Clinical evaluation of Gac extract (Momordica cochinchinensis) in an antiwrinkle cream formulation.

The objective of this work was to evaluate the antioxidant and antityrosinase activities of Gac (Momordica cochinchinensis) extract and to clinically evaluate a Gac-containing antiwrinkle cream formulation. Gac extract exhibited higher antioxidant activity than vitamin C or E, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH, 41.25 ± 0.34 mg TEAC/ml extract), 2, 2'-azinobis 3-ethylbenzothialine-6-sulfonic acid (ABTS, 47.70 ± 0.18 mg TEAC/ml extract), and ferric reducing antioxidant power (FRAP, 105.03 ± 2.326 mg TEAC/ml extract) scavenging. The antioxidant activity of Gac extract was 5.85- and 11.75-fold higher than that of vitamin E in the DPPH and ABTS assays, respectively. The FRAP assay indicated that the antioxidant activity of Gac extract was 2.91-fold higher than that of vitamin C. Gac extract also exhibited high tyrosinase inhibition (62.83% ± 1.99%). The tyrosinase inhibition activity of Gac extract was 1.51- and 2.06-fold greater than that of vitamins C and E, respectively. The safety and efficacy of the formulated Gac extract cream as an antiwrinkle agent and its promotion of skin moisturization and smoothness were investigated. Evaluation of acute skin tolerance indicated nonirritating properties. A clinical study revealed increases in cutaneous hydration. Average roughness was decreased, while smoothness was increased. Skin overlook analysis indicated skin roughness relief. These results indicate that the formulated Gac extract product is an effective antiwrinkle cream.

Determination of steroids adulterated in liquid herbal medicines using QuEChERS sample preparation and high-performance liquid chromatography.

QuEChERS sample preparation was optimized using solvent extraction with acetonitrile and dispersive-solid phase extraction with primary and secondary amine sorbents, and validated for high-performance liquid chromatographic determination of nine steroids commonly used to adulterate herbal medicines: such as triamcinolone, prednisolone, hydrocortisone, methylprednisolone, betamethasone, dexamethasone, beclomethasone, fludrocortsone acetate and cortisone acetate. Satisfactory extraction recoveries of 91-113% for all nine steroids were obtained, along with an acceptable precision in extraction recoveries shown by R.S.D. of ≤4.6 and 3.2% for intraday and interday, respectively. The QuEChERS sample preparation developed here allows the reliable detection of adulterated steroids with the limits of detection in the range of 0.06-0.17ppm. Adulterated steroids in three out of six real commercial liquid herbal medicines were found, such as 1.6 and 8.8ppm dexamethasone and 0.43ppm prednisolone.

In vitro study of the effects of plaunotol on oral cell proliferation and wound healing.

Plaunotol is an acyclic diterpene alcohol extracted from a medicinal plant called plau-noi, Croton stellatopilosus Ohba, and has been widely used for the treatment of gastric ulcers in Japan. The aim of this study was to examine the effects of plaunotol on human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs). To assess the cytotoxic effect, HGFs and HOKs were treated with plaunotol. Subsequently, the morphology of cells was recorded and cells were subjected to MTT assay. To investigate cell proliferation effect, cells were treated with plaunotol and counted with a haemocytometer. To determine wound healing effect, the number of cells repopulated into the wounded areas in monolayer culture and in fibroblast-populated collagen lattice (FPCL) was measured. The results showed that 10 and 1 µg/ml (33 and 3.3 µmol/l) plaunotol induced toxicity in HGFs and HOKs, respectively. However, 0.1 µg/ml (0.33 µmol/l) plaunotol promoted HGF proliferation and wound healing in monolayer and FPCL models. In contrast, 0.1 µg/ml plaunotol could not induce HOK proliferation nor in vitro wound healing using monolayer culture, but it induced wound healing in a modified FPCL model. Our data suggested that plaunotol could promote oral cell proliferation and wound healing in vitro and may have an implication on oral wound healing.

Hemagglutinating activity of proteins from Parkia speciosa seeds.

