Quantitative determination of formononetin and its metabolite in rat plasma after intravenous bolus administration by HPLC coupled with tandem mass spectrometry

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• 作者：Sheelendra Pratap Singh ; Wahajuddin ; Dinesh K. Yadav ; Preeti Rawat ; Rakesh Maurya ; Girish Kumar Jain
• 关键词：Formononetin ; Daidzein ; Rat plasma ; Validation ; LC– ; MS/MS ; Pharmacokinetics
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2010
• 出版时间：1 February 2010
• 年：2010
• 卷：878
• 期：3-4
• 页码：391-397
• 全文大小：703 K

文摘

A new simple, rapid, sensitive and accurate quantitative detection method using liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) for the measurement of formononetin (FMN) and daidzein (DZN) levels in rat plasma is described. Analytes were separated on a Supelco Discovery C18 (4.6 × 50 mm, 5.0 μm) column with acetonitrile: methanol (50:50, v/v) and 0.1 % acetic acid in the ratio of 90:10 (v/v) as a mobile phase. The method was proved to be accurate and precise at linearity range of 5–100 ng/mL with a correlation coefficient (r) of ≥0.996. The intra- and inter-day assay precision ranged from 1.66–6.82 % and 1.87–6.75 % , respectively; and intra- and inter-day assay accuracy was between 89.98–107.56 % and 90.54–105.63 % , respectively for both the analytes. The lowest quantitation limit for FMN and DZN was 5.0 ng/mL in 0.1 mL of rat plasma. Practical utility of this new LC–MS/MS method was demonstrated in a pharmacokinetic study in rats following intravenous administration of FMN.