title: HPLC-MS/MS method for the simultaneous determination of MB07133 and its metabolites, cytarabine and arabinofuranosyluracil, in rat plasma.
abstract: MB07133 is an intravenously administered cytarabine mononucleotide (araCMP) prodrug, for the treatment of hepatocellular carcinoma (HCC). A simple, selective and sensitive HPLC-MS/MS method using high pressure liquid chromatography (HPLC) coupled to triple-quadrupole mass spectrometer, was developed and validated for the detection of prodrug MB07133 and its metabolites, cytarabine (araC) and arabinofuranosyluracil (araU) in rat plasma. Protein precipitation using 3% trichloroacetic acid (TCA) was employed to extract analytes from 100μL rat plasma. Adequate separation of araC and araU from their endogenous compounds was achieved on the Synergi(®) fusion-RP column (150mm×4.6mm, 4μm) by a gradient-elution with a mobile phase consisting of ammonium formate (1mM) and methanol at a flow rate of 1mL/min. Multiple reaction monitoring mode (MRM) was applied in the detection of MB07133, araC, araU and Ganciclovir (internal standard) with ion pairs 441.2/330.2, 244.2/112.2, 245.2/113.2 and 256.1/152.2, respectively. The assays were validated with respect to specificity, linearity (100-50000ng/mL for MB07133, 2-1000ng/mL for araC and araU), accuracy and precision, extraction recovery, matrix effect and stability. The validated method has been successfully applied to an intravenous bolus pharmacokinetic study of MB07133 in male Sprague-Dawley rats (18mg/kg i.v.).
authors.affiliation: Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210009 China.
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.affiliation: Department of Clinical Pharmacy, School of Pharmacy, China Pharmaceutical University, Nanjing 210009 China.
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Xiao
authors.affiliation: Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210009 China.
authors.forename: Wanqiu
authors.initials: W
authors.lastname: Yang
authors.affiliation: Department of Phase I Clinical Trials, Affiliated Nanjing University Medical College, Nanjing 210008 China. Electronic address: qwguyi708@126.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Qian
authors.affiliation: Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210009 China. Electronic address: yjcpu@yahoo.com.
authors.forename: Jin
authors.initials: J
authors.lastname: Yang
mesh_terms: D000818:Animals; D001086:Arabinofuranosyluracil / Q000032:analysis* / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003561:Cytarabine / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C497722:4-amino-1-(5-O-(2-oxo-4-(4-pyridyl)-1,3,2-dioxaphosphorinan-2-yl)arabinofuranosyl)-2(1H)-pyrimidinone; D003561:Cytarabine; D001086:Arabinofuranosyluracil
keywords: Arabinofuranosyluracil; Cytarabine; HPLC-MS/MS; MB07133; Prodrug