Proteins from Parkia speciosa Hassk. (Fabaceae) seeds were extracted and stepwise precipitated using ammonium sulfate. Proteins precipitated with 25% ammonium sulfate were separated by affinity chromatography on Affi-Gel Blue gel followed by protein liquid chromatography on Superdex 200. The protein Gj, which was identified as a protein similar to putative aristolochene synthase, 3'-partial from Oryza sativa L. (Poaceae), had hemagglutinating activity of 0.39 mug/muL. Moreover, fraction C2 from the proteins precipitated with 60% ammonium sulfate, separated by lectin-specific adsorption chromatography using Con A Sepharose, had hemagglutinating activity of 1.17 mug/muL. Using gel electrophoresis, two proteins C2a and C2b were separated, having molecular weights of 45 kDa and 23 kDa, respectively. From protein identification, C2a was found to be similar to the hypothetical protein B1342F01.11 from Oryza sativa, and C2b was similar to the hypothetical protein At1g51560 from Arabidopsis thaliana (L.) Heynh. (Brassicaceae).

Quantitative thin layer chromatographic analysis of the saponins in tea seed meal.

INTRODUCTION: The defatted seed meal of Camellia oleifera Abel., which contains saponins, is used extensively in aquaculture to eliminate unwanted fish and harmful insects during the cultivation of prawns, and as a molluscicide to control Pomacea canaliculata Lamarck. For the quality control of such tea seed meal products, a method for the determination of saponin content is required. OBJECTIVE: To develop a simple, sensitive and specific method for the quantitative determination of saponins in tea seed meal using high-performance thin layer chromatography and densitometry. METHODOLOGY: Powdered tea seed meal samples were extracted with methanol, filtered, evaporated to dryness and the residue taken up in methanol. Samples, together with methanolic solutions of saponin standard, were analysed by HPTLC on silica gel layers eluted with ethyl acetate:methanol:water (6:3:1.5, v/v/v) and detected at 214 nm. The amounts of saponins were determined from the respective calibration curves obtained by plotting the concentrations of saponin standard against peak area. RESULTS: Tea saponin peaks were detected at about R(f )0.40. Good linearity was observed in the range of 0.9-22.2 microg/spot with a correlation coefficient of 0.9978. The limits of detection and quantification were 0.87 and 2.90 microg/spot, respectively. The content of tea saponins in 10 tested samples was found to be between 13.1 and 21.1% w/w. CONCLUSION: The assay proved to be a specific, sensitive and inexpensive method for the quality control of saponins in tea seed meal.

Capillary zone electrophoresis for separation and analysis of hydroxycitric acid and hydroxycitric acid lactone: application to herbal products of Garcinia atroviridis Griff.

Capillary zone electrophoresis (CZE) was developed for quantitative determination of hydroxycitric acid and hydroxycitric acid lactone in herbal products of Garcinia atroviridis Griff. Resolution optimization was investigated by varying type, concentration and pH of buffers. Using the pH 9.2 buffer containing 30 mM Na(2)B(4)O(7), 90 mM NaH(2)PO(4) and 0.5 mM tetradecyltrimethyl ammonium bromide, baseline resolution (R(s)>1.5) was found for all analytes. Advantages of the developed CZE method include simple sample preparation, fast analysis time within 5 min and high accuracy and precision.

Cassane furanoditerpenoids from the seed kernels of Caesalpinia bonduc from Thailand.

Three new cassane furanoditerpenoids ( 1- 3), together with 13 known cassane diterpenes ( 4- 16), were isolated from the EtOAc extract of the seed kernels of Caesalpinia bonduc. Their structures were elucidated on the basis of spectroscopic analysis, mainly NMR and MS. Compounds 1- 3 exhibited good antimalarial activity against the multidrug-resistant K1 strain of Plasmodium falciparum, while compound 4 was inactive. None of the compounds were cytotoxic against any of the tumor cell lines tested.

Furanocembranoids from the stem bark of Croton oblongifolius.

Four novel furanocembranoids (1-4) were isolated from the stem bark of Croton oblongifolius. Their structures were elucidated on the basis of spectroscopic analysis, mainly NMR and MS. Compounds 1, 3, and 4 exhibited good cytotoxicity against several human tumor cell lines.

