HPLC-MS/MS法测定血浆中苯扎贝特浓度及其药代动力学研究

HPLC- MS/MS method for determination of bezafibrate concentration in human plasma and its pharmacokinetics in healthy volunteers

目的建立高效液相色谱与质谱联用法测定人血浆中苯扎贝特浓度,并用于苯扎贝特两种制剂的药代动力学研究。方法将6例健康受试者分成3组,3个周期内单次口服受试制剂或参比制剂400 mg。用色谱柱:Welch Ultimate C_(18)(2.1 mm×50 mm,5μm),流动相:乙腈-0.02%甲酸水溶液,梯度洗脱进行分离。用Q-TRAP质谱仪多反应检测模式进行分析测试。用DAS 3.2.7计算主要药代动力学参数。结果苯扎贝特在50.0~1.5×10~4ng·mL^(-1)内线性关系良好(r=0.996 9),定量下限为50.0 ng·mL^(-1),日内、日间精密度RSD分别为1.21%~6.36%、5.73%~7.62%,回收率为67.56%~77.42%。单次空腹口服受试制剂主要药代动力学参数:C_(max)为(14.52±4.78)μg·mL^(-1),t_(max)为(3.00±0.89)h,t_(1/2)为(2.49±0.84)h,AUC_(0-t)为(52.36±6.23)μg·mL^(-1)·h,AUC_(0-∞_为(52.48±61.91)mg·mL^(-1)·h。根据服用两次参比制剂方差分析结果计算得出的变异系数估计样本含量为18例。结论本试验建立了一种快速、高效、灵敏的分析方法,适用于临床上苯扎贝特血药浓度检测及药代动力学研究。

Objective To establish a HPLC- MS / MS method for determination of human plasma concentrations of Bezafibrate,and for the study of pharmacokinetic characteristics of bezafibrate（ two formulations）.Methods Chromatographic separation was using Welch Ultimate C（18）column（ 5 μm,2. 1 mm × 50 mm） with gradient elution（ A： acetonitrile,B： water with 0. 02% formic acid）. Quantification was performed using Q- TRAP MS with multiple- reaction monitoring. Six health subjects were divided into three groups and received single 400 mg dose of test preparation or reference preparation in three period. Plasma samples were collected at different time. The concentrations of bezafibrate were determined by LC- MS / MS. Pharmacokinetic parameters and related statistics analysis were calculated by DAS 3. 2. 7. Results The linear range is 50. 0- 1. 5 × 10^4ng·mL^-1,and had a good linear relationship（ r = 0. 996 9）. The lower limit of quantitation of bezafibrate was 50ng·mL^-1. The intra- and inter- day precisions were 1. 21%- 6. 36%and 5. 73%- 7. 62%,respectively. And the absolute recoveries of bezafibrate were range from 67. 56% to 77. 42%. The main pharmacokinetic parameters of single oral test preparation were： C（max）was（ 14. 52 ± 4. 78）μg·mL^-1,t（max） was（ 3. 00 ± 0. 89） h,t（1 /2）was（ 2. 49 ± 0. 84） h,AUC（0- t）was（ 52. 36 ± 6. 23） μg · mL^（-1）·h,AUC（0- ∞）was（ 52. 48 ± 6. 19） μg·mL^（-1）·h. According to the coefficient of variation to estimate sample size was 18 cases. Conclusion This study established a rapid,efficient and sensitive method