title: Development and validation of a liquid chromatography-tandem mass spectroscopy method for simultaneous determination of (+)-(13aS)-deoxytylophorinine and its pharmacologically active 3-O-desmethyl metabolite in rat plasma.
abstract: CAT ((+)-(13aS)-deoxytylophorinine) is a novel anticancer drug belonging to phenanthroindolizidine alkaloids. A sensitive and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of CAT and its pharmacologically active 3-O-desmethyl metabolite (S-4) was developed and validated in rat plasma using rotundine as the internal standard (IS). CAT, S-4 and IS were extracted by acetonitrile protein precipitation and separated on an Eclipse XDB-C18 column (1.8 μm, 4.6 mm × 50 mm) with acetonitrile-water (27:73, v/v) mobile phase containing 0.1% formic acid at a 0.4 mL/min flow rate. Positive ion electrospray ionization in multiple reaction monitoring mode was employed to measure CAT, S-4 and IS by monitoring the transitions m/z 364.2→70.1 for CAT, 350.1→70.1 for S-4 and 356.2→192.2 for IS. Good linear correlation (r(2)>0.991) was achieved for CAT and S-4 over the range of 0.214-128.16 and 0.044-11.00 ng/mL, respectively. The lower limit of quantification was 0.214 ng/mL for CAT and 0.044 ng/mL for S-4, using 50 μL rat plasma samples. The intra- and inter-day precisions were not exceed 15% and the accuracy ranged between 94.80% and 108.22%. The average extraction recoveries of both analytes were greater than 94.62%. The method was successfully applied to the pharmacokinetic study of CAT and S-4 in rats after oral administration.
authors.affiliation: Department of Pharmaceutics, Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
authors.forename: Feifei
authors.initials: F
authors.lastname: Yu
authors.affiliation: Department of Pharmaceutics, Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
authors.forename: Haining
authors.initials: H
authors.lastname: Lv
authors.affiliation: Department of Pharmaceutics, Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China. Electronic address: dwujun@imm.ac.cn.
authors.forename: Wujun
authors.initials: W
authors.lastname: Dong
authors.affiliation: Department of Pharmaceutics, Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Ye
authors.affiliation: Department of Pharmaceutics, Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
authors.forename: Huazhen
authors.initials: H
authors.lastname: Hao
authors.affiliation: Department of Pharmaceutics, Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
authors.forename: Shuanggang
authors.initials: S
authors.lastname: Ma
authors.affiliation: Department of Pharmaceutics, Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
authors.forename: Shishan
authors.initials: S
authors.lastname: Yu
authors.affiliation: Department of Pharmaceutics, Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China. Electronic address: ylliu@imm.ac.cn.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D000470:Alkaloids / Q000737:chemistry; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D054836:Indolizidines / Q000097:blood* / Q000737:chemistry*; D008297:Male; D010618:Phenanthrolines / Q000097:blood* / Q000737:chemistry*; D010949:Plasma / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: C569808:(13aS)-deoxytylophorinine; D000097:Acetonitriles; D000470:Alkaloids; D054836:Indolizidines; D010618:Phenanthrolines; C032159:acetonitrile
keywords: (+)-(13aS)-Deoxytylophorinine; LC–MS/MS; Metabolite; Pharmacokinetics; Rat plasma