Hemagglutinating activity of Curcuma plants.

Crude proteins obtained by Mg/NP-40 extraction from Thai medicinal plants of the Curcuma species exhibited agglutination activity against rabbit erythrocytes. A crude extract from Salingalinthong, a Thai Curcuma specie, exhibited the strongest hemagglutinating activity, 2 x 10(-5) mg/ml.

Cytotoxic constituents from Butea superba Roxb.

A carpin (3-hydroxy-9-methoxypterocarpan) (Medicarpin) (1) and four isoflavones, 7-hydroxy-4'-methoxy-isoflavone (Formononetin) (2); 7,4'-dimethoxyisoflavone (3); 5,4'-dihydroxy-7-methoxy-isoflavone (Prunetin) (4) and 7-hydroxy-6,4'-dimethoxyisoflavone (5) were isolated from the tuber roots of Butea superba Roxb. Compounds 2 and 4 showed moderate cytotoxic activity on KB cell lines with IC(50) (microM) values of 37.3+/-2.5 and 71.1+/-0.8 and on BC cell lines with IC(50) (microM) values of 32.7+/-1.5 and 47.3+/-0.3, respectively.

Novel intermediates of acenaphthylene degradation by Rhizobium sp. strain CU-A1: evidence for naphthalene-1,8-dicarboxylic acid metabolism.

The acenaphthylene-degrading bacterium Rhizobium sp. strain CU-A1 was isolated from petroleum-contaminated soil in Thailand. This strain was able to degrade 600 mg/liter acenaphthylene completely within three days. To elucidate the pathway for degradation of acenaphthylene, strain CU-A1 was mutagenized by transposon Tn5 in order to obtain mutant strains deficient in acenaphthylene degradation. Metabolites produced from Tn5-induced mutant strains B1, B5, and A53 were purified by thin-layer chromatography and silica gel column chromatography and characterized by mass spectrometry. The results suggested that this strain cleaved the fused five-membered ring of acenaphthylene to form naphthalene-1,8-dicarboxylic acid via acenaphthenequinone. One carboxyl group of naphthalene-1,8-dicarboxylic acid was removed to form 1-naphthoic acid which was transformed into salicylic acid before metabolization to gentisic acid. This work is the first report of complete acenaphthylene degradation by a bacterial strain.

Inhibitory effects of selected Thai medicinal plants on Na+,K+-ATPase.

Extracts of ten Thai indigenous medicinal plants having ethnomedical application in the treatment of dysuria were tested for their Na(+),K(+)-ATPase inhibitory activity. The hexane extracts of Cyperus rotundus and Orthosiphon aristatus showed high potent inhibitory activity on crude enzyme Na(+),K(+)-ATPase from rat brain.

Microemulsion electrokinetic chromatography for separation and analysis of curcuminoids in turmeric samples.

Microemulsion EKC (MEEKC) was developed for quantitative analysis of curcuminoids, such as curcumin (C), demethoxycurcumin (D), and bis-demethoxycurcumin (B). MEEKC separation of curcuminoids was optimized, and a change in resolution was explained using a modified equation for resolution in MEEKC without electroosmosis. The suitable MEEKC conditions for separation of curcuminoids were obtained to be the microemulsion buffer containing 50 mM phosphate buffer at pH 2.5, 1.1% v/v n-octane as oil droplets, 180 mM SDS as surfactant, 890 mM 1-butanol as cosurfactant, and 25% v/v 2-propanol as organic cosolvent; applied voltage of -15 kV; and separation temperature 25 degrees C. Achieved baseline resolution of C:D and D:B was obtained with R(s) -2.4 and analysis time within 18 min. In addition, high accuracy and precision of the method were obtained. This MEEKC method was used for quantitative determination of individual curcuminoids in medicinal turmeric capsules and powdered turmeric used as coloring additive in food, with simple sample preparation such as solvent extraction, dilution, and filtration, and without cleaning up by SPE.

