HPLC-MS/MS测定比格犬血浆中非诺贝酸浓度及其药代动力学研究  

Determination of Fenofibric Acid in Beagle Plasma by HPLC-MS/MS and the Study of its Pharmacokinetics

作　　者：马飞[1,2] 邢春宇[1] 吴卓娜[1] 孟志云[1] 甘慧[1] 朱晓霞[1] 顾若兰[1] 窦桂芳[1] 王德才[2] 

机构地区：[1]军事医学科学院野战输血研究所药物代谢重点实验室,北京100850 [2]泰山医学院,山东泰安271016

出　　处：《解放军药学学报》2013年第3期196-199,202,共5页Pharmaceutical Journal of Chinese People's Liberation Army

基　　金：国家科技重大专项课题-"临床前药代动力学技术平台"资助;No.2012ZX09301003-001-007

摘　　要：目的建立快速有效的HPLC-MS/MS方法测定比格犬血浆中非诺贝特的代谢物非诺贝酸,并探讨其在比格犬体内的药代动力学。方法 4只健康雄性比格犬随机分为两组,在禁食状态下口服给药〔非诺贝特片(纳米)和对照药Tri-cor〕,HPLC-MS/MS测定非诺贝酸浓度。流动相为甲醇-水(0.1%甲酸),在200μl.min-1流速下梯度洗脱,地西泮为内标,色谱柱为Thermo Syncronic-C8,柱温:20℃。质谱测定利用多离子监测扫描模式,电离方式为ESI正离子模式,检测离子反应非诺贝酸为m/z 319.10→232.86,地西泮为m/z 285.04→193.04。血药浓度数据经Origin Pro 7.5与WinNonlin Phoenix软件处理,计算主要药动学参数。结果本方法中非诺贝酸在5～1000 ng.ml-1范围内线性良好(r2>0.98),定量下限为5 ng.ml-1。方法的精密度准确度,专属性,绝对回收率,稳定性和基质效应均符合要求。非诺贝特纳米片和Tricor在比格犬体内的主要药动学参数:tmax为(1.50±1.16)和(1.94±2.48)h、t1/2为(8.58±3.41)和(8.02±2.08)h、Cmax为(5857.20±3563.44)和(6697.56±3912.92)ng.ml-1、AUC为(25 328.13±16 009.09)和(26 379.89±17 392.47)ng.h.ml-1。结论建立了准确、灵敏的HPLC-MS/MS检测方法用于比格犬血浆中非诺贝特代谢产物非诺贝酸的含量测定;研究了非诺贝酸在比格犬体内的药代动力学,结果显示非诺贝酸在比格犬体内的代谢动力学过程呈单室模型,该结果为不同来源非诺贝特片的生物等效性研究奠定了基础,为非诺贝特临床安全合理用药提供参考依据。Objective To develop a quick and effective HPLC-MS/MS method for the determination of fenofi- bric acid in Beagle plasma and to study its pharmacokinetics in Beagle dogs. Methods A 145 mg dose of fenofibrate nano tablets or tricor was given to four healthy male beagles in a randomized two-way crossover design. The plasma concentration of the drug was assayed by HPLC-MS/MS with the internal standard diazepam. Chromatographic sepa- ration was performed on a Thermo Syncronic-Cs column （2.1 mm ×50 mm, 5 μm） with the mobile phase consisting of water （0.1% formic acid）-methanol at 20 ℃. The flow-rate of gradient elution was 200 μl ·min-1. Detection and quantification were performed by mass spectrometry in the multiple reaction monitoring mode with positive electrospray ionization m/z at 319.10→232.86 for fenofibric acid and 285.04→193.04 for diazepam （internal standard）, respec- tively. Origin Pro 7.5 and WinNonlin Phoenix software were used to calculate the pharmacokinetic parameters. Re- suits The assay was linear in the range of 5 -1000 ng .m1-1 with a correlation coefficient （ r 2） of 0.98. The limit of quantification was 5 ng .m1-1. In addition, the specificity,extraction recovery,stability and matrix effect met the re- quirement of analysis methodology. The main pharmacokinetic parameters of fenofibrate nano tablets and Tricor wereas follows： t max （1.50 ±1.16） and （1.94 ±2.48） h, t 1/2 （8.58±3.41） and （8.02 ±2.08 ） h , C max （5857.20 ± 3563.44） and （6697.56 ± 3912.92） ng .ml-1, AUC （25 328.13 ±16 009.09） and （26 379.89± 17 392.47） ng ·h ·ml -1 , respectively. Conclusion A sensitive and specific HPLC-MS/MS method was established for determina- tion of the concentration of fenofibric acid, which was the metabolite of fenofibrate in Beagle dogs. The metabolism was in line with the one-compartment mode. This study will further evaluation of bioequivalenee between different fenofi- brate preparations and offer reference for clinical application of

关 键 词：非诺贝特纳米片 非诺贝酸 HPLC-MS MS 药代动力学 

分 类 号：R969.1[医药卫生—药理学]