Objective: To develop a thin-layer chromatography (TLC) densitometric method for the determination of oxyresveratrol content in Artocarpuslakoocha heartwood and in the traditional drug ‘Puag-Haad’. Materials and Methods: Sample solution of A. lakoocha heartwood was prepared by Soxhlet extraction of the plant material in ethanol, whereas the Puag-Haad solution was obtained by dissolving the drug in methanol. Analysis of each sample solution was performed on a Silica gel 60 F254 TLC plate (20 × 10 cm) with methylene chloride/methanol (85:15) as the mobile phase. After development, the TLC plate was examined with a TLC scanner in the absorbance mode at 254 nm. The newly developed analytical method was validated using an authentic sample of oxyresveratrol previously isolated from A. lakoocha heartwood, and was used to analyze the oxyresveratrol content in samples of A. lakoocha heartwood and the traditional drug Puag-Haad. Results: A sensitive and reliable TLC densitometric method was successfully developed. The method was validated in terms of accuracy (99.11–102.60%) and precision (1.66–4.23% coefficient of variation). The limits of detection and quantitation were 15.6 and 52 ng/spot, respectively. The amounts of oxyresveratrol in 3 samples of A. lakoocha heartwood collected from its natural habitat were 49.0–182.3 mg/g, whereas those in 11 commercial samples were in the range of 23.4–69.6 mg/g. The oxyresveratrol contents in 2 samples of traditional drug Puag-Haad were 780.1 and 837.5 mg/g. Conclusion: The TLC densitometric method developed in this study is a simple, convenient, sensitive and reliable procedure. It was an effective analytical tool for the evaluation of oxyresveratrol content in both A. lakoocha heartwood and the traditional drug Puag-Haad.
Backgound: Guaiacum officinale L. is an alien species to Thailand. It is used as anti-arthritis and anti-rheumatoid agents in Indian folklore medicine. Objective: The present study was aimed to investigate total phenolic contents and free radical scavenging activity of the extracts from Guaiacum officinale L. (Zygophyllaceae). Methods: The plant parts including bark, twig and leaf were extracted using different solvents (ethyl acetate, water and ethanol). Total phenolic contents were determined by Folin-Ciocalteu Colorimetry method while free radical scavenging activity of the extracts was investigated by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity assay. Results: The results show that the highest total phenolic content is found in the ethyl acetate twig extracts (20.3±0.0031 µg GAE/1 µg extracts). The ethanolic twig extracts had the highest free radical scavenging activity with IC 50 of 0.45±0.0188 mg/ml. Conclusion:The extracts from Guaiacum officinale L. exhibit good anti-oxidant activity and may be suitable for development as drugs and supplementary food. MATERIALS AND METHODS ChemicalsGallic acid (Sigma-Aldrich), ascorbic acid (SigmaAldrich), sodium carbonate (Na 2 CO 3 ), 2,2-diphenyl-1-picrylhydrazyl (DPPH) (Sigma-Aldrich), Folin-Ciocalteu (Sigma-Aldrich), ethanol AR grade (Merck) and ethy acetate AR grade (Merck) Plant preparation and extractionBark, twig and leaf of Guaiacum officinale L. were collected from campus of Suan Sunandha Rajabhat University, Bangkok, Thailand. Voucher specimens were deposited and identified at herbarium of National Park, Wildlife and Plant Conservation Department, Ministry of Natural Resources and Environment, Bangkok, Thailand. Plant parts were dried in hot air oven at 60 °C until absolutely dry. They were then ground as fine powder. 600 ml of different solvents (ethyl acetate, water and ethanol) were added to 30 g plant powder (bark, twig and leaf). The plant powders were then extracted by Soxhlet extraction apparatus at 60 °C for 4 hours. The extracts were evaporated in water bath at 60 °C and frozen dry in order to obtain crude extracts. The crude extracts were kept in dark container at room temperature until uses. Determination of total phenolic contents Total phenolic contents were determined by Folin-Ciocalteu Colorimetry method (adapted from Amin et al., 2006). 7 Gallic acid is used as standard (concentration of 12.5-100 mg/ml). The crude extracts were dissolved in methanol. Folin-Ciocalteu reagent was added to 0.5 ml of different extracts (1 mg/ml) and left for 5 minutes. 2 ml Na 2 CO 3 was then added to the solution. The solutions were mixed and adjusted volume as 5 ml by distilled water and left at room temperature for 2 hours. The solutions were then measured the absorbance at 760 nm wavelength. Total phenolic contents in the extracts were determined by comparison of standard curves between mg of Gallic acid equivalent/g extract. DPPH free radical scavenging activity studyFree radical scavenging activity of the extracts was investiga...
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