title: Quantification of 2'-deoxy-2'-β-fluoro-4'-azidocytidine in rat and dog plasma using liquid chromatography-quadrupole time-of-flight and liquid chromatography-triple quadrupole mass spectrometry: Application to bioavailability and pharmacokinetic studies.
abstract: 2'-Deoxy-2'-β-fluoro-4'-azidocytidine (FNC) is a novel pyrimidine analog that inhibits not only the replication of the hepatitis B virus (HBV), hepatitis C virus (HCV) and HIV but also the replication of lamivudine-resistant HBV, 4'-azidocytidine or 2'-β-methylcytidine-resistant HCV, and nucleoside reverse-transcriptase inhibitor-resistant HIV variants. The present study was undertaken to evaluate the absolute oral bioavailability of FNC in rats and the pharmacokinetic properties of FNC after intragastric administration of single and multiple doses in rats and dogs. A sensitive high-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (HPLC/Q-TOF MS) method and a reliable high-performance liquid chromatography tandem triple quadrupole mass spectrometry (HPLC/QqQ MS/MS) method were established for the determination of FNC in the rat and dog plasmas, respectively. The sample preparation involved a protein-precipitation method with methanol after the addition of lamivudine as an internal standard. FNC was analyzed by LC using a YMC-Pack Pro C18 column (150mm×4.6mm, 3μm) with methanol (containing 0.3% formic acid): 10mM ammonium acetate (containing 0.3% formic acid, pH 2.8) (35:65, v/v) as the mobile phase. Both mass spectrometers were equipped with an electrospray ionization interface in the positive-ion mode. The linear range was from 2.00 to 2000.00ngmL(-1) in rat plasma and 0.50 to 400.00ngmL(-1) in dog plasma. The intraday and interday precision were less than 10.55%, and the accuracy was in the range of -5.86 to 5.13%. The mean recoveries were greater than 82.70% and 82.97% for FNC and IS, respectively. The HPLC/Q-TOF MS and HPLC/QqQ MS/MS methods were both successfully applied in the pharmacokinetic studies of FNC in rats and dogs.
authors.affiliation: Henan Key Laboratory for Pharmacology of Liver, Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450052, China; College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001, China.
authors.forename: Youmei
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Pharmaceutical College of Henan University, Kaifeng 475004, China.
authors.forename: Tiefeng
authors.initials: T
authors.lastname: Cheng
authors.affiliation: Henan Key Laboratory of Fine Chemicals, Zhengzhou 450001, China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Dong
authors.affiliation: Henan Key Laboratory for Pharmacology of Liver, Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450052, China.
authors.forename: Yuhai
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Henan Key Laboratory for Pharmacology of Liver, Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450052, China; School of Pharmaceutical Science, Zhengzhou University, Zhengzhou 450001, China.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Chen
authors.affiliation: Henan Key Laboratory for Pharmacology of Liver, Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450052, China.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Henan Key Laboratory for Pharmacology of Liver, Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450052, China; School of Pharmaceutical Science, Zhengzhou University, Zhengzhou 450001, China.
authors.forename: Jingmin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Henan Key Laboratory of Fine Chemicals, Zhengzhou 450001, China.
authors.forename: Xiaohe
authors.initials: X
authors.lastname: Guo
authors.affiliation: School of Pharmaceutical Science, Zhengzhou University, Zhengzhou 450001, China.
authors.forename: Mintong
authors.initials: M
authors.lastname: Guo
authors.affiliation: College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001, China. Electronic address: changjunbiao@zzu.edu.cn.
authors.forename: Junbiao
authors.initials: J
authors.lastname: Chang
authors.affiliation: Henan Key Laboratory for Pharmacology of Liver, Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450052, China. Electronic address: wangqd@zzu.edu.cn.
authors.forename: Qingduan
authors.initials: Q
authors.lastname: Wang
mesh_terms: D000818:Animals; D001682:Biological Availability; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003562:Cytidine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D004285:Dogs; D005260:Female; D019259:Lamivudine / Q000097:blood / Q000737:chemistry; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D010949:Plasma / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards
chemical_list: C000596970:2'-deoxy-2'-fluoro-4'-azidocytidine; D003841:Deoxycytidine; D019259:Lamivudine; D003562:Cytidine; C508950:4'-azidocytidine
keywords: 2′-Deoxy-2′-β-fluoro-4′-azidocytidine; Bioavailability; High-performance liquid chromatography-quadrupole time-of-flight mass spectrometry; High-performance liquid chromatography-triple quadrupole mass spectrometry; Pharmacokinetics