title: Development and validation of a sensitive LC-MS/MS method for simultaneous quantification of sinotecan and its active metabolite in human blood.
abstract: Sinotecan is a camptothecin analog, currently under clinical testing as an antitumor medication. We developed and validated a rapid, specific and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of sinotecan and its active metabolite, 7-hydroxyethyl-camptothecin (7-HEC), in human blood. Aliquots (200μL) of heparinized blood samples were processed by deproteinization with 400μL acetonitrile each. Chromatographic analyte separation used an Agilent Zorbax SB C8 column (4.6mm×150mm, 5μm) and methanol/10mM ammonium acetate/formic acid (70/30/0.14, v/v/v) as mobile phase, at a flow rate of 0.60mL/min. A Thermo Finnigan TSQ Quantum Ultra tandem mass spectrometer was operated in multiple-reaction monitoring mode. The precursor-to-product ion transitions m/z 493→m/z (331+375) for sinotecan, m/z 393→m/z (233+261) for 7-HEC, and m/z 396→m/z 352 for d3-SN38 (IS) were used for quantification. The method was validated for 1.0-500ng/mL for sinotecan and 0.5-250ng/mL for 7-HEC using 200μL of blood sample. Total time for each chromatograph was ∼6.0min. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels exhibited relative standard deviations (RSD)<13.8% and the accuracy values ranged from -5.3% to 2.4%. The method was successfully applied to a pharmacokinetic study of sinotecan in cancer patients.
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China; Department of Clinical Pharmacy, Bethune International Peace Hospital, 398 West Zhongshan Road, Shijiazhuang 050082, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhan
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Chen
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China. Electronic address: dfzhong@mail.shcnc.ac.cn.
authors.forename: Dafang
authors.initials: D
authors.lastname: Zhong
mesh_terms: D002166:Camptothecin / Q000031:analogs & derivatives* / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D005260:Female; D006801:Humans; D008297:Male; D009369:Neoplasms / Q000097:blood / Q000188:drug therapy; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002166:Camptothecin
keywords: 7-Hydroxyethyl-camptothecin; LC-MS/MS; Pharmacokinetics; Sinotecan