title: Simultaneous determination of pradefovir, PMEA and tenofovir in HBV patient serum using liquid chromatography-tandem mass spectrometry and application to phase 2 clinical trial.
abstract: Pradefovir, a prodrug of PMEA, is under phase 2 clinical trial in China to evaluate its pharmacokinetic and pharmacodynamics after multiple-dose study, with adefovir dipivoxil and tenofovir disoproxil fumarate as positive control. A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of pradefovir, PMEA and tenofovir in HBV patient serum. Serum samples were pretreated via simple protein precipitation with methanol and entecavir was used as internal standard. Chromatographic separation was carried out on a Synergi(®) fusion-RP column (150mm×4.6mm) by gradient elution with methanol and 0.1% formic acid in water (v/v) at a flow rate of 1mL/min. The analytes were detected in multiple reaction monitoring mode with positive ion electrospray ionization at m/z 424.1/151.0, 274.1/162.2, 288.1/176.1, and 278.1/152.2for pradefovir, PMEA, tenofovir and IS, respectively. The assays were validated according to current bioanalytical guidelines including specificity, linearity (2.0-500ng/mL for pradefovir and PMEA, 4.0-1000ng/mL for tenofovir), accuracy and precision, extraction recovery, matrix effect and stability. The validated method has been successfully applied to the pharmacokinetic study of pradefovir, adefovir dipivoxil and tenofovir disoproxil fumarate in a set of HBV patients.
authors.affiliation: Department of Clinical Pharmacy, China Pharmaceutical University, No.24 Tong Jia Xiang, Nanjing 210009, China.
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Xiao
authors.affiliation: Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, No.24 Tong Jia Xiang, Nanjing 210009, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.affiliation: Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, No.24 Tong Jia Xiang, Nanjing 210009, China.
authors.forename: Wanqiu
authors.initials: W
authors.lastname: Yang
authors.affiliation: Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, No.24 Tong Jia Xiang, Nanjing 210009, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, 321 Zhong Shan Road, Nanjing 210008, China. Electronic address: yitao_ding@163.com.
authors.forename: Yitao
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, No.24 Tong Jia Xiang, Nanjing 210009, China. Electronic address: yjcpu@yahoo.com.
authors.forename: Jin
authors.initials: J
authors.lastname: Yang
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005260:Female; D006801:Humans; D016014:Linear Models; D008297:Male; D008875:Middle Aged; D063065:Organophosphonates / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D009943:Organophosphorus Compounds / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D011355:Prodrugs; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000068698:Tenofovir / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics
chemical_list: D063065:Organophosphonates; D009943:Organophosphorus Compounds; D011355:Prodrugs; C053001:adefovir; D000068698:Tenofovir; C497721:pradefovir; D000225:Adenine
keywords: LC–MS/MS; PMEA; Pradefovir; Tenofovir; Validation