title: Determination and validation of hupehenine in rat plasma by UPLC-MS/MS and its application to pharmacokinetic study.
abstract: In this work, a sensitive and selective ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determination of hupehenine in rat plasma was developed and validated. After addition of imperialine as an internal standard (IS), protein precipitation by acetonitrile-methanol (9:1, v/v) was used to prepare samples. Chromatographic separation was achieved on a UPLC BEH C18 column (2.1 × 100 mm, 1.7 µm) with 0.1% formic acid and acetonitrile as the mobile phase with gradient elution. An electrospray ionization source was applied and operated in positive ion mode; multiple reaction monitoring mode was used for quantification using target fragment ions m/z 416.3 → 98.0 for hupehenine, and m/z 430.3 → 138.2 for IS. Calibration plots were linear throughout the range 2-2000 ng/mL for hupehenine in rat plasma. Mean recoveries of hupehenine in rat plasma ranged from 92.5 to 97.3%. Relative standard deviations of intra-day and inter-day precision were both <6%. The accuracy of the method was between 92.7 and 107.4%. The method was successfully applied to a pharmacokinetic study of hupehenine after either oral or intravenous administration. For the first time, the bioavailability of hupehenine was reported as 13.4%.
authors.affiliation: Laboratory Animal Centre, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Congcong
authors.initials: C
authors.lastname: Wen
authors.affiliation: The Laboratory of Clinical Pharmacy, The People's Hospital of Lishui 323000, Wenzhou Medical University, Lishui, China.
authors.forename: Shuanghu
authors.initials: S
authors.lastname: Wang
authors.affiliation: Analytical and Testing Center, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Xueli
authors.initials: X
authors.lastname: Huang
authors.affiliation: Laboratory Animal Centre, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Zezheng
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Laboratory Animal Centre, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Laboratory Animal Centre, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Suping
authors.initials: S
authors.lastname: Yang
authors.affiliation: Analytical and Testing Center, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Jianshe
authors.initials: J
authors.lastname: Ma
authors.affiliation: The Laboratory of Clinical Pharmacy, The People's Hospital of Lishui 323000, Wenzhou Medical University, Lishui, China.
authors.forename: Yunfang
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Analytical and Testing Center, Wenzhou Medical University, Wenzhou, 325035, China.
authors.forename: Xianqin
authors.initials: X
authors.lastname: Wang
mesh_terms: D000470:Alkaloids / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016014:Linear Models; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000470:Alkaloids; C532925:hupehenine
keywords: UPLC-MS/MS; hupehenine; pharmacokinetics; rat