UPLC-MS/MS Method for Determination of Khasianine in Mouse Blood: Application for Its Pharmacokinetic Study

Author(s): Lianguo Chen, Qinghua Weng, Yijing Lin, Xiaojie Lu, Zuoquan Zhong, Jianhua Xiong, Xianqin Wang*

Journal Name: Current Pharmaceutical Analysis

Volume 16 , Issue 6 , 2020

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Graphical Abstract:


Background: The aim of this study was to determine the concentrations of khasianine in mouse whole blood sample and its application for the pharmacokinetics by a rapid, selective and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method.

Methods: The blood samples were preprocessed by one-step protein precipitation with acetonitrile. The study was performed on an ACQUITY I-Class UPLC system with a UPLC BEH column. Lannaconitine (internal standard, IS) and khasianine were gradient eluted by a mixture of acetonitrile and water with 0.1% formic acid at a flow rate of 0.4 mL/min. The mass spectrometer was equipped with an Electrospray Ionization (ESI) source in positive mode. The quantitative detection was performed in a multiple reaction monitoring modes at transitions m/z 722.4→70.7 for khasianine and m/z 585.3→119.9 for the corresponding IS.

Results: The calibration curve was of good linearity ranging from 0.5 to 1000 ng/mL (r > 0.995). The Lower Limit of Detection (LLOD) and Lower Limit of Quantitation (LLOQ) were 0.2 and 0.5 ng/mL, respectively. The inter-day and intra-day precision (RSD%) were both less than 14%, and the accuracy ranged from 86.6% to 108.3%. The matrix effects were between 98.0% and 103.7%, and the average recovery was better than 67.4%.

Conclusion: This assay established a sensitive, rapid, selective UPLC-MS/MS method which was successfully used for the pharmacokinetic study of khasianine in mouse blood, and the absolute availability of khasianine was 0.78% which exhibited a poor oral absorption.

Keywords: Khasianine, mouse, UPLC-MS/MS, determination, pharmacokinetics, bioavailability, chinese herbal medicine, pharmacology.

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Year: 2020
Published on: 30 June, 2020
Page: [705 - 711]
Pages: 7
DOI: 10.2174/1573412915666190220101658
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