title: A rapid and sensitive UPLC-MS/MS method for determination of HZ08 in rat plasma and tissues: application to a pharmacokinetic study of liposome injections.
abstract: Overexpression of P-glycoprotein leads to tumor multidrug resistance (MDR). HZ08, a novel tetrahydro-isoquinoline derivate, was discovered to inhibit the MDR in the cancer cell lines of MCF-7/ADM, K562/ADM and KBV in our previous studies. A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometric method (UPLC-MS/MS) was developed and validated for determination of HZ08 in rat plasma and tissues after intravenous administration of HZ08 liposome injection at different doses. The analytes were extracted from plasma and tissues using protein precipitation by acetonitrile with clotrimazole as internal standard. The chromatographic separation was performed on a Thermo BDS HYPERSIL C18 column (100 mm × 4.6mm, 2.4 μm) at a flow rate of 0.7 ml/min using 0.2% ammonium acetate solution (containing 0.1% formic acid) and methanol as mobile phase. The total run time was 4 min. The tandem mass detection was applied with electrospray ionization in positive ion selected reaction monitoring mode. The ion transitions monitored were m/z 523.5 to 342.3 for HZ08 and 277.1 to 165.1 for the internal standard, respectively. The calibration curves obtained were linear in different matrices, and the lower limit of quantification (LLOQ) achieved was 1 ng/ml for rat plasma and 0.25 ng/ml for rat tissues, respectively. The RSDs for intra- and inter-day precision were less than 15%. Extraction recovery, matrix effect and stability were satisfactory in rat plasma and tissues. The developed method was successfully applied to a pharmacokinetic study of HZ08 liposome injection following intravenous administration of 1, 3, 10mg/kg to Sprague-Dawley rats. The data profiles revealed that HZ08 had linear pharmacokinetic properties at the tested doses, and was rapidly distributed into the systemic circulation with wide distribution throughout the body followed by a rapid elimination phase. The major distribution tissues of HZ08 in rats were lung, spleen and liver. These results provided constructive contribution to support the clinical evaluation.
authors.affiliation: Department of Pharmacy, China Pharmaceutical University, Nanjing, PR China.
authors.forename: Fang
authors.initials: F
authors.lastname: Yan
authors.affiliation: Department of Pharmacy, China Pharmaceutical University, Nanjing, PR China.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Sun
authors.affiliation: Department of Pharmacy, China Pharmaceutical University, Nanjing, PR China. Electronic address: hangtj@cpu.edu.cn.
authors.forename: Taijun
authors.initials: T
authors.lastname: Hang
authors.affiliation: Department of Pharmacy, China Pharmaceutical University, Nanjing, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Pharmacy, China Pharmaceutical University, Nanjing, PR China.
authors.forename: Xia
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Center of Drug Discovery, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, PR China.
authors.forename: Xin
authors.initials: X
authors.lastname: Deng
authors.affiliation: Department of Pharmacy, China Pharmaceutical University, Nanjing, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Ge
authors.affiliation: Center of Drug Discovery, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, PR China.
authors.forename: Hai
authors.initials: H
authors.lastname: Qian
authors.affiliation: Department of Pharmacy, China Pharmaceutical University, Nanjing, PR China.
authors.forename: Ding
authors.initials: D
authors.lastname: Ya
authors.affiliation: Center of Drug Discovery, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, PR China.
authors.forename: Wenlong
authors.initials: W
authors.lastname: Huang
mesh_terms: D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007267:Injections; D007546:Isoquinolines / Q000008:administration & dosage / Q000493:pharmacokinetics*; D008081:Liposomes; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014018:Tissue Distribution
chemical_list: D007546:Isoquinolines; D008081:Liposomes; C524509:N-cyano-1-((3,4-dimethoxyphenyl)methyl)-3,4-dihydro-6,7-dimethoxy-N'-octyl-2(1H)-isoquinoline-carboximidamide
keywords: HZ08; Pharmacokinetics; Rat plasma; Rat tissues; UPLC–MS/MS