Application of a sensitive and specific LC‐MS/MS method for determination of wilforine from Tripterygium wilfordii Hook. F. in rat plasma for a bioavailability study |
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Authors: | Meng‐Xiang Su Min Song Bin Di Tai‐jun Hang |
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Institution: | 1. Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing, China;2. Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing, China |
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Abstract: | A highly selective and specific LC‐MS/MS method was developed and validated for the determination of wilforine in rat plasma. The analyte was separated from plasma matrix by using methyl tertiary butyl ether liquid–liquid extraction with bulleyacinitine A as internal standard (IS). The analysis was carried out on a Sepax GP‐Phenyl column using a mixture of methanol and 10 mmol/L ammonium formate buffer solution containing 0.1% formic acid (75:25, v/v) as the mobile phase pumped at a flow rate of 1.0 mL/min. The detection was operated using a triple‐quadrupole mass spectrometer in multiple selected reaction monitoring with the parent‐to‐product quantifier transitions M + H]+ m/z 867.6 →206.0 for wilforine and 664.1 →584.1 for IS. The main advantage of this method was the high sensitivity (a lower limit of quantification of 0.02 ng/mL) and the small amount of sample (0.1 mL plasma per sample). The method was fully validated to be accurate and precise with a linear range of 0.02–100 ng/mL, and successfully applied to a bioavailability study of wilforine in rats after intravenous and oral administration. The oral absolute bioavailability of wilforine in rats was estimated to be 84%. Copyright © 2014 John Wiley & Sons, Ltd. |
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Keywords: | wilforine Tripterygium wilfordii LC‐MS/MS bioequivalence rat |
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