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Wirod Meerungrueang,Pharkphoom Panichayupakaranant 한국생약학회 2015 Natural Product Sciences Vol.21 No.3
An antibacterial benzoquinone, 2,6-dimethoxy-1,4-benzoquinone, isolated from Ficus foveolata stems was used as a standard marker for establishment of quantitative HPLC analysis for the stem extracts of F. foveolata . The method utilized a TSK-gel ODS-80Ts column (5 μm, 4.6 × 250 mm) with the mixture of methanol and 5% acetic acid in water (24:76, v/v) as the mobile phase at a flow rate of 1 mL/min, and quantitative detection at 289 nm. The parameters i.e. linearity, intraday and interday precision, accuracy, specificity and sensitivity of the method were evaluated for method validation. The recoveries of the method were 99.5 - 103.6% and good linearity (R2 ≥ 0.9999) was obtained. A high degree of specificity, sensitivity as well as repeatability and reproducibility (RSD less than 2 and 5%, respectively) were also achieved. Chloroform was served as the most suitable solvent for extraction of 2,6-dimethoxy-1,4-benzoquinone. The optimised sample preparation and HPLC method can be practically used in the routine quality control process of F. foveolata stem extracts.
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Abdi Wira Septama,Pharkphoom Panichayupakaranant 한국생약학회 2016 Natural Product Sciences Vol.22 No.2
A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of three antibacterial flavonoids, artocarpanone, artocarpin, and cycloartocarpin in ethyl acetate extracts from Artocarpus heterophyllus heartwoods. Separation was achieved using a TSK-gel ODS-80Tm column (5 μm, 4.6 × 150 mm) at 25 ℃ with a gradient elution system of methanol and water as follows: 0-8 min, 60:40; 8-27 min, 80:20; 27-35 min, 60:40, v/v, at a flow rate of 1 mL/min, and a quantitative UV detection at 285 nm. The method was validated by measuring the key parameters, including specificity, linearity, sensitivity, accuracy, repeatability and reproducibility. A high degree of specificity and sensitivity was achieved. The calibration curves for all three flavonoids showed good linearity with a coefficient of determinations (R2) of ≥ 0.9995. The recoveries of the method were from 98-104%, with good reproducibility and repeatability (RSD values of less than 2%) were also achieved. Ethyl acetate was the best solvent for extraction of these three flavonoids using the heat reflux conditions for 1 h. This optimized sample preparation and HPLC method can be practically used for a routine standardization process of the extracts from the A. heterophyllus heartwoods.
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Panupong Puttarak,Adelheid Brantner,Pharkphoom Panichayupakaranant 한국생약학회 2016 Natural Product Sciences Vol.22 No.1
Pentacyclic triterpenes, mainly, asiatic acid, madecassic acid, asiaticoside, and madecassoside are the active constituents of Centella asiatica. A pentacyclic triterpene enriched C. asiatica extract (PRE) was prepared and standardized to contain a total pentacyclic triterpenes not less than 65% w/w. This work was focused on determination of antiinflammatory, antioxidant, and tyrosinase inhibitory activities of PRE and its stability. The PRE exhibited a satisfactory nitric oxide inhibitory effect, with an IC50 value of 64.6 μg/mL. In addition, the PRE inhibited tyrosinase enzyme activity with an IC50 value of 104.8 μg/mL. In contrast, the PRE possessed only weak antioxidant activity. The PRE was stable over a period of four months when stored as a dried powder but only in a well-closed container protected from light at 4 oC. An aqueous alcoholic solution of the PRE was stable at pH values of 5.8 and 7.0, but was not stable at a pH of 8.2. Preparations of the PRE in an aqueous solution should be performed in acidic or neutral conditions.
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