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INTERNATIONAL JOURNAL OF PHARMACEUTICAL RESEARCH

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Published by : Advanced Scientific Research
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0975-2366
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IJPR 9[3] July - September 2017 Special Issue

July - September 9[3] 2017

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Simultaneous Determination Of Phyllanthin, Hypophyllanthin, And Niranthin In Rabbit Plasma By Lc-Ms/Ms

Author: PHAM SON, TRAN LONG THAI, CHUONG NGOC NAI, PHAM THI THANH THAO, NGUYEN DUC TUAN, TRAN VIET HUNG, NGUYEN NGOC VINH
Abstract: Phyllanthus amarus Schum. et. Thonn. (Eurphorbiaceae) has hepatoprotective and anti-hepatitis B virus (HBV) activities. A simple, rapid, sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed and fully validated to simultaneously quantify phyllanthin, hypophyllanthin, and niranthin in rabbit plasma using diazepam as an internal standard (IS). LC-MS/MS with an electrospray ionization (ESI) interface performed under the multiple reaction monitoring (MRM) mode was used to detect analytes. Simple liquid-liquid extraction was applied to extract analytes from rabbit plasma using n-hexane as an agent. The reconstituted samples were separated on a C18 column using the mobile phase composed of methanol-5 mM ammonium formate (63:37), which was isocratically delivered at a flow rate of 0.3 ml/min. A detailed validation of the method was performed in accordance with the guidelines of the US FDA and EMA. The calibration curves were established as linear in the range of 1–1,000 ng/ml for all analytes. The results of the intra- and inter-day precision and accuracy tests were well within the acceptable limits. Moreover, all analytes were stable under the test conditions. The validated bioanalytical method can be applied in bioavailability studies in rabbits after administration of Phyllanthus amarus.
Keyword: Hypophyllanthin, LC-MS/MS, Niranthin, Phyllanthin, Phyllanthus amarus.
DOI: https://doi.org/10.31838/ijpr/2020.12.04.281
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