A rapid and sensitive liquid chromatography–tandem mass spectrometric method for the determination of timosaponin B-II in blood plasma and a study of the pharmacokinetics of saponin in the rat |
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Authors: | Fei Cai Lianna Sun Shouhong Gao Yang Yang Qi Yang Wansheng Chen |
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Affiliation: | aDepartment of Pharmacy, Changzheng Hospital, Second Military Medical University, No. 415 Fengyang Road, Shanghai 200003, PR China;bDepartment of Pharmacognosy, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China |
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Abstract: | A rapid, sensitive and selective liquid chromatography–tandem mass spectrometric (LC–MS/MS) method was developed and validated for the determination of Timosaponin B-II (TB-II), a pharmacologically active constituent isolated from Anemarrhena asphodeloides. This method was used to examine the pharmacokinetics and bioavailability of TB-II in rats using ginsenoside Re as an internal standard. After simple protein precipitation of the plasma samples with acetonitrile, the analytes were separated on an ODS column (150 mm × 2.1 mm i.d., 5 μm) with the mobile phase of acetonitrile–water (35:65, v/v) containing 0.05% formic acid and detected by electrospray ionization mass spectrometry in the negative multiple reaction monitoring (MRM) mode with a chromatographic run time of 3.0 min. The calibration curves were linear over the range of 5–15,000 ng/ml and the lower limit of quantification (LLOQ) was 5 ng/ml in rat plasma. In this range, relative standard deviations (R.S.D.) were <7.4% for intra-day precision and <9.0% for inter-day precision. The accuracy was within the range of 97.7–107.3%. The method was successfully applied to assess the pharmacokinetics and oral bioavailability of TB-II after intravenous and oral administration in rats, with the oral bioavailability being only 1.1%. |
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Keywords: | Timosaponin B-II Saponin Pharmacokinetics Rat plasma LC– MS/MS |
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