 

enzymatic hydrolysis. The apparent permeability coefficient P app of 

compound 1 at 53 μM and 106 μM were 13.94 (0.60) x 10 -6 and 

14.33 (0.17) x 10 -6 cm/s respectively, while those of compound 2 

were 9.45 (0.29) x 10 -6 and 10.08 (0.32) x 10 -6 cm/s, respectively. 

P app values of compounds 3 were 7.48 (0.31) x 10 -6 cm/s at 53 μM 

and 8.60 (0.55) x 10 -6 cm/s at 106 μM. P app values of the parent 

compound (compound D), i.e. (E)-4-(3,4-dimethoxyphenyl) but-3en-1-ol 

were 8.53 (0.83) x 10 -6 cm/s at 53 m M and 16.38 (0.61) x 10 - 

6 cm/s at 106 μM. The ester hydrolysis of compounds, 1, 2 and 3 by 

porcine liver esterase was monitored by HPLC and the hydrolysis 

reactions were completed within 10 minutes. Using the Caco-2 cell 

monolayer model, the present study finds that compounds (E)-4-(3,4dimethoxyphenyl) 

but-3-en-1yl linoleate (1), (E)-4-(3,4dimethoxyphenyl)but-3-en-1-yl 

oleate (2) and (E)-4-(3,4dimethoxyphenyl)but-3en-1-yl 

palmitae (3) originated from Prasaplai 

preparation (a Thai herbal formula) may be transported through a 

facilitated mechanism and serve as prodrugs to increase the compound 

D level in the blood. 

(Published in Chinese Medicine 2009, 4:19 doi:10.1186/1749-8546- 

4-19) 

COMPARISON OF BIOACTIVE COMPOUNDS 

CONTENT, FREE RADICAL SCAVENGING AND 

ANTI-ACNE INDUCING BACTERIA ACTIVITIES 

OF EXTRACTS FROM THE MANGOSTEEN 

FRUIT RIND AT TWO STAGES OF MATURITY 

(NO. 0861) 

Werayut Pothitirat 1 , Mullika Traidej Chomnawang 2 , 

Roongtawan Supabphol 3 , Wandee Gritsanapan 1 


University; 2 Department of Microbiology, Faculty of Pharmacy, 

Mahidol University; 3 Department of Physiology, Faculty of 

Medicine, Srinakarintharawirot University. E-mail : 


Key words : Garcinia mangostana, antioxidant acitivyt, Acne 

vulgaris 

Contents of bioactive components, free radical scavenging 

and anti-acne producing bacteria activities of young and mature fruit 

rind extracts of mangosteen were compared. The young fruit rind 

extract contained significantly higher contents of phenolics and 

tannins and promoted higher free radical scavenging activity than 

the mature fruit rind extract, while the later extract contained higher 

contented higher contents of flavonoids and α-mangostin xanthone 

and gave higher anti-acne producing bacteria activity than the young 

fruit ring extract. Thus, the young and mature stages of mangosteen 

fruit rind should be beneficial for further development of antioxidant 

and anti-acne pharmaceutical preparations, respectively. 

(Published in Fitoterapia 80 (2009) 442-447) 

QUANTITATIVE ANALYSIS AND TOXICITIES 

DETERMINATION OF ARTIFACTS ORGINATED 

IN A THAI TRADITIONAL MEDICINE PRASALAI 

(NO. 0862) 

Prasan Tangyuenyongwatana and Wandee Gritsanapan 


Univeristy, Bangkok, Thailand; E-mail : pywgs@mahidol.ac.th 

Key words : Artifact formation, cytotoxicity, Prasaplai 

Prasaplai is a Thai traditional medicine for relieving 

dysmenorrheal and adjusting the menstrual cycle. Three fatty acid 

esters, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1 yl linolete (1), (E)-4- 

(3,4-dimethoxyphenyl) but-3-en-1-yl oleate (2) (E)-4-(3,4dimethoxyphenyl) 

but-3-en-1-yl palmitate (3) are formed during

320 

storage from the reaction of chemical components in two herbs i.e. 

fatty acids in Nigella sativa (L.) (Ranunculaceae) and (E)-4-(3, 4dimethoxypheyl) 

but-3-en-1-o1 (compound D) in Zingiber 

cassumunar (Roxb.) (Zingiberaceae). The formations of these 

artifacts were monitored for 1 year and their amounts were analyzed 

by HPLC at certain periods of time. The results showed that artifacts 

formation was saturated after 73 days of storage. The amount of 

each artifact in the saturation period ranged from 3.93 + 0.06 to 4.30 

+ 0.18% w/w for compound 1, 1.69+0.08 to 1.9 + 0.13% w/w for 

compound 2 and and 0.09 + 0.003 to 0.1 + 0.005% w/w for compound 

3. Cytotoxicyt of the artifacts was evaluated using NCI-H187, KB, 

and BC cancer cell lines and found that the IC 50 of all artifacts in all 

tests were higher than 20 μg/mL. For acute toxicity in mice, the 

LD 50 of each artifact was more than 300 mg/kg. 

(Published in Pharmaceutical Biology, 2009) 

FREE RADICAL SCAVENGING AND ANTI-ACNE 

ACTIVITIES OF MANGOSTEEN FRUIT RIND 

EXTRACTS PREPARED BY DIFFERENT EXTRAC- 

TION METHODS (NO. 0863) 

Werayut Pothitirat 1 , Mullika Traidej Chomnawang 2 , 

Roongtawan Supabphol 3 , and Wandee Gritsanapan 1 

1 Department of Pharmacognosy, 2 Department of Microbiology, 

Faculty of Pharmacy, Mahidol University, Bangkok, Thailand; 

3 Department of Physiology, Faculty of Medicine, 

Srinakarintarawirot University, Bangkok, Thailand. E-mail : 


Key words : Acne vulgaris, anti-acne, Garcinia mangostana 

The ethanol extracts of mangosteen fruit rinds prepared 

by several extraction methods were examined for their contents of 

bioactive compounds, DPPH-scavenging activity, and anti-acne 

producing bacteria against Propionibacterium acnes and 

Staphylococcus epidermidis. The dried powder of the fruit rind was 

extracted with 95% ethanol by maceration, percolation, Soxhlet 

extraction, ultrasonic extraction, and extraction using a magnetic 

stirrer. Soxhlet extraction promoted the maximum contents of crude 

extract (26.60% dry weight) and α-mangostin (13.51% w/w of crude 

extract), and also gave the highest anti-acne activity with MIC 7.81 

and 15.63 μg/mL and MBC 15.53 and 31.25 μg/mL against P. acnes 

and S. epidermidis, respectively. Ethanol 70% and 50% (v/v) were 

also compared in Soxhlet extraction. Ethanol 50% promoted the 

extract with maximum amounts of total phenolic compounds (26.96 

g gallic acid equivalents/tract) and total tannins (46.83 g tannic acid 

equivalent/100 g extract), and also exhibited the most effective DPPHscavenging 

activity (EC 50 12.84 μg/mL). Considering various factors 

involved in the process, Soxhlet extraction carried a low cost in terms 

of reagent sand extraction time. It appears to be the recommended 

extraction method for mangosteen fruit rind. Ethanol 50% should 

be the appropriate solvent for extracting free radical-scavenging 

components, phenolic compounds, and tannins, while 95% ethanol 

is recommended for extraction of α-mangostin, a major anti-acne 

component from this plant. 

(Published in Pharmaceutical Biology, 2009) 


HIGH PERFORMANCE LIQUID CHROMATO- 

GRAPHIC ANALYSIS OF ANTHRAQUINONE 

COMPOUNDS IN THE Laurera benguelensis (NO. 0864) 

N. Manojlovic 1 , Z. Markovi 2 , W. Gritsanapan 3 , and K. 

Boonpragob 4 

1 Department of Pharmacy, Medical Faculty, University of 

Kragujevac, S. Markovi 69, 34000 Kragujevac, Serbia; 

2 Department of Biochemical and Medical Sciences, State 

Universtiy of Novi Pazar, Vuka Karad I a bb, Novi Pazar 36300, 

Serbia; 3 Faculty of Pharmacy, Mahidol University, 10400 

Bangkok, Thailand; 4 Faculty of Science, Ramkhamhaeng 

University, Bangkok, Thailand. E-mail : ntm@kg.ac.yu 

Key words : Laurere benguelensis, anthraquinone, HPLC 

A high performance liquid chromatographic (HPLC) 

method has been developed for the characterization of anthraquinone 

metabolites in extracts of the lichen Laurera benguelensis. With 

this method four anthraquinone derivatives 1,8-dihydroxy-3methoxy-6-methylanthraquinone,1,8-dihydroxy-3-formyl-6methoxyanthraquinone,1,8-dihydroxy-3-hydroxymethyl-6-methoxyanthraquinone 

and 1,3,8-trihyroxy-6-methylanthraquinone can be 

analyzed. Components of lichen were detected by characteristic 

ultraviolet spectra and relative retention times. This is first report of 

phytochemical analysis of L. benguelensis. Importance of this 

research is in recognizing some new source (lichen and its extracts) 

as a natural emplacement of antioxidants because oxidation with free 

radicals or auto-oxidation is big problem for preservation of food 

products. 

(Published in Russian Journal of Physical Chemistry a, 2009, 83(9), 

1554-1557) 

N-ACETYLCYSTEIN INHIBITS PROLIFERATION, 

ADHESION, MIGRATION AND INVATION OF 

HUMAN BLADDER CANCER CELLS (NO. 0865) 

Athikom Supabphol 1 , Verasing Muangman 2 , Warinthorn 

Chavasiri 3 , Roongtawan Supabphol 4 , Wandee Gritsanapan 5 

1 Interdisciplinary Programme in Clinical Sciences, Faculty of 

Medicine Siriraj Hospital, Mahidol University, Bangkok; 

2 Department of Surgery, Vichaiyut Hospital and Medical Center, 

Bangkok, 3 Department of Chemistry, Faculty of Sciences, 

Chulalongkorn University, Bangkok; 4 Department of Physiology, 

Faculty of Medicine, Srinakharinwirot University, Bangkok; 


University, Bangkok, Thailand. 

Key words : N-acetylcystein, urinary bladder, neoplalsms 

Bladder cancer is not only a major public health and 

economically burden for the patients but also a major clinical impact 

for Thai urlogists. The authors’ aim was to study the anti-metastic 

effect of N-acetylcysteine (NAC), one of the cheap, safe and widely 

used over-the-counter-drugs in Thailand, on the human bladder cancer 

cells. Effects of NAC at various concentrations on the growth, 

adhesion, migration and invasion of the human bladder cancer cell 

line were assessed in vitro. NAC at the concentrations of 5, 10, 20


and 30 mM could directly and significantly inhibit the growth, 

adhesion, migration, and invasion of the human bladder cancer cells 

in a dose-dependent manner. The 50% inhibitory concentration (IC 50 ) 

value for cell viability was 33.33 + 0.78 mM. The inhibitory effects 

on migration, invasion and adhesion properties of the cancer cells 

were dramatically observed at the concentrations of > 10, >20, and 

>30 mM respectively. NAC has an anti-metastatic effect on the 

human bladder cancer cells by inhibiting their growth, adhesion, 

migration, and invasion properite. This implies the high possibility 

that the urologists may apply the results to use it intravesically before, 

during and after the transurethral resection of bladder tumour, in 

addition to its conventional usage by oral and parenteral routes. 

(Published in J Med Assoc Thai 2009; 92(9): 1171-1177.) 

HPLC QUANTITATIVE ANALYSIS METHOD FOR 

THE DETERMINATION OF αα 

α-MANGOSTIN α 

IN 

MANGOSTEEN FRUIT RIND EXTRACT (NO. 0866) 

W. Pothitirat and W. Gritsanapan 



ac.th 

Key words : Garcinia mangostana, HPLC, mangostin 

The fruit rind extract of mangosteen (Garcinia 

mangostana Linn.) of which a major active componet is α-magnostin 

has been popularly used in food supplements and herbal cosmetic 

preparations. α-Mangostin is used as a marker quantitative analysis 

and standardization of the raw materials and preparation from this 

plant. The precise method for analysis of plant constituents is 

normally a reverse-phase high performance liquid chromatographic 

(RP-HPLC). The aims of this study were to develop and validate a 

RP-HPLC method for determination of α-mangostin content in the 

extracts of mangosteen fruit rind. Chromatographic separation was 

carried out on a Hypersil@BDS C-18 column (4.6 x 250 mm, 5μm) 

at room temperature using a gradient mobile phase consisting of 70- 

80% acetonitrile in 0.1% v/v ortho phosphoric acid at the flow rate 

of 1 mL min-1 with a UV detection at 320 nm. The method was 

validated for linearity, precision, accuracy, limit of detection (LOD), 

and limit of quantitation (LOQ). The linearity of the proposed method 

was found in the range of 10-200 mg mL -1 with regression coeficient 

0.9999. Intraday and interday precision studies showed the relative 

standard deviation less than 2%. Accuracy of the method was 

determined by a recovery study conducted at 3 different levels, and 

the average recovery was 100.01% . The LOD and LOQ were 0.06 

and 0.17 μg mL -1 , respectively. Two samples of mangosteen fruit 

rind were separately extracted and analyzed using validated HPLC 

method. The contents of α-mangostin in the crude extracts and dried 

powder were within the ranges of 8.36 – 10.04 and 1.84 – 2.47 %w/ 

w, respectively. This developed HPLC method was proven to be 

precise, specific, sensitive, and accurate for routine quality assessment 

of raw material of mangosteen fruit rind, its extracts, and products. 

(Published in Journal Agricultural Science 2009, 42(1):7-12.) 

IN VITRO ANTIGONOCOCCAL ACTIVITY OF 

Coscinium fenestratum STEM EXTRACT (NO. 0867) 

Mullika Traidej Chomnawang 1 , Chutimaporn Trinapakul 2 , 

Wandee Gritsanapan 2 

1 Department of Microbiology, Faculty of Pharmacy, Mahidol 

University 10400, Bangkok, Thailand; 2 Department of 

Pharmacogonosy, Faculty of Pharmacy, Mahidol University, 

Bangkok 10400, Thailand. 

Key words : Neisseria gonorrhoeae, antigonococcal activity, 

Coscinium fenestratum 

Gonorrhea is a sexually transmitted disease (STD), which 

originates from bacteria, Neisseria gonnorrhoeae. It is still one of 

the major health problems worldwide in both men and women. Many 

medicinal plants have been recorded in Thai folk medicine for 

relieving STD but there is no scientific report of these plants for 

antigonococcal activity. This study was conducted to evaluated 

antigonococcal activity of 22 Thai medicinal plants selected from 

the plants popularly used in Thai folk medicine for treatment of 

gonorrhea. The study was performed by disc diffusion and agar 

dilution methods against Neisseria gonorrhoeae. Active compound 

was investigated by bioautographic assay. Among the selected plants, 

Coscinium fenestratum (Gaertn.) Colebr. Extract showed the most 

effective activity against Neisseria gonorrhoeae ATCC 49226 with 

MIC value of 47.39 μg/mL. Bioautographic assay revealed that 

berberine was the active compound of Coscinium fenestratum against 

Neisseria gonorrhoeae. The average MIC values of purified berberine 

against Neisseria gonorrhoeae ATCC 492226 and 11 clinical isolates 

were 13.51 and 17.66 μg/mL, respectively while average MIC value 

of the crude extract of Coscinium fenestratum against all clinical 

isolates was about 56.39 μg/mL. There was no acute toxicity detected 

at the dose of 5 g of Coscinium fenestratum crude extract per 

kilogram. These results provide theoretical support for 

ethnopharmacological relevance of antigonococcal activity of 

Coscinium fenestratum and its active compound. 

(Published in Ethnopharmacology 122 (2009) 445-449) 

EXTRACTION METHOD FOR HIGH FREE 

RADICAL SCAVENGING ACTIVITY OF SIAMESE 

NEEM TREE FLOWERS (NO. 0868) 

Worarat Chaisawangwong and Wandee Gritsanapan 


University, Bangkok 10400, Thailand. 

Key words : antioxidant, Siamese neem flower, Azadirachta indica 

Siamese neem tree (Azadirachta indica A. Juss. Var. 

siamensis Valeton) is a medicinal plant found in Thailand. Young 

leaves and young flowers of this plant are commonly consumed as a 

bitter tonic vegetable. The flowers are also used for treatment of 

fever. The flower extract has been reported to exhibit in vitro free 

radical scavenging activity and can inhibit lipid peroxidation of 

bronchogenic cancer cell line. Active compounds in the flowers are 

flavonoids such as rutin and quercetin. The content of these 

compounds in the crude extract depends on the method of extraction.

322 

Therefore, the appropriate extraction method promoting high yield 

of total flavonoids and high free radical scavenging activity was 

investigated in this study. Six different extraction methods, i.e. 

maceration, percolation, decoction, soxhlet extraction, ultrasonic 

extraction (UE), and microwave assisted extraction (MA) were carried 

out for extracting dried powder of Siamese neem tree young flowers. 

The solvent used for maceration, percolation, and soxhlet extraction 

was 50% ethanol, while distilled water was used for decoction and 

MA, and both solvents were used for UE. The content of crude extract, 

free radical scavenging activity, and total flavonoids content of each 

extract were investigated and compared. Comparing the various 

extraction methods, decoction provided and extract containing a high 

amount of total flavonoids (17.54 mgRE/g extract) and promoting 

the highest scavenging activity at EC 50 11.36 μg/mL. Decoction is 

also simple, cheap, and convenient and could be used in developing 

countries. Thus, it should be the recommended extraction method 

for the flowers of Siamese neem tree for further development of 

antioxidant pharmaceutical preparations. 

(Published in Songklanakarin J. Sci. Technol. 31 (4), 419-423, Jul. 

– Aug 2009) 

AN APPROPRIATE SOLVENT FOR THE PREPA- 

RATION OF THAI TRADITIONAL MEDICINE 

PRASAPLAI EXTRACT (NO. 0869) 

Prasan Tangyuengyongwatana and Wandee Gritsanapan 


University, Bangkok 10400, Thailand. 

Key words : Prasaplai, artifact, Thai traditional medicine 

Prasaplai is a Thai traditional formulation for relieving 

dysmenorrheal and adjusting the cycle of menstruation. Three fatty 

acid esters, (E)-4-(3,4-dimethoxyphenyl) but-3-en-1yl linoleate (1), 

(E)-4-(3,4-dimethoxyphenyl) but-3-en-1-yl oleat (2) and (E)-4-(3,4dimethoxyphenyl)but-3-en-1-yl 

palmitate (3), were formed during 

storage by the interaction of componets in the preparation. The 

recommended dose (1.0 g) of Prasaplai was separately extracted by 

exhaustive sonication with three different solvents; which are hexane, 

40% ethanol and distilled water, and yielded 26.70 + 0.11 mg (2.7% 

w/w), 33.96+0.05 mg (3.40% w/w), and 49.83 + 0.30 mg (4.98% w/ 

w), respectively. The crude extracts were analyzed by HPLC for 

contents of the four major compounds i.e. (E)-1-(3, 4dimethoxyphenyl) 

but-3-en-1-o1 (compound D), (E)-1-(3,4dimethoxyphenyl) 

butadiene (DMPBD), piperine, β-asaroen, and 

three artifacts. The results showed that only the hexane extract 

contained the artifacts while the 40% ethanol extract contained the 

maximum amounts of the major active anti-inflammatory components 

and water extract contained only compound D. These results suggest 

that the 40% ethanol extracts should be the appropriate extract for 

the preparation of Prasaplai in modern dosage forms due to the high 

content of active anti-inflammatory agents in the extract. 

(Published in Songklanakarin Journal of Science and Technology 

31(5), 527-531, 2009) 


ANTI-ACNE INDUCING BACTERIA ACTIVITY 

AND α-MANGOSTIN CONTENT OF Garcinia 

mangostana FRUIT RIND EXTRACTS FROM 

DIFFERENT PROVENIENCE (NO. 0870) 

Werayut Pothitirat 1 , Mullika Traidej Chomnawang 2 and Wandee 

Gritsanapan 1 

1 Department of Pharmacognosy, 2 Department of Microbiology, 

Faculty of Pharmacy, Mahidol University, Bangkok 10400, 

Thailand. E-mail pywgs@mahidol.ac.th 

Key words : μ-mangostin, mangosteen, Propionibacterim acnes 

The fruit rind of Garcinia mangostana Linn. has been 

traditionally used for treatment of skin infection, wounds, dysentery, 

and diarrhea . α-Mangostin, a major constitutent of the fruit rind, 

was reported to possess a strong inhibitory effect against 

Propionibacterium acnes and Staphylococcus epidermindis, which 

is involved in acne development. This study was conducted to 

quantitative analyze the content of α-mangostin in the fruit rind of 

this plant collected from 13 locations in the South and East of Thailand 

by validated TLC-densitometric method. Antibacterial activity against 

P. acnes and S. epidermindis of the extracts was also determined. α- 

Mangostin contents in the fruit rinds and in the 95% ethanolic extracts 

were found in the ranges of 3.39-5.68 and 11.83-23.11 % dry weight, 

respectively. The samples from the South showed higher contents of 

α-mangostin (average 17.64 % w/w in the extract and 4.85% w/w in 

the dried powder) than the eastern samples. The MIC values of all 

extracts against P. acnes and S. epidermidis were in the range of 

7.81 – 15.63 and 15.63-31.25 μg/mL, respectively, while the MBC 

values were in the range of 15.63-31.25 and 62.50-125.00 μg/mL, 

respectively. The antibacterial activity and α-mangostin content of 

the samples from different locations were significantly different 

(p


one of the plants recommended to be used in primary health care in 

Thailand and is included in the Thai traditional household drug list 

for laxative and antifungal herbal drugs. It is also listed in the Lists 

of National Herbal Drugs of Thailand. The leaves contain 

anthraquinones both aglycone and glycoside forms. The content of 

anthraquinones in the leaves are varied mainly according to 

cultivating locations and harvesting period. It is necessary to control 

the quantity of antraquinones in the leaves and leaf extracts of this 

plant for good quality of laxative and antifungal raw materials. This 

work was conducted to standardize 80% ethanolic extracts prepared 

by the appropriate maceration method of S. alata leaves collected 

from 10 different locations in 4 major parts i.e. the North, North- 

East, Central and Southe of Thailand. The extract ratio was 3-4:1, 

while the characteristics of the extracs were dark brown semi-solid 

with characteristic odour. The extract gave a positive Bontrager’s 

test and its TLC-fingerprints showed 2 major anthraqinone 

components as rhein and aloe-emodin. Total anthraquinones content 

in the extract analyzed by UV-vis spectrophotometric method was 

not lees than 2% w/w (average 2.48% w/w) calculated as rhein. Loss 

on drying of the extract should not more than 6% w/w while the 

extract was slightly soluble in water but freely soluble in 95% ethanol. 

For heavy metals contamination, the extracts should not contain 

arsenic, while lead, mercury and cadmium should not be found more 

than 2, 0.1 and 0.2 ppm, respectively. Total aerobic bacteria count 

and total yeast and mold count should not exceed 100 and 1000 cfu/ 

g, respectively while none of pathogenic bacteria was found in the 

extract. Standardized S. alata leaf extracts could be used as a good 

quality raw material containing anthraquinones for laxative and 

antifungal pharmaceutical preparations. 

(Published in J Health Res 2009, 23(2), 59-64). 

IDENTIFICATION OF STEMONA BY MICRO- 

SCOPIC CHARACTERIZATION (NO. 0872) 

Sumet Kongkiatpaiboon 1 , Sopida Chidchenchey 2 , Vichien 

Keeratinijakal 2,3 , Wandee Gritsanapan 1 


Universityl; 2 National Center for Agricultural Biotechnology, 

Kasetsart University; 3 Agronomy Department, Faculty of 

Agriculture, Kasetsart University, Bangkok 10900, Thailand; Email 


Key words : Stemona, microscopy, characterization 

Stemona plants are widely used for insecticides and 

antitusive remedy Stemona in Thailand comprises 9 known species 

which can be separated into 2 main groups according to their 

morphological characteristics and bioactive components, i.e. tuberose 

group (S. tuberose and S. phyllantha) and non-tuberosa group (S. 

curtisii, S. collinsiae, S. aphylla, S. burkillii, S. Kerrii, S. 

cochinchinensis and and S. pierrei). There are confusion when the 

powder of Stemona is used. This study was investigated the characters 

of each group of Stemona for separation of its species. Cross-sectional 

histology of fresh root samples of 6 species of Stemona, i.e. S. 

burkillii, S. cochinchinensis, S. curtisii, S. kerrii, S. phyllantah and 

S. tuberose were examined. Powdered drug characteristics were 

studied under a microscope using mounting reagents Cross-sectional 

showed that tuberosa group had a non-lignified pith, while the nontuberosa 

group had a smaller lignified one. Powder drug of both 

groups appeared as creamish-yellow color containing vessels, fibers, 

stracth grains and parenchyma cells. Tuberosa group can be 

discriminated from the others by numerous of parenchyma cells. 

Cross-sectional histology and powder drug charactristics of varius 

Stemona species growing in Thailand showed that tuberose group 

had a non-lignified pith and contained numerous parenchyma cells, 

while non-tuberosa group had smaller lignified pith and less abundant 

of parenchyma cells. These microscopic charactrs can be used as a 

tool to identify Stemona groups, and this is the first report on these 

useful characterization. 

(This Manuscript was presented at the Sixth Indochina Conference 

on Pharmaceutical Sciences, Hue, Vietnam, December 15-18, 2009) 

DEVELOPMENT OF ANTIDANDRUFF HERBAL 

SHAMPOO (NO. 0873) 

Phoyphad Chotipattomwan, Sasikan Panyadee, Wandee 

Gritsanapan, Mansuang Wuthi-udomlert 

Faculty of Pharmacy, Mahidol University, Bangkok, Thailand. 

E-mail : pywgs@mahidol.ac.th 

Key words : antidandruff, herbal shampoo, lemongrass 

Lemongrass oil shows a good antifungal activity, Citral, 

which is the main component in lemongrass oil, is potential to inhibit 

Malassezia furfur, a main fungus causing dandruff problem in human. 

This experiment focuses on the development of a good stability and 

efficacy of lemongrass oil antidandruff shampoo. The lemongrass 

oil was analyzed by TLC compared with citral reference standard. 

The minimal inhibitory concentration (MIC) against M. furfur of 

lemongrass oil was determined by broth dilution method. The 1, 2, 

3, 4 and 5% w/w lemongrass oil shampoo were formulated to 

determine the good stability and effective formula. The stability test 

was performed by storage the preparations at room temperature (28- 

30 o C) and accelerated condition (45 o C) for 6 weeks. The Rf value 

(0.38) of lemongrass oil and of citral was similar. The lemongrass 

oil could inhibit M. furfur’s growth at MIC 6.3 μl/ml. The 2% w/w 

lemongrass oil shampoo showed the best stability with good 

appearance, clear, yellow color, not separated. The MICs of the freshly 

prepared shampoo, the shampoo kept at room temperature, and at 

accelerated condition for 6 weeks were similar. The in vivo study in 

human is under determination. Considerig on the good efficacy, 

stability, appearance, and low cost, the 2% w/w lemongrass oil 

shampoo should be the appropriate antidandruff preparation. 

(The Six Indochina Confereence on Pahrmaceutical Sciences, Hue, 

Vietnam, December 15-18, 2009) 

DEVELOPMENT OF DIETARY SUPPLEMENT 

FROM HAIRY BASIL (NO. 0874) 

Pimporn Somkittithum, Ratanaporn Nilanont, Wandee 

Gristsanapan, Mansuang Wuthi-udomlert 



Key words : dietary supplement, hairy basil, Ocinum basilicum 

The husk of hairy basil (Ocinum basilicum L. f. citratum 

Back.) contains fibers that can absorb water 25 times of its dry weight. 

It has been used for laxative and weight control. This study was

324 

aimed to develop instant drink form the mucilage of this plant for a 

dietary supplement. Swollen hairy basil seeds were prepared by 

soaking the seeds in water at different temperatures. The mucilage 

was separated from the seeds and dried in an oven. The dried mucilage 

was determined for it swell volume, loss on drying, total ash and 

acid insoluble ashes according to USP XXXI. The dried mucilage 

was then prepared as 3 formulae instant drink by mixing 0.5, 1 and 

1.5 g of the mucilage with 30 g of Ovantine . The mixture was 

mixed with hot water and the instant drinks were tasted by 30 subjects 

to find out the appropriated formulation. The best condition to make 

the mucilage of hairy basil swollen well (88.6 mL/g) was soaking the 

seeds in the hot water at 60 o C for 90 mins, while the best condition 

for drying the mucilange was 90 o C for 6 hrs. The results showed that 

the swell volume of the dried mucilage was 200 mL/g whiel loss on 

drying, total ash and acid insoluble ash were 7.23, 4.16 and 0.3% w/ 

w, respectively. From all subjects, the most preference formula 

contained hairy basil dried mucilage 0.5 g + 30 Ovantine . Hairy 

basil seeds could be prepared as an instant drink for a dietary 

supplement. The appropriate conditions were soaking the seeds in 

water 60 o C for 90 mins, and the swollen mucilage was dried at 90 o C 

for 6 hrs. The dried mucilage 0.5 g was a suitable amount to mix 

with the drink for laxative and weight control preparation. 

(The Sixth Indochina Conference on Pharmaceutical Sciences, Hue, 

Vietnam, December 15-18, 2009.) 

TURMERIC CREAM : IRRITATION TEST ON 

HUMAN SKIN (NO. 0875) 

Gritsanapan W. 1 , Pitakvongsaporn P. 1 , Sivayathorn A. 2 


University, Bangkok 10400, Thailand; 2 Faculty of Medicine 

Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. 


Key words : Tumeric, Curcuma longa, irritation test 

Turmeric (Curcuma longa Linn.) is a medicinal plant in 

Zingiberaceae family. The rhizome of this plant has been used in 

traditional medicines for treatment of carminatives, gastric ulcer, 

inflammation, and fungal and bacterial skin diseases. The main 

components promoting anti-inflammatory and antimicrobial activities 

are curcuminoids and essential oil, respectively. In this study, turmeric 

oil and crude curcuminoids were extracted from dried rhizomes of 

C. longa and incorporated in a cream base. Five formulations 

consisting of cream base, 0.026% crude curcuminoid cream (CC), 

6% turmeric creams with and without curcuminoids (6TC + CC, 

6TC) and 20% turmeric cream with curcuminoids (20TC+CC), were 

tested for human skin irritation. A 21-day cumulative irritation 

methods was used for irritation test in 22 volunteers (10 women aged 

21-35 and 12 men aged 21-32). The irritation reaction of 20TC+CC, 

6TC+CC and 6TC could be visually observed from day 7, 14 and 10 

on, respectively while their IT 50 was found to be 14.5, 17.2 and 15.9 

days, respectively. It was found that 6TC+CC and 6TC casued 

irritancy potential significantly (p


glycosides, calculated as rhein, by UV-vis spectrophotometric method. 

The contents of rhein and aloe-emodin were also determined by 

HPLC. The leaf extracts contained total anthraquinones 1.13-7.96% 

w/w (average 4.55% w/w), total anthraquinone glycosides 0.62 – 

2.01% w/w (average 1.52% w/w), rhein 0.76 – 3.37% w/w (average 

1.71% w/w) and aloe-emodin 0.01 – 0.15% w/w (average 0.07% w/ 

w). In dried leaves, total anthraquinones, total anthraquinone 

glycosides, rhein, and aloe-emodin contents were found to be 0.16 – 

2.12 % w/w (average1.12% w/w), 0.09 – 0.63% w/w (average 0.36% 

w/w), 0.16-1.06% w/w (average 0.41% w/w), and 0.01 – 0.05% w/w 

(average 0.02% w/w), respectively. Compared with C. angustifolia 

leaf officially in European Pharmacopoeia, which contains not less 

than 2.5% w/w of total hydroxyanthracene glycosides, anthraquinone 

glycosides in the leaves of C. fistula are about 7 times less than in 

the leaves of C. angustifolia, while the C. fistula leaf extract contained 

less hydroxyanthracene glycosides than the leaf extract of C. 

angustifolia about 3.5 times. Thus, in case that C. fistula leaf extract 

would be used as an alternative source of laxative drugs, it has to be 

used for about 3-4 times more than the amount of C. angustifolia 

leaf extract. 

(The 57 th International Congress and Annual Meeting of the Society 

for Medicinal Plant and Natural Product Research, Geneva, August 

16-20, 2009.) 

STANDARDIZATION OF Garcinia mangostana 

FRUIT RIND EXTRACT (NO. 0878) 

Pothitirat W. 1 , Pithayanukul P. 2 , Chomnawang MT. 3 , and 

Gritsanapan W. 1 


University; 2 Department of Pharmacy, Faculty of Pharmacy, 

Mahidol University; 3 Department of Microbiology, Faculty of 

Pharmacy, Mahidol University, Bangkok 10400, Thailand. E-mail 


Key words : Garcinia mangostana, standardization, mangosteen 

The extract of Garcinia mangostana fruit rind has been 

popularly used in food supplement and herbal cosmetics. Therefore, 

it is necessary to establish the specification for quality assessment of 

the extract. The 95 % thanolic extract of G. mangostana fruit rind 

collected from 15 different locations in Thailand were prepared by 

soxhlet extraction. The content of α-mangostin, which is a major 

component, was analysed by HPLC method [1]. Loss on drying, 

solubility, TLC fingerprint and chemical identification of each extract 

was determined. The heavy mental, pesticide and microbial 

contamination were also determined. The extract ratio (crude drug : 

extract) was 2- 4:1, while the contents of α-mangostin in dried powder 

and in the extract were in the range of 1.71-2.70 and 5.79 – 11.12, 

respectively. The extracts were yellow-brown powder with 

characteristic odor, while loss on drying was in the range of 4.52 – 

9.27 % w/w. All extracts were soluble in ethanol, propylene glycol 

and polyethylene glycol but insoluble in water. The extract gave a 

positive with Shinoda’s test, gelatin salt test and promote green-color 

with ferric chloride solution. TLC-fingerprints of all extracts showed 

similar pattern with band of a major α-mangostin. The extract 

contained arsenic < 0.05 ppm, lead < 0.25 ppm, cadmium < 0.02 

ppm and pesticide contamination was not detected. The extracts 

contained total aerobic bacteria count and total yeast mold count not 

more than 5 x 10 5 and 5 x 10 3 cfu/gram, respectively and no 

pathogenic bacteria was found. This information will be useful for 

quality assessment of G. mangostana fruit rind extract used as a raw 

material in pharmaceutical and herbal cosmetic products. 


for Medical Plant and Natural Product Research, Geneva, August 

16-20, 2009.) 

(Planta Medica 2009: 75(9): 991) 

IN VITRO ANTI-ACNE INDUCING BACTERIA 

ACTIVITY OF MANGOSTEEN FRUIT RIND 

EXTRACT GEL (NO. 0879) 

Pothitirat W. 1 , Pithayanukul P. 2 , Chomnawang MT. 3 , and 

Gritsanapan W. 1 


University, Bangkok 10400, Thailand; 2 Department of Pharmacy, 


Thailand; 3 Department of Microbiology, Faculty of Pharmacy, 

Mahidol University, Bangkok 10400, Thaiand. 

Key words : Garcinia mangostana, mangosteen, anti-acne 

The extract of fruit rind of mangosteen (Garcinia 

mangostana Linn.) was reported to possess a strong inhibitory effect 

against Propionibacterium acnes which is a major bacteria involves 

in acne development [1]. Therefore, it is interesting for developing 

anti-acne gel preparation from this using Soxhlet extraction. Hydroalcohol 

gel preparation containing 1% w/w of the extract was 

formulated. The antibacterial activity of the extract and its preparation 

against P. acnes was evaluated using broth microdilution method 

[2]. Based on MIC and MBC values, the extract promoted a good 

inhibitory effect against P. acnes (MIC = 7.81 μg/ml, MBC = 15.63 

μg/ml), while the 1% w/w mangosteen fruit rind gel showed similar 

anti-acne activity with standard 2.5% benzoyl peroxide antiacne gel 

(Pan-Oxyl 2.5 gel) at MIC and MBC 1.56 mg/ml. The results showed 

that anti-acne gel with mangosteen fruit rind extract promoted good 

effect against acne inducing bacteria. The stability of this preparation 

is being investigated 


for Medicinal and Natural Product Research, Geneva, August 16- 

20, 2009.) 

(Planta Medica 2009: 75(9): 1022.) 

CYCLOOXYGENASE-2 ASSAY AND MODELING 

STUDY OF ARTIFACTS FROM PRASAPLAI, A THAI 

TRADITIONAL MEDICINE (NO. 0880) 

Prasan Tangyuenyongwatana 1 , Nipa Jongkon 2 , Chak Sangma 2 

and Wandee Gritsanapan 1 


University, Bangok 10400, Thailand; 2 Department of Chemistry, 

Faculty of Science, Kasetsart University, Bangkok 10900, 

Thailand. 

Key words : Prasaplai, dysmenorrheal, cyclooxygenase-2

326 

The rhizome of Zingiber cassumunar Roxb. (Zingiberaceae) and the 

seed of Nigella sativa L. (Ranunculaceae) are components in 

Prasaplai, a Thai traditional medicine used for relieving dysmenorrhea 

and adjusting the menstrual cycle. The rhizome of Z. cassumunar 

containes (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-o1 (compound D) 

as a major active anti-inflammatory component while the seed of N. 

sativa contains fatty acids which are linoleic, oleic and palmetic acids. 

The powdered mixture of these two plants produced three artificial 

fattyacid esters i.e. (E)-4-(3,4-dimethoxy-phenyl)but-3-en-1-yl 

linoleate (1), (E)-4-(3,4-dimethoxy-phenyl-but-3-en-1-yl oleate (2), 

and (E)-4-(3,4-dimethoxy-phenyl)but-3-en-1-yl palmitate (3) after 

storage. The objectives of this study were to examine the 

cyclooxygenase-2(COX-2) inhibitory activity of these artificial fatty 

acid esters in vitro and to prodict their bindings with COX-2 by using 

molecular docking comparing to compound D. The immortalized 

mouse PGH-1 and PGH-2 null cells were used as the COX-1 and 

COX-2 deficient cell lines, respectively to investigate the inhibition 

activities and selectivity of these compounds. From the experimental 

results, all three artifact showed the inhibition potencies lessthan 

50% suggested that they were inactive compound which corresponded 

to the poor binding results from molecular docking with AutoDock 

3.0.5 program. These results suggest that these artifact might act as 

prodrugs of compound D which were not active directly to COX-2 

and they would be hydrolyzed after absorption in intestinal tract and 

release the parent active anti-inflammatory compound (compound 

D). 

(The 3 rd Asian Pacific Regional ISSX Meeting Bangkok, May 10-12, 

2009). 

VALIDATED HPLC FOR ANALYSIS OF FLAVO- 

NOID CONTENT IN SIAMES NEEM YOUNG 

FLOWERS (NO. 0881) 

Worarat Chaisawangwong, Wandee Gritsanapan 



ac.th 

Key words : HPLC,. flavonoid, Siamese neem tree 

Siamese neem tree (Azadirachta indica a. Juss. var. 

siamensis Valeton) is a medicinal plant found in every part of 

Thailand. The young flower is commonly consumed with sweet sauce 

as bitter tonic vegetable. Moreover, it has been used as element tonic 

and for treatment of fever. The flower extract was reported to exhibit 

in vitro free radical scavenging activity and can ihbibit lipid 

peroxidation of bronchogenic cancer cell line. Active compounds in 

the flowers are flavonoids such as rutin and quercetin. To evaluate 

the quantity of active components in Siamese neem young flowers, a 

high performance liquid chromatographic (HPLC) method was 

developed for the assessment of two bioactived flavonoids: rutin and 

quercetin in the aqueous extract of this plant. A hypersil BDS C 18 

column (250 x 4.6 mm, 5 μm particle size) was used. The elution 

was carried out with isocratic solvent systems with flow rate of 1 ml/ 

min at ambient temperature. The mobile phase was consisted of 

0.5% formic acid in methanol:water (3:7) (A) and methanol (B). 

Quantitative analysis of rutin and quercetin waws programmed at A 

: B(8.5:1.5). The wavelength of UV-vis detector was set at 254 nm. 

Separation of the two compound achieved by the proposed method 


showed good linearity in the range of 15.62 to 500 and 0.49 to 31.25 

μg/ml for rutin and quercetin, respectively. The correlation coefficient 

was higher than 0.999. Intra- and interday precisions showed relative 

standard deviation (%RSD) less than 2%. Accuracy of the method 

was determined by a recovery study conducted at 3 different levels, 

and the average recoveries of rutin and quercetin were 101.97% and 

99.05%, respectively. The LOD and LOQ of rutin were found to be 

0.6518 and 1.9752 μg/ml, while of quercetin were 0.0357 and 0.1081 

μg/ml, respectively. This proposed HPLC method was sensitive, 

accurate and precise for quantitative analysis of flavonoid content in 

the extracts of Siamese neem flowers. The method should be useful 

for routine analysis of flavonoids in the flower raw materials, extracts 

and their preparations of this plant. 


2009.) 

COMPARISON OF ANTI-ACNE INDUCTING 

BACTERIA ACITVITY AND α αα-MANGOSTIN 

α 

CONTENT OF MANGOSTEEN FRUIT RIND 

EXTRACTS PREPARED USING DIFFERENT 

SOLVENTS (NO. 0882) 

Werayut Pothitirat 1 , Mullika Traidej Chomnawang 2 and Wandee 

Gritsanapan 1 


University, Bangkok, Thailand; 2 Department of Microbiology, 


Key words : α-mangostin, Garcinia mangostana, anti-acne 

Mangosteen (Garcinia mangostana Linn.) of the family 

Guttiferae is one of the important exported fruits of Thailand and 

known as ‘Queen of Fruit’. The fruit rind of this plant has been used 

in Thai traditional medicine for the treatment of skin infections, 

wounds diarrhea, dysentery. The extract of the fruit rind has 

demonstrated various biological effects especially antimicrobial 

activity. It contains high amount of xanthones, especially αmangostin 

which is a major constitutent and responsible for 

antibacterial activity. The purpose of this study is to compare antiacne 

inducing bacteria activity and α-mangostin content of 

mangosteen fruit rind extracts which were prepared using different 

solvents i.e., hexane, dichloromethane, ethanol and water. All extracts 

were tested for antibacterial against bacteria inducing acne, including 

Propionibacterium acnes and Staphylococcus epidermidis, using 

broth microdilution method. The dichloromethane extract exhibited 

the strongest antibacterial effect with MIC values for both bacterial 

species at 3.91 μg/ml while MBC values against P. acnes and S. 

epidermidis were 3.91 and 15.63 μg/ml, respectively. TLC 

autobiography indicated that α-mangostin in all extracts, except the 

water extract, was a major activecomponent against both P. acnes 

and S. epidermidis. By HPLC, the dichloromethane extract yielded 

the highest content (46.21 % w/w) of α-mangostin followed by the 

ethanol extract (18.03% w/w), the hexane extract (17.21% w/w) and 

the water extract (0.54% w/w), respectively. These findings suggest 

that the dichloromethane extract of G. mangostna fruit rind should 

be a potential source of natural anti-acne inducing bacteria for further 

development of anti-acne pharmaceutical preparations. 


2009.)


VARIATION OF ANTHRAQUINONE GLYCOSIDES 

IN CASSIA FISTULA LEAVES IN THAILAND 

Aurapa Sakulpanich and Wandee Gritsanapan 

(NO. 0883) 



pywgs@mahido.ac.th 

Key words : Cassia fistula, anthraquinone, UV-spectrophotometry 

The pod of C. fistula L. has been use as a laxative drug in 

Thai tradition medicine for a long time. The pods and leaves contain 

anthraquinone aglycones and anthraquinone glycosides, which are 

the active laxative form while rhein is a major component. Degree of 

laxative potency is depended on the content of anthraquinone 

glycosides. This study determined the content of total anthraquinone 

glycosides in leaves C. fistula collected in early summer (February- 

March, 2008) from 10 provinces in the North, North-East, Central, 

and South of Thailand. The leaves were extracted by decoction which 

was found to be a suitable method for extraction of anthraquinone 

glycosides from C. fistula. All extracts were analyzed using a UVvisible 

spectrophotometric method. The contents of total 

anthraquinone glycosides in the decoction extracts were 0.62-2.01% 

dry weight (average 1.56% dry weight) while in the dried leaves were 

0.09-0.63% w/w (average 0.36% w/w) calculated as rhein. The leaves 

collected from the Central and the North-East area, where the weather 

are warm in summer, contained high amount (average 0.46 and 0.45% 

dry weight of total anthraquinones glycosides while the samples from 

the South, where the weather are cool and raining all year, contained 

lower amount (average 0.13% dry weight). According to standard of 

ASEAN Herbal Medicine, The central and north-eastern leaf samples 

of C. fistula which contained about 0.5% of total anthraquinone 

glycosides might be used as a sources of laxatives as same as the ripe 

pods. The decoction extract of the leaves containing average total 

anthraquinone glycosides 1.56% w/w could be used as a source of 

raw material for various laxative preparations. 

(International Herbal Conference-2009: Herbal medicine- 

Evaluation of quality, Efficacy and Safety”Bangalore, India, 

February 26-28, 2009.) 

VARIABILITY OF FREE RADICAL SCAVENGING 

ACTIVITY AND TOTAL FLAVONOID CONTENT 

IN YOUNG FLOWERS OF SIAMESE NEEM TREE 

IN THAILAND (NO. 0884) 

Worarat Chaisawangwong and Wandee Gritsanapan 



ac.th 

Key words : flavonoid, Siamese neem tree, free radical scavenging 

activity 

Siamese neem tree (Azadirachta indica A. Juss var. 

siamensis Valeton) is a Thai medicinal plant of which young leaves 

and young flowers have been consumed as a bitter tonic vegetable. 

The leaf and flower extracts exhibited in vitro free radical scavenging 

activity and can inhibit peroxidation in cell line experiment. The 

leaves were reported for the content of their active constituents and 

antioxidant activity but not yet in the young flowers. Therefore, 

young flowers of this plant are interesting for determination of free 

radical scavenging activity and total flavonoid content. Free radical 

scavenging activity determined by DPPH scavenging assay total 

flavonoid content determined by aluminium chloride colorimetric 

method of the flower aqueous extracts of Siamese neem young flowers 

collected from different locations in Thailand were performed in this 

experiment. The crude extracts were purified by paritition with 

butanol. Butanol fraction extract was assayed and found that Siamese 

neem young flower extracts contained total flavonoids in a range of 

418 to 844 mg rutin equivalent (RE) per 100 g dried powder. EC 50 

values obtained from DPPH scavenging assay were within a range of 

9.37 to 46.97 μg/ml. The butanol fraction extract from 

Nakhonpathom province in the central part of Thailand promoted 

both strongest antioxidant activity (EC 50 , 9.37 μg/ml) and the highest 

total flavonoid content (844 mg RE/100 g extract). Thin layer 

chromatographic fingerprints of the extracts of Siamese neem young 

flower collected from several locations showed similar pattern. The 

young flowers promoted high free radical scavenging activity 

supporting ethnomedical uses of this plant as a bitter tonic promoting 

good health. The ranges of total flavonoid content and EC 50 values 

will be useful as guidances for standardization of this plant extract 

for medical and health supplement for antioxidant purposes. 

(International Herbal Conference-2009 : Herbal medicine- 

Evaulation of quality, Efficacy and Safety” Bangalore, India, 

February 26-28, 2009.) 

RAPID CATALYTIC HYDROGENATION FOR 

PREPARATION OF TETRAPHYDROCURCUMIN 

AND ZINGERONE (NO. 0885) 

Prasan Tangyuenyongwatana 1 , and Wandee Gritsanapan 3 

1 Faculty of Oriental Medicine, Rangsit University, Patumthani 

12000; 2 Department of Pharmacognosy, Facult of Pharmacy, 

Mahidol University, Bangkok 10400, Thailand. 

Key words : tetrahydrocurcumin, zingerone , Zingiber officinale 

A simple catalaytic hydrogenation at atmospheric pressure 

using ammonium formate as a source of hydrogen gas and Pd/C as 

catalysis has been apllied to synthesized tetrahydrocurcumin and 

zingerone. The present study offers the simple synthesis methods of 

these two interesting compounds with high yield for further 

pharmacological studies. For THC, the target compound was obtained 

by allowing curcumin to react with ammonium formate (3.5 

equivalent) in methanol and using 10% Pd/C as catalyst in reflux 

condition for 30 min. For zingerone, the intermediate (3E)-4-(4hydroxy-3-methoxyphenyl)but-3-en-2-one 

which was synthesized 

from vanillin and acetone by aldol condensation reaction was reacted 

with ammonium formate(2.0 equivalent) in methanol and using 10% 

Pd/C as catalyst in reflux condition for 20 min. In both conditions, 

the top of the condenser was cover with ballon. In both catalytic 

hydrogenation methods, the reaction mixtures were performed by 

immersion the reaction flask into a pre-heated oil bath at 100 o C and 

the mixture was allowed to reflux without stirring for 20-30 min. 

For THC synthesis, we obtained THC as major product in 65% yield 

and hexahydrocurcumin as minor product (30%). Curcumin normally 

stay in the equilibrium of keto-enol forms. In keto-form, two double

328 

bonds were reduced to obtain THC while in enol-form all three double 

bond were reduced to obtaine hexahydrocurcumin. For the catalytic 

hydrogenation of (3E)-4-(4-hydroxy-3-methoxy phenyl)but-3-en-2one 

with ammonium formate andPd/C, zingerone was obtained in 

90.56% yield. The reaction is very clean and high yield. The catalytic 

hydrogenation method with ammonium formate as hydrogen gas 

source and Pd/C as catalyst in this study is a simple and effective 

method for reducing double bond. It was applied to synthesized 

valuable compounds such as THC and zingerone. This method can 

be used as an alternative hydrogenation method without using high 

pressure apparatus or complicated glassware. It can be used to reduce 

double bond in natural product synthesis or structural modification. 

(The 26 th Annual Research Conference in Pharmaceutical Sciences, 

2009, Bangkok, Thailand, December 4, 2009.) 

EFFECTS OF CHANGES IN PATIENTS’ NUTRI- 

TIONAL STATUS UPON COMPLICATIONS, 

CLINICAL OUTCOMES, LENGTH OF HOSPITAL 

STAY AND HOSPITAL EXPENSE (NO. 0886) 

Busba Chindavijak 1 , Surat Komindr 2 , Woraluck Ritsri 1 , Preecha 

Montakantikul 1 

1 Faculty of Pharmacy, Mahidol University, Bangkok, Thailand; 

2 Faculty of Medicine Ramathibodi Hospital, Mahdiol University, 

Bangkok, Thailand. E-mail : pybcd@mahidol.ac.th 

Key words : subjective global assessment, nutritional status, 

malnourished patients 

Purpose : Complications, clinical outcomes, length of 

hospital stay (LOS), and hospital expense, in relation to changes in 

patients’ nutritional stuatus were investigated. Procedures. Nutritional 

status of each patient was studied with the use of subjective global 

assessment at the time of admission, every 7 days during 

hospitalization, and at discharge. Episode of complications, clinical 

outcomes, LOS, and hospital expense, which did not include health 

accessories and salary, were recorded. Findings : The total of 145 

patients were assessed for their nutritional status Declined nutritional 

status assessed at discharge comparing to admission was found in 44 

patients (30.3%), maintained nutritional status was found in 100 

patients (69.0%), while 1 (0.7%) went home with improved nutritional 

status. Complications occurred in 22 patients with declined 

nutritional status (50.0%) and in 20 patients with maintained 

nutritional status (20.0%). Eight patients with declined nutritional 

status died (18.2%) while there was 1 dead patient in the group with 

maintained nutritional status (1.0%). LOS and hospital expense of 

patients with declined nutritional status were longer and higher than 

those who had maintained nutritional status. Conclusion : The 

declining nutritional status of patients during their hospitalization 

was associated with higher incidence of complications and death, 

longer length of hospital stay and higher hospital expense 

(Thai Journal of Clinical Nutrition 2009; 3(2):81-5.) 

QUALITY OF COLD STORAGE DRUGS TRAN- 

SPORTATION AND DELIVERY TO THAI 

HOSPITALS (NO. 0887) 

Cha-oncin Sooksriwong 1 , Woranut Bussaparoek 2 

1 Social and Administrative Pharmacy Research Unit, Faculty of 

Pharmacy, Mahidol University, Thailand; 2 Wangnoi Hosptial, 

Phra Nakhon Si Ayutthya, Thailand. 

Key words : cold storage drugs, logistics management, drug delivery, 

drug transportation, problem 

In Thailand there is no regulation and standard guideline 

to control the quality of drug transportation, including cold storage 

drugs. Objective : To explore the condition of cold storage drugs 

transportation and delivery to general public hospitals, community 

hospitals and private hospitals. Methods : This is a cross-sectional 

survey research. There were 301 questionnaires returned from 720 

questinnaries sent (41.8% response rate) Results : The serious 

problems of cold storage drugs on delivery were 1) Cold storage drugs 

(2C -8C) were delivered to the hospitals without controlled 

temperature boxes, was found in private, public and community 

hospitals at the rate of 46.7%, 48.3% and 72.9% respectively. 2) 

Cold storage drugs (-20C,) i.e. polio vaccine came to the hospitals 

with a temperature higher than 8C or ice melting in the box, was 

found in private, public and community hospitals at the rate of 22.9%, 

12.7% and 35.0% respectively. Conclusion : There are differences 

in quality of cold storage drug transportation and delivery among 

types of hospitals. It is highly recommended that Thai FDA should 

develop a standard and implement the control system for logistics 

management of cold storage drugs. 

(J Med Assoc Thai, 2009; 92(12):1685-5.) 


MEDICINE PRICES IN THAILAND : A RESULT OF 

NO MEDICINE PRICING POLICY (NO. 0888) 

Cha-oncin Sooksriwong 1 , Worasuda Yoongthong 2 , Siriwat 

Suwattanapreeda 3 , Farsai Chanjarupong 4 

1,3 Social and Administrative Pharmacy Excellence Research Unit, 

Faculty of Pharmacy, Mahidol University, Thailand; 2.4 Drug 

Control Division, the Office of Food and Drug Administration, 

Ministry of Public Health, Thailand. 

Key words : Medicine pricing, medicine price policy, Thailand 

Objectives : The main goal of this study was to document 

the situation of medicine prices in public and private health sectors 

for policy recommendation. Methods : A field study of measure prices 

of selected medicines was undertaken in Thailand using a 

standardized methodology developed by the World Health 

Organization (WHO) and Health Action International (HAI). Prices 

of 43 medicines were measured in health facilities and pharmacies 

in the capital city and three districts in different parts of Thailand. 

Medicine prices were expressed as the ratios relative to a standard 

set of international reference prices (median price ratio or MPR). 

Results : The public sector procured generics and innovator brands 

at 1.46 and 3.3 MPR while patients paid 2.55 and 4.36 MPR, 

respectively. Private pharmacies procured lowest price generics at


1.48 MPR and innovator brands at 9.67 MPR. Because of no 

medicine pricing policy in Thailand, it was found that between public 

and private sectors, among different public hospitals, and among 

different private pharmacies, the same generic products were procured 

and sold to patients at different prices. The median mark-up for 

innovator brands were 31% in the public sector and 22% in the private 

sector. For lowest priced generics, the median mark-up were 80% in 

the public sector and 96% in the private sector. Different prices for 

the identical product were problems to the health insurance 

organizations in terms of reimbursement, and to patients in term of 

fairness. Conclusion : The results highlight priority areas for action 

by the Ministry of Public Health and others in improving the drug 

pricing systems. The price regulation system should be implemented 

at every level of drug supply chain and appropriate pricing strategies 

should be employed. 

(Souther Med Review, 2009; 2(2):10-4.) 

PHARMACIST PARTICIPATION IN THE REDUC- 

TION OF PREVENTABLE ADVERSE DRUG 

REACTIONS IN THAI HOSPITALISED PATIENTS 

(NO. 0889) 

Pramote Tragulpiankit 1,2 , Sming Kaojarern 3 , Chalermsri 

Pummangura 4 , Winai Wananukul 3 , and David Luscombe 2 

1 Faculty of Pharmacy, Mahidol University, Bangkok, Thailand, 

2 Centre for Patient Safety Research, Welsh School of Pharmacy, 

Cardiff University, Cardiff, United Kingdom, 3 Faculty of 

Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, 

Thailand; 4 Faculty of Pharmacy, Siam University, Bangkok, 

Thailand: E-mail : pyptg@mahidol.ac.th 

Key words : Adverse drug reaction, Clincial pharmacist, Thailand 

Objectives : Assessing whether a research clinical 

pharmacist’s intervention could reduce or prevent adverse drug 

reactions (ADRs) from occurring at a large teaching hospital. Method 

: The study was of a non-randomized prospective design carried out 

in two general medical wards during a 10 month pre-intervention 

(control) period and then 10 month intervention (after) period when 

a clinical pharmacist was recruited to the medical care team. The 

probability and preventability of ADRs occurring during both periods 

were assessed using the Roussel Uclaf Causality Assessment Method 

(RUCAM) and Schumock and Thornton criteria, respectively. The 

rate of preventable ADRs occurring in hospitalized patients which 

were determined before and after periods of study was a primary 

outcome measure. The acceptance of a clinical pharmacist’s 

interventions for preventing an ADR was a secondary measure. 

Results : A total of 1,548 patients (765 male, 783 female) were 

monitored for ADRs during the control period with 985 (514 male, 

471 female) patients being studied in the intervention period. It was 

found that the rate of preventable ADRs was 5.20 per 1,000 patientdays 

(95% confidence interval, 5.60-4.80) during the control period 

compared with 1.72 patient-days (95% confidence interval, 2.24- 

1.20) during the intervention period. Thus, the pharmacist’s 

interventions resulted in a reduction in the number of preventable 

ADRs by 66.9% (p < 0.001). In total, the clinical pharmacist made 

143 recommendations to the medical care team resulting in avoidance 

of an ADR. These recommendations included dosage modification 

(48.3%), prevention of a possible drug-drug interaction (18.9%), 

inappropriate medication for the disease under treatment (13.9%), 

requests for therapeutic drug monitoring (13.3%) and the 

indentification of a history-related drug-induced allergy (5.6%). Eight 

percent of all the pharmacist’s recommendations were accepted by 

the physician and implemented by the medical care team. Conclusion 

: The data from the present study demonstrate the value of providng 

a clinical pharmacy service in reducing the number of drug-induced 

ADRs in hospitalized patient resulting in improved patient safety to 

the benefit of both patients and the health service. 

(Journal of Applied Therapeutic Research 2009; 7(1):3-10.) 

AN EFFCT OF ONE-DAY TRAINING COURSES 

ON THE IMPROVEMENT OF ADR REPORT’S IN 

THAI HOSPITALS (NO. 0890) 

Tragulpiankit P. 

The Assocication of Hospital Pharmacy, Faculty of Pharmacy, 

Mahidol University, Bangkok, Thailand. 

Key words : adverse drug reaction reporting, training course, quality 

Background : In Thailand although the number of 

individual case safety (ICSRs) or national spontaneous ADR report 

has been increased, the quality of their ADR report are still challenge 

and should be improved. Adverse drug reaction’s Community of 

Pharmacy Practice (AdCoPT) under the auspices of the Association 

of Hospital Pharmacy (Thailand) provided one-day training courses 

for government hospital pharmacists in order to improve quality of 

ADR report. Objectives : To investigate the effect of one-day training 

courses on the quality of ADR report. Methods : The one-day training 

courses which were conducted for four times at four distinct 

geographical regions of Thailand: the North, the North-East, the 

Central region (in Bangkok), and the South. The course included 

WHO criteria of ADR report’s quality and how to correctly report of 

ADR. The quality of ADR report was compared between before and 

after pharmacist’s training group. The reports were stratified and 

randomized according to the geographical regions based on place of 

training courses. The quality of report was classified by WHO criteria 

which were divided into the zero, 1 st , 2 nd and 3 rd level. The high rank 

level is likely high quality. Results : Thirty-two hospitals and six 

hundred and seventy-seven reports were included in the study. The 

number of reports by before training group was 378 reports while 

after training group was 299 reports. The number of the reports in 

the before training group which were classified as the 1 st level, 2 nd 

level, and 3 rd level accounted by 22.8%, 73.5%, and 2.9%, 

respectively. In the after training group, the number of the reports 

which were classified as the 1 st level, 2 nd level, and 3 rd were 19.4%, 

74.6%, and 5.0%, respectively. The reports which were classified as 

greater than the 1 st level in the after training group (79.6%) were 

more than in the before training group (76.4%). The difference was 

statistical significant difference (p < 0.001; McNemar’s test). 

Conclusions : The one-day training courses were a process of some 

effect for the improvement of ADR report’s quality. 

(The 4 th Asian Conference on Pharmacoepidemiology; 2009 October 

23-25; Tainan, Taiwan.)

330 

IMPACT OF PHARMACITS’S INTERVENTIONS 

ON ADVERSE DRUG EVENTS REDUCTION IN 

OUTPATIENTS WITH RHEUMATOID ARTHRITIS 

(NO. 0891) 

P. Tragulpiankit 1 , S. Chulavatnatol 1 , T. Limsuwan 2 , S. 

Janwityanujit 2 , S. Somjarit 1 , U. Sirikhedgon 1 

1 Department of Pharmacy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand; 2 Department of Medicine, Faculty 

of Medicine, Ramathibodi Hospital, Bangkok, Thailand. E-mail 

: pyptg@mahidol.ac.th 

Key words : Adverse drug events, rheumatoid arthritis, pharmacist 

0Instroduction : Adverse drug events (ADEs) are common 

in rheumatoid arthritis (RA) patients. Many studies have proved 

that ADEs were reduced in hospitalized patients by pharmcists but 

relatively few studies have been performed in outpatients. Aim : To 

study whether pharmacist’s interventions could reduce ADEs in 

outpatient with RA at rheumatology clinic. Ramathibodi Hospital, 

Thailand. Methods : Prospective controlled study was performed 

during April 30 th and August 30 th 2007. ADEs were detected before 

(the 1 st visit) and after intervention period (the 2 nd visit) in study 

group while no interventions were provided in control group. The 

pharmacist’s interventions consisted of detection and management 

of prescribing error and providing pateint’s counseling. All causality 

assessment and preventability of ADE were validated by 

rheumatologists. Rates of preventable ADE were the primary 

outcome. The differences in the rates at the 2 nd visit and the 1 st visit 

were compared between the study and the control group. P value < 

0.05 was defined as statistical significance using Chi-squared test. 

Results : The number of patients, ADEs, preventable ADEs, and 

preventable ADE rates in both study and control groups at the 1 st and 

2 nd visit are presented in Table I. The difference in the rates of the 

study and control group at the 2 nd visit was 10.7% (0.05 

0.10) and 10.0% (p > 0.10), respectively. 

Table I. Preventable adverse drug event (ADE) rates. 

Study group Control group 

1 st Visit 2 nd Visit 1 st Visit 2 nd Visit 

Number of patients 72 72 70 70 

All ADEs, events 22 25 33 38 

Preventable ADEs, events 12 16 16 23 

Preventable ADE rates (%) 16.7 22.2* 22.9 32.9* 

*0.05


ANTI-NECROSIS POTENTIAL OF POLYPHENOLS 

AGAINST SNAKE VENOMS (NO. 0893) 

Jiraporn Leanpolchareanchai 1 , Pimolpan Pithayanukul 1 , 

Rapepeol Bavovada 2 


University, Bangkok, Thailand. E-mail : pypph@mahidol.ac.th; 

2 Department of Pharmaceutical Botany, Faculty of 

Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 

Thailand. 

Key words : Anti-snake venom; necrosis, plant polyphenols 

Polyphenols from the extracs of Areca Catechu L. and 

Quercus infectoria Oliv. Inhibited phospholipase A(2), proteases, 

hyaluroniase and L-amino acid oxidase of Naja naja kaouthia Lesson 

(NK) and Calloselasma rhodostoma Kuhl (CR) venoms by in vitro 

tests. Both extracts inhibited the hemorrhagic activity of CR venom 

and the dermonecrotic activity of NK venom by in vivo tests. The 

inhibitory activity of plant polyphenols against local tissue necrosis 

induced by snake venoms may be caused by inhibition of 

inflammatory reactions, hemorrhage, and necrosis. The results 

implies the therapeutic potential of plant polyphenols against necrosis 

in snakebite victims. 

(Immunopharmacol Immunotoxicol 2009; 31(4): 556-62.) 

ANTIOXIDANT AND HEPATOPROTECTIVE 

ACTIVITIES OF THAI MANGO SEED KERNEL 

EXTRACT (NO. 0894) 

Saruth Nithitanakool 1 , Pimolpan Pithayanukul 1 , Rapepol 

Bavovada 2 

1 Department of Pharmacy, Faculty of Pharmacy, Mahdiol, 

Bangok, Thailand. E-mail : pypph@mahidol.ac.th; 2 Department 

of Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, 

Chulalongkorn University, Bangkok, Thailand. 

Key words : Mangifera indica L., Anacardiaceae, antioxidant 

Three polyphenolic principles, 1,2,3,4,6-penta- O-galloylbeta-D-glucopyranose 

(PGG), methyl gallate (MG), and gallic acid 

(GA), were isolated from the ethanolic extract of seed kernels of Thai 

mango (MSKE) (MANGIFERA INDICA L.cv “Fahlun”) and 

quantified using a TLC scanning densitometric method. The MSKE 

and its isolates were investigated by studying their antioxidant 

capacities using four different methods, by determining their IN 

VITRO anti-inflammatory activities, and by evaluating their 

hepatoprotective potential against liver injury in rats induced by 

carbon tetrachloride (CCI (4)). The hepatoprotective effect of MSKE 

is clearly supported by its polyphenolic nature of the main principle, 

PGG, which exhibited potent antioxidant and anti-inflammatory 

activities 

(Planta Med, 2009; 75(10):1118-23) 

HEPATOPROTECTIVE POTENTIAL OF EXTRACTS 

FROM SEEDS OF ARECA CATECHU AND 

NUTGALLS OF QUERCUS INFECTORIA. (NO. 0895) 

Pimolpan Pithayanukul 1 , Saruth Nithitanakool 1 , Rapepol 

Bavovada 1 




Pharmaceutical Sciences, Chulalongkorn Univesity, Bangkok, 

Thaialnd. 

Key words : Anti-inflammatory, antioxidant, hepatoprotective 

activity 

Aqueous extracts from seeds of Areca catechu L. 

(Arecaceae) (AC) and nutgalls of Quercus infectoria Oliv. (Fagaceae) 

(QI) were investigated for their hepatoprotectivge potential by 

studying their antioxidant capacity using four different methods, by 

determining their in vitro anti-inflammatory activity against 5lipoxygenase, 

and by evaluating their hepatoprotective potential 

against liver injury induced by carbon tetrachloride (CCI(4)) in rats. 

AC and QI extracts exhibited potent antioxidant and antiinflammatory 

activities. Treatment of rats with AC and QI extracts 

reversed oxidative damage in hepatic tissues induced by CCI(4). It 

is suggested that extracts rich in either condensed or hydrolysable 

tannins and known for their potent antioxidant and anti-inflammatory 

activites, may potentially confer protection against oxidative stressinduced 

liver injury. These data should contribute to evidence-based 

traditional medicines for anti-inflammatory and hepatoprotective 

effects of both extracts. 

(Molecules, 2009; 14(12):4987-5000) 

INHIBITORY EFFECT OF TEA POLYPHENOLS 

ON LOCAL TISSUE DAMAGE INDUCED BY 

SNAKE VENOMS (NO. 0896) 

Pimolpan Pithayanukul 1 , Jiraporn Leanpolchareanchai 1 , 

Rapepol Bavovada 2 





Thailand. 

Key words : Anti-snake venoms, Camellia sinensis L., plant 

polyphenols 

The methanolic extract of fresh tea leaves of Camellia 

sinesis L. (Theaceae) (CS) was assayed for its potential to inhibit 

enzymes with hydroltytic activity in Naja naja kaouthia Lesson 

(Elapidae) and Calloselasma rhodostoma Kuhl (Viperidae) venoms. 

These snake venom enzymes are responsible for the early effects of 

envenomation, such as local tissue damage and inflammation. The 

CS extract inhibited phospholipase A(2), proteases, hyaluronidase 

and 1-amino acid oxidase in both venoms by in vitro neutralization 

and inhibited the hemorrhagic and the dermonecrotic activities of 

the venoms in vivo. It is suggested that the inhibitory potential of

332 

the CS extract against local tissue damage induced by snake venoms 

may be attributed to complexation and chelation between the venom 

proteins and the phenolic contents of the extract. 

(Phytother Res, 2009; 24(S1)):S56-S62) 

MOLECULAR DOCKIGN STUDIES AN ANTI- 

ENZYMATIC ACTIVITIES OF THAI MANGO 

SEED KERNEL EXTRACT AGAINST SNAKE 

VENOMS (NO. 0897) 

Jiraporn Leanpolchareanchai 1 , Pimolpan Pithayanukul 1 , 

Rapepol Bavovada 2 , Patchreenart Saparpakorn 3 

1 Department of Pharamcy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand. E-mail : ry_110@yahoo.com; 



Thailand; 3 Department of Chemistry, Faculty of Science, 

Kasetsart University, Bangkok, Thailand. 

Key words : Calloselasma rhodostoma, Mangrifera indica L., 

Molecular docking study 

The ethanolic extract from seed kernels of Thai mango 

(MSKE) Mangifera indica L. cv. ‘Fahlun’ (Anacardiaceae) and its 

major phenolic principle (pentagalloyl glucopyranose) exhibited dosedependent 

inhibitory effects on enzymatic activities of phospholipase 

A(2) (PLA(2)), hyaluronidase and L-amino acid oxidase (LAAO) of 

Calloselasma rhodostoma (CR) and Naja naja kaouthia (NK) venoms 

by in vitro tests. The anti-hemorrhagic and anti-dermonecrotic 

activities of MSKE against both venoms were clearly supported by 

in vivo tests. Molecular docking studies indicated that the phenolic 

molecules of the MSKE could selectively bind to the active sites or 

their proximity, or modify conserved residues that are critical for the 

catalysis of PLA(2), and selectively bind to the LAAO binding pocket 

of both CR and NK venoms and thereby inhibit their enzymatic 

activities. The results imply a potential use of MSKE against snake 

venoms. 

(Molecules, 2009; 14(4):1404-22) 

MOLECTULAR DOCKING STUDIES AND ANTI- 

SNAKE VENOM METALLOPROTEINASE 

ACTIVITY OF THAI MANGO SEED KERNEL 


Pimolpan Pithayanukul 1 , Jiraporn Leanpolchareanchai 1 , 

Patchreenart Saparpakorn 2 



pypph@mahidol.ac.th; 2 Department of Chemistry, Faculty of 

Sciences, Kasetsart University, Bangkok, Thailand. 

Key words : anti-snake venom metalloproteinase activity; 

Calloselasma rhodostoma; Mangrifera indica L., Molecular docking 

study 

Snakebite envenomations cause severe local tissue 

necrosis and the venom metalloproteinases are thought to be the key 

toxins involved. In this study, the ethanolic extract from seed kernels 

of Thai mango (Mangifera indica L. cv. ‘Fahlun”) (Anacardiaceae) 

and its major phenolic principle (pentagalloyglucopyranose) exhibited 

potent and dose-dependent inhibitory effects on the caseinolytic and 

fibrinogenolytic activities of Malayan pit viper and Thai cobra venoms 

in invitro tests. Molecular docking studies revealed that the binding 

orientations of the phenolic principles were in the the binding pockets 

of snake venom metalloproteinases (SVMPs). The phenolic principles 

could form hydrogen bonds with the three histidine residues in the 

conserved zinc-binding motif and could chelate the Zn(2+) atom of 

the SVMPs, which could potentially result in inhibition of the venom 

enzymatic activities and thereby inhibit tissue necrosis. 

(Molecules, 2009; 14(9): 3198-213.) 

MOLECULAR DOCKING STUDIES AND ANTI- 

TYROSINASE ACTIVITY OF THAI MANGO 

SEED KERNEL EXTRACT (NO. 0899) 

Saruth Nithitanakool 1 , Pimolpan Pithayanukul 1 , Rapepol 

Bavovada 2 , Patchreenart Saparpakorn 3 



saruth_pipe@yahoo.com; 2 Department of Pharmaceutical 

Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn 

University, Bangkok, Thailand; 3 Department of Chemistry, 

Faculty of Sciences, Kasesart University, Bangkok, Thailand. 

Key words : Mangrifera indica L., Molecular docking study; 

polyphenols, Tyrosinase inhibitor 

The alcoholic extract from seed kernels of Thai mango 

(Mangifera indica L. cv. ‘Fahlun’) (Anacardiaceae) and its major 

phenolic principle (pentagalloyglucopyranose) exhibited potent, dosedepent 

inhibitory effects on tyrosine with respect to L-DOPA. 

Molecular docking studies revealed that the binding orientations of 

the phenolic principles were in the tyrosinase binding pocket and 

their orientations were located in the hydrophobic binding pocket 

surrounding the binuclear copper active site. The results indicated a 

possible mechanism for their anti-tyrosinase activity which may 

involve an ability to chelate the copper atoms which are required for 

the catalytic activity of tyrosinase. 

(Molecules, 2009; 14(1): 257-65) 


EFFECT OF TRIPOLYPHOSPHATE ON PHYSICAL 

AN ENZYMATIC STABILITIES OF INSULIN 

LOADED NANOPARTICLES OF N-TRIMETHYL 

CHITOSAN (NO. 0900) 

Anchalee Jintapattanakit 1,2 , Thomas Kissel 2 and Varaporn 

Buraphacheep Junyaprasert 1 


University, Bangkok 10400, Thailand. E-mail : pyvbp@mahidol. 

ac.th; 2 Department of Pharmaceuticds and Biopharmacy, 

Philipps-Universitat Ketzerbach 63, D-35032 Marburg, 

Germany.


Key words : N-trimethyl chitosan, nanoparticles, insulin, 

tripolyphosphate, trypsin, stabil 

The aim of the present work was to elucidate the influence 

of tripolyphosphate (TPP) on the colloidal and insulin stabilities. 

Using a trimethyl chitosan (TMC) with quaternization degree of 40% 

insulin loaded nanoparticles were prepared by ionotropic gelation 

with TPP crosslinker. The nanoparticles were characterized for size, 

zeta potential, insulin loading, process yield, colloidal stability and 

the protection capabilities of insulin against enzymatic degradation 

of trypsin. The result showed that insulin nanoparticles were in the 

range of 200- 260 nm with spherical or oval morphology. The highest 

insulin loading efficiency of nanoparticles with narrow size 

distribution was achieved when using TPP:TMC:insulin mass ration 

of 0.4:1:1. The colloidal stability was TPP concentration dependent. 

The presence of TPP accelerated the degradation of free insulin and 

insulin loaded nanoparticle which increased with increasing TPP 

concentration. Therefore, the preparation of nanoparticles by 

ionotropic gelation with TPP would not be suitable for the 

development of insulin deliver system. 

(MU J Pharm Sci, 2008;35(1-4):1-7. (Published in 2009) 

BIOADHESION AND ORAL ABSORPTION OF 

ENOXAPARIN NANOCOMPLEXES (NO. 0901) 

Wei Sun 1 , Shirui Mao 1 , Yanjun Wang 1 , Varaporn B. 

Junyaprasert 2 , Tingting Zhang, Lidong Na 2 , and Juan Wang 1 

1 School of Pharmacy, Shenyang Pharmaceutical University, 103 

Wenhua Road, Shenyang 110016, China, E-mail : 

shiruimao156@hotmail.com; 2 Department of Pharmacy, Faculty 

of Pharmacy, Mahidol University, Bangkok 10400, Thailand. 

Key words : Chitosan, Thiolated chitosan, Trimethyl chitosan; 

PEGylation, Enoxaparin, Polyelectrolyte complex, Bioadhesion, Oral 

absorption 

Polyelectrolyte complexs (PEC) formed between chitosan 

derivaties and enoxaparin were prepared by a self-assembly process 

and were characterized in terms of particle size and surface charge. 

The morphology was observed by atomic force microscopy (AFM). 

The colloidal stability and bioadhesion of the PEC were characterized 

by dynamic light scattering (DLS). The absorption of enoxaparin in 

rats was evaluated by activated partial thromboplastin time (APTT) 

assay. It was shown that the prepared PEC had a spherical shape 

with positive charge and a mean diameter in the range of 200-600 

nm. An increase in temperature led to a decrease in particle size (ca. 

10%) with an increased kcps value (ca. 10-20%) for the PEC studied, 

depending on the polymer structure. Thiolation and methylation of 

chitosan could significantly improve the corresponding PEC’s 

bioadhesion and hence the oral absorption of enoxaparin. A good 

relationship between bioadhesion and in vivo absorption was 

established. However, PEC of PEGylated chitosan did not display a 

significantly enhanced permeation of enoxaparin compared with 

unmodified chitosan. In conclusion, the oral bioavailability of 

enoxaparin can be enhanced by improving the bioadhesive properities 

of PEC via the chemical modification of chitosan employed. 

(Int J Pharm 2009; (Article in Press) 

EFFECT OF LIPID TYPES ON PHYSICOCHE- 

MICAL CHARACTERISTICS STABILITY 

AND ANTIOXIDANT ACTIVITY OF GAMMA- 

ORYZANOL-LOADED LIPID NANOPARTICLES 

(NO. 0902) 

Uracha Ruktanonchai 1 , Usawadee Sakulkhu 1 , Piyawan 

Bejrapha 1 , Praneet Opanasopit 2 , Nuntavan Bunyapraphatsara 3 , 

Varaporn Junyaprasert 3 , Satit Puttipipatkhachorn 3 

1 National Nanotechnology Center, National Science and 

Technology Development Agency, Thailand Science Park, 

Thailand; 2 Faculty of Pharmacy, Silpakorn University, Nakhon 

Pathom, Thailand; 3 Faculty of Pharmacy, Mahidol University, 

Bangkok, Thailand. 

Key words : Antioxidation activity, Gamma-oryzanol, Nanoemulsion, 

Solid lipid nanoparticle, Stability 

In the present study gamma-oryzanol, an antioxidant, was 

incorporated into three different types of solid lipid, wax, triglycerides, 

a mixture of glycerides as solid lipid nanoparticles (SLN) and liquid 

lipid (Miglyol 812) as nanoemulsion (NE). Instability was found 

only from NE due to its significant increase in particle size and 

decreased entrapment efficiency (EE) at a storage temperature of 

45 o C. Solid lipid type in SLN plays an important role only on EE, 

but not chemical stability. A decrease in crystallinity of SLN was 

observed with the incorporation of gamma-oryzanol and low 

recrystallization index were found with two glycerides-based SLN. 

The in vitro release studies demonstrated that a biphasic release 

pattern fitted well with the Higuchi model of SLN formulations. In 

comparison, nearly constant release was observed in NE comprised 

of similar compositon. Wax-based SLN demonstrated the lowest 

cytotoxicity. NE, was-based SLN and a mixture of glycerides-based 

SLN were considered to enhance the antioxidant activity of gammaoryzanol. 

(J Microencapsul, 2009; 26(7):614-626.) 

MICROEMULSIONS AND NANOEMULSIONS : 

NOVEL VEHICLES FOR WHITENING COSMECEU- 

TICALS (NO. 0903) 

Prapaporn Boonme 1 , Varaporn B. Junyaprasert 2 , N. Suksawad 1 , 

S. Songkro 1 

1 Department of Pharmaceutical Technology, Faculty of 

Pharmaceutical Sciences, Prince of Songkla University, Songkhla 

90112, Thailand; 2 Department of Pharmacy, Faculty of 

Pharmacy, Mahidol University, Bangkok 10400, Thailand. 

Key words : Cosmeceuticals, Micromulsions, Nanoemulsions, Skin 

penetration 

For Asian women, white skin is preferable. During the 

last decade, skin whitening products appear to be the largest and 

continually growing segment in skin-care market in Asia and have 

an impaction of economic worth. Skin whitening or lightening agents 

are as cosmetics which act as a drug-like benefit since melanin 

producing process of the skin is disturbed and can be classified as 

cosmeceuticals. To increase efficiency, novel vehicles are necessary

334 

for skin penetration enhancement of these agents. Microemulsions 

and nanoemulsions are one of useful nanocarriers for skin application 

in view of achieving efficiency of the active substances. Moreover, 

they can be formulated with ease for active ingredient incorporation, 

high stability and good appearance. In this review article, applications 

of well-known whitening or lightening agents were summarized. In 

addition, the use of microemulsions and nanoemulsions as novel 

vehicles for whitening products were discussed. 

(J Biomed Nanotech, 2009; 5(4):373-383.) 

PHYSICOCHEMICAL CHARACTERISTICS, CYTO- 

TOXICITY, AND ANTIOXIDANT ACTIVITY 

OF THREE LIPID NANOPARTICULATE FORMULA- 

TIONS OF ALPHA-LIPOIC ACID (NO. 0904) 

Uracha Ruktanonchai 1 , P. Bejrapha 1 , U. Sakulkhu 1 , P. 

Opanasopit 2 , Nuntavan Bunyapraphatsara 3,4,5 , Varaporn 

Junyaprasert 3,4,5 , Satit Puttipipatkhachorn 3,4,5 

1 National Nanotechnology Center, National Science and 

Technology Development Agency, Paholyothin Road, 

Pathumthani 12120, Thailand; 2 Faculty of Pharmacy, Silpakorn 

University, Nakohpathom, Thailand; 3 Department of 

Pharmacognosy, Faculty of Pharmacy, Mahidol University, 

Bangkok 10400, Thailand; 4 Department of Pharmacy, Faculty 

of Pharmacy, Mahidol University, Bangkok 10400, Thailand; 

5 Department of Manufacturing Pharmacy, Faculty of Pharmacy, 

Mahidol University, Bangkok 10400, Thailand. 

Key words : Alpha-lipoic acid; Antioxidatin activity, Nanoemulsion, 

Nanostructure lipid carriers, Solid lipid nanoparticles 

Exogenously suppled alpha-lipoic acid (LA) has proven 

to be effective as an antioxidant. In an effort to develop a watersoluble 

formulation for topic administration, LA was formulated in 

the form of solid lipid nanoparticles (SLN), nanostructure lipid 

carriers (NLC), and nanoemulsion (NE) and characterized in terms 

of physical and biological properties. Mean particle size of 113, 110, 

and 121 nm were obtained for NE, NLC, and SLN, repectively, with 

narrow size distribution. Zeta potential was approximately in the range 

of -25 to -40 mV. Disc and spherical structures of nanoparticles were 

observed by cryo-scanning electron microscopy. Entrapment 

efficiency of LA in three formulations was found to be more than 

70%. After 120 days of storage at 25 o C, physical stability of all 

formulations remained unchanged whereas the entrapment efficiency 

of SLN and NLC could be maintained, suggesting relative long-term 

stability. Prolonged relase of LA formulation following the Higuchi 

model was found where a faster release was observed from NE 

compared with that of SLN and NLC. More than 80% of cell survivals 

were found up to 1 μM of LA concentrations. Antioxidant activity 

analysis demonstrated that all LA-loaded formulations expressed 

antioxidant activity at a similar magnitude as pure LA. These results 

suggest that chosen compositions of lipid nanoparticles play an 

important role on drug loading, stability,and biological activity of 

nanoparticles. Both SLN and NLC demonstrated their potential as 

alternative carriers for aqueous topic administration of LA. 

(AAPS PharmSciTech 2009; 10(1):227-234.) 

Q10-LOADED NLC VERSUS NANOEMUL- 

SIONS. STABILITY RHEOLOGY AND IN VITRO 

SKIN PERMEATION (NO. 0905) 

Varaporn B. Junyaprasert 1 , Veerawat Teeranachaideekul 1,2 , 

Eliana B. Souto 3 , Prapaporn Boonme 4 , Rainer H. Muller 2 

1 Department of Pharmacy, Facult of Pharmacy, Mahidol 

University, Thailand. E-mail : pyvbp@mahidol.ac.th; 

2 Department of Pharmaceutic, Biopharmaceutic and Quality 

Management, Free University of Berlin, Bermany; 3 Department 

of Pharmaceutical Technology, Faculty of Health Sciences, 

Fernando Pessou University, Portugal; 4 Department of 

Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, 

Prince of Songkla University, Thailand. 

Key words : Nanostructured lipid carriers, NCL, Q 10 , Chemical 

stability 

In this study, nanoemulsions (NE) of medium chain 

triacylglycerols (MCT) and nanostructured lipid carriers (NLC) of 

cetyl palmiate/MCT were produced to load coenzyme Q(10) (Q10) 

and characterized for their stability before and after incorporation 

into xanthan gum hydrogels. After storage at 4, 25 and 40 degrees 

C, the particles remained in the nanosize range for 12 months, with 

zeta potential higher than 40 mV. Similar results were found in 

xanthan gum-based hydrogels containing NE or NLC. The 

crystallinity index of Q(10)-loaded NLC increased after being 

incorporated into hydrogels. The Q(10) entrapped in NLC and NE 

remained higher than 90% at all temperatures for 12 months but 

dramatically decreased when exposed to light. From the rheological 

studies, both NLC and NE dispersions possessed pseudoplastic flow 

having more liquid characteristics, whereas NLC and NE hydrogels 

exhibited plastic flow with thixothopy, showing more elastic rather 

than viscous properties. The occurrence of a spatial arrangement of 

lipid molecules was observed in the matrix of NLC when entrapped 

into hydrogels. From in vitro permeation studies, it could be stated 

that the amount of Q(10) released and occlusiveness were major keys 

to promote the deep penetratin of Q(10) into the skin. 

(Int J Pharm 2009; 377(1-2):207-14.) 


THE ROLE OF MUCOADHESION OF TRIMETHYL 

CHITOSAN AND PEGYLATED TRIMETHYL 

CHITOSAN NANOCOMPLEXES IN INSULIN 

UPTAKE (NO. 0906) 

Anchalee Jintapattanakit 1,2 , Varaporn Buraphacheep 

Junyaprasert 1 , Thomas Kissel 2 


University, Bangkok 10400, Thailand; 2 Department of 

Pharmaceutics and Biopharamacy, Philipps-Univesitaty, 

Germany. 

Key words : mucoadhesin, PEGylatin, trimethyl chitosan, insulin 

nanocomplexes 

The aim of this work was to investigate the role of 

mucoadhesion in the insulin uptake of nanocomplexes (NC) based 

of trimethyl chitosan (TMC) and poly(ethylene glycol) (PEG)-graft-


TMC copolymes. Self-assembled insulin NC were prepared by 

polyelectrolyte complexation. The effects of PEGylation and positive 

charge density on mucoadhesion were assessed using a mucin assay 

and mucus-secreting HT29-MTX-E12 (E12) monolayers. The 

behaviors of corresponding isulin NC after adhesion to E12 were 

also established. All PEGylated TMC copolymers showed 

significantly higher levels of adhesion to mucus than unmodified 

TMC. The copolymer composed of 298 PEG chains per TMC 

macromolecules exhibited the highest level of mucoadhesin, being 

3.4 times higher than TMC. The higher mucoadhesive properties of 

PEGylated TMC copolymers resulted from the synergistic effects of 

interpenetration of PEG chains into the mucus and electrostatic 

interaction between positive charged TMC and anionic glycoproteins 

present in the mucus layer. Compared to TMC, insulin NC based on 

PEGylated TMC copolymers demonstrated no evidence of insulin 

uptake improvement due to complete release of insulin from NC after 

adhering to mucus. CLSM revelaed the localization of TMC and its 

corresponding insulin NC at cell surface membranes of E12. 2009 

Wiley-Liss, Inc. and the American Pharmacists Association. 

(J Pharm Sci, 2009; 98(12): 4818-30) 

INVESTIGATION OF NIOSOMAL PREPARATION 

OF POORLY SOLUBLE PHYTOCHEMICAL 

ELLAGIC ACID (NO. 0907) 

Pratyawadee Singhsa, Dougdaw Chantasart, Varaponr B. 

Junyaprasert 

Department of Pharmacy, Faculty of Pharmacy, Mahidol 

University, Bangkok 10400, Thailand. E-mail : pyvbp@mahidol. 

ac.th 

Key words : niosomes, ellagic acid, reverse phase evaporation, 

solubilization 

Ellagic acid (EA) is a potent antioxidant phytochemical 

substance. Its pharmacological activity has been reported in wide 

variety of diseases. In contrary, the use of EA has limitation due to 

its poor biopharmaceutical properties, low aqueous solubility and 

low permeability. The aim of the present work was to develop EA 

loaded niosomes, a novel system for encapsulation poorly water 

soluble phytochemical EA which is sparing soluble or insoluble in 

the general solvents used for preparation. The EA niosomes were 

prepared by the reverse phase evaporation (REV) method usin 1:1 

mole ratio of surfactants and cholesterol as vesicle forming agents. 

The mole ratio of surfactants, Span 60 and Tween 60 was varied 

from 1:0, 2:1, 1:1 to 0:1, respectively. Cholesteryl poly-24oxyethylene 

ether (Solulan C24) at 5 mol% of total lipids was added 

as a membrane additive. In this study, methanol, polyethylene glycol 

400 and propylene glycol were used as a cosolvent to solubilize EA 

in niosomal preparations. During preparation, 20 and 30% v/v 

methanol was added into the organic phase containing EA, ortherwise 

10 and 15 % v/v polyethylene glycol 400 or propylene glycol was 

added into the aqueous phase containing EA. In addition, the amounts 

of EA at 1, 3 and 5 mol% of total lipid were loaded and investigated. 

The influence of the process variables on the physicochemical 

properties, including vesicle size, entrapment efficiency and stability 

of the obtained vesicles was studied. The results indicated that the 

EA niosomes prepared using Span 60 and Tween 60 at 2:1 mole ratio 

and 20% v/v methanol possessed the best physicochemical properties 

and stability and had the highest entrapment efficiency. 

(Proceeding in the 6 th Indochina Pharamcy Conference, December 

15-17, 2009, Hue, Vietnam.) 

SYNTHESIS OP PEG-POLYCAPROLACTONE 

GRAFTED WITH NICOTINIC ACID FOR DRUG 

DELIVERY (NO. 0908) 

Varaporn B. Junyaprasert 1 , Jiraphong Suksiriworapong 1 , 

Kittisak Sripha 2 

1 Department of Pharamcy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand. E-mail Pyvbp@mahidol.ac.th; 


Mahidol University, Bangkok, Thailand. 

Key words : polycaprolactone, PEG-polycaprolactone, biodegradable 

polymer, nicotinic acid, click chemistry 

Modification of biodegradable polymers has gained much 

attention to increase the efficacy of drug carriers. The aim of this 

study was to prepare polyethylene glycol and polycaprolactone (PEG- 

PCL) polymer to increase hydrophilicity of this polyester polymer, 

and to graft nicotinic acid onto the polymer by using mild conditions. 

The copolymer of α−chloro-ε-caprolactone (αClεCL) and εCL 

containing 30% αClεCL units was fabricated by ring opening 

polymerization initiated by PEG 4000 with tin (II) octanoate as a 

catalyst. Consequently, the chloride pendants of the copolymer were 

converted to azide by the reaction with sodium azide at the amount 

equivalent to 1.02 mole of chloride. Finally, the azide on the 

copolymer was replaced with nicotinic acid by addition of but-3ynyl 

nicotinate using Huisgen’s1,3-dipolar cycloaddition or click 

reaction. From H-NMR spectroscopy, it is revealed that the fraction 

of αClεCL was 27% and 13% with respect to εCL unit and the 

polymer chain, respectively. In addition, the chloride group was 

completely converted to azide, and the entire replacement of nicotic 

acid at the azide was observed. In conclusion, the PEG-PCL grafted 

with nicotinic acid was successfully prepared by click reaction wihc 

is a mild and short-time reaction without the degradation of the 

copolymer. 

COMPARISON OF IN VITRO DRUG RELEASE AND 

IN VITRO SKIN PERMEATION OF Q10-LOADED 

NLC (NO. 0909) 

Veerawat Teeranachaideekul 1 , Varaporn Junyaprasert 1 , 

Prapaporn Boonme 2 , Rainer Muller 3 and Eliana Souto 4 

1 Department to Pharmacy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand; 2 Department of Pharmaceutical 

Technology, Faculty of Pharmaceutical Sciences, Price of Songkla 

University, Thailand; 3 Department of Pharmaceutical 

Technology, Biotechnology & Quality Management, Free 

University of Berlin, Berlin, Germany; 4 Department of 

Pharmaceutical Technology, Faculty of Health Sciences, 

Fernando pessoa University, Portugal Porto, Portugal. 

Even thoug several advantages of solid lipid nanoparticles 

(SLN) such as protection nof the incorporated sensitive drug 

molecules due to the solid matrix around drug molecules have been 

shown, some drawbacks of SLN have been reported, for example, 

low drug payload, drug expulsion during storage, and unpredictability

336 

of gelation phenomenon. The concept of SLN with the less ordered 

structure of lipids matrix within solid particles so-called 

nanostructured lipid carriers (NLC), therefore, has been introduced 

around the beginning of the millennium. NLC can be produced by 

mixing various different lipid molecules, i.e. blending solid lipid with 

liquid lipid (oils). In a previous study, the localization of oil molecule 

was elucidated by means of 1H-NMR. The aim of this study was to 

evaluate the effect on oil content on the localization of oil molecule 

within the lipid nanoparticles. Cetyl palmitate (CP) and medium 

chain triglycerides (MCT) were chosen as solid liquid lipid (oil) of 

NLC, respectively. Q10-loaded NLC were prepared with different 

ratios of CP and MCT by high pressure homogenization (HPH) 

technique. The 1H-NMR of Q10-loaded NLC was compared to that 

of Q10-loaded nanoemulsions (NE). It was found that Q10-loaded 

NLC showed a broader signal between 0.9 ppm and 2.5 ppm in 

comparison to that of Q10-loaded NE depending on the MCT loading 

content. This indicates the MCT are strongly restricted to and/or 

incorporated into the solid matrix of CP, especially Q10-loaded NLC 

containing the low ratios of MCT and CP. 

(3 rd Asian Pacific Regional Meeting, May 10-12, 2009, The Imperial 

Queen’s Park Hotel.) 

DEVELOPMENT OF CAPSAICIN GEL (NO. 0910) 

Orapha Thanasutthikarn, Anyarat Chuenjitkulthaworn 

Department of Pharmacy, Faculty of Pharmacy, Mahdiol 

University. 

Key words : capsaicin gel capsaicin, topical gel, pain relief 

Capasaicin gel is the drug recipe in the second group (Drug 

recipe developed from herb) of the Essential Drug List (Herbal 

medicine) Capsaicin gel containing 0.025% capsaicin is topically 

used for pain relief of join, muscle and neuropathic pain from diabetes. 

The purpose of this special project was to develop the capsaicin gel 

having good physical properties and consisting of the proper amount 

of the active substance according to Essential Drug List. The 

experiment wereperformed to quantitatively analyze capsaicin in the 

chili extract by high pressure liquid chromatography (HPLC) method 

using N-vanillyl-n-nonamide as a reference standard and to study 

the solubility of capsaicin extract in different concentrations of alcohol 

and in different solvent systems of alcohol and cosolvent, such as 

propylene glycol, glycerin, PEG 4000 and PEG40-hydrogenated 

coster oil (PH). The results showed that the chilli extract in the solvent 

comprised of 22.5% alcohol, 7% glycerin, 17% PEG 4000 and 1% 

PHC showed a clear brow solution without precipitate. The obtained 

solution was used to prepare the capsaicin gel by using two kinds of 

gelling agents which were Carbopol 940 and hydroxyethylcellulose, 

Physical properties of the capsaicin gel such, viscosity and pH were 

evaluated and the results showed that the for mulation of Carbopol 

940 1% gave a clear gel with good physical properties. The 

quantitative analysis of the capsaicin gel by HPLC revealed that the 

contained 0.030% capsaicin. Therefore, the developed gel possed 

physical and chemical properties which should be further studied 

for its stability. 

(The 6 th Indochina Pharamcy Conference, December 15-17, 2009, 

Hue, Vietnam.) 


ANTITUBERCULOTIC DRUGS – CAN DISSO- 

LUTION GUIDE THE BIOEQUIVALENCE 

DECISION? (NO. 0911) 

Stefanie Strauch 1 , Ekarat Jantratid 1,2 , Matthias Stahl 3 , Sabine 

Kopp 3 , Lembit Rago 3 , Jennifer B. Dressman 1 

1 Institute of Pharmaceutical Technology, Johann Wolfgang 

Goethe University, Frankfurt am Main, Germany; 2 Department 

of Pharamcy, Faculty of Pharamcy, Mahidol University, Bangkok, 

Thailand; 3 World Health Organization, Geneva, Switzerland. 

Key words : biowaiver, antituberculotic drugs dissolution 

The aim of this work was to evaluate whether the 

biowaiver-based bioequivalence (BE) approval can be applied to 

antituberculotic products In vitro dissolution data for approved 

antituberculotics were generated according to biowaiver methods and 

the BE decision on this basis was compared to that based on in vivo 

pharmacokinetic (PK) data. The dissolution characteristics of a 

selection of drug products from the WHO Prequalificatin Programme 

including three ethambutol dihydrochloride(ETB), two isoniazid 

(INH) and one pyrazinamide (PYR) products were investigated 

according to the methods given in the WHO Guideline. Then, (a) 

the dissolution data of these products were compared with those 

obtained from the recommended comparator product, (b) PK data 

provided bythe manufacturers were compared to those of the 

recommended comparator product using BE criteria and (c) the results 

of (a) and (b) were compared and evaluated. Only one investigated 

ETB product was able to meet the WHO criteria for “very rapidly 

dissolving”. The other ETB test products (including the 

recommended comparator suggested by the WHO) failed to fulfill 

the WHO criteria for either “very rapidly dissolving” or “rapidly 

dissolving” products. All tested INH and PYR products (including 

their recommended/accepted comparators suggested by the WHO) 

met the WHO criteria for “very rapidly dissolving”. Evaluation of 

the in vivo PK data demonstrated that two ETB test produts are 

bioequivalent to the recommended comparator, but one is not. All 

INH and PYR test products demonstrated BE to their recommended 

comparators. 

(Poster presentation at the 2009 AAPS Annual Meeting and 

Exposition, November 8-12, 2009. Los Angeles Convention Center, 

Los Angeles, CA, USA.) 

BIORELEVANT DISSOLUTION MEDIA SIMU- 

LATING CANINE GASTROINTESITNAL MILIEU 

(NO. 0912) 

Ekarat Jantratid 1,2 , Marcel Arndt 2 , Hitesh Chokshi 3 , Kin Tang 3 , 

Neil J. Parrot 4 , Christos Reppas 5 , Jennifer B. Dressman 2 


Universtiy, Bangkok, Thailand; 2 Institute of Pharmceutical 

Technology, Johann Wolfgang Goethe University, Frankfurt am 

Main, Germany; 3 Hoffmann-la Roche Inc., Nutley, New Jersey, 

USA, Pharamceuticals Division, Pharma Research Non-Clinical 

Development, Non-Clinical Drug Safety, F. Hoffmann-la Roche 

Ltd., Basel, Switzerland; 4 Faculty of Pharmacy, National and 

Kapodistrain University Athens, Athen, Greece


Key words : canine biorelevant media, prandial states, preclinical 

This study was aimed to design canind biorelevant 

dissolution media with composition and characteristics reflecting the 

conditions generated in the proximal gastrointestinal tract of the dogs 

current standard preclinical study protocols. Two dissolution media 

simulating the fasted canine stomach were designed to cover low 

and high gastric pH conditions (FaSSGF c /2-pH 2.0 and FaSSGF c /6pH 

6.0). To mimic the fed stategastric conditions. FeSSGFc/3.5, an 

equal volume mixture of full-fat (3.5%) UHT milk and buffer pH 3.0 

was designed for the study protocol using high fat meal, and FeSSGF c / 

1.5, an equal volume mixture of low-fat (1.5%) UHT milk and buffer 

pH 3.0 for the study protocol using standard meal. For the fasted 

state upper small intestine, the previous version of human FaSSIF, 

pH 6.5 appears to be appropriate. FeSSIF c , pH 4.5 was designed as a 

medium representing the upper small intestinal conditions in a dog 

after meal intake. The characteristics of media were assessed upon 

preparation and under standard dissolution test as well as ambient 

storage conditions. All canine biorelevant media were designed to 

have physicochemical properties close to those of aspirates obtained 

fom dogs. Further, the media were shown to be stable under ambient 

conditions for 72 h and standard dissolution test conditions for at 

least 8 h. 


Exposition, November 8-12, 2009, Los Angeles Convention Center, 


IN VITRO DISSOLUTION PERFORMANCE OF A 

NEW HYPROMELLOSE CAPSULE – VCAPS* 

PLUS IN BIORELEVANT MEDIA (NO. 0913) 

Murat Kilic 1 , Ekarat Jantratid 1,2 , Dominique Cade 3 , Hassan 

Benameur 3 , Jennifer B. Dressman 1 



of Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok, 

Thailand; 3 Capsugel, a Division of Pfizer, Colmar, France. 

Key words : Vacps Plus, gelatin capsules, biorelevant dissolution 

The objective of this study was to compare the in vitro 

dissolution performance of a new hypromellose capsule, Vcaps Plus, 

with hard gelating capsules in various biorelevant media. Caffeine, 

a highly soluble marker compound, was used to enable maximum 

discrimination of capsule shell performance. The test conditions 

consisted of USP Apparatus 2, paddle speed of 75 rpm, and a media 

volume of 500 mL.Biorelevant media to simulate the gastric 

conditions, including (i) Fasted state Simulated Gastric Fluid 

(FaSSGF), (ii) Fed State Simulated Gastric Fluid (FeSSGF), and to 

simulate the upper small intestinal conditions,(iii) Fasted State 

Simulated Intestinal Fluid (FaSSIF), and the updated version (iv) 

FaSSIF-V2, (v) Fed State Simulated Intestinal Fluid (FeSSIF) and 

the updated version (vi) FeSSIF-V2, were applied. A lag time of about 

5 min was observed in release of caffeine from Vcaps Plus capsules. 

However, the dissolution performance of both capsule shells, Vcaps 

Plus and gelatin, was generally slower and more variable in 

biorelevant media than in the compendial media. During the tests in 

biorelevant media gelling of the Vcaps Plus shells (and also of the 

gelatin capsules to some extent) was observed, hindering the release 

of caffeine. The results suggest that complex components of the 

media, especially in the fed state e.g. bile salt/ lipids, tend to impede 

the dissolution performance of Vcaps Plus shells. Nevertheless, 

independent of the media used, the dissolution profiles generally 

became comparable to the of gelatin capsules within 30 min. The 

results in biorelevant media indicate that Vcaps Plus capsules would 

be unlikely to exhibit significant differences in terms of mean 

pharmacokinetic parameters compared with the gelatin capsules. 


Exposition, November 8-12, 2009. Los Angeles Convention Center, 


MODELLING OF PLASMA PROFILES FOR 

MICRONIZED AND NANOSIZED FENOFIBRATE 

(NO. 0914) 

Daniel Juenemann 1 , Ekarat Jantratid 1,2 , Jennifer B. Dressman 1 



of Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok, 

Thailand. 

Key words: fenofibrate, micronization, nanonization 

This study was conducted to predict human plasma profiles 

of micronized and nanosized fenofibrate using an in silico modeling 

approach coupled with biorelevant dissolution testing. Initial 

dissolution rates of nanosized fenofibrate (Lipidil 145 ONE , Solvay, 

Germany) were obtained in FaSSGF and FaSSIV-V2, media 

simulating the preprandial intestinal conditions. Syringe filters with 

nominal pore sizes of 0.1 μm(Anotop 25 plusTM, Whatman, 

England), and 0.2 μm (Minisart RC 25TM, Sartorius, Germany) were 

used. Dissolution results were used to generate plasma profiles using 

Stella 9.1 (Iseesytems, NH USA) software using two different 

hypotheses: 1. sink conditions prevail in the GI-tract, 2. absorption 

of fenofibrate can be permeability-limited. The simulated and in 

vivo plasma profiles in the fasted state were compared using ratios 

of C max , T max and AUC. Additionally, simulated plasma profiles of 

nanosized fenofibrate were compared to simulated plasma profiles 

of micronized fenofibrte (Lipidil Ter , Solvay, Germany). When the 

model which assumed sink conditins in the GI tract was applied to 

dissolution data generated with the 0.1 and 0.2 μm filters, inaccurate 

predictions of C max and T max in the fasted state (irrespective of filter 

pore size, ratios: C max -164, T max -0.34, AUC-0.93) were obtained. 

When the same dissolution data were coupled with the model 

assuming a permeability limitation to absorption, use of data 

generated with 0.2 μm filters continued to overestimate drug 

dissolution and lead to inaccurate plasma profile simulation (ratios: 

C max -1.19, T max -0.69, AUC-0.93). However, when data generated with 

0.1 μm filters were used, the simulated plasma profiles were very 

similar to the in vivo profiles (ratios :C max -0.94, T max -0.86, AUC-0.93). 

This model also predicted the in vivo plasma profiles for the 

micronized formulation accurately. 

(Poster presentation at the 2009 AAPS Annaul Meeting and 

Exposition, November 8-12, 2009, Los Angeles Convention Center, 

Los Angeles, CA, USA.)

338 

ECONOMIC EVALUATIONS OF HEPATITIS B 

VACCINATION FOR DEVELOPING COUNTRIES 

(NO. 0915) 

Hong-Anh T Tu, Heman J Woerdenbag, Sumit Kane, Arthorn 

Riewpaiboon, Marinus van Hulst, Maarten J Postma 

Unit of PharmacoEpidemiology & PharamcoEconomics (PE2), 

Department of Pharmacy, University of Groningen, Groningen, 

The Netherlands, E-mail : h.a.tu@rug.nl 

Key words : health-economci study, hepatistis B vaccine, developing 

country 

Economic evaluations, in particular cost-effectiveness, are 

important determinants for policy makers and stakeholders involved 

in decision-making for health interventions. Up until now, most 

evaluations of cost-effectiveness of hepatitis B vaccination have been 

performed in developed countries. Appropriate health-economci 

studies on this topic specifically targeted at the developing world are 

essential in order to justify adding another vaccine into the existing 

Expanded Program on Immunization in these countries. We present 

a systematic review of economic evaluations of vaccinatioin against 

HBV for developing and less-developed countries. Vaccine price, 

the discout rate, incidence and prevalence of HBV infection were 

found to be major drivers of cost-effectiveness. Data accuracy and 

reliability were also major issues, with major potentials for 

improvement in studies of these countries. The choice between 

monovalent or combination vaccines (diphtheria, tetanus and poliohepatitis 

B) poses new challenges to cost-effectiveness analysis. It 

is concluded that for many developing countries implementation of 

universal immunization against HBV to reduce the level of endemicity 

of hepatitis B is an appropriate strategy, and probably cost effective 

in many settings. Given their limited financial resources, developing 

countries should properly plan how to achieve this. Further countryspecific 

economic evaluations and related gathering of high-quality 

data must be conducted in developing countries in order to raise 

both public awareness of the effectiveness and economic 

attractiveness of universal immunization against HBV. 

(Expert Rev Vaccines, 2009; 8(7):907-20.) 

ECONOMIC VALUATION INFORMAL CARE IN 

ASIA : A CASE STUDY OF CARE FOR DISABLED 

STROKE SURVIVORS IN THAILAND (NO. 0916) 

Arthorn Riewpaiboon 1 , Wachara Riewpaiboon 2 , Kanyarat 

Ponsoongnern 1 , and Bernard Van den Berg 3,4 

1 Division of Social and Administrative Pharamcy, Faculty of 

Pharamcy, Mahidol University, Bangkok, Thailand. E-mail : 

pyarp@mahidol.ac.th; 2 Health Promotion Program for People 

with Disability, Sirindhorn National Medical Rehabilitation 

Center, Ministry of Public Health, Thailand; 3 Department of 

Health Sciences, VU University Amsterdam, The Netherlands; 

4 Department of Economics and Business and CRES, Universittat 

Pompeu Fabra, Barcelona, Spain. 

Key words : Informal care, Economic value, Cost of illness, Stroke, 

Disabled persons, Caregivers, Thaialnd. 

This study values informal care for disabled stroke survivors in 

Thailand. It applies the conventional recommended apportunity cost 

method to value informal care in monetary terms. Data were collected 

by means of face-to-face interviews conducted during 2006. The 

sample consisted of 101 disabled persons who had suffered a stroke 

at least six months prior to the interview, and who had a functional 

status score of less than 95 as measured by the Barthel Index. Average 

monthly time spent on informal care was 94.6 hours, and the major 

source of opportunity cost was forgone unpaid work (43.5%). The 

averge monthly monetary value of informal care was 4642.6 baht, 

based on 2006 prices. This study shows that providing informal care 

involves a substantial apportunity cost, implying a hidden value to 

Thai society. 

(Social Science & Medicine, 2009; 69(4):648-653.) 

HOW THE BLIND COPE WITH PROBLEMS 

OF MEDICINE UTILIZATION : A STUDY IN 

BANGKOK, THAILAND (NO. 0917) 

Arthorn Riewpaiboon 


Department of Pharmacy, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand. E-mail : pyarp@mahidol.ac.th 

Key words : Blind, Medicine utilization, Pharmaceutical services, 

Visually impaired persons 

Purpose : This project is aimed at studying medicine 

utilization among blind people in Bangkok, Thailand. Methods : 

The study was designed as a cross-sectional descriptive research. 

The study group comprised visually impaired persons, aged at least 

18 years, and living in Bangkok. Data were collected via face-toface 

interviews in 2007. Results : The study included 86 people. 

The majority of the subjects were 31-50 years old (65%) and male 

(61%). During the two weeks before the interview, the most frequently 

used medicines were anti-inflammatory drugs and muscle relaxants 

(26%), followed by antipyretics. Most of the subjects received their 

medicines from drugstores. During the three months before the 

interview, 75% of the subjects received pharmacy information through 

various cannels. The major source was drugstores, followed by radio. 

With regard to procurement of medication, 79% of the subjects went 

to hospitals and 57% acquired the medicines by themselves. 

Approximateloy 20% of the subjects received medicines with 

touchable-differentiated packaging. For purposes of self-medication, 

the subjects were able to differentiate types, doses, and durations of 

drug use based on memory. Regarding pharmaceutical services, 81% 

of the subjects required special services intended for the disabled, as 

well as further explanation of the medicine’s use, including drug 

labeling. Conclusions : Based on the study results, most of the blind 

did not receive assistance regarding medicine use. Their use of 

medicine was based on memory, and they were in need of improved 

pharmaceutical services. 

(Pharmacoepidemiol Drug Saf. 2009; 18(8):708-12.)


POTENTIAL COST-EFFECTIVENESS OF ROTA- 

VIRUS IMMUNIZATION PROGRAM IN RURAL 

CHINA (NO. 0918) 

Xuan-Yi Wang 1,5 , Arthorn Riewpaiboon 6 , Lorenz von Seidlein 5 , 

Xing-Bao Chen 2 , Paul E. Kilgore 5 , Jing-Chen Ma 3 , Sun-Xiang 

Qi 3 , Zhi-Yong Zhang 4 , Zhi-Yong Hao 4 , Ji-Chao Chen 4 , and Zhi- 

Yi Xu 5 

1 Institutes of Biomedical Sciens and 2 Department of Health 

Economics, School of Public Health, Fudan University, Shanghai, 

3 Center for Disease Control and Prevention, and 4 Zhengding 

Center for Disease Control and Prevention, Hevei, China; 

5 Division of Translational Research, International Vaccine 

Institute, Seoul Koea; and 6 Department of Pharmacy, Faculty of 

Pharmacy, Mahidol University, Bangkok, Thailand. 

Background : To assess the incidence and economic 

burdenof rotavirus diarrhea and the potential cost-effectiveness of a 

rotavirus immunization program in rural zhengding County in Hebei 

Province, China. Method : Population-based surveillance was 

conducted during the peak season for diarrhea among children who 

were

340 

ROLE OF CLINICAL PHARMACIS IN MEDI- 

CATION RECONCILIATION AT GENERAL 

MEDICAL WARDS, RAMATHIBODI HOSPITAL, 

THAILAND (NO. 0921) 

T. Paiboonvong 1 , Preecha Montakantikul 2 , S. Chansirikarnjana 3 , 

P. Tragulperngit 2 

1Department of Pharmacy, Ramathibodi Hosptial, Thailand; 

2Faculty of Pharmacy, Mahidol University, Thailand; 

3Department of Medicine, Faculty of Medicine, Ramathidodi 

Hospital, Thaialnd. 

Key words : Hospital Pharmacy, Medication reconciliation 

Background : Clinical pharmacists have an important role 

to identify and resolve drug related problems (DRPs) in medication 

reconciliation process. The objectives of this study were to determine 

DRPs and evaluate effectiveness of clinical pharmarcist in medication 

reconciliation. Setting : At general medical wards, Ramathidobi 

Hospital, a teaching care hospital affiliated with Mahidol University, 

Thailand. Method : A prospective descriptive study, the patients were 

enrolled at the time of admission between February and April 2008. 

The clicinal pharmacist interviewed each patient to obtain medication 

history and reconciled medication lists with medication orders. All 

patients were followed to identify and resolve DRPs at the admission 

through discharge Results : One hundred and seven patients were 

included in the study. Most patients were elderly who had the mean 

age of 57.3 years old, and current medications before admission were 

approximately 6 items per patient. The clinical pharmacist was able 

to identified DRPs in 45.8% of all patients at the time of admission 

through discharge. At admission, the clinical pharmacist found DRPs 

in 36.4% of all patients and identified 89.6% of all DRPs which 

were accepted by the physician in 86.7% of all interventions. Most 

DRPs were need for addition drug therapy. In addition, groups of 

medications commonly causing DRPs were antimicrobial agents and 

ophthalmic drugs by 14.3% and 12.5%, respectively. Only seven 

patients were transferred to other care units within the hospital. 

During transfer, the pharmacist identified DRPs in two patients and 

the physician accepted for all interventions. At discharge, DRPs 

were identified in 18 out of 100 patients (18.0%). The clinical phar 

macist identified 95.5% of all DRPs which were accepted by the 

physician in 85.7% of all interventions. Most DRPs were need for 

additional drug therapy. Moreover, group of medications commonly 

causing DRPs was antihyperglycemic drugs (33.3%). Conclusion : 

We found DRPs occurring in medication reconciliation process. Most 

patients needed an additional drug therapy at the time of admission 

and discharge. Importantly, the clinical pharmacist was able to 

improve the quality of care by identifying and resolving DRPs. 

(World Congress of Pharamcy and Pharmaceutical Sciences 2009, 

69 th International Congress of FIP, September 3-8, 2009, Istanbul, 

Turkey.) 

INHIBITION OF GASTRIC ACID SECRETION BY 

YA-HOM IN ISOLATED MOUSE WHOLE 

STOMACH (NO. 0922) 

Duangmate Chantharangsikul 1 , Suwan Siriphaisarnpipat 

Thirawarapan 1 , Nuntavan Bunyapraphatsara 2 , Wisuda 

Suvitayavat 1 

1 Department of Physiology, 2 Department of Pharmacognosy, 


Thailand. E-mail : pyvsv@mahidol.ac.th 

Key words : gastic acid secretion, isolated mouse stomach, Ya-hom 

Background : Ya-hom, the traditional Thai formula for 

abdominal discomfort treatment has been reported to partially inhibit 

gastric acid secretion in gastric fistula rats. However, the mechanism 

underlying its action remains unclear. Objective : To investigate the 

gastric acid inhibitory action of Ya-hom and its mechanism of action 

by using isolated mouse whole stomach model. Methods : The gastric 

acid secretion of isolated mouse whole stomach was stimulated by 

histamine (5.0 μM) or bethanechol (10 or 100 μM) after adding the 

inhibitors (atropine 1 μM, ranitidine 10 μM, indomethacin 0.1 μM 

or L-NAME 300 μM) and/or Ya-hom to the serosal solution. The 

effluent perfusate was collected continuously in 10 minute fractions 

for 120 minutes after stimulation. Results : Redissolved lyophilized 

Ya-hom extract at doses of 2.5, 5.0, 10.0 and 20.0 mg/mL inhibited 

histamine-stimulated gastric acid secretion in a dose-dependent 

manner. Inhibition of Ya-hom (10mg/mL) was also observed in the 

presence of atropine (1 μM), which was used to eliminate effects of 

endogenous acetylcholine. Ya-hom inhibited bethanechol-stimulated 

gastric acid secretion in the presence and absence of ranitidine. While 

the inhibitory action of Ya-hom on histamine-stimulated gastric 

secretion was not affected by indomethacin, it was attenuated by 

concomitant treatment with the nitric oxide synthase inhibitor (L- 

NAME). Conclusion : Ya-hom did not stimulate gastric acid secretion 

in the isolated mouse whole stomach. Ya-hom significantly inhibite 

gastric acid secretion after this was stimulated via histamine or 

bethanechol. Nitric oxide stimulation plays an important role in the 

inhibitory action of Ya-hom. 

(Asian Biomedicine 2009, Vol.3, No. 6:663-673.) 


AQUEOUS EXTRACT OF Abutilon indicum Sweet 

INHIBITS GLUCOSE ABSORPTION AND STIMU- 

LATES INSULIN SECRETION IN RODENTS 

(NO. 0923) 

Chutwadee Krisanapun 1 , Penchom Peungvicha 1 , Rungravi 

Temsiririrkkul 2 , Yuvadee Wongkrajang 1 

1 Department of Physiology, 2 Department of Pharmaceutical 

Botany, Faculty of Pharmacy, Mahidol University, Bangkok 

10400, Thailand. E-mail : pyppv@mahidol.ac.th 

Key words : Abutilon indicum Sweet, Antidiabetic activity, Insulin 

secretion 

The objective of this study was to evaluate the antidiabetic 

effects of the aqueous extract derived from the Thai Abutilon indicum 

Sweet plant and to explore its effects on intestinal glucose absorption 

and insulin secretion. The authors hypothesized that the plasma 

glucose level could be reduced through the inhibition of glucose 

absorption and/or the enhancement of insulin secretion. 

Administration of the extract (0.5 and 1 g/kg body weight) in an oral 

glucose tolerance test led to a significant reduction in plasma glucose 

levels in 30 minutes after the administration in moderately diabetic 

rats, as compared with untreated rats (P < .05), and this was at a 

faster rate than the use of an antidiabetic drug, glibenclamide. The 

inhibition of glucose absorption through the small intesting was


investigated using an everted intestinal sac. The results showed that 

the extract at concentrations of 0.156 to 5 mg/mL caused a reduction 

of glucose absorption in a dose response manner. The maximum 

response was noted at a dose of 2.5 mg/mL. The promotion of the 

extract on insulin secretion was confirmed by incubating β cell of 

pancreatic islets and INS-1E insulinoma cells with the extract at 1 to 

1000 μg/mL. These observations suggest that the aqueous extract 

from the A indicum plant has antidiabetic properties, which inhibited 

glucose absorption and stimulated insulin secretion. Phytochemical 

screening also revealed that the extract contained alkaloids, 

flavonoids, tannins, glycosides, and saponins that could account for 

the observed pharmacologic effects of the plant extract. 

(Nutr Res, 2009; 29(8):579-587.) 

ANTI-INFLAMMATORY, ANALGESIC AND 

WOUND HEALING ACTIVITIES OF THE 

LEAVES OF Memecylon edule Roxb. (NO. 0924) 

Somsak Nualkaew 1 , Kwanchai Rattanamanee 2 , Suchitra 

Thongpraditchote 3 , Yuwadee Wongkrajang 3 , Adolf Nahrstedt 4 

1 Department of Pharmaceutical Sciences, Faculty of Pharmacy, 

Mahasarakham University, Mahasarakham 44150, Thailand; Email 

: nualkaew@yahoo.com; 2 Department of Pharmacy Practice, 

Faculty of Pharamceutical Sciences, Naresuan Univesity, 

Pitsanulok 65000, Thailand; 3 Department of Physiology, Faculty 

of Pharmacy, Mahidol Universtiy, Bangkok 10400, Thailand; 

4 Institute for Pharmazeutische Biologie and Phytochemie, 

Westifalishche Wilhelms-Universitat, D-48149 Munster, 

Germany. 

Key words : Analgesic, Anti-inflammatory activity, Memecylon edule 

Aim of the study : To determine the anti-inflammatory, 

analgesic and antioxidant activities of the leaves of Memecylon edule 

Roxb. used traditionally in Thailand. Materials and methods : Hexane, 

(Hex) ethyl acetate (EtOAc), methanol (MeOH) and 50% methanol 

(MeOH50) fractions of the dry leaves were tested in vitro for their 

interleukin-10 production; the most active fraction was further studied 

in vivo for its anti-inflammatory and analgesic activities using the 

ethylphenylpropiolate (EPP)-induced mouse ear edema and the 

writhing test with mice. All fraction except Hex were tested for their 

radical scavenging activity towards 1’-diphenhl-2-picrylhydrazyl 

radical (DPPH{radical dot}). Results : The EtOAc showed the highest 

stimulation for interleukin-10 production. In the EPP test, fraction 

was significantly active 30 min after topical application at all doses 

used (0.5, 1.0, 2.0 mg/ear); after 4 h and at 1.0 mg/ear EtOAc was 

slightly less active (inhibition 47.8%) than the reference, 

indomethacin, at the same dose (62.4%). At 200 mg/kg orally, the 

EtOAc caused a significant inhibition of the writhing response by 

56.6% which was like indomethacin at 10 mg/kg. EtOAc, MeOH 

and MeOH50 exhibited radical scavenging activity. The order of IC 50 

values was : ascorbic acid (9.1 μg.mL) > trolox (11.6 μg/mL) > MeOH 

(46.9 μg/mL) > MeOH50 (152.1 μg/mL) > EtOAc (1742.2 μg/mL). 

Conclusion : The results provide support for the traditional use of 

Memecylon edule leaves in relieving inflammation and pain. 

(J Ethnopharmacol, 2009; 121(2):278-281.) 

ANTIOXIDANT ACTIVITY AND LIPID-LOWERING 

EFFECT OF ESSENTIAL OILS EXTRACTED FROM 

Ocimum sanctum L. LEAVES IN RATS FED WITH A 

HIGH CHOLESTEROL DIET (NO. 0925) 

Thamolwan Suanarunsawat 1 , Watcharaporn Devakul Na 

Ayutthaya 2 , Thanapat Songsak 3 , Suwan Thirawarapan 4 , and 

Somlak Poungshompoo 5 

1 Physiology Unit, 2 Pharmacology and Toxicology Unit, 

Department of Medical Science, Faculty of Science, 3 Department 

of Pharmacognosy and Pharmaceutical Analysis. Faculty of 

Pharmacy, Pathumtani, Rangsit University, Thailand. 

Key words: high fat diet, Ocimum sanctum L., antioxidant 

It has been reported that Ocimum sanctum L. (OS) leaves 

decrease serum lipid profile in normal and diabetic animals. No 

experimental evidences support the antihyperlipidemic and 

antioxidative actions against hypercholesterolemia. Morever the 

identity of the specific chemical ingredients in OS leaves responsible 

for these pharmacological effects are unknown. Since OS leaves are 

rich in essential oil (EO). Therefore the present study was conducted 

to investigate the anti-hyperlipidemic and antioxidative activities of 

EO extracted from OS leaves in rats fed with high cholesterol (HC) 

diet. EO was extracted by the hydrodistillation method and the 

chemical constituents were then indentified by GC-MS. The 

experiment was performed in Male Wistar rats fed with 2.5 g% (w/ 

w) of cholesterol diet for seven weeks. During the last 3 weeks, rats 

were daily fed with EO. The results showed that phenyl propanoid 

compounds including eugenol and methyl eugenol were the major 

constituents of EO. EO suppressed the high serum lipid profile and 

atherogenic index as well as serum LDH and CK-MB without 

significant effect on high serum levels of AST, ALT and alkaline 

phosphatase in rats fed with HC diet. In addition, EO was found to 

decrease the high levels of TBARS, GPx and SOD without impacting 

CAT in the cardiac tissue while in the liver, it decreased high level of 

TBARs without significantly effecting GPx, SOD and CAT. 

Histopathological results confirmed that EO preserved the myocardial 

tissue. It can be concluded that EO extracte from OS leaves has 

lipid-lowering and antioxidative effects that protect the heart against 

hypercholesterolemia. Eugenol that is contained in EO likely 

contribute to these pharmacological effects. 

(J Clin Biochem Nutr, 2009; 45: 1-8.) 

GASTRIC ULCER PROTECTIVE EFFECT OF 

YA-HOM IN RATS (NO. 0926) 

Wisuda Suvitayavat, Pathawee Intayoong, Suwan Thirawarapan 

Department of Physiology, Faculty of Pharmacy, Mahidol 


pyvsv@mahidol.ac.th 

Key words : gastric ulcer, Ya-hom 

Gastric ulcer can be induced by several factors including 

an imbalance between aggressive and defensive factors, such as in 

creasing gastric secretion and decreasing gastric mucus secretion, 

Ya-hom, a Thai traditional medicine, has been shown to inhibit gastric

342 

secretion and stimulate gastric mucus secretion which supports its 

use for gastric ulcer protection and treatment. However, the effect of 

Ya-hom on gastric ulcer protection has not been reported. Thus, this 

study aims to assess gastric ulcer protective activity of Ya-hom. The 

gastric lesion protective effect of Ya-hom was evaluated by oral 

administration of Ya-hom (1, 2 and 4 g/kg) before induction of gastric 

ulcer by hydrochloric acid (0.6 M HCI, 6 ml/kg), aspirin (ASP, 200 

mg/kg) and water immersion restraint stress (WIR, 16 + 2 o C) in 

comparison to cimetidine (0.1 g/kg). Four, six and five after HCI-, 

ASP- and WIR-gastric ulcer induction, respectively, the rats were 

sacrificed for determination of gastric ulcer. The results showed that 

Ya-hom (1, 2 and 4g/kg) inhibited HCI-, ASP- and WIR-induced 

gastric ulcer in a dose dependent manner with the maximum inhibition 

of 93.4, 54.5 and 61.8%, respectively. Cimetidine inhibited HCI-, 

ASP- and WIR-induced gastric ulcer with the inhibition of 78.4, 76.3 

and 48.0%, respectively. This data indicates that Ya-hom can protect 

gastric ulcer induced by acid, aspirin and stress. 

(3 rd Asian Pacific Regional Meeting 10-12 May 2009, The Imperial 

Queen’s Park Hotel) 

IN VITRO STUDY ON ANTI-INFLAMMATORY 

ACTIVITY OF THAI MULBERRY EXTRACT 

(NO. 0927) 

Suchitra Thongpraditchote 1 , Omboon Luanratana 2 , Kwanchai 

Rattanamanee 3 , Narongcai Pongpan 2 , and Yuvadee 

Wongkrajang 1 

1 Department of Physiology, 2 Department of Pharmacognosy, 


Thailand; 3 Department of Pharmacy Practice, Faculty of 

Pharmaceutical Sciences, Naresuam University, Pitsanulok 

65000, Thailand. E-mail : pystp@mahidol.ac.th 

Key words : anti-inflammation, mulberry, interleukin 

The anti-inflammatory effect iof the crude extracts from 

different parts of Thai Mulberry was determined. The stem bark, 

stem wood, root bark and root wood were extracted by soxhlet 

extraction using hexane, dichloromethane and methanol as the 

extraction solvent. All extracts were tested in vitro for their inhibition 

on interleukin-1 beta (IL-1β) production and their promotion of 

interleukin 10 (IL-10) production. All methanolic extracts inhibited 

the production of IL-1β at a dose of 0.1 mg/ml. The extract from 

stem wood showed the best activity (58.2%), followed by root wood 

(57.9%), root bark (45.6%) and stem bark (36.1%). The stimulation 

of antiinflammatory cytokine, IL-10, was also determine in this study. 

The percentage of stimulation of IL-10 was 58% and 67.3% for the 

stem wood and the root bark extracts at a dose of 0.01 mg/ml, 

respectively. These results indicated the anti-inflammatory activity 

of Thai Mulberry extracts from the stem wood and the root. These 

findings support the traditional use of Mulbery stem and root in the 

treatment of inflammation. 

(The 3 rd Asian Pacific Regional Meeting May 10-12, 2009, Bangkok, 

Thailand.) 

ANALGESIC, ANTIOXIDANT AND ACUTE 

TOXICITY PROPERTIES OF GINGER RHIZOMES 


Suchitra Thongpraditchote 1 , Yuvadee Wongrajang 1 , Arunporn 

Itharat 2 , Wisuda Suvittayavat 1 , Suwan Thirawarapan 1 

1 Department of Physiology, Faculty of Pharmacy, Mahidol 

University, Bangkok, Thailand; 2 Applied Thai Traditional 

Medicine, Faculty of Medicine, Thammasart University, 

Thailand. E-mail : pystp@mahidol.ac.th 

Key words : analgesic, antioxidant ginger 


Ginger (Zingiber officinale Roscoe, Zingiberacae) is one 

of the most commonly used as food additive and speices in many 

part of the world, especially in the South-Eastern Asian countries. 

Ginger is also a medicinal plant that has been widely used in Thaiand 

as antimotion sickenss and antiflatulence. The present study was 

undertaken to investigate the analgesic, antioxidant and oral acut 

toxicity properties of ethanolic extract of ginger rhizomes. The 

analgesic effect of the ginger extract was examined in mice using 

acetic acid-induced writhing responses while the antioxidant effect 

of the extract was tested in vitro by 1,1- diphenyl-2 picrylhydrazyl 

(DPPH) assay. Oral administration of the ginger rhizome extract 

(50, 100 and 200 mg/kg) inhibited the writhing behavior in a dosedependent 

manner. The extract exhibited a strong radical scavenging 

activity with an IC 50 value of 3.80 μg/ml. In acute toxicity study, the 

median lethal dose (LD 50 ) of the extract was greater than 2 g/kg for 

oral administration in rats and mice and no deaths or abnormalities 

in clinical signs were observed, suggesting a slightly toxic property 

of the extract. These findings support the traditional use of ginger 

rhizomes in the treatment of painful condition and oxidative stress. 

(The fifth International Symposium on the Family Zingiberaceae 

July 6-9, 2009, Xishuangbana, China.) 

ANTIOXIDANT PROPERTY OF A THAI TRADI- 

TIONAL FORMULA FOR LONGEVITY (NO. 0929) 

Yuvadee Wongkrajang 1 , Rungravi Temsiririrkkul 2 , Penchom 

Peungvicha 1 , Somjai Nakornchai 3 , Siripen Luanchoy 1 , Sirirat 

Tiangkul 1 


Botany, 3 Department of Pharmacology, Faculty of Pharmacy, 

Mahidol University, Bangkok 10400, Thailand. E-mail : 

pyywk@mahidol.ac.th 

Key words : antioxidant, Thai traditional formula, longevity 

Antioxidant properties of a Thai traditional formula for 

longevity, which is composed of 6 herbs as follows: Albizia procera, 

Diospyros rhodocalyx, Tinospora crispa, Cyprus rotundus, Streblus 

asper and Piper nigrum were studied. Each herb including the 

formula was extracted by 95% ethanol and concentrated by using 

vacuum evaporator. The antioxidant properties were detected by 

DPPH method. Vitamin C and Trolox were used as reference standard. 

The in vitro oxidative hemolysis of sheep red blood cells was used as 

a model to study the free radical-induced damage of biological 

membranes by AAPH. From DPPH method, Albizia procera extract


possessed the most potent antioxidant properties (IC 50 44.34 μg/ml) 

while vitamin C and Trolox had IC 50 at 17.47 μg/ml and 22.75 μg/ml 

respectively. From AAPH hemolysis method, Albizia procera, the 

formula for longevity, Cyperus rotundus, Diospyrosrhodocalyx, Piper 

nigrum, Tinospora crispa and Streblus asper extracts at the 

concentration of 5 mg/ml could prolong the time of 50% hemolysis 

from 78 minutes to 157,142, 126, 114, 108, 101, 100 minutes, 

respectively, while the time of 50% hemolysis of trolox at the 

concentration of 0.5 mg/ml was 160 minutes. The phytochemical 

screening tests showed the presence of phenolic compounds, tannins 

and flavonoids in the formula and Cyperus rotundus, Albizia procera; 

phenolic compounds and flavonoid in Piper nigrum, Diospyros 

rhodocalyx and Streblus asper. Phenolic compounds was found in 

Tinospora crispa extract. 

(The 57 th International congress and Annual Meeting of the Society 

for Medical Plant Research August 16-20, 2009, Geneva, 

Switzerland.) 

ANTIOXIDANT ACTIVITY OF Moringa oleifer Lam. 

(NO. 0930) 

Yuvadee Wongkrajan 1 , Rungravi Temsiririrkkul 2 , Penchom 

Peungvicha 1 , Somjai Nakornchai 3 , Rattiya Maksub 1 , Sasiwimon 

Wichairam 1 


Botany, 3 Department of Pharmacology, Faculty of Pharmacy, 

Mahidol University, Bangkok 10400, Thailand. E-mail : 

pyywk@mahidol.ac.th 

Key words : Morinfga oleifera Lam, antioxidant 

Background & Objective : Nowadays in Thailand, various 

parts Moringa oleifera Lam wer used for health promotion due to 

the claim of many activity such as antitumor, antipyretic, antiinflammatory, 

antiulcer, hepatoprotective and antibacterial activities. 

Therefore it was interested to study antioxidant properties of Moringa 

oleifera Lam.: leaves bark and young fruits in Thailand because 

radical scavenging or antioxidant properties is the basic property of 

the preventive effects of diseases. Methods : Each part was extracted 

by 80% methanol or water. The antioxidant properties were detected 

by DPPH method. The in vitro oxidative hemolysis of sheep red 

blood cells was used as a model to study the free radical induced 

damage of biological membranes by using AAPH. Results : From 

DPPH method it was showed that leaves extract by water possessed 

the most potent properties IC 50 (179.22 μg/ml), the range of IC 50 of 

other parts were 218.93 – 302.93 μg/ml while IC 50 of Vitamin C and 

Trolox were 16.17 and 27.85 mg/ml respectively. From AAPH 

hemolysis method it was found that leaves extract by water at the 

concentration of 2 mg/ml also possessed the most potent it could 

prolong the time of 50% hemolysis from 82 minutes in control group 

to 175 minutes, the range of the time of 50% hemolysis of ther parts 

were 127-101 minutes while the time of 50% hemolysis of 0.5 mg/ 

ml trolox was 176 minutes. Conclusions : Water extract of the leaves 

had the most antioxidant activity in both DPPH method and AAPH 

hemolysis method. From DPPH method the extract showed the 

activity about 10 time less than vitamin C and Trolox whereas in 

AAPH hemolysis method, it had the property with 4 times less than 

trolox. 

(The 2009 DMS Annual Scientific Conference and the 10 th National 

Cancer Conference : A Joint Meeting November 2-4, 2009, Bangkok, 

Thailand.)

Faculty of Pharmacy - Mahidol University

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?