Elongation of C16:0 to C18:0 fatty acids in methylotrophic yeast Hansenula polymorpha CBS 1976 and fatty acid auxotrophic mutants.

Fatty acid elongation defective mutant was isolated from the ethyl methanesulfonate treated Hansenula polymorpha based on the growth ability. Using biochemical and genetic approaches, the mutant was characterized. When compared with the fatty acid phenotype of the parental strain, the differences in profile and content of fatty acids in V1 mutant were found. In this V1 mutant, polyunsaturated fatty acids, linoleic and alpha-linolenic acids, could not be detected with a corresponding increase in the content of mono-unsaturated fatty acids. The ratio of C16/C18 fatty acids revealed that the accumulation of C16 fatty acids was increased significantly. The experiments on fatty acid supplementation indicated that the mutant required C18:0 for the proper growth. The results of genetic complementation with the elongase genes of Saccharomyces cerevisiae confirmed that the lesion was occurred at least in the extension of C16:0 to C18:0 of V1. The H. polymorpha mutant obtained in this work will be used as a useful tool for unraveling the pathway of fatty acid synthesis and the role of fatty acids on biological processes.

New halimane diterpenoids from Croton oblongifolius.

Three new halimane-type diterpenoids, crotohalimaneic acid ( 1), crotohalimoneic acid ( 2) and 12-benzoyloxycrotohalimaneic acid ( 3), were isolated from the stem bark of Croton oblongifolius Roxb. Their structures were established on the basis of spectroscopic and X-ray analysis. Compounds 1 and 2 showed non-specific strong cytotoxicity against a panel of human tumor cell lines; whereas 3 was inactive.

Diterpenoids from the stem barks of Croton robustus.

Three compounds were isolated from the stem barks of Croton robustus. Their structures were elucidated as trachyloban-19-oic acid, trachyloban-19-ol and poilaneic acid by spectroscopic analysis. Among them, trachyloban-19-ol and methyl trachyloban-19-oate exhibited weak cytotoxic activity against gastric carcinoma and colon carcinoma with ED50 of 9.2, 9.6 and 8.3, 9.1 microg/mL, respectively.

Inhibition of Na+,K+-ATPase activity by (-)-ent-Kaur-16-en-19-oic acid and its derivatives.

The diterpene, (-)- ent-kaur-16-en-19-oic acid, from Croton oblongifolius was identified as an Na +,K +-ATPase inhibitor. This compound exhibits an IC 50 of 2.2 x 10( -5 )M against crude enzyme Na +,K +-ATPase from rat brain. The semi-synthetic derivatives, (-)-methyl kaur-16-en-19-oate, (-)-kaur-16-en-19-ol, (-)-16beta,17-epoxykauran-19-oic acid and (-)-17-hydroxykaur-15-en-19-oic acid were also tested and their IC 50 were 5.5 x 10 (-4), 5.0 x 10( -4), 4.8 x 10( -4) and 6.0 x 10 (-4) M, respectively.

Cytotoxic activity of natural labdanes and their semi-synthetic modified derivatives from Croton oblongifolius.

Labda-7,12( E),14-triene-17-oic acid, previously isolated from Croton oblongifolius, and its derivatives were investigated for cytotoxicity against five human tumor cell lines. Six of these compounds, labda-7,12( E),14-triene-17-al, 17-hydroxylabda-7,12( E),14-triene, 17-acetoxylabda-7,12( E),14-triene, 15-hydroxylabda-7,13( E)-diene-17,12-olide, labda-7,13( E)-diene-17,12-olide, and 12,17-dihydroxylabda-7,13( E)-diene showed non-specific, moderate, cytotoxicity against all cell lines, whereas the other compounds were inactive.

Croblongifolin, a new anticancer clerodane from Croton oblongifolius.

A new furoclerodane, croblongifolin, together with one known clerodane, crovatin and one known labdane, nidorellol, were isolated from the stem bark of Croton oblongifolius. Structures were established based on spectroscopic and X-ray analysis. Croblongifolin showed a significant cytotoxicity against various human tumor cell lines including HEP-G2, SW620, CHAGO, KATO3 and BT474.