title: Simultaneous determination of GFA and its active metabolites in human plasma by liquid chromatography electrospray ionization mass spectrometry and its application to pharmacokinetic studies.
abstract: A sensitive and rapid liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) method has been developed and validated for simultaneous quantification of guanfu base A (GFA) and its metabolites guanfu base I (GFI) and guanfu alcohol-amine (AA) in human plasma with phenoprolamine hydrochloride (DDPH) as the internal standard. The analytes were extracted from human plasma by using liquid-liquid extraction with ethyl acetate and the LC separation was performed on a Diamonsil C(18) analytical column (150 mm x 2.1 mm i.d., 5 microm). The MS acquisition was performed in selected ion monitoring (SIM) mode of positive ions. Analysis was carried out in SIM mode at m/z 430.25 for GFA [M+H](+), m/z 388.25 for GFI [M+H](+), m/z 346.25 for AA [M+H](+) and m/z 344.20 for the IS DDPH [M+H](+). The calibration curves were linear over the range of 50-5000 ng/mL for GFA and 5-1000 ng/mL for GFI and AA, with coefficients of correlation above 0.999. The lower limit of quantification for GFA was 1 ng/mL, while for GFI and AA were both 5 ng/mL. The intra- and inter-day precisions (CV) of analysis were within 9%, and the accuracy ranged from 91% to 108%. The overall recoveries for GFA, GFI and AA were about 94.2%, 87.8% and 80.6%, respectively. The total LC-MS run-time was only 5.5 min. This quantitation method was successfully applied to the simultaneous determination of GFA and its metabolites in human plasma for the metabolic study and pharmacokinetic evaluation.
authors.affiliation: Key Laboratory of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210038, Jiangsu Province, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Huang
authors.forename: Guang-Ji
authors.initials: GJ
authors.lastname: Wang
authors.forename: Jian-Guo
authors.initials: JG
authors.lastname: Sun
authors.forename: Ji-Ye
authors.initials: JY
authors.lastname: A
authors.forename: Wei-Bin
authors.initials: WB
authors.lastname: Zha
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jing-Wei
authors.initials: JW
authors.lastname: Zhang
authors.forename: Bei
authors.initials: B
authors.lastname: Yan
authors.forename: Sheng-Hua
authors.initials: SH
authors.lastname: Gu
authors.forename: Hong-Can
authors.initials: HC
authors.lastname: Ren
authors.forename: Lin-Sheng
authors.initials: LS
authors.lastname: Liu
mesh_terms: D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006576:Heterocyclic Compounds, 4 or More Rings / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D006801:Humans; D011786:Quality Control; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D006576:Heterocyclic Compounds, 4 or More Rings; C040615:Guan-fu base A