molecular modeling investigations of pyrimethamine...

©Pure and Applied Chemistry International Conference 2014 NMC-1

Natural Products, Chemical Biology and Medicinal ChemistryNMC-OR-01

Molecular Modeling Investigations of Pyrimethamine Analogues Binding to Mycobacterium tuberculosis Dihydrofolate Reductase

Pimonluck Sittikornpaiboon1*, Ubolsree Leartsakulpanich2, Chawanee Thongpanchang2, Pisanu Toochinda1,

Luckhana Lawtrakul1

1School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology,

Thammasat University, 99 Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand 2National Center for Genetic Engineering and Biotechnology, National Science and Technology Development

Agency, 113 Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand *E-mail: [email protected]

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis which remains a major public health problem and cause ill-health among millions of people each year. The resistance of bacteria to the drugs is still increasing. Therefore, the finding of new anti-tuberculosis agent is an importance issue. In this study, 24 antimalarial agents of pyrimethamine analogues were used to derive as anti-tuberculosis agents through molecular docking and QSAR techniques. Docking studies exploring possible binding affinities and modes of two main clusters of pyrimethamine analogues on M. tuberculosis dihydrofolate reductase (mtbDHFR) targets. Our docking results suggest that some of pyrimethamine analogues, antimalarial agents, can be used as anti-tuberculosis agents which inhibit the function of mtbDHFR. On the basis, these analogues showing high binding affinities to the target in silico. 2D- and 3D-QSAR models were developed using MOE and Sybyl programs to identify the key structural features requirements for binding to mtbDHFR. Keywords Tuberculosis; Dihydrofolate reductase; Pyrimethamine; QSAR; CoMFA



Development of Thai Silk Fibroin/Hyaluronic Acid Microspheres by Water in Oil Emulsion Method

Piyarat Sungkhaphan1, Sorada Kanokpanont1, Siriporn Damrongsakkul1*

1 Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn University, Phatumwan

Bangkok, 10330 Thailand *E-mail: [email protected]

Hyaluronic acid (HA) is a high molecular weight linear polysaccharide, naturally presented in extracellular matrix in many parts of living body. It is widely used as a lubricant biomaterial with excellent biocompatibility. Silk fibroin (SF), from domesticated silkworm, Bombyx mori, is a fibrous protein that is well-known for its biocompatibility and good mechanical properties. Recently it has been widely explored for medical applications. In this study, silk fibroin extracted from domesticated Thai silk cocoons (Nangnoi Srisaket 1) and hyaluronic acid were employed to develop microspheres aimed for a controlled release application. After degumming the cocoons, SF solution was obtained and blended with HA solution at different weight blending ratios (SF/HA=30/70, 50/50 and 70/30). SF/HA microspheres were fabricated by water-in-oil emulsion method. Aqueous SF/HA solution and vegetable oil were used as water and oil phases, respectively. To stabilize the microspheres, crosslinking with 0.15 %(v/v) glutaraldehyde was used prior to freeze-drying. Effects of SF/HA blending ratios on the morphology and characteristics of obtained microspheres were investigated. Electrophoretic mobility (µe) of SF and HA solutions at different pH were investigated to understand the most appropriate region for the SF/HA solution blending. Morphology of SF/HA microspheres was observed using a scanning electron microscope (SEM). The results showed that at pH 6 and above, homogeneous blend solutions could be obtained though the charge of both SF and HA solutions were negative. The microsphere formation could possibly be the result of hydrophobic interaction. SEM micrographs revealed that SF/HA (70/30) microspheres were spherical with a size of 40-110 µm. and smooth surface. The SF/HA blended microspheres were expected to be applied as alternative carriers for controlled release application. Keywords: Silk Fibroin; Hyaluronic acid; Microsphere



Chemical Constituents from the Twigs of Caesalpinia furfuracea

Ittipon Siridechakorn* and Surat Laphookhieo

Natural Products Research Laboratory, School of Science, Mae Fah Luang University, Chiang Rai 57100,

Thailand *E-mail: [email protected]

The first phytochemical investigation of Caesalpinia furfuracea twigs led to the isolation and identification of four new compounds together with four known compounds (1-4). Their structures were elucidated by intensive spectroscopic analysis. The antibacterial activity against Escherichia coli TISTR 780, Salmonella typhimurium TISTR 292, Staphylococcus aureus TISTR 1466, and methicillin-resistant S. aureus (MRSA) SK1 of some compounds was also evaluated.

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Keywords Caesalpinia furfuracea; Leguminosae; Isopimarane diterpene; Stilbene; Antibacterial activity



Bio-inspiration from Asian Green Mussel Shells: Security Markers Based on Dye-Loaded Stacked Aragonite

Microcrystals

Chutiparn Lertvachirapaiboon, Thiluksakorn Jirapisitkul, Prompong Pienpinijtham, Chuchaat Thammacharoen, and Sanong Ekgasit*

Sensor Research Unit, Department of Chemistry, Faculty of Science, Chulalongkorn University,

254 Phayathai Rd., Pathumwan, Bangkok 10330, Thailand *E-mail: [email protected]

Perna viridis (Asian green mussel) shells are an assembly of alternate-stratified bilayers of aragonite calcium carbonate and protein with thickness range of 200-500 nm and 20-30 nm, respectively. The protein in the shell structure could be removed by consecutive alkaline/thermal treatment. The decrease of protein content from 6.9 %wt. to 2.1 %wt. as determined by thermal gravimetric analysis indicated a removal of inter-aragonite crystals binding protein. After removal, the protein layers were replaced by vacant space while the geometrical structure of the shell was still intact. The newly generated air gaps were loaded with dyes such as red dye, blue dye, and fluorescent dye (Rhodamine 6G).The incorporated dye provided multimodal security observation under bright and dark filed illuminations. Under bright filed illumination, the regular optical microscope (OM) image of aragonite/air stratified bilayers was observed. The incorporated dye within the structure could be visualized under dark field illumination. The bio-inspired material demonstrated the unique potential applications for forgery of valuable protection such as brand name products, legal documents, contracts and banknotes. Keywords Stratified bilayers; Perna viridis; Security markers



Benzophenones and Xanthones from Cratoxylum sumatranum Roots

Cholpisut Tantapakul*, Wisanu Maneerat, Tawanun Sripisut, and Surat Laphookhieo

Natural Products Research Laboratory, School of Science, Mae Fah Luang University, Tasud, Muang,

Chiang Rai 57100, Thailand

*E-mail: [email protected] The chemical investigation of Cratoxylum sumatranum roots led to isolation of two new benzophenones and three new xanthones together with nine known compounds. Their structures were elucidated on the basis of spectroscopic methods. Some compounds were evaluated for their cytotoxicity against three human cancer cell lines including human epidermoid carcinoma of oral cavity (KB) and breast adenocarcinoma (MCF-7).

Keywords Cratoxylum sumatranum; Clusiaceae; cytotoxicity



Fluorinated Isatin Sulfonamides: Putative PET-Compatible Radiotracers for Molecular Imaging of Apoptosis

Panupun Limpachayaporn1,2,3, Stefan Wagner4, Klaus Kopka4, Otmar Schober4,

Sven Hermann5, Michael Schäfers5, Günter Haufe1,5,*

1 Organisch-Chemisches Institut, Westfälische Wilhelms-Universität Münster, Corrensstraße 40, D-48149 Münster, Germany

2 International NRW Graduate School of Chemistry, Westfälische Wilhelms-Universität Münster, Wilhelm-Klemm-Straße 10, D-48149 Münster, Germany

3 Department of Chemistry, Faculty of Science, Silpakorn University, Sanamchandra Campus Nakhon Pathom 73000, Thailand

4 Klinik für Nuklearmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A1, D-48149 Münster, Germany

5 European Institute for Molecular Imaging (EIMI), Westfälische Wilhelms-Universität Münster, Mendelstraße 11, D-48149 Münster, Germany

*E-mail: [email protected], [email protected] Molecular imaging of apoptosis (programmed type-I cell death) by tracking executing caspases (cysteinyl aspartate-specific proteinases) using Positron Emission Tomography (PET) allows visualizing and localizing as well as monitoring pathological conditions relating to a wide variety of diseases associated with dysregulation of the cell death programme. Herein, we have been developing (S)-5-[1-(2-methoxymethylpyrrolidinyl)sulfonyl]isatin (1) which is a potent non-peptidic inhibitor of caspases-3 and -7 to function as PET imaging agent, the so called “radiotracer”, for apoptosis imaging. In total, 74 structurally modified compounds including fluorinated analogues of 1 were synthesized and in vitro inhibitory activities towards caspases-1, -3, -6 and -7 were evaluated. The purpose is to find the fluorinated derivatives with high binding affinities, which are putative radiotracers for the imaging. As a result, the synthesized fluorinated analogues exhibited good to excellent activities with selectivity against caspases-3 and -7 having IC50 in nanomolar to subnanomolar ranges in most cases. Furthermore, 18F-radiolabeling of the selected N-fluorohydroxybutyl analogues were carried out successfully by cyclic sulfate ring-opening strategy using non carrier added [18F]fluoride in the presence of Kryptofix®. The labeling process was followed by acidic hydrolysis leading to furnishing the desired tracers with satisfactory yields having high specific radioactivity. Finally, in vivo biodistribution studies in a healthy mouse using PET/CT imaging after tail vein injection showed fast clearance of the tracer. Keywords Caspase inhibitor; Radiofluorination; Radiotracer; Molecular imaging; Positron Emission Tomography (PET)


Natural Products, Chemical Biology and Medicinal ChemistryNMC- OR-07

Phytochemistry and Bioactivities Evaluation of Oroxylum indicum (L.) Vent.

Kulsiri Yossathera, Siripat Suteerapataranon, Suwanna Deachathai*

School of Science, Mae Fah Luang University, Tasud, Muang, Chiang Rai, 57100, Thailand

*Email: [email protected]

The phytochemistry of leaves, roots, stems, and twigs of Oroxylum indicum resulted in the isolation of five flavonoids: 5,7-dihydroxy-6-methoxyflavone (oroxylin A, 1), 5,7-dihydroxyflavone (chrysin, 2), 5,7,4 -trihydroxy-6-methoxyflavone (hispidulin, 3), 5,7,4 -trihydroxyflavone (apigenin, 4), and 5,6,7,4 -tetrahydroxyflavone (scutellarein, 5), a coumarin derivertive: 9-[(3,3-dimethyloxiran-2-yl)methoxy]furo[3,2-g]chromen-7-one (heraclenin, 6), three xanthones: 5 -demethoxycadensin G (7), 1,3,6-trihydroxy-7-methoxy-2-(3,7-dimethyl-2,6-octadienyl)xanthone (cowaxanthone, 8), and 1,7-dihydroxyxanthone (euxanthone, 9), and two steroids: stigmasterol (10) and β-sitosterol (11). Their structures were established on the basis of spectroscopic analysis. All extracts exhibited weak to moderate antibacterial activities against gram-positive bacteria (Bacillus cereus TISTR 687, Staphylococcus aureus TISTR 1466, Methicillin-resistant Staphylococcus aureus SK1) and gram-negative bacteria (Escherichia coli TISTR 780, Pseudomonas aurenginosa TISTR 781, and Salmonellae typhimurium TISTR 292). Compound 3 exhibited significant antibacterial activity against MRSA SK1 with an MIC value of 32 μg/mL, whereas compound 7 exhibited against B. cereus with an MIC value of 16 μg/mL. Evaluation of antioxidation activity of all extracts and isolated compounds using a flow injection-spectrophotometric system were performed. The acetone extracts of the leaves and roots exhibited strong antioxidation activity with IC50 values of 8.01±0.29 and 9.81±0.09 µg/mL, respectively. Whereas all isolated compounds showed weak antioxidation activity.

Keywords Oroxylum indicum; Antibacterial activity; Antioxidant activity; Flow Injection-Spectrophotometric System


Natural Products, Chemical Biology and Medicinal ChemistryNMC- OR-08

Bioactive Compounds from the Roots of Garcinia cowa Roxb.

Ngampuk Tayana, Siripat Suteerapataranon, Suwanna Deachathai*

School of Science, Mae Fah Luang University, Tasud, Muang, Chiang Rai 57100, Thailand

*E-mail: [email protected] Chemical investigation of acetone extract from the roots of Garcinia cowa Roxb. has resulted in the isolation of twenty-two known compounds, cochinchinone E (1), 1,3,6-trihydroxy-7-methoxy-2,5-bis(3-methy-2-butenyl)xanthone (2), α-mangostin (3), β-mangostin (4), cowagarcinone B (5), cochinchinone A (6), cowanin (7), cowanol (8), cochinchinone G (9), 7-geranyloxy-1,3-dihydroxyxanthone (10), cowaxanthone (11), xanthone VI (12), macluraxanthone (13), 9,10-di-O-methylmacluraxanthone (14), 10-O-methylmacluraxanthone (15), isocudraniaxanthone B (16), euxanthone (17), formoxanthone C (18), pyranojacareubin (19), cratoxycochinchinone C (20), stigmasterol (21), and friedelin (22). Their structures were elucidated on the basis of spectroscopic data. Four isolated compounds, 3, 4, 7, and 8 exhibited strong antibacterial activity against Methicillin-resistant staphylococcus aureus SK1, Bacillus cereus, Staphylococcus aureus and Salmonella typhimurium with MIC range of 0.5-4 μg/mL. Compounds 9, 10 and 16 showed moderate antibacterial activity against Staphylococcus aureus with an MIC value of 8 μg/mL. Evaluation of antioxidant activity of crude acetone extract and all isolated compounds using flow injection analysis (FIA) method were performed. The crude extract found to possess moderate activity with an IC50 value of 35.86 ± 0.14 µg/mL. Compounds 12, 13, 16, and 18 were effective in the DPPH assay with IC50 values of 19.70 ± 0.39, 35.86 ± 0.15, 19.75 ± 0.39, and 35.55 ± 0.43 μg/mL, respectively. Keywords Garcinia cowa Roxb.; Xanthone; Antibacterial activity; Antioxidant activity; Flow injection analysis.


Natural Products, Chemical Biology and Medicinal ChemistryNMC-P-001

Characterization of Crustin Antimicrobial Peptide from the Black Tiger Shrimp Penaeus monodon

Suwattana Visetnan2, Premruethai Supungul2,3, Anchalee Tassanakajon2, Vichien Rimphanitchayakit2, Suchao

Donpudsa1*

1Department of Chemistry, Faculty of Science, Srinakharinwirot University, Bangkok 10110, Thailand

2Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phyathai Road, Bangkok 10330, Thailand

3National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani 12120, Thailand

*E-mail: [email protected] A crustin gene, crustinPm4, previously found in the hemocyte cDNA library of black tiger shrimp (Penaeus monodon), contains the open reading frames of 807 bp encoding a putative protein of 269 amino acid residues. The putative signal peptides of the crustinPm4 were identified using the online SignalP 3.0 with predicted cleavage sites between Ala18-Gly19, resulting in 251 residue mature protein with calculated molecular mass of 22.5 kDa and predicted pI of 8.4. This crustin contains a Gly-Pro rich region at the amino-terminus and a single whey acidic protein (WAP) domain at the carboxyl-terminus. In order to characterize their properties and biological activities, the recombinant crustin protein was produced in the Escherichia coli expression system. Antimicrobial assays showed that the growth of Bacillus megaterium was inhibited by this recombinant crustin with MIC of about 3.13-6.25 µM. Keywords crustin; Penaeus monodon; antimicrobial activity



Investigation of Antioxidant Activity of Active Compounds in the Crude Methanol Extract from Paederia foetida Linn. Stem

Rawee Pithakvarin1, Sattawat Dueansawang1, Noppadol Seeja1, Amornrat Boon-oad1 and

Boonjira Rutnakornpituk1*

Department of Chemistry and Center of Excellence in Biomaterials, Faculty of Science,

Naresuan University, Pitsanulok, 65000, Thailand * E-mail : [email protected]

This research reported the antioxidant activity of active compounds in the crude methanol extract from the stem of Paederia foetida Linn. by using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) free scavenging method. Antioxidant activity of the crude methanol extract was determined and the percentage of inhibition was 93.80%, indicating a good antioxidant activity. The crude methanol extract was then separated by column chromatography over silica gel to yield 4 fractions (1B-4B). The result showed that fraction 2B showed highest percentage about 95.13% in the inhibition. The functional group and structure of the compound in this fraction was then analyzed via Fourier Transform Infrared Spectroscopy (FT-IR), Nuclear Magnetic Resonance (NMR) spectroscopy and Thin Layer Chromatography (TLC). These evidences implied that the fraction containing active compounds possibly composed of flavonoid derivatives which have hydroxyl groups as substituent group. Keywords ; 2,2-diphenyl-1-picrylhydrazyl free scavenging; Antioxidant; Paederia foetida Linn.

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Total Lipid and Fatty Acid Composition of Spirogyra spp. in Northern Part of Thailand

Thitiphan Chimsook1*

1Department of Chemistry, Faculty of Science, Maejo University, Chiang Mai, 50290, Thailand

*E-mail: [email protected] The macroalga, Spirogyra spp., were collected from freshwater in northern part of Thailand and extracted with methanol. The extract was determined by GC to reveal the presence of saturated and unsaturated fatty acids. The former acids were slightly larger in proportion (46.55%) than latter ones (53.42%). Linolenic (18.43%) and oleic (10.84%) acids were found in high proportion, while palmitic acid occurred in small proportion (4.87%). Furthermore, two sterols, isodecortinol and dinosterol, were also obtained from the extract. The biodiesel potential was evaluated by extraction of algal oil using both soxhlet extraction and the Bligh and Dyer method to yield saturated and unsaturated fatty acids 5.28% and 7.96% respectively. The study showed that Spirogyra spp. could be better used as a food supplements than strains for bioethanol and biodiesel. Keywords Spirogyra spp.; Fatty acids; GC-MS



Diphenyl Ether and Methyl Benzoate Derivatives from the Seagrass-derived Fungus Pestalotiopsis theae PSU-ES148

Kanyapat Lumyong1, Jirawat Junlapan1, Souwalak Phongpaichit2, Preuttiporn Supaphon2, Jariya Sakayaroj3,

Vatcharin Rukachaisirikul1*

1Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand

2Department of Microbiology, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand

3National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand Science Park, Klong Luang, Pathumthani 12120, Thailand

*E-mail: [email protected] Plant endophytic fungi are a well-known source of bioactive compounds. In our ongoing search for bioactive metabolites from the endophytic fungi, the fungus Pestalotiopsis theae PSU-ES148 was isolated from the seagrass Enhalus accoroides. Purification of the broth and mycelial extracts from this fungus by various chromatographic techniques led to the isolation of four known compounds. Both broth and mycelial EtOAc extracts yielded (R)-penipratynolene, pestheic acid and pestalotether B while the mycelial hexane extract gave pestalotether A. The structures were characterized by analysis of the spectroscopic data and by comparison of the 1H and 13C NMR data with those previously reported. The absolute configuration of (R)-penipratynolene was assigned on the basis of the similar optical rotation to that of the known one. Keywords Seagrass-derived fungus; Pestalotiopsis theae; Diphenyl ether; Methyl benzoate



Pimarane Diterpenes and Cytochalasins from the Unidentified Endophytic Fungus PSU-H267

Thippaya Kongprapan1, Souwalak Phongpaichit2, Wimarak Poonsuwan 2, Jariya Sakayaroj3,





*E-mail: [email protected] Endophytic fungi are a rich source of novel organic compounds with interesting biological activities and a high level of biodiversity. The unidentified endophytic fungus PSU-H267 was isolated from a leaf of Hevea brasiliensis collected in Songkhla province, Thailand. The broth EtOAc extract was active in anti-cancer activity against KB-oral cavity cancer cell lines with an IC50 value of 1.63 µg/mL. In addition, the extract displayed antioxidant activity in DPPH• assay with the IC50 value of 0.066 mg/mL. Upon chromatographic separations, seven known compounds were isolated from the broth extract of this fungus including diaportheins A and B, 11-deoxydiaporthein A, scoparasins A and B, tyrosol and 2-hydroxypropanoic acid. The structures were determined by spectroscopic data and confirmed by comparison of the 1H and 13C NMR data with those previously reported. The relative configuration was established by the NOEDIFF data and comparison of their specific rotations with those of the known ones. Keywords Endophytic fungi; Hevea brasiliensis; Diaportheins; Cytochalasins; Scoparasins



Meroterpenoids from the Seagrass-derived Fungus Talaromyces sp. PSU-ES194

Jiraporn Arunpanichlert1, Souwalak Phongpaichit2, Orathai Supaphon2, Jariya Sakayaroj3,


1Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand

2Department of Microbiology, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand

3National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand Science Park, Klong Luang, Pathumthani, 12120, Thailand

*E-mail: [email protected] Fungi in the genus Talaromyces have been reported to be a rich source of bioactive natural products. They are capable of producing various types of secondary metabolites that show significant biological activities. In our ongoing search for bioactive metabolites from the seagrass-derived fungi, the fungus Talaromyces sp. PSU-ES194 was isolated from the seagrass Enhalus accoroides. Chemical investigation of the broth ethyl acetate extract led to the isolation of four known meroterpenoids named austin, dehydroaustin, dehydroaustinol and acetoxydehydroaustin. Their structures were elucidated based on the spectroscopic data and confirmed by comparison of the 1H and 13C NMR data with those previously reported. Keywords Meroterpenoids; Seagrass-derived fungus; Talaromyces sp.; Enhalus accoroides



Metabolites from the Soil Fungus Aspergillus sclerotiorum PSU-RSPG178

Patima Phainuphong1, Souwalak Phongpaichit2, Kawitsara Borwornwiriyapan2, Jariya Sakayaroj3, Vatcharin Rukachaisirikul1*

1Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science,

Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand 2Department of Microbiology, Faculty of Science, Prince of Songkla University,

Hat Yai, Songkhla 90112, Thailand 3National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand Science Park,

Klong Luang, Pathumthani 12120, Thailand *E-mail: [email protected]

Soil fungi in the genus Aspergillus which belong to the Ascomycetes play an important role in a degradation process in an ecosystem. They are capable of producing a wide range of bioactive secondary metabolites with significant biological activities. In our ongoing search for bioactive metabolites from soil fungi, the broth ethyl acetate extract of the fungus Aspergillus sclerotiorum PSU-RSPG178 displayed interesting antimicrobial activity against Staphylococcus aureus ATCC25923, Cryptococcus neoformans ATCC90113 and Microsporum gypseum with MIC values of 128, 64 and 128 μg/mL, respectively. It also possessed the activities against Plasmodium falciparum, KB-oral cavity cancer and MCF-7 breast cancer cell lines with IC50 values of 2.39, 6.70 and 11.30 μg/mL, respectively. The investigation of the broth ethyl acetate extract led to the isolation of four known compounds, including lovastatin, α,β−dehydrolovastatin, 3-methoxy-4-methyl-2,4-dienpentanoic acid and dihydropenicillic acid. Their structures were elucidated by spectroscopic methods and confirmed by comparison of the 1H and 13C NMR data with those previously reported. The absolute configurations of the lovastatin derivatives were established by NOEDIFF data as well as optical rotations. Keywords Soil fungus; Aspergillus sclerotiorum; Anticancer; Antimalarial; Antimicrobial



Eremophilane Sesquiterpenes from the Soil Fungus Penicillium copticola PSU-RSPG138

Charuwan Daengrot1, Souwalak Phongpaichit2, Kawitsara Bowornwiriyapan2,

Jariya Sakayaroj3,Vatcharin Rukachaisirikul1*




*E-mail: [email protected]

Six eremophilane sesquiterpenes were obtained from the broth ethyl acetate extract of the soil fungus Penicillium copticola PSU-RSPG138. Based on analysis of spectroscopic data, their structures were elucidated as sporogen-AO 1, dihydrosporogen-AO 1, 3-acetyl-13-deoxyphomenone, petasol, 7-hydroxypetasol and JBIR-27. The absolute configuration was assigned on the basis of the NOEDIFF data and by comparison of optical rotations with those previously reported. Among six isolated metabolites, sporogen-AO 1 exhibited the best antimalarial (Plasmodium falciparum, K1 strain), anticancer (KB-oral cavity cancer cells) and cytotoxic (African green monkey kidney cells) activities with IC50 values of 0.38, 4.43 and 1.10 μg/mL, respectively. Keywords Eremophilane sesquiterpenes; Penicillium copticola; Antimalarial; Anticancer; Cytotoxicity



Essential Oil Compositions of Kaempferia larsenii Sirirugsa and Kaempferia marginata Carey

Rhizomes from Thailand

Orawan Theanphong1*, Chanida Palanuvej2, Nijsiri Ruangrungsi2,3, Kanchana Rungsihirunrat2 and Witchuda Thanakijcharoenpath1

1 Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences,

Chulalongkorn University,Bangkok 10330, Thailand 2 College of Public Health Sciences, Chulalongkorn University, Bangkok 10330, Thailand

3Faculty of Pharmacy, Rangsit University, Pathumthani 12000, Thailand

* Corresponding author. Email:[email protected]

The hydrodistilled essential oils obtained from Kaempferia larsenii Sirirugsa and K. marginataCarey from Thailand were analyzed by GC-MS. A total of 53 compounds were identified. Major components of the essential oil from K. larsenii were found to be epi-13-manool (21.93%), caryophyllene oxide (12.66%) and camphene (9.71%) whereas those from K. marginata included 2-propenoic acid, 3-(4-methoxyphenyl)-, ethyl ester (49.00%), E-ethyl cinnamate (20.41%) and pentadecane (10.28%). Keywords Kaempferia larsenii; Kaempferia marginata; Zingiberaceae; essential oil



Zearalenone Derivatives from the Seagrass-derived Fungus Fusarium sp. PSU-ES123

Praphatsorn Saetang1, Souwalak Phongpaichit2, Preuttiporn Supaphon2, Jariya Sakayaroj3,





*E-mail: [email protected] Seagrasses are marine plants which are a source of endophytic fungi with capability of producing interesting bioactive compounds. Fungi were isolated from the seagrass Enhalus accoroides and their broth ethyl acetate extracts were screened for biological activities. We found that the broth ethyl acetate extract of the fungus Fusarium sp. PSU-ES123 displayed interesting antimicrobial activity against Staphylococcus aureus and Cryptococcus neoformans with MIC values of 64 and 128 μg/mL, respectively. Purification of this extract led to the isolation of four known compounds, including zearalenone, 7'-dehydrozearalenone, 8'-hydroxyzearalenone and β-zearalenol. The structures were characterized by analysis of the spectroscopic data and by comparison of the 1H and 13C NMR data with those previously reported. The absolute configurations of the isolated compounds were assigned on the basis of the similar optical rotations to those of the known ones. Keywords Seagrass-derived fungus; Fusarium sp.; Zearalenone; Antimicrobial activity



Cytotoxicity of Chemical Constituents from the Fruits of Derris indica

Phatharapoll Decharchoochart, Chavi Yenjai*

Natural Products Research Unit, Department of Chemistry and Center of Excellence for Innovation in

Chemistry, Faculty of Science, KhonKaen University, KhonKaen, 40002, Thailand. *E-mail: [email protected]

Investigation of the chemical constituents from the fruits of Derris indica (Luguminosae) yielded three known flavonoids, karanjin (I), karanjachromene (II), demethoxykanugin (III) and four known chalcones, tunicatachalcone (IV), pongamol (V), obovatachalcone (VI), ovalichalcone (VII), and. The structures of all isolated compounds were elucidated by 1D (1H, 13C-NMR), 2D NMR (COSY, HMBC and HMQC), IR and MS spectroscopic analysis. Pongamol (V) showed moderate cytotoxicity against KB cell line with an IC50 value 8.94 µg/ml and compounds IV and V also exhibited moderate cytotoxicity against this cell line with IC50 values 14.74 and 18.34 µg/ml, respectively. Keyword: Derris indica, flavonoids, chalcones



Effect of Phenolic Attractive Substances on the Regeneration Efficiency in a Thai Rice Cultivar

Sutee Chutipaijit1*, Kanokporn Sompornpailin1,2

1College of Nanotechnology, King Mongkut’s Institute of Technology Ladkrabang, Bangkok 10520, Thailand

2Thailand Center of Excellence in Physics, CHE, Ministry of Education, Bangkok 10400, Thailand *E-mail: [email protected]

Improvement of high frequency in indica rice regeneration is still essential investigation especially Thai rice cultivars. Oxidized phenolic compounds are a strong inhibition of the somatic embryogenesis and organogenesis in tissue culture or in vitro system. The objective of this research was to study a media composition by using different types and concentrations of antiphenolic compounds (silver nitrate, ascorbic acid and cysteine). Mature seeds of the Thai rice cultivar KDML105 were used as a source of the explants. The results showed that the regeneration frequency was significantly different in various antiphenolic compounds. Optimum green spots, regeneration frequency and the ratio of seedlings and callus were observed on NB medium (macroelements of N6 medium + microelements and vitamins of B5 medium) supplemented with a combination of antiphenolic compounds (5 mg L-1 silver nitrate + 5 mg L-1 ascorbic acid + 10 mg L-

1 cysteine). The shoot buds showed good proliferation and healthy plantlets on NB medium without hormone. These results indicated that the type and concentration of antiphenolic compounds influenced plant regeneration efficiency in the Thai rice cultivar. Keywords Antiphenolic compounds; Indica rice; Plant regeneration; Tissue culture



Chemical Constituents from the Combretum griffithii

Pattamaporn Worsawat, Panawan Moosophon*

Natural Products Research Unit, Department of Chemistry and Center of Excellence for Innovation in

Chemistry, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand *E-mail: [email protected]

Five known compounds, (2S)-4'-hydroxy-7,3'-dimethoxyflavan (1), 1-(2-hydroxy-4-methoxyphenyl)-3-(4-hydroxy-3-methoxyphenyl)propane (2), Griffitthane E (3) and a mixture of stigmasterol (4) and β-sitosterol (5) were isolated from the EtOAc extract of the roots of Combretum griffithii. Their structures were established on the basis of spectroscopic data. Compounds 1-3 showed cytotoxic activity against three cancer cell lines (KB, MCF-7 and NCI-H187). Keywords Combretum griffithii; Combretum; Cytotoxicity; Diarylpropane; Flavan



Isolation and Structure Elucidation of Bioactive Metabolites from Astraeus asiaticus

Preeyanuch Pimjuk1, Cherdchai Phosri2, Wiyada Mongkolthanaruk3, Sirirath Sodngam1*

1Natural Products Research Unit, Department of Chemistry, Faculty of Science, Khon Kaen University,

Khon Kaen, 40002, Thailand 2Department of Biology, Faculty of Science and Technology, Pibulsongkram Rajabhat University, Pitsanulok,

65000, Thailand 3Department of Microbiology, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand

*E-mail: [email protected] The isolation and purification of Astraeus asiaticus crude extracts by chromatographic methods yielded eight compounds. Their structures were elucidated by spectroscopic data, 1D (1H and 13C NMR), 2D NMR and compared to the data reported in literature. The results showed that the yielded compounds were ergosterol (1), astraodorol (2), hypaphorine (3), 5-hydroxyhypaphorine (4), triterpenes derivatives (5-7) and an ergosterol derivative (8). The preliminary bioactivities screening showed that compound 5 exhibited weak cytotoxicity against KB cancer cell line with an IC50 value of 38.78 µg/mL, however, it was also toxic to Vero cells with an IC50 value of 17.95 µg/mL.

Keywords Astraeus asiaticus; Cytotoxicity; KB cancer cell line



Chemical Constituents from the Roots of Melodorum fruticosum

Siriwat Hongnak, Santi Tip-pyang

Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand

*E-Mail: [email protected]

The genus Melodorum belongs to the Annonaceae family and comprise 55 species. Melodorum fruticosum Lour., locally known as “Lumduan”, is shrub and medium tree distributed in many parts of Thailand. In Thai traditional medicine recipe known as “Geasorn Thung Gao” which are used as tonic stimulant and mild cardiotonic. Previous phytochemical study on this plant revealed the presences of various flavonoids, triterpenoids and heptenes. The phytochemical investigation of the dichloromethane crude extract from the roots of M. fruticosum afforded four flavonoids, pinocembrin(1), chamanetin(2), isochamanetin(3), dichamanetin(4), a triterpenoid, polycarpol(5), and three heptenes, melodorinol(6), acetylmelodorinol(7) and (4Z)-6-benzoyloxy-7-heptadien-4-olide(8). Their structures were identified on the basis of spectroscopic evidence as well as comparison with the previous data. In addition, all of these compounds will be evaluated for their biological activity.

Keywords Melodorum fruticosum; Annonaceae; flavonoid; heptene; triterpenoid



Chemical Constituents from the Roots of Glycosmis pentaphylla

Saowapa Jitphusa1, Vatcharin Rukachaisirikul1, Potchanapond Graidist 2, Yaowapa Sukpondma1*


Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand 2Department of Biomedical Sciences, Faculty of Medicine, Prince of Songkla University,

Songkhla 90112, Thailand *E-mail: [email protected]

Phytochemical investigation of Glycosmis pentaphylla ’s roots led to the isolation of six known alkaloids: glycozolidine (1), glycozoline (2), O-methylglycocitrine II (3), aborinine (4), O-methylglycosolone (5) and atanine (6). Their structures were characterized by analysis of spectroscopic data and comparison of the 1H and 13C NMR data with their previously reported. Keywords Glycosmis pentaphylla; Rutaceae; Anticancer



Chemical Constituents and Antioxidant Activities of Hexane and Ethyl Acetate Extract from Leaves of Dodonaea viscosa

Nawara Samritsakulchai1, Sirima Naksongkeaw1, Norman Mangnall2, Stefan Kohlbacher1

and Nipawan Pongprom1*

1Department of Chemistry and Centre of Excellence for Innovation in Chemistry, Faculty of Science, Ubon Ratchathani University, Ubon Ratchathani, 34190, Thailand

2Faculty of Agriculture, Ubon Ratchathani University, Ubon Ratchathani, 34190, Thailand *E-mail: [email protected]

Dodonaea viscosa is an evergreen woody herb found in many part of the world, most notably in Australia. Recent phytochemical studies have confirmed a positive correlation between several groups of active constituents and the traditional usage. In this work, we report the studies of D. viscosa planted in Ubon Ratchathani, Thailand. The leaves were collected in May 2013 and dried in the oven at 70 oC for 12 h. We primarily investigated the antioxidant activities of tea from D. viscosa including total phenolic contents and total flavonoid content in comparison with the conventional teas. Moreover, the dried leaves were successively extracted with hexane and ethyl acetate at room temperature over 1 week. The crude extracts were evaporated under reduced pressure to afford green hexane and ethyl acetate extracts in 8.9 g and 81.9 g, respectively. Then, antioxidant activities of crude extracts were evaluated including total phenolic contents, DPPH Radical Scavenging Activities, ABTS Cation Radical Scavenging Activities and Ferric Reducing Antioxidant Power (FRAP). The results showed that tea from D. viscosa exhibited remarkable antioxidant activities. The crude extracts were then separated mainly by column chromatography on silica gel and subsequently eluted with gradient of three solvent (hexane, EtOAc, MeOH) by gradually increasing the polarity of the elution solvents system. The investigation of the hexane extract let to glyceryl trilinoleate. Moreover, the ethyl acetate extract let to some triterpenoids and viscosol (1).

O

O

HO

MeO

OH

OMe

OMe

(1)

Keywords Dodonaea viscosa; Antioxidant activities; Flavonoids



Cationic Lipids with Di(hydroxyethyl)amino Core Structure: An efficient synthesis and application for gene delivery

Nattisa Niyomtham1, Nuttapon Apiratikul2, and Boon-ek Yingyongnarongkul1*


Ramkhamhaeng University, Bangkapi, Bangkok 10240, Thailand 2Department of Chemistry, Faculty of Science, Siam University, Bangkok, 10160, Thailand

*E-mail: [email protected] Lipid-mediated delivery of DNA into cells holds great promise both for gene therapy and basic research applications. To development of potent synthetic non-viral vectors for gene therapy applications, the design and syntheses of new lipid systems with alternative structure types are essential. This paper describes the solid phase synthesis of symmetric cationic lipids processing di(hydroxyethyl)amino core structure with diverse polar heads and hydrophobic tails. The obtained cationic lipids were tested for gene delivery efficiency into mammalian cell alone and in combination with helper lipid using plasmid DNA encoding beta-Galactosidase as a reporter gene. The cationic lipid bearing spermine polar head and dodecyl tails showed highest transfection efficiency. The highest transfection activity of HeLa cells were achieved when liposome formulation with helper lipid, DOPE. The DNA/liposome weight ratio of 1:20 was optimal. In comparison to the commercially available transfection agent, LipofectamineTM2000, the di(hydroxyethyl)amino-based cationic lipid exhibited comparable transfection efficiency and lower toxicity to the tested cell. In conclusion, cationic liposomes with di(hydroxyethyl)amino core structure containing spermine head group demonstrated promising potential as gene carriers. Keywords Cationic lipids; Liposomes; Gene delivery; Solid phase synthesis



Chemical Constituents from the Roots of Dalbergia parviflora

Supornrat Maneechot1, Morakot Kaewpet1*, Vatcharin Rukachaisirikul1

1 Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science,

Prince of Songkla University, Songkhla 90112, Thailand * E-mail: [email protected]

Purification of the methanol extract of the roots of Dalbergia parviflora by chromatographic techniques afforded four known isoflavonoids, (3R)-mucronulatol (1), (3R)-5′-methoxyvestitol (2), tectorigenin (3) and isotectorigenin (4), together with two known cinnamylphenols, isomucronustyrene (5) and 4-cinnamyl-3-methoxycatechol (6). Their chemical structures were elucidated by spectroscopic methods, especially 1D and 2D NMR, and confirmed by comparison of their spectroscopic data with those previously reported. The absolute configuration of compounds 1 and 2 was established by comparison of the optical rotations with those previously reported. Keywords Dalbergia parviflora; Isoflavonoids; Cinnamylphenols



Phenalenone and Anthraquinone Derivatives from the Soil Fungus Penicillium herquei PSU-RSPG93

Athip Maha1, Thippaya Kongprapan1, Chittreeya Tansakul1, Souwalak Phongpaichit2,

Kawitsara Borwornwiriyapan2, Jariya Sakayaroj3, Vatcharin Rukachaisirikul1*




*E-mail: [email protected] Soil fungi have been recognized as a prolific source of biologically active metabolites. With an effort to search for bioactive compounds from the soil fungi, we have conducted our research with the fungus Penicillium herquei PSU-RSPG93 isolated from a soil sample collected from the Rajjaprabha Dam, Suratthani Province, Thailand. Chemical investigation of the broth and mycelial extracts from this fungus by various chromatographic techniques provided four known phenalenone derivatives, including herqueinone, deoxyherqueinone, sclerodin and lactone 8, together with one known anthraquinone, physcion. The structures were determined by analysis of spectroscopic data, especially 1D and 2D NMR data. The relative configurations were assigned using NOEDIFF data while the absolute configurations were identified on the basis of the similar optical rotations to the previously reported compounds.

Keywords Soil fungus; Penicillium herquei; Phenalenone derivatives; Anthraquinone



Calibration Model for Enhancing Efficiency of Pharmaceutical Quality Control in Production Scale by Near Infrared

Spectroscopy (NIRS) Technique

Tatiya Kittikhunpong1, Anongnat Somwangthanaroj1*, Nathaporn Hongsrisuk2

1Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok, 10330, Thailand

2Research and Development Institute the Government Pharmaceutical Organization 75/1 Rama VI Road, Ratchathewi, Bangkok 10400 Thailand

* E-mail: [email protected] The pharmaceutical industrial quality control includes quality control of raw materials, manufacturing processes and products. This study was using Near Infrared Spectroscopy (NIRS) technique to establish calibration model for controlling the quality of blend homogeneity, holding time and tablet products. Partial Least Squares (PLS) Cross-Validated regression which was used in calibration model showed a correlation with the reference values of active pharmaceutical ingredients (API) without lubricant sample set (R2) = 0.9979, standard error of cross validation (SECV) = 0.1011 and correlation with the reference values of API with lubricant sample set (R2)= 0.9938, SECV = 0.1512. In addition, the paired t-test at the 95% confident level was used to validate this model and compared the NIRS technique with high performance liquid chromatography (HPLC) technique. The results did not indicate any differences between the validation results by two methods. Therefore, the calibration model was accurate and suitable for using in the pharmaceutical quality control.

Keywords Near Infrared Spectroscopy; Partial Least Squares; Pharmaceutical blend



Bioactive Alkaloids from Apocynaceae Plants: Alstonia macrophylla and Kopsia aborea

Sarot Cheenpracha1,*, Thunwadee Ritthiwigrom2, Achara Raksat3, Surat Laphookhieo3

1School of Science, University of Phayao, Maeka, Muang, Phayao 56000, Thailand

2Department of Chemistry, Faculty of Science, Chiang Mai University, Sutep, Muang, Chiang Mai 50200, Thailand

3Natural Products Research Laboratory, School of Science, Mae Fah Luang University, Tasud, Muang, Chiang Rai 57100, Thailand

*E-mail: [email protected] Chemical investigation of alkaloidal extracts of Alstonia macrophylla bark and Kopsia aborea twigs led to the isolation and identification of two new nitrogenous derivatives, alstoniaphyllines A (1) and B (2), a new indole alkaloid, alstoniaphylline C (4), and ten known alkaloids. Alstonisine exhibited antiplasmodial activity against Plasmodium falciparum, with an IC50 of 7.6 μM.

Keywords: Alkaloid; Alstonia macrophylla; Kopsia aborea; Antiplasmodial activity



Inhibition of Acetylcholinesterase Activity by the Indole Alkaloid, Alstonerinal

Manutchaya Komek1,*, Sunisa Tongseenak1, Waraporn Takampanya1,Thunwadee Ritthiwigrom2,

Surat Laphookhieo3, Sarot Cheenpracha1

1School of Science, University of Phayao, Maeka, Muang, Phayao 56000, Thailand 2Department of Chemistry, Faculty of Science, Chiang Mai University, Sutep, Muang, Chiang Mai 50200,

Thailand 3Natural Products Research Laboratory, School of Science, Mae Fah Luang University, Tasud, Muang,

Chiang Rai 57100, Thailand *E-mail: [email protected]

Seven indole alkaloids: alstonerinal (1), alstophyllal (2), talcarpine (3), macrocarpine B (4), alstonisine (5), Nb-demethylalstophylline oxindole (6) and 10-methoxycathafoline (7), isolated from the barks of Alstonia macrophylla, were evaluated to their inhibitory properties for AChE enzyme. The structures of these alkaloids were determined on the basis of their spectroscopic data. Acetylcholinesterase activity was measured using TLC bioautography and a colorimetric method in the presence or absence of the extracts. Compound 1 exhibited weak in vitro acetylcholinesterase inhibitory activity (1 μg), in comparison with the standard compound galantamine. Keywords Indole alkaloids; Alstonia macrophylla; Acetylcholinesterase activity



Surface pH of Novel Chitosan Modified Glass-Ionomer Cement During Setting

Sareepah Manmoo1*, Ureporn Kedjarune-Leggat1, Chanothai Hengtrakool2

1Department of Oral Biology and Occlusion, Faculty of Dentistry, Prince of Songkla University, Songkhla, 90112, Thailand

2Department of Conservative Dentistry, Faculty of Dentistry, Prince of Songkla University, Songkhla, 90112, Thailand

*E-mail:[email protected] Glass ionomer cement (GICs) have been used for a range of applications such as using for a variety of purposes in dental restorative procedures, bone cement and also for bone reconstruction. The modification of GIC with chitosan in attempt to have cement being able to sustain release protein with its retained bioactivity. However, this modification may alter acid–base setting reaction of cement. This study aimed to investigate surface pH of novel chitosan modified glass-ionomer cement during setting. GIC used in this study was a conventional type (Lining cement, GC®). The powder of this novel cement was composed of calcium fluoroaluminosilicate glass incorporated thoroughly (w/w) with 15% of chitosan selected from two molecular weights either 545 kDa(F) or 62 kDa(K) and 5% bovine serum albumin(A). The liquid part was contained 40% poly(acrylic acid). Five groups according to the different compositions of the powder were assigned as following: G, GF, GFA, GK, and GKA. Four disc specimens per group (10 mm in diameter, 1 mm in height) were prepared. A flat-headed combination electrode was used to measure the pH of each specimen throughout the experiments. The electrode was placed on the surface of the setting cement and recorded every minute for up to 5 min and at 10, 20, 30, 60, 90, 120 min. The pattern of pH changes of all specimens was the same which means of pH at 1 min was 3.46±0.17, 3.42±0.07, 3.25±0.35, 3.21±0.15, 3.06±0.22 in G, GF, GFA, GK, GKA respectively. The pH of all specimens tends to increase rapidly since 6 min after setting. There was no significant difference of pH among different GICs groups. (ANOVA with repeated measure, P>0.05) This study suggested that the modifications of GIC by adding chitosan and albumin may not have effect on setting reaction of the cements. Keywords Surface pH; Glass-ionomer cement; Chitosan



Chemical Constituents from the Ripe Fruits of Garcinia cowa

Teerayut Sriyatep* and Surat Laphookhieo

Natural Products Research Laboratory, School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand


Phytochemical investigation of Garcinia cowa ripe fruits led to the isolation and identification of seven known xanthones including fuscaxanthone C (1), β-mangostin (2), α-mangostin (3), butyraxanthone A (4), 9-hydroxycalabaxanthone (5), dulcisxanthone F (6) and 3-O-methylmangostenone D (7). Their structures were elucidated by the combination of spectroscopic methods as well as comparison with previous literature data.

Keywords Garcinia cowa, Clusiaceae, Xanthone



Purification of Recombinant Human Epidermal Growth Factor (rhEGF) by Ni-NTA Resin Technique for Primary Human

Gingival Keratinocytes Proliferation

Supreya Wanichpakorn1*, Ureporn Kedjarune-Laggat 1, Wilaiwan Chotigeat 2, Panchalika Deachamag2

1 Department of Oral Biology and Occlusion, Faculty of Dentistry, Prince of Songkla University, Hat Yai,

Songkhla, 90112, Thailand. 2 Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla

University, Hat Yai, Songkhla, 90112, Thailand. *E-mail: [email protected]

Human epidermal growth factor (hEGF) is a growth factor which has a mitotic activity. This growth factor is found in kidney, liver and is also essential for the epithelial cells proliferation in vivo and in vitro. Nowadays, recombination human epidermal growth factor (rhEGF) can be cloned and using to stimulate oral mucosal cells proliferation. Thus, the culture of primary keratinocytes from oral tissue should be supplemented with optimal conditioned medium which included rhEGF in culture medium. The objectives of this study were to purify rhEGF with Ni-NTA resin technique and studying its bioactivity. The rhEGF was cloned in pET-28 and transformed in Escherichi coli BL21 (DE3). The rhEGF was purified by Ni-NTA resin, the amount of rhEGF was quantitated by ELISA technique and the bioactivity of this protein was assessed using MTT assay in primary human gingival keratinocytes (PGK). The results showed that rhEGF has 11 kDa due to the attached 6 histidine amino acids (rhEGF-His). This method gave 79.66 µg/ml of rhEGF-His from 131.21 µg/ml of total protein which was 60.70 % yield. From MTT assay, rhEGF-His has the same bioactivity or can proliferate PGK as the commercial rhEGF.

Keywords Epidermal growth factor (EGF); recombinant human epidermal growth factor (rhEGF); Ni-NTA resin; gingival keratinocytes



Phloroglucinols from the Leaves of Rhodomyrtus tomentosa

Asadhawut Hiranrat*, Wanrudee Hiranrat and Chamaiporn Tongkamkeaw

Natural Products Research Laboratory, Faculty of Science, Thaksin University, Phatthalung, 93110, Thailand

*E-mail: [email protected] This research was focused on the isolation and structural determination of the chemical constituents from the hexane extract of Rhodomyrtus tomentosa leaves. Chromatographic investigation of this extract had led to the isolation of two phloroglucinols. These structures were exhaustedly elucidated using spectroscopic data especially 2D NMR techniques.

Keywords Phloroglucinol; Myrtaceae; Rhodomyrtus tomentosa; Spectroscopic data



In silico Study of Curcumin Analogs as HER2 Kinase Inhibitors

Wannarat Yim-im1, Apichart Suksamrarn2, Orathai Sawatdichaikul3*, Kiattawee Choowongkomon4*

1Interdisciplinary Graduated Program in Genetic Engineering, Graduated Program, Kasetsart University, Bangkok,10900,Thailand

2Department of Chemistry, Faculty of Science, Ramkhemhaeng University, Bangkok, 10240, Thailand 3Institute of Food Research and Product Development, Kasetsart University, Bangkok, 10900, Thailand

4Department of Biochemistry, Faculty of Science, Kasetsart University, 10900, Thailand *E-mail: [email protected] (O. Sawatdichaikul), [email protected] (K. Choowongkomon)

HER2 overexpression can be found in many types of cancer such as lung or breast cancer. As HER2-positive cancer is considered aggressiveness and poor prognosis, direct inhibitors against this protein have been used for cancer treatment. Recently, curcumin has shown the anticancer property by inhibiting a wide range of proteins including kinases involved in cell proliferation and cell survival. In previous study, high dose usage of curcumin has no negative effect on kidney and liver, which will be beneficial to be an anticancer drug with low side effects. This study investigated how curcumin and curcumin analogs interacted with HER2 kinase by using molecular docking. The results showed that two benzene rings of curcumin could form hydrophobic interactions with the hydrophobic pocket and adenine region. The carbonyl group at the linkage chain could form a hydrogen bond with the key residue Thr862. With crucial interactions with HER2 kinase, curcumin could be a potential candidate for HER2 inhibitor. Because of its flexibility, the modified linkage chain of curcumin analogs could promote H-bond formation between the hydroxyl groups and Ser728, Ser783, Cys805 or Asp808, which might increase affinity of protein-ligand interactions. Therefore, this suggested that to enhance binding affinity HER2 inhibitors should have benzene rings for hydrophobic interactions and hydroxyl groups for H-bond formations. The further in vitro and in vivo studies are still required to evaluate the efficacy of curcumin and curcumin analogs against HER2 kinase. keywords HER2 kinase inhibitor; Curcumin; Molecular docking



Chemical Constituents from the Roots of Cratoxylum formosum ssp. pruniflorum

Achara Raksat,1,* Wisanu Maneerat,1 Sarot Cheenpracha,2 Surat Laphookhieo1

1Natural Products Research Laboratory, School of Science, Mae Fah Luang University,

Chiang Rai 57100, Thailand 2School of Science, University of Phayao, Maeka, Muang, Phayao 56000, Thailand

*E-mail: [email protected] Chemical investigation from the roots of Cratoxylum formosum ssp. pruniflorum led to the isolation and identification of 11 known compounds: one anthraquinone, vismiaguinone A (1), nine xanthones: pruniflorone K (2), formoxanthone A (3), formoxanthone B (4), 1,7-dihydroxy-8-methoxyxanthone (5), 1,7-dihydroxyxanthone (6), 3-hydroxyblancoxanthone(7), cratoxyarborenone F (8), 7-hydroxy-1,2,3,8-tetramethoxyxanthone (9) 1,2,3,4,5-pentamethoxyxanthone (10), and one flavonoid: 5,7-dihydroxyflavanone (11). The structures were elucidated by spectroscopic methods.

Keywords Cratoxylum formosum; Clusiaceae; Xanthones; Anthraquinone; Flavonoid



The Recovery Effect of The Vascular Endothelial Growth Factor in 2-Hydroxy-Ethyl Methacrylate-treated Human Dental

Pulp Cells.

Chunyanut Wongkhum1*, Ureporn Kedjarune-Leggat2, Wilaiwan Chotigeat1

1Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University,Songkhla, 90112, Thailand

2Department of Oral Biology and Occlusion, Faculty of Dentistry, Prince of Songkla University, Songkhla, 90112, Thailand

*E-mail: [email protected] 2-Hydroxy-ethyl methacrylate (HEMA) is one of the major residual monomers released from resin-based dental restorative materials. HEMA is cytotoxic and causes cells apoptosis. This study examined the protective effect of vascular endothelial growth factor (VEGF) against apoptosis from HEMA in human dental pulp cells (HDPCs) using MTT assay. Primary HDPCs were cultured from dental pulp tissue by enzymatic method. The cell suspensions were seeded at 8 x 103 cells per well in 96-well plates in humidified atmosphere of 5% CO2 at 37°C. After 24 h, the HDPCs were exposed to various concentrations of VEGF, ranging from 50ng/ml to 50µg/ml, which had already been mixed in culture medium (œMEM with supplements) for 48 h before MTT assay. The viable cells of HDPCs after exposed to VEGF at 100 ng/ml showed the highest percentages (113.99%) when compared with the control (no VEGF). This concentration was selected to be the optimal concentration to determine the protective effect of VEGF against HEMA. The HDPCs were treated with 4mM HEMA mixed in culture medium for 1 h prior the medium supplemented with 100ng/ml of VEGF was replaced and left for 23 h. After that, the fresh media was replaced and incubated for 48 h before the MTT assay. The results revealed that HEMA-treated HDPCs in medium supplemented with VEGF had a significantly higher number of viable cells (137.64%) than the HEMA-treated cells alone (15.55%, one way-ANOVA, P<0.05). This study suggests that VEGF has an ability to recover the cytotoxic effect of HEMA-treated pulp cells. Keywords 2-Hydroxy-ethyl methacrylate (HEMA); Vascular endothelial growth factor (VEGF) ; Resin modified glass ionomer cement (RMGIC) ; Human dental pulp cells (HDPCs)



Effective of Plant Extract on Inhibition of Contaminated Fungi in Tissue Culture Laboratory, Science Department, RMUTSV,

Thailand.

Jarin Poodngam*, Waida Praju, and Amornpan Kaewsrinuan

Department of Science, Faculty of Science and Technology, Rajamangala University of Technology, Nakhonsithammarat, 80110, Thailand.


Fungi contamination is a constant problem, which often compromise development of all in vitro technique. This study aimed at investigating the source of fungi contamination in tissue culture laboratory in Science Department of RMUTSV, Thailand and used plant extract for control them. Three kinds of contaminated fungi were isolated from PTC room include : WAR 556, WSR 114 and WSR 115 strain found at preparation room, transfer room and culture room. The laboratory wall, the laboratory indoor air and table also harbored most of the contaminating fungi. Based on the observation all of them were contamination in culture bottle too. The plant crude extracts of Zingiber zerumbet (L.) Smith., Piper sarmentosum Roxb., and Eupatorium odoratum L., at the concentration of 1,000, 2,000, 3,000, 4,000, 5,000 and 6,000 ppm were used to control 3 contaminated fungi. Results found that the crude extract of P. sarmentosum Roxb at 2,000 ppm showed highest inhibition activity against three fungal contaminated at 57, 31 and 39 %, respectively. Keywords : Plant tissue culture; Fungal contamination; Anti fungi



Antioxidant and Ant Repellent Activities of Thai Propolis from Stingless Bees

Isara Mongkolpichayarak1,2, Amphawan Pa-Toh1, Jenjeera Banthoengchai1, Panyapon Muangma1, Orawan Duangphakdee3, Oraphin Chantarasriwong1*

1Department of Chemistry, Faculty of Science, King Mongkut’s University of Technology Thonburi,

Bangmod, Thungkru, Bangkok 10140, Thailand 2Program in Petrochemistry and Polymer Science, Faculty of Science, Chulalongkorn University, Bangkok

10330, Thailand 3King Mongkut’s University of Technology Thonburi, Ratchaburi Campus, Bangkok 10140, Thailand

*E-mail: [email protected] Propolis from stingless bees, Trigona apicalis, was collected from Ratchaburi Province, Thailand. It was extracted with different polarity solvents: hexane, dichloromethane (CH2Cl2), ethyl acetate (EtOAc) and methanol (MeOH) to evaluate their antioxidant and ant repellent activities. The DPPH radical scavenging assay indicated that all extracts exhibited potent antioxidant activity (IC50 1.94-5.96 mg/mL) with the MeOH extract showing the highest antioxidant activity. In the Folin-Ciocalteau assay, the MeOH extract showed the highest total phenolic content value (621.04 ± 9.7068 mg gallic acid equivalent (GAE)/1 g sample) followed by EtOAc extract (561.86 ± 6.5107 mg GAE/g), CH2Cl2 extract (398.58 ± 1.6127 mg GAE/g) and hexane extract (332.47 ± 4.7380 mg GAE/g). In addition, all extracts displayed 100% repellency against the weaver ants, Oecophylla smaragdina, at 50 mg of extract used. These results indicated that propolis is a good source of natural antioxidant and ant repellent agents. Keywords Propolis; Stingless bees; Trigona Apicalis; Antioxidant activity; Ant repellent



Anti-Cholangiocarcinoma of Alkaloids and Their Analogues from Tiliacora triandra

Archawin Nakaew,1 Somkid Janeklang,2 Sopit Wongkham2 and Apichart Suksamrarn1*

1Department of Chemistry and center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok 10240, Thailand.

2Department of Biochemistry, Liver Fluke and Cholangiocarcinoma Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.


Cholangiocarcinoma (CCA) is a fatal cancer arising from bile duct epithelia. CCA is poor prognosis with a very low five-year survival. The cancer has low response to chemotherapeutics and recurrence of cancer is commonly found in CCA patients after hepatic resection. The effects of plant derived compounds, such as biscoclaurine alkaloid (cepharanthine), xanthone (gambogic acid), polyphenolic compound (curcuminoids), have been reported for CCA chemotherapy and chemoprevention. In this study, the cytotoxic effect of bisbenzylisoquinoline alkaloids with structurally related to cepharanthine, was investigated. Bisbenzylisoquinoline alkaloids were extracted and purified from a Thai medicinal plant, Tiliacora triandra (Colebr.) Diels or “Ya nang” of the Menispermaceae family. It is of interest to investigate whether the analogues obtained from chemical modification of the isolated alkaloids would inhibit more potent anti-cholangiocarcinoma activity. Biological evaluation of the isolated alkaloids and their analogues revealed that a number of these compounds exhibited high anti-cholangiocarcinoma activity. Keywords Tiliacora triandra; Bisbenzylisoquinoline alkaloids; Structural modification; Anti-cholangiocarcinoma



In Vitro Total Phenolics and Antioxidant Activity of Various Organic Extracts from Leaves, Branches, Fruits and Seeds of

Cynometra cauliflora L.

Wanrudee Hiranrat*, Asadhawut Hiranrat and Nipaporn Khunmuang

Natural Products Research Laboratory, Faculty of Science, Thaksin University, Phatthalung, 93110, Thailand

*E-mail: [email protected] The in vitro total phenolics and antioxidant capacity of various organic extracts from leaves, branches, fruits and seeds of Cynometra cauliflora L.were studied. The dry powder of all samples were extracted with hexane, dichloromethane, methanol, 80% methanol and 50% methanol. The twenty extracts, namely hexane, dichloromethane, methanol, 80% methanol and 50% methanol extract of leaves (LH, LD, LM, LM80, LM50), branches (BH, BD, BM, BM80, BM50), fruits (FH, FD, FM, FM80, FM50) and seeds (SH, SD, SM, SM80, SM50), respectively. The total phenolic contents and antioxidant activity of all extracts were evaluated using the Folin-Ciocalteu method and DPPH assay, respectively. The phenolic contents of different extracts as gallic acid equivalents were found to be highest in LM extract (392.24 mg/g) followed by LM80, BM, BM80, LM50, BM50, FM, FM80, DM, SM80, FM50, SM50, LD, BD, FD, SD, LH, BH, FH, and SH. The antioxidant activity using butylated hydroxytoluene (BHT) as a standard showed the relative results with the phenolic contents. It was found that LM and BM extracts exhibited activity with IC50 values of 1.07 and 12.22 μg/mL, respectively, stronger than the standard BHT (IC50 13.89 μg/mL). This study suggests that the methanolic extract of leaves can potentially be used as a readily accessible source of natural antioxidants. Keywords Total phenolic content; Antioxidant activity; Cynometra cauliflora L.



Chemical Constituents and Structural Characterization of Compounds from Makwaen Fruits

(Zanthoxylum limonella Alston)

Pornthip Bubpawan, Anusorn Vorasingha, Chaturong Suppharpprom, Vijitr Udeye *

Department of Chemistry. Faculty of Science, Naresuan University, Phitsanulok 65000, Thailand *E-mail: [email protected]

Zanthoxylum is a group of plants belonging to Rutaceae Family. There are over 200 species including Makwaen (Zanthoxylum limonella Alston) which was found in the northern and the central part of Thailand. Makwaen was widely used in food product because of its medicinal properties such as anti-inflamatory, anti-fungal, anti-bacterial and anti-oxidant. In this work, the fruits of Makwaen were extracted with ethanol to get crude extract in 14.6 %. Then, the crude was purified by chromatography techniques using various materials such as sephadex LH-20, silica gel and C-18 on HPLC. The collected fraction was characterized with NMR, IR and UV- Vis spectroscopy. The preliminary results from 1H-NMR showed characteristic of terpenoid as the major components. Keywords Makwaen; Anti-inflamatory; Anti-bacterial; Anti-fugal; Anti-oxidant; Rutaceae;Terpenoid



Solid Phase Synthesis and DNA Binding Affinities of Novel Synthesized Cationic Lipids

Widchaya Radchatawedchakoon1*, Nattisa Niyomtham2, Nuttapon Apiratikul3,

Uthai Sakee1, Boon-ek Yingyongnarongkul2


Mahasarakham University, Maha Sarakham, 44150, Thailand 2Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science,

Ramkhamhaeng University, Bangkok, 10240, Thailand 3Department of Chemistry, Faculty of Science, Siam University, Bangkok, 10160, Thailand

*E-mail: [email protected]; [email protected] Cationic lipid is a nonviral vector for DNA delivery. Because of the cationic lipid has low toxicity and immunogenicity, many researchers interest to discover new high efficient carriers. The solid phase synthesis of eight novel cationic lipids (A-B)(W-Z) having amide and carbamate bond is presented. The structure of synthesized lipids was characterized by IR, 1H NMR and MS techniques. DNA binding affinities of the synthesized lipids were determined by gel retardation assay. The results indicated that the synthesized lipids bound efficiently with DNA when the weight ratio of DNA/lipids (1:5, 1:10, 1:20) was increased. These lipids may have potential to be used as nonviral vector to deliver DNA into mammalian cells.

OHW

Wang resin

OW NH

O HN

NH2

OW NH

ON

NH

R1OO

NNH2

R2O

NNH

Lipids (A-B)(W-Z)

R1OO

N

R2O

NH2 H2N

R1

A

B

R2

OW

X

Y

Z

O

Keywords Solid phase synthesis; Cationic lipid; Nonviral vector



Effect of Chitosan as Antifungal on Para Rubber Sheet in Nakhon Si Thammarat Thailand

Duenpen Artthaisong1*, Jarin Poodngam1 and Kwanruethai Boonsong2

1Department of Science, Faculty of Science and Technology,

Rajamangala University, Nakhon Sri Thammarat 80110, Thailand 2Department of Rubber and Polymer technology, Faculty of Science and Technology,

Rajamangala University, Nakhon Sri Thammarat 80110, Thailand *Corresponding author: [email protected]

The fungi was isolated from air dry para rubber sheet and smoked para rubber sheet in Nakhon Si Thammarat, with a total of 56 isolates. The 37 isolates were found in air dry para rubber sheet and 19 isolates in smoked para rubber sheet. Among the 56 isolates, Aspergillus, Rhizopus, Mucor, Penicillium and Telaromyces were the most frequently noticed. This study used chitosan extracted from the skrimps’ skeleton. The study aimed to examine the effectiveness of the chitosan as an antifungal at 0.0, 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 % of concentration. The result found that 2.0 % chitosan could inhibit the investigated fungus; Aspergillus, Rhizopus, Mucor, Penicillium and Telaromyces. Keywords : Antifungal; Chitosan; Para Rubber Sheet



Chemical Constituent from Casearia graveolens Twigs

Pornphimol Meesakul*, Wisanu Maneerat, Surat Laphookhieo

Natural Products Research Laboratory, School of Science, Mae Fah Luang University, Chiang Rai, 57100 Thailand


The first phytochemical investigation of Casearia graveolense twigs led to the isolation and identification of three known clerodane diterpenes including, casearvestrin B (1) casearvestrin C (2) and casearlucin G (3). Their structures were elucidated by extensive 1D and 2D NMR spectroscopy.

Keywords Casearia graveolense; Flacoutiaceae; clelodane diterpene



Toxicity of Alpinia galanga (Zingiberaceae) Rhizome Extracts Against Spodoptera litura (Lepidoptera: Noctuidae)

Anupap Puangsomchit1, Vasakorn Bullangpoti2, Wanchai Pluempanupat1,*

1Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand

2Department of Zoology, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand *E-mail: [email protected]

The tropical armyworm (Spodoptera litura) is an economically pest in Thailand. Chemical pesticides have been used for this pest management; however, they have been great concern due to toxicity to human, environmental residue and bioaccumulation through food chain. The objective of the study is to develop an alternative strategy by using botanical pesticide for the control of the pest. The rhizomes of Alpinia galanga were extracted sequentially by soaking in hexane, dichloromethane, ethyl acetate and methanol, respectively by soaking at room temperature for seven days. Each crude extract was examined the toxicity against the second instars of S. litura using topical application method. Dichloromethane crude extract showed the most effective (LD50 3177 and 2099 ppm after treatment 24 and 48 h, respectively). Keywords; Alpinia galanga; Toxicity; Spodoptera litura; Insecticidal activity; Botanical pesticide



Dihydrocinnamic Acid Derivatives and Coumarin Glycosides from Micromelum minutum

Poolsak Sahakitpichan1, Waraporn Tanpatanan1, Somsak Ruchirawat1, Nitirat Chimnoi1, Tripetch

Kanchanapoom1,2*

1Chulabhorn Research Institute, Kamphaeng Phet 6, Lak Si, Bangkok, 10210, Thailand

2Department of Chemistry, Faculty of Pharmaceutical Science, Khon Kaen University, Khon Kaen, 40002, Thailand

*E-mail: [email protected] Three new dihydrocinnamic acid derivatives (micromelumosides A-C) and a new coumarin glycoside (micromelumoside D) were isolated from the aerial portions of Micromelum minutum (Family Rutaceae) together with scopolin, haploperioside, (S)-peucedanol 7-O-β-glucopyranoside, (R)-peucedanol 7-O-β-glucopyranoside, vanilloloside, citroside B, kaempferol 3-O-rutinoside, kaempferol 3-O-sophoroside. Structure determinations of these compounds were based on analyses of chemical and spectroscopic data, including 1D and 2D NMR. Further investigations are in progress. Keywords Micromelum minutm; Rutaceae; dihydrocinnamic acid derivatives; coumarin glycoside



Chemical Constituent from Aerial Parts of Sphearanthus Senegalensis

Watchara Sangsopha, Keeratika Meesuk, Kitiyawadee Sittiworadet, Ratsami Lekphrom*

Natural Products Research Unit, Department of Chemistry, and Center of Excellence for innovation in

Chemistry, Faculty of Science, Khonkaen University 4002, Thailand *E-mail: [email protected]

Chemical investigation of extracts from the aerial parts of Sphearanthus seneqalensis (Asteraceae), named "Matomsuea" in Thai. Their structures were identified by spectroscopic methods and compared with those reported in the literature. Our research resulted in the isolation of six compounds, stigmasterol (I), 7-hydroxyfrullanolide (II), ilicic acid (III) and frullanolide (IV) while compound V and VI are in the progress. Keywords Asteraceae; Eudesmanolide; Frullanolide



The Efficiency of Crude Enzymes for Cell Lysis and Polyhydroxyalkanoate Recovery

Kanokphorn Sangkharak*, Tewan Yunu, Nisa Pichid and Piyaporn Vangsirikul

Research Center of Energy and Environment, Department of Chemistry, Faculty of Science, Thaksin

University, Phatthalung 93110, Thailand *E-mail: [email protected]

The partial purified enzymes including bromelain, cellulase, lysozyme and papain were isolated from agricultural sources and utilized for cell lysis and recovery of polyhydroxyalkanoate (PHA). Among four enzymes, bromelain gave high efficiency in cells lysis (54% of cell lysis). Therefore, bromelain was selected for the PHA recovery process. The recovery and purification of PHA was determined in Cupriavidus necator TISTR1095. Crude bromelain was suitable for PHA purification and gave high PHA purity (96.5%). The results were also compared with commercial bromelain. Crude enzyme gave lower efficiency in PHA recovery process, but the value was not significant different. Therefore, high PHA purity means the partial purified bromelain had a great ability to recover and purify of PHA. In addition, the purify process need a simple method and low operation cost make it a promising method for industrial applications. Keywords Bromelain; Crude enzyme; PHAs; Polyhydroxyalkanoates; Recovery process



The Extraction of Polyhydroxybutyrate from Alcaligenes eutrophus by Pineapple Juice

Piyaporn Vangsirikul, Tewan Yunu, Nisa Pichid and Kanokphorn Sangkharak*

Research Center of Energy and Environment, Department of Chemistry, Faculty of Science, Thaksin

University, Phatthalung 93110, Thailand *E-mail: [email protected]

The study of polyhydroxybutyrate (PHB) extraction from Alcaligenes eutrophus by pineapple juice was investigated. The result revealed that pineapple juice from different age (100-200 days of cultivation) had protein content and bromelain activity in the range of 98.7-123.46 mg and 1.41-1.64 unit/mL, respectively. However, pineapple age did not affected on protein content and bromelain activity. Therefore, pineapple juice was utilized for PHB extraction from A. eutrophus cultivated in Nutrient broth (NB) contained glucose (40 g/L) as carbon source. The extraction PHB by pineapple juice was optimized. The optimum condition contained 1 g of bacterial cell with 3 mL of pineapple juice under 12 h of incubation time. The highest PHB purity (86.18%) was obtained under suitable condition. The structure of isolated polymer was confirmed by Infrared spectroscopy compared with commercial PHB. The result indicated that isolated polymer was PHB. The thermal properties were also observed. A glass transition point (Tg) was 1.02 ºC, a crystallization temperature on heating (Thc) was 35.683 ºC, crystallization temperature on cooling (Tcc) was 94.316 ºC and melting temperature (Tm) was 168.683 ºC. Keywords Alcaligenes eutrophus; Bromelain; FTIR; Nutrient Broth; Polyhydroxybutyrate



Antioxidant Activity and Total Flavonoid Contents of Garcinia schomburgkiana Extracts

Imron Meechai1, Worrapong Phupong1*, Warangkana Chunglok2 and Puttinan Meepowpan3

1School of science, Walailak University, Thasala, Nakhonsi Thammarat, 80161 2School of Allied Health Sciences and Public Health, Walailak University, Nakhonsithamrat, 80161

3Department of Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai, 50200 *E-mail: [email protected]

Antioxidant activities of 12 extracts from Garcinia schomburgkiana (Ma-dan) were performed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and their total flavonoid contents were measured by the aluminum chloride colorimetric method. Their antioxidant activities and total flavonoid contents were expressed as vitamin C equivalent antioxidant capacity (VCEAC) in mg Vitamin C /g extracts and quercetin equivalent in mg quercetin /g extracts, respectively. The branches acetone extract revealed the best antioxidant activity with VCEAC 114.114 ± 0.510 mg/g, and the high total flavonoid contents with quercetin equivalents at 414.486 ± 15.991 mg/g. The positive correlation between the total flavonoid contents and VCEAC values of the extracts (r = 0.6268) revealed that the flavonoid contents played an important role in the radical scavenging. It is an interesting subject for further phytochemical investigation. This result will provide the information of the antioxidant benefit of Ma-dun, Thai traditional fruit, for the consumers Keywords Garcinia schomburgkiana; Anti-oxidant activity; Total flavonoid contents



Chemical Composition and Antioxidant Capacity of An Ethanolic Extract from Ripe Fruits of Namdang (Carissa

carandas Linn.)

Pakinee Kangvalnarakul, Kwanpicha Suphapthai, Kanok-on Rayanil, Oranart Suntornwat*

Department of Chemistry, Faculty of Science, Silpakorn University, Nakornpathom, 73000, Thailand *E-mail: [email protected]

Carissa carandas is one of the widely used medicinal plants belonging to the family Apocyanaceae which have yielded numerous pharmacologically active compounds including antidiabetics and antioxidants. Most of the previously available data from namdang were obtained from leaves and unripe fruits. In this study, an ethanolic extract of ripe fruits was prepared and was subsequently separated sequentially into hexane, ethyl acetate, n-butanol, and water fractions. The fractions were then analysed by siliga gel thin layer chromatography. Many phytochemicals including alkaloids, flavonoids and terpenoids were detected especially in the hexane and ethyl acetate fractions. Antioxidant capacity and phenolic content were detected in all fractions tested with the highest level in the ethyl acetate fraction (Trolox equivalent = 588.23±0.01 mg/g extract) and the water fraction (40.70±0.01 mg gallic acid/g extract), respectively. Anthocyanin was mainly found in the n-butanol and water fractions. Further fractionation of the hexane fraction by Flash column chromatography afforded four bioactive compounds identified as β-sitosterol, stigmasterol, lupeol and betulinate ester. This work is part of an ongoing study to resolve the nature and extent of, in a particular, the antioxidant properties of Carissa carandas Keywords: Carissa carandas; antioxidant capacity; anthocyanin



Steroids from the Fungus Gymnoascus sp.

Sasiwimon Kitchawalit,1 Kwanjai Kanokmedhakul,1 Somdej Kanokmedhakul,1,* Kasem Soytong2

1Natural Products Research Unit, Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand

2Department of Plant Pest Management, Faculty of Agricultural Technology, King Mongkut’s Institute of Technology Ladkrabang, Bangkok 10520, Thailand

*e-mail: [email protected]

Eight steroids, (22E,24R)-ergosta-4,7,22-triene-3-one (1), ergosta-4-6,8(14),22-tetraene-3-one (2), ergosterol (3), cyathisterone (4), 24(R)-5a,8a-epidioxyergosta-6-22-diene-3β-ol (5), herbarulide (6), calvasterol A (7), and calvasterol B (8) were isolated from the fungus Gymnoascus sp.. Structures of these isolate compounds were identified based on spectroscopic method. Keywords Gymnoascus sp., steroids, isolation



Lanostane Triterpenoid Lactones from Ganoderma sp. KM01

Waranya Lakornwong,1 Kwanjai Kanokmedhakul,1 Somdej Kanokmedhakul,1,* Phoutthasone Sibounnavong,2 Kasem Soytong3

1Natural Products Research Unit, Department of Chemistry and Center for Innovation in Chemistry, Faculty

of Science, Khon Kaen University, Khon Kaen 40002, Thailand 2Faculty of Agricultural Technology, National University of Laos, Na Bong, Vientaine, Lao PDR

3Department of Plant Production Technology, Faculty of Agricultural Technology, King Mongkut’s Institute of Technology Ladkrabang, Ladkrabang, Bangkok 10520, Thailand


Investigation of the chemical constituents from culture fungal biomass of mushroom Ganoderma sp. KM01 afforded five known lanostane triterpenoid lactones. Structures of these compounds were established on the basis of spectroscopic evidence as colossolactone I (1), schisanlactone A (2), colossolactone B (3), colossolactone E (4), colossolactone IV (5), and ergosterol (6). Keywords Ganoderma sp; colossolactones; schisanlactone A



Chemical Constituents from Barks of Akschindlium godefroyanum

Nattapong Chaipukdee, Kwanjai Kanokmedhakul,* Somdej Kanokmedhakul and Ratsami Lekphrom

Natural Products Research Unit, Department of Chemistry, and Center of Excellence for Innovatioแn in Chemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand

*E-mail: [email protected] Phytochemical investigation of bark of Akschindlium godefroyanum (Kuntze) H. Ohashi (Fabaceae), a Thai herbal medicine, resulted in the isolation of eight known compounds comprising : one triterpenoid, lupeol (1); one steroid, stigmasterol (2); one steroid glycoside, stigmasterol-3-O-β-D-glucoside (3); one flavonol, geraldol (4); three flavanonols, (+)-taxifolin (5), (+)-fustin (6), aromadendrin 5-methyl ether (7) and one catechin, (–)-epigallocatechin (8). Their structures were identified by spectroscopic methods and data comparison with those in literature. Keywords Akschindlium godefroyanum (Kuntze) H. Ohashi, flavanonol, flavonol, catechin



Synthesis and Antibacterial Activity of Cholic Acid-diamine Conjugates

Kanyarat Chanchang1, Sukanlaya Leejae2, Saowanee Kumpun3, Supayang P. Voravuthikunchai2,

and Boon-ek Yingyongnarongkul1 *

1Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok, 10240, Thailand

2Department of Microbiology and Natural Product Research Center of Excellence , Faculty of Science, Prince of Songkhla University, Songkla, 90112, Thailand

3Department of Chemistry, Faculty of Science and Technology, Suansunandha Rajabhat University, Bangkok 10300, Thailand

*E-mail: [email protected] Nitrogen containing steroids have been studied on a variety of biological properties including antibacterial and anticancer activities. Cholic acid is one of the steroidal templates that used for chemical modification. In this study, a series of new cholic acid-diamine conjugates has been synthesized. The structures of newly synthesized compounds were established on the basis of spectral data. The in vitro antibacterial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria were investigated. The results indicated that cholic acid conjugated with 1,6-diaminohexane via carbamate linkage showed highest antibacterial activity against S. aureus with minimum inhibitory concentration (MIC) / minimum bactericidal concentration (MBC) values of 4/8 μg/ml. However, this compound was less potent than the reference drug, vancomycin. Two derivatives bearing 1,2-diaminoethane and 1,6-diaminohexane exhibited antibacterial activity against Gram-negative bacteria with MIC/MBC values of 8/8 μg/ml. The results showed that the spacer of diamino groups of cholic acid-diamine conjugates had significant effects on the antibacterial activity. Keywords Cationic steroid antibiotic; Cholic acid; Antibacterial activity



Depsidones from the Fungus Pilobolus hetrosporous

Oue-artorn Rajachan,1 Kwanjai Kanokmedhakul,1 Somdej Kanokmedhakul,1,* Kasem Soytong2

1Natural Products Research Unit, Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand

2Department of Plant Production Technology, Faculty of Agricultural Technology, King Mongkut’s Institute of Technology Ladkrabang, Bangkok 10520, Thailand

*E-mail: [email protected] Chemical investigation from the fungus Pilobolus hetrosporous, resulted in the isolation of five depsidones (1-5). These structures were established on the basis of spectroscopic data as auranticin A (1), auranticin B (2), 11H-dibenzo[b,e][1,4]dioxepin-11-one,3,8-dihydroxy-4-(hydroxyl methyl)-9-methyl-1,6-bis[(1E)-1-methyl-1-propen-1-yl] (3), 11H-dibenzo[b,e][1,4]dioxepin-4-carbox-aldehyde,3,8-dihydroxy-9-methyl-1,6-bis[(1E)-1-methyl-1-propen-1-yl]-11-oxo (4), 11H-dibenzo [b,e][1,4]dioxepin-11-one,3,8-dihydroxy-4-(methoxymethyl)-9-methyl-1,6-bis[(1E)-1-methyl-1-propen-1-yl (5)

Keywords Pilobolus hetrosporous; Depsidone; Antimalarial; Cytotoxic



6-n-Pentyl-2H-pyran-2-one from Trichoderma asperellum F009

Chotika Jeerapong1, Warin Intana2, Patoomratana Tuchinda3 and Worrapong Phupong1*

1School of science, Walailak University, Thasala, Nakhonsi Thammarat, 80161 2School of agricultural technology, Walailak University, Thasala, Nakhonsi Thammarat, 80161 3Department of Chemistry, Faculty of Science, Mahidol University, Phayathai, Bangkok, 10400

*E-mail: [email protected] A crude ethyl acetate extract was obtained from a culture filtration of Trichoderma asperellum F009. It showed significant antifungal activities against Colletotrichum gloeosporioides with 76.5 % inhibition by mixed agar technique. The major constituent, 6-n-Pentyl-2H-pyran-2-one, which was isolated for the first time from Trichoderma asperellum F009, was characterized by spectroscopic methods, mainly by NMR techniques and compared with the values reported in literature. Keywords Trichoderma asperellum; Colletotrichum gloeosporioides; 6-n-Pentyl-2H-pyran-2-one



Pharmaceutical Cocrystallization and Crystal Characterization of Theophylline and p-aminobenzoic Acid

Weenawan Somphon1*, Kenneth J. Haller2

1Department of Chemistry, Faculty of Liberal Arts and Science, Kasetsart University,

Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand 2School of Chemistry, Institute of Science, Suranaree University of Technology,

Nakhon Ratchasima 30000, Thailand *E-mail: [email protected]

Theophylline (TP) is an oral bronchodilator medicine, used to treat respiratory diseases. Problems with TP include low aqueous solubility, slow release tablets, and solid state interconversion between anhydrous and monohydrate forms as a function of relative humidity. Physicochemical properties of this medicine, including solubility, humidity sensitivity, and chemical and physical stability can be improved by cocrystallizing with another compound. This work presents the cocrystallization of TP and p-aminobenzoic acid (PA), vitamin B, that form noncovalent interactions between TP and PA thereby improving the physicochemical properties of TP. Mixtures of TP and PA were screened by neat grinding and melt methods, as well as by slow evaporation of solutions at room temperature with varying solvents, molar ratios, and pH stability studies. The products were characterized by SEM, XRD, FT-IR/FT-Raman, and DSC techniques. FT-IR results show that the screening products are almost identical to the products from slow evaporation. The nature of interactions between two components in the lattices was obtained by FT-IR and Raman. XRD spectra show that new compounds of 1:1 and 1:2 TP-PA form from ethanol and the mixed solvent water-methanol. The cocrystals of 1:2 TP-PA in water-methanol depend on pH. The amorphous background was decreasing when adjusted pH to 2 and 4 but transforms to another phase at pH 7. FT-IR found a new O-H stretching band, and DSC exhibited an endothermic peak corresponding to the loss of a water molecule at 80 °C, suggesting that a new structural form of 1:2 TP-PA formed at pH 7 from water-methanol solutions. Keywords theophylline; pharmaceutical cocrystal; p-aminobenzoic acid; crystallization; physical property



Preparation and Physical Characterization of 1:1 Theophylline Cocrystallized with 1,3,5-Benzenetricarboxylic Acid

Weenawan Somphon1*, Butsanee Jaturaphitsamajan1, Kenneth J. Haller2

1Department of Chemistry, Faculty of Liberal Arts and Science, Kasetsart University,

Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand 2School of Chemistry, Institute of Science, Suranaree University of Technology,

Nakhon Ratchasima 30000, Thailand *E-mail: [email protected]

Pharmaceutical cocrystals consist of an active pharmaceutical ingredient (API) and coformers, used to improve the physicochemical properties of an API such as solubility, hygroscopicity, permeability, compressability, and flowability. Theophylline (TP) is a bronchodilator used for treatment of respiratory diseases such as asthma. TP is known to exist in a monohydrate and three anhydrous forms (I, II, and III), and converts reversibly between monohydrate and anhydrous forms as a function of relative humidity. In this work, formation of a 1,3,5-benzentricarboxylic acid (BTC) adduct with TP was studied. The 1:1 TP-BTC adduct was screened by solvent-drop grinding, and by crystal growth by slow evaporation (at unadjusted as-mixed pH and at pH adjusted to 2 with HCl). The products were characterized by XRD, FT-IR/FT-Raman, and DSC/TGA. The cocrystals of TP-BTC and pure anhydrous TP were stored at room temperature (34±2 °C) with 58±2% relative humidity for one month and their stability examined by FT-IR. Keywords Theophylline; 1,3,5-benzenetricarboxylic acid; Theophylline cocrystal; Cogrinding



Chemical Constituents of the Extract of Zingiber ottensii Valeton Rhizome

Suwapitcha Srisakul, Pornthip Bubpawan, Anusorn Vorasingha, Chaturong Supharpprom, Vijitr Udeye*

Department of Chemistry, Faculty of Science, Naresuan University, Phitsanulok 65000, Thailand

*E-mail: [email protected] The objective of this study was to investigate chemical constituents with antioxidant and antifungal activity from the Zingiber ottensii Valeton rhizome (Plai dam). Hexane, dichloromethane and ethanol were used for solvent extraction. The crude extract was purified by column chromatography using Sephadex LH-20 and silica gel and characterized structure by spectroscopy (Nuclear magnetic resonance, Infrared spectroscopy and UV spectroscopy). The phenolic compound was investigated the antioxidant activity with by DPPH radical scavenging method and antifungal activities (Pityrosporum orbiculare and Dermatophyte) by disc diffusion method. Keywords Zingiber ottensii Valeton rhizome; Antioxidant; Antifungal



Chemical Constituents of the Stems of Dracaena loureiri. Structural Modification of Resveratrol and Cytotoxicity

Evaluation of the Analogues

Anek Lekky1, Nattapon Apiratikul2, Boon-ek Yingyongnarongkul1, Ciro Isidoro3, and Apichart Suksamrarn1*

1Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok 10240, Thailand

2Department of Chemistry, Faculty of Science, Siam University, Bangkok 10160, Thailand 3Department of Health Sciences, University of Piemonte Orientale “Amedeo Avogadro”,

Novara 28100, Italy *E-mail: [email protected]

Investigation of the MeOH extract of the stems of Dracaena loureiri led to the isolation of four dihydrochalcones, loureirin B (1), 2,4′-dihydroxy-4,6-dimethoxydihydrochalcone (2), loureirin C (3), and 2,4,4′-trihydroxy-6-methoxydihydrochalcone (4), and one stilbene, resveratrol (5). Resveratrol derivatives (6-15) and resveratrol dimers (16 and 17) were synthesized and identified. All natural compounds and synthetic analogues were evaluated for cytotoxicity against a number of cancer cell lines. The structure-cytotoxic activity relationship study of these compounds will be presented.

Keywords Dracaena loureiri; Resveratrol; Resveratrol dimer; Structural modification; Cytotoxicity



Anti-oxidation Capacity of Baccaurea macrocarpa Hull Extracts

Worrapong Phupong*, Imron Meechai, Chotika Jeerapong

School of science, Walailak University, Thasala, Nakhonsi Thammarat, 80161 *E-mail: [email protected]

Baccaurea macrocarpa, commonly known in Thailand as Lung-kae, is a species in genus Baccaurea belonging to the family Phyllanthaceae. Its edible fruit contains citrus-tasted pulp with a delicious tangy flavor. Several reports from past studies of the plants in this genus have revealed interesting biological activities such as antioxidant, anticancer, antibacterial, anti-inflammatory, etc. A preliminary exploration of the Lung-kae fruit on the antioxidant property was carried out through DPPH radical assay (expressed as vitamin C equivalent capacity (VCEAC)) and nitric oxide scavenging determination (expressed as IC50). The VCEAC values of crude Hexane, CH2Cl2, and MeOH hull extracts were 4.135 ± 1.485, 7.884 ± 0.227 and 176.161 ± 4.351, respectively. The IC50 values of all extracts were more than 1,000 µg/mL. Keywords Baccaurea macrocarpa; Anti-oxidant activity



Chemical Constituents from the Stems of Dalbergia parviflora

Warita Lempan1, Morakot Kaewpet1*

1 Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science,

Prince of Songkla University, Songkhla 90112, Thailand * E-mail: [email protected]

Chemical investigation of the crude MeOH extract from the stems of Dalbergia parviflora (Leguminosae) led to the isolation of five known compounds: 4-hydroxy-3-methoxybenzaldehyde (1), methyl 4-hydroxybenzoate (2), 4-hydroxy-3-methoxycinnamaldehyde (3), 7-hydroxy-4′-methoxyisoflavone (4) and 4′,5,7-trihydroxy-6-methoxyisoflavone (5). Their structures were determined by analysis of spectroscopic data, particularly 1D and 2D NMR spectroscopic data, and confirmed by comparison of the 1H and 13C NMR data with those previously reported in the literature. Keywords Dalbergia parviflora; Phenols; Cinnamaldehyde; Isoflavone



Chemical Constituents and Antioxidant Activities of Methanol and Water Extract from Leaves of Dodonaea viscosa

Nutida Singjam1, Amornrat Chalee1, Norman Mangnall2, Stefan Kohlbacher1

and Nipawan Pongprom1*

1Department of Chemistry and Centre of Excellence for Innovation in Chemistry, Faculty of Science, Ubon Ratchathani University, Ubon Ratchathani, 34190, Thailand

2Faculty of Agriculture, Ubon Ratchathani University, Ubon Ratchathani, 34190, Thailand *E-mail: [email protected]

Dodonaea viscosa is an evergreen hopbush found mostly in Australia. Leaves were found to contain sufficient levels of flavonoid constituents to account for purported antioxidant and wound healing actions. In this work, antioxidant activities of tea from dry leaves of D. viscosa including Ferric Reducing Antioxidant Power (FRAP) and ABTS Cation Radical Scavenging Activities in comparison with the conventional teas have been preliminary investigated. Methanol extract was prepared from extraction of 500 g dry leaves of D. viscosa by 3 L methanol at room temperature for 7 days, whereas water extract was prepared from 20 g in 150 mL water at reflux temperature for 3 h. The crude extracts obtained after solvents removal under vacuum. The result showed that tea from D. viscosa exhibited remarkable antioxidant activity. Evaluation of antioxidant activities of methanol and water extracts using DPPH assay, FRAP assay, ABTS assay and total phenolic content showed high antioxidant capacities. In addition, the methanol and water extracts were then separated mainly by column chromatography on silica gel to afford glyceryl trilinoleate, flavonoids and disaccharides. Biological testing of pure substance is in progress.

O

O

HO

MeO

OH

OMe

OMe

Keywords: Dodonaea viscosa; Antioxidant activities; Flavonoids



Preliminary Phytochemical Screening, Antioxidant Activity and Total Phenolic Contents in Selected Plants.

Panya Buahongsa, Ruengrit Sripaco, Chanvit Srihathaikool, Juthamat Tadee, Weerapan Chonboon,

Adisak Chaisoerm and Wipaporn Kitisriworaphan*

The Chemistry Program, Faculty of Science, Udon Thani Rhajabhat University, Udon Thani, 41000, Thailand

*E-mail: [email protected] This study was conducted to investigate phytochemical screening, antioxidant activity and total phenolic contents of selected plants. The rhizomes of Zingiber ottensii Valeton., flowers of Jerusalem artichoke (Helianthus tuberosus L.), stems and seeds of Oenanthe stolonifera Wall. were extracted by marceration with hexane, ethyl acetate and methanol, respectively. The phytochemicals were determined using basic method. Antioxidant activity was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) as Vitamin C equivalent antioxidant capacity (VEAC). Total phenolic contents were determined using a spectrophotometric method, based on the Folin-Ciocalteau reagent. Phytochemical screening of all extracts showed the presence of flavonoids and tannins. The IC50 values of all extracts were obtained in the range 26-671.ppm. The results showed that the methanol extract of Helianthus tuberosus L. had the highest antioxidant capacity, which VEAC was 266 mg vitamin C/g extract. The total phenolic content ranged from 125-939 mg gallic acid/g extract, Helianthus tuberosus L. had the highest total phenolic content followed by Zingiber ottensii Valeton. and Oenanthe stolonifera Wall. respectively (939, 710.5, and 470.25 mg gallic acid/g extract). The results indicated that the amount of phenolic compounds has affected to free radical scavenging activity. Keywords Zingiber ottensii Valeton.; Oenanthe stolonifera Wall; Helianthus tuberosus L.; Phytochemicals; Antioxidant



The Study of Bioactive Compounds of Some Medicinal Plant Extracts in Udon Thani

Wipaporn Kitisriworaphan*

The Chemistry Program, Faculty of Science, Udon Thani Rhajabhat University, Udon Thani,

41000, Thailand *E-mail: [email protected]

This study was carried out to investigate the characteristic of bioactive compound from extract of medicinal plants in Udon Thani. Ancistrocladus tectorius was collected from Phandon - Paco National Forest, Udon Thani. Diospyros ehretioides Wall. ex G. Don, Ellipanthus tomentosus Kurz var. tomentosus and Ochna integerrima (Lour.) Merr were collected from Pha Sing, Nong Wua So district, Udon Thani. Medicinal plants were macerated with hexane, ethyl acetate, methanol and 70% v/v ethanol, respectively. Screening of crude extract for antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans by agar well diffusion techniques. Isolation of crude extract by chromatography techniques using bioassay-guided fractionation and recrystallization. The bioactive compound was isolated from ethyl acetate extract of Ochna integerrima (Lour.) Merr as an orange solid. The result shows that an inhibitory effect of bioactive compound on S. aureus with an inhibition zone was 19.33±0.29 mm in the concentration 2 mg/L. The IR spectrum indicates that the compound was flavonol. Moreover antioxidant activity assay by 2,2-diphenyl-1-picryl hydrazyl (DPPH) showed that percent inhibition was 85.18 in the concentration 100 mg/L. Keywords Ochna integerrima (Lour.) Merr; Bioactive compound; Bioassay-guided fractionation



Preliminary Phytochemical Screening and Antioxidant Activity of Crude Extracts from Nymphaea lotus L. var. pubescens

Hook.f.&Thomson

Nutthamon Prajudtasri and Wipaporn Kitisriworaphan*

The Chemistry Program, Faculty of Science, Udon Thani Rhajabhat University, Udon Thani, 41000, Thailand

*E-mail: [email protected] The aims of the study were to screen for the presence of phytochemicals and determine antioxidant activity of crude extracts from Nymphaealotus L. var. pubescens Hook.f.&Thomson or Bua Daeng. The pollen, petal and stem of Bua Daeng were extracted by macerated with methanol and partition with petroleum ether, chloroform, ethyl acetate and n-butanol, respectively. The antioxidant activity of the extracts were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging assay and ferric reducing antioxidant power (FRAP) assay. The DPPH radical scavenging was in the range of 17.57-99.35%, which were between those of two commercial antioxidants, ascorbic acid and vitamin E. Ferric Reducing abilities of crude extract were in the range 0.008-0.268 mmol Fe2+/g extract. The results indicated that Bua Daeng is rich in natural antioxidant compounds. Keywords Phytochemical screening; Antioxidant activity; Nymphaea lotus L. var. pubescens Hook.f.&Thomson



Hydrolysis of Antidiabetic Drug Metformin by Decavanadate Anion Catalysis

Aungkana Chatkon1,2*, Alexa Barres2, Nuttaporn Samart2,3, Kenneth J. Haller3, and Debbie C. Crans2

1Chemistry Program, Faculty of Science and Technology, Nakhon Ratchasima Rajabhat University, Nakhon Ratchasima 30000 Thailand

2Department of Chemistry, Colorado State University, Fort Collins CO 80523 USA 3School of Chemistry, Institute of Science, Suranaree University of Technology,

Nakhon Ratchasima 30000 Thailand *E-mail: [email protected]

A new material (HGU+)4(HMet+)2(V10O28

6−)·2H2O was prepared from two antidiabetic agents, metformin and vanadate, and its characterization demonstrated that the salt consisted of decavanadate and two types of cations, metfornium and protonated guanylurea. The new compound was obtained after a mixture of metformin and decavanadate was heated in aqueous solution at pH 6.5 at 60 °C for 25 hours. The title compound crystallizes in triclinic space group Pī; a = 10.5330(4); b = 10.9952(3); c = 12.9375(5) Å; α = 84.7947(15)°; β = 67.1390(19)°; γ = 87.1139(15)°; V = 1374.75 A3; Z = 1; Dcalc = 2.013 Mg m−3; λMoKα = 0.71073 Å; µ = 1.73 cm−1; T = 100 K; 35924 data collected, Rint = 0.0332 for 9070 unique data; R1 = 0.0309 for 6798 Io > 2σ(Io); ρmax = 0.39(8). The structures of the two cations and the decavanadate anion correspond to those reported previously. The decavanadate anion (V10O28

6−) lies on an inversion center and charge is balanced by four guanylurea (HGU+) cations around the center axis and two metformium (HMet+) cations capping the ends of the polyoxometalate cluster. These ions and two waters of solvation engage in an extensive multiple H-bonded network with the HGU+ bound at the strongest H-bond acceptor sites of the V10O28

6− and the HMet+ at lesser sites. The hydrolysis of metformin to form guanylurea in the presence of a decavanadate catalyst was confirmed in solution studies using 13C NMR spectroscopy. Keywords Decavanadate; Metforminhydrolysis; Guanylurea; X-ray crystallography



A colorimetric Aptasensor for the Diagnosis of Malaria based on Cationic Polymers and Gold Nanoparticles

Seonghwan Lee, Hunho Jo, Seong-Kyeong Kim, Changill Ban*

Department of Chemistry, Pohang University of Science and Technology, Pohang, Gyeongbuk, 790-784,

Republic of Korea *E-mail: [email protected]

Malaria, a major burden of disease caused by parasites of the genus Plasmodium, is widely spread in tropical and subtropical regions. Here, we have successfully developed a diagnostic technique for malaria. The proposed method is based on the interaction among the Plasmodium lactate dehydrogenase (pLDH), which is a biomarker for malaria, and pL1 aptamer against Plasmodium vivax lactate dehydrogenase (PvLDH) and Plasmodium falciparum lactate dehydrogenase (PfLDH). In addition, the cationic polymers, poly(diallyldimethylammonium chloride) (PDDA) and poly(allylamine hydrochloride) (PAH), aggregate gold nanoparticles (AuNPs) that should be possible to observe the change in color from red to blue, which depends on the concentration of pLDH. Using this aptasensor, pLDH proteins were successfully detected with low detection limits. Moreover, the specificity test proved that the aptasenor is very specific in targeting proteins over other interfering proteins. In addition, the pLDH from infected blood samples of the two main species of malaria were also detected. The limits of detection for P. vivax were determined as 80 parasites/μl for PDDA and 74 parasites/μl for PAH. The aptasenor has great advantages that can simply and rapidly diagnose malaria. Thus, the developed aptasensor for detection of pLDH can offer an effective and sensitive diagnosis of malaria. Keywords Malaria; Aptamer; Biosensor; Cationic polymer; Gold nanoparticle



Electrochemical Aptasensor towards Cardiac Troponin I for the Early Diagnosis of Acute Myocardial Infarction

Changill Ban*, Seonghwan Lee, Hunho Jo, Seong-Kyeong Kim

Department of Chemistry, Pohang University of Science and Technology, Pohang, Gyeongbuk, 790-784,

Republic of Korea *E-mail: [email protected]

In this study, we developed single-stranded DNA (ssDNA) aptamers against cardiac troponin I (cTnI), which is a diagnostic biomarker for acute myocardial infarction (AMI), using magnetic beads-based systematic evolution of ligands by exponential enrichment (SELEX). The aptamers were selectively bound to both cTnI and the cardiac troponin complex. The binding affinities of the cTnI aptamers were measured using surface plasma resonance (SPR), and their dissociation constants were lower than the cTnI antibody. In order to detect cTnI, electrochemical aptasensor was fabricated, and ferrocene-modified silica nanoparticles were employed as electrochemically active probes. cTnI was successfully detected at 2.3 pM (55.2 pg/mL) using a proposed label-free electrochemical method under the buffered conditions. Furthermore, the specificity tests using the cTnI aptamers clearly demonstrated that the aptamers were bound only to cTnI. Additionally, cTnI was successfully detected in the human serum albumin (HSA) solution and a mimic specimen was detected using the calibration plot. Therefore, the cTnI aptamers could be applied instead of an antibody to various biosensors for diagnosing AMI. Furthermore, the proposed label-free cTnI aptasensor has a major advantage: it can simply and rapidly detect cTnI, providing the possibility of diagnosing AMI. Keywords Aptamer; SELEX; Cardiac troponin I; Acute myocardial infarction; Aptasensor



Effect of Drying Temperature on Antioxidant Activity of White Holy Basil

Nasreeya Saleah, Supamas Intarit

Department of Industrial Chemistry Faculty of Science and Technology, Rajamangala University

of Technology Srivijaya, Nakhonsithammarat 80110, Thailand *E-mail: [email protected]

Study effect of drying temperature on antioxidant of white holy basil from ethanol extract, the antioxidant activity were tested by using DPPH radical scavenging assay, compared with standard BHT. The results were found that, the ethanol extract from white holy basil drying temperature at 60 oC showed that was higher than in antioxidant capacity than temperature at 70 oC and 80 oC

respectively. So, the drying of extract from holybasil at lower temperature help to keep ability in antioxidant was better than using high temperature. Keywords Holy Basil; Antioxidant; Drying



Antibacterial activity of Thai Native Herb Extracts against Spoilage Bacteria

Nasjarin Limsaree, Chutimon Satirapipathkuls*

Chemical Engineering Research Unit for Value Adding of Bioresource, Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok 103330, Thailand

*E-mail: [email protected] The antibacterial activities of ethanol, dichloromethane and hexane extracts of five Thai herbs (Syzygium cumini, Cyperus rotundus, Entada phaseoloides, Albizzia Odoratissima and Croton oblongifolius) against Escherichia coli and Staphylococcus aureus were determined by Disc diffusion method and the broth dilution assay. The ethanol extracts of all herbs displayed activity against both of the bacteria tested. Staphylococcus aureus was more sensitive than E. coli to all solvent extracts. The ethanol extract of Entada phaseoloides showed highest activity against both pathogens. The extracts of the Thai native herbs examined in this study have potential for application in foods. Keywords Antibacterial; Herbs; Spoilage bacteria



Screening of Northern Thai Plants for Antioxidant and Antimicrobial Activities.

Wasana Prapalert1*, Jantraporn Panyo1, Watisa Tuykhiaw1

1Faculty of Science and Technology, School of Chemistry,,Chiang Mai Rajabhat University, Chiang Mai,

50200 Thailand *E-mail: [email protected]

Screening of hexane, ethyl acetate and aqueous extracts obtained from the aerial parts of 9 Northern Thai plants for antioxidant and antimicrobial activities have been investigated. The antioxidant activity was evaluated by measuring the scavenging activity of the extracts on DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals. Camellia simenssis, Clausena wallichii, Piper ribesoides, and Chromolaena odorata extracts exhibited significant antioxidant activity. Screening for antimicrobial activity has been conducted using the agar disc diffusion method against Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae. The extracts of 7, 6 and 3 species were effective against E. coli, S. aureus and K. pneumoniae, respectively. Keywords Antioxidant; Antimicrobial; Northern Thai plants



Quantitative Thin-Layer Chromatographic Method for

Determination of Andrographolide using Image Analysis Software

Sinchai Chomngam, Palangpon Kongsaeree*

Department of Chemistry and Center for Excellence in Protein Structure and Function,

Faculty of Science, Mahidol University, Rama VI Rd, Bangkok 10400, Thailand *e-mail: [email protected]

Andrographolide is a major active component in the medicinal plant Andrographispaniculata, with a broad range of pharmacological effects such as suppression of various cancer cells and anti-inflammatory properties. A simple and accurate thin-layer chromatographic (TLC) method for quantitative determination of andrographolide using ImageJ analysis software has been developed and validated. This method is fast, easy and inexpensive. After dried A. paniculata powder was extracted, the quantity of andrographolide was quantified by TLC with suitable conditions then subsequently analyzed with ImageJ graphic software. The quantity of andrographolide on TLC has been validated with linear regression better than 0.95. This newly developed quantification method will be useful to various groups especially the production of A. paniculata herbal products. Keywords Andrographolide; Thin-layer chromatography; ImageJ; TLC quantitation



Screening of Antimicrobial Activity of Hevea brasiliensis Latex Serum

Thitima Rujiralai1*, Wilairat Cheewasedtham2, Kanda Panthong1

1Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Prince

of Songkla University, Songkhla, 90112, Thailand 2Chemistry Division, Department of Science, Faculty of Science and Technology, Prince of Songkla

University, Pattani, 94000, Thailand *E-mail: [email protected]

The aim of this work was to screen antibacterial activity against Bacillus cereus, antifungal activity against Candida albicans and antiphytopathogenic fungal activities against Magnaportha grisea and Curvularia lunata of Hevea brasiliensis latex serum. Skim latex serum used in this work was treated by two methods. For the first method, serum was dried to obtain the dried rubber sheet which was then extracted with water and ethanol. The second method involved the ultrafiltration of serum. The result showed that only the ethanol extract possessed antimicrobial activities against B. cereus and C. albicans with the percentage inhibition of 55.35 and 10.29, respectively. The aqueous extract, ethanol extract and ultrafiltrated sample showed antiphytopathogenic activity against M. grisea at 31.65, 44.99 and 43.36%, respectively but not C. lunata. These results indicated that the ethanol extract showed the best activity when compared to the aqueous extract and ultrafiltrated sample. Keywords antimicrobial activity; Hevea brasiliensis; latex serum



Effect on 5α-Reductase Inhibitory Activity of Red and Black Native Pigmented Rice for Preventing Hair Loss

Natthawut Thitipramote 1,2*, Junniphaphorn Nimkamnerd1, Prinyaporn Pradmeeteekul1,

Phanuphong Chaiwut2, Nisakorn Saewan2, Punyawatt Pintathong2

1 Excellent Center for Cosmetics and Wellness, Mae Fah Luang University, Chiang Rai, 57100, Thailand 2 School of Cosmetic Science, Mae Fah Luang University, Chiang Rai, 57100, Thailand

*E-mail: [email protected] The aims of this study were to determine bioactive compounds and to investigate the 5α-reductase (5αR) inhibitory activity of native pigmented rice (Oryza sativa L.) for preventing hair loss. Four native pigmented rice, two red rice (Mun-Pu: MP and SangYod: SY) and two black rice (Black sticky: KA and HomNil: HN) were extracted with 70% ethanol at ratio sample: solvent (1:10) using shaking method 150 rpm at room temperature for 6 hours. The bioactive compounds, the total phenolic (TPC) and proanthocyanidin contents, and 5α-reductase (5αR) inhibitory activity of each extract were determined. The results showed that the total phenolic and proanthocyanidin contents were significantly different between red, black and white rice (p<0.05) that the MP and SY extracts had the higher phenolic compound (0.182 and 0.190 mg GAE/g sample, respectively) than those of black rice. Moreover, the proanthocyanidin compound was significantly greater content in the red rice (MP: 0.089 and SY: 0.076 mg ECE /g sample) than black KA and HN rice extracts (0.013 and 0.003 mg ECE/ g sample, respectively) (p<0.05). Consistency, the tendency of 5αR inhibitory activity was similar to the amount of bioactive compound that the extracts of red native pigmented rice (MP and SY) had the significantly higher 5αR inhibitory activity (11.526 and 18.484 mg finasteride (FAE) equivalent/ g sample) than those of black rice (KA: 9.290 and HN: 2.825 mg FAE/ g sample) (p<0.05). In conclusion, Mun-Pu rice extracts showed the most active in 5α-reductase inhibition when compared with other pigmented rice. The results suggested that the native pigmented rice, especially red rice, might be used as active gradients in natural for preventing hair loss product. Keywords 5-α Reductase; Black rice; Hair loss; Proanthocyanidin; Red rice



Isolation and Structure Modification of Durantoside I from the Aerial Parts of Citharexylum spinosum L.

Patamawadee Silalai, Anan Athipornchai, Sakkasem Kasemsook and Rungnapha Saeeng*

Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Burapha

University, Sansook, Chonburi 20131, Thailand. *E-mail: [email protected]

Citharexylum spinosum L. is distributed widely throughout many gardens in Thailand, it is an evergreen tree, known as Bu-nga-sa-ri. From the EtOAc extract of the aerial parts of C. spinosum L., a known iridoid glucoside, named durantoside I was isolated and characterized. The resulting durantoside I from the plant was modified by silylation and alkylation of the hydroxyl groups, leading to the preparation of new iridoid glucoside analogues. Pharmacological activities of natural and new synthetic analogues will be evaluated. Keywords Citharexylum spinosum L.; Durantoside I; Iridoid glucoside



Synthesis and α-Glucosidase Inhibitory Activity Evaluation of 17-Amino-epi-isoandrographolide Analogues

Sakkasem Kasemsuk1,2, Rada Bunthawong2, Uthaiwan Sirion2, Pawinee Piyachaturawat3,

Apichart Suksamrarn4, Rungnapha Saeeng2*

1 Department of Chemistry, Faculty of Science and Technology, Rambhai Barni Rajabhat

University,Chanthaburi 22000, Thailand. 2Department of Chemistry and Center for Innovation in Chemistry, Faculty of Science, Burapha University,

Chonburi 20131, Thailand. 3Department of Physiology, Mahidol University, Bangkok 10400, Thailand.

4Department of Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok 10240, Thailand. *E-mail: [email protected]

Andrographolide is a major component isolated from Andrographis paniculata and has been shown to exhibit a wide range of pharmacological activities. In the present study, a series of 17-amino-epi-isoandrographolide analogues were synthesized from naturally occurring andrographolide by epoxidation reaction at C-8 to obtain C-8-epoxy-andrographolide. Ring opening of epoxide ring at C-8 followed by intramolecular cyclization reactions with various anilines gave 17-amino-epi-isoandrographolides. Twenty-five synthetic analogues were obtained in good yields and were evaluated for in vitro α-glucosidase inhibitory. New synthetic compound, C-17-bomoaniline derivative showed higher affinities than the reference acarbose. This compound can be considered as lead molecule for further study. Keywords 17-amino-epi-isoandrographolide; Andrographis paniculata; α-glucosidase inhibitory activity



Scanning Electron Microscopic Study on a Hyperthermophilic Strain FC 302 Belonging to Order Thermotogales,

Formation of Giant Toga

Witoon Wattananit1*, Porranee Keawram2, Wirojne Kanoksilapatham2

1Scientific and Technological Equipment Centre, Faculty of Science, Silpakorn University ,

Nakhon Pathom, 73000, Thailand 2Department of Microbiology, Faculty of Science, Silpakorn University, Nakhon Pathom, 73000, Thailand

*E-mail: [email protected] Members of order Thermotogales are ancient bacteria that have been phylogenetically positioned in close proximity to hyperthermophilic Archaea. Members belonging to genera Thermotoga spp. and Fervidobacterium spp. were reported to grow optimally at temperatures between 70-100°C. They are obligately anaerobic heterothrophs. Because of their ability to survive in extremely hot environments, they are considered as important sources of thousands of gene encoding thermozymes, including cellulases, xylanases, amylases, and ß-glycosidases. Thermotrogales are gram-negative, nonendospore forming rods. Cells are usually encapsulated in an outer membrane envelope named “a toga”, an unconventional balloon-like structure presenting at one or both poles of the cells. The envelope forms a periplasmic space and serves as a barrier to its external surroundings. In addition, its surface contains many polysaccharide hydrolases, allowing the bacteria to hydrolyze insoluble carbon sources. Keywords Scanning electron microscope (SEM); Preparation technique; Thermotogales



Chemical Composition and Antimicrobial Activity of Kaempferia galanga Extracts and Essential Oil

Theeraphan Machan1*, Kittichai Phata2, Sunita Chamyuang2, Angkana Chaipuang1,

Chalermporn Thongpoon3, Chalida Phungpanya1

1Program of Applied Chemistry, School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand 2Program of Biological Science, School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand

3Program of Chemistry, Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phitsanulonk 65000, Thailand

*E-mail: [email protected] In this report, two extracts (hexane and acetone-hexane) and essential oil of Kaempferia galanga collected from Phitsanulok, Thailand were screened for their chemical components and antimicrobial activity. The chemical composition of the extracts and the oil were identified by gas chromatography-mass spectrometry (GC-MS). Major types of volatile compounds found in the extracts were cinnamic acid derivatives, fatty acid methyl ester, and n-alkanes, while mono terpenes and cinnamic acid derivatives were found to be the major components of the essential oil. According to chemical investigation, the major components of the hexane extract were ethyl-p-methoxycinnamate (52.5%), pentadecane (17.2%), ethyl cinnamate (15.0%), methyl-p-methoxycinnamate (7.0%), and methyl cinnamate (1.83%). The most prominent compounds found in acetone-hexane extract were ethyl-p-methoxycinnamate (71.0%), ethyl cinnamate (18.0%), pentadecane (5.6%), methyl linoleate (1.2%), and methyl palmitate (1.0%). The main identified compounds found in the essential oil were ethyl-p-methoxycinnamate (56.6%), ethyl cinnamate (30.0%), δ-3-carene (4.3%), 1,8-cineol (1.2%), borneol (1.2%) and camphene (1.0%). Antimicrobial activities of the extracts and the oils were evaluated against different bacterial and fungal strains by risazurin microtitre assay. Preliminary testing showed that hexane, acetone-hexane extracts and essential oil of K. galanga had significant antifungal activity against Candida albicans with MIC values of 25.0, 12.5 and 0.4 mg/mL, respectively. In vitro studies revealed that the essential oil showed strong antibacterial activity against eight bacterial strains, Bacillus subtilis, Micrococcus luteus, Staphylococcus epidermidis, Staphylococcus aureus, Bacillus Cereus, Escherichia coli, Pseudomonas fluorescens, and Samonella typhimurium, with MIC values of 6.2, 3.1, 12.5, 6.2, 6.2, 3.1, 0.4, and 6.2 mg/mL, respectively, whereas the extracts showed no antibacterial activity against the bacterial strains. Keywords Chemical composition; Antimicrobial activity; Kaempferia galanga; Extract; Essential oil



Extraction of Phenolic Antioxidants from Fermented Tea Leaves Product

Punyawatt Pintathong*, Kittipong Kaewdusit, Nont Thitilertdecha, Natthawut Thitipramote, Sarita Sangthong, Phanuphong Chaiwut

School of Cosmetic Science, Mae Fah Luang University, Chiang Rai, 57100, Thailand

Email: [email protected]

This study aimed to extract and evaluate phenolic antioxidants from fermented tea leaves (miang). The fermented tea leaf extracts obtained from the application of different kinds of solvents and solvent combinations were determined their extractable phenolic content (EPC), extractable flavonoid content (EFC), DPPH free radical scavenging activity and ferric reducing antioxidant power (FRAP). The results showed that the fermented tea leaves contained phenolic compounds that can provide antioxidant activity. The extract of fermented tea leaf powder using methanol significantly provided the highest values of all parameters. The EPC, EFC, DPPH radical scavenging activity and FRAP values were 29.81 ± 0.62 mg GAE/g dw, 76.90 ± 5.85 mg QE/g dw, and 124.20 ± 6.70 mg TEAC/g dw and 61.30 ± 0.71 mg TEAC/g dw, respectively. Moreover, the application of combination of different solvents for extraction could also increase the efficiency of phenolic antioxidant extraction from the fermented tea leaves. The solvent mixture of methanol-DI water-propylene glycol significantly exhibited an excellent extraction outcome providing the EPC, EFC, DPPH free radical scavenging activity and FRAP value of 101.11 ± 4.79 mg GAE/g dw, 119.25 ± 2.37, 240.21 ± 4.84 mg TEAC/g dw and 168.42 ± 0.95 mg TEAC/g dw, respectively. In conclusion, miang which was considered as a traditional food product can be used as an alternative natural source of phenolic antioxidant for further applications as an active ingredient in cosmetic, pharmaceutical and food products. Keywords Antioxidant; Extraction; Miang; Phenolics; Solvent combination



Three Liquid Phase Extraction of Antioxidant Glycosides from Acacia concinna

Phanuphong Chaiwut*, Rossukon Sukkantasawat, Sarita Sangthong,

Nont Thitilertdecha, Natthawut Thitipramote, Punyawatt Pintathong

School of Cosmetic Science, Mae Fah Luang University, Chiang Rai, 57100, Thailand *Email: [email protected]

This study investigated the extraction of antioxidant glycoside from Acacia concinna by three liquid phase (TLP) techniques. Optimization of the extraction was performed using response surface methodology (RSM) and experimental data minimization was employed by Box-Behnken design (BBD). Microwave was used to assist extraction. Three levels of factors of ammonium sulfate (45, 50 and 55% w/v), ethanol concentration (60, 80 and 100% v/v) and microwave power (360, 630 and 900 Watt) were used to study their effect on extraction potential of antioxidant glycoside. Ethanol-rich medium phase contained higher antioxidant capacity and carbohydrate content than those of petroleum ether-rich top phase and salt-rich bottom phase. The results also revealed that linear term of all variables were significantly affecting to the DPPH radical scavenging and carbohydrate content. The quadratic regression of ethanol and salt concentration were also influenced. The highest antioxidant capacity and carbohydrate content of 4,086 µg TEAC/g sample and 150.6 mg GE/g sample, respectively, were found in experimental run consisting of 50% w/v ammonium sulfate, 100% v/v ethanol and 900 W of microwave power combination. This research confirmed that TLP method could be effectively used for bioactive compound extraction. Keywords: Acacia concinna; Antioxidant; Glycoside; Response surface methodology; Three liquid phase

molecular modeling investigations of pyrimethamine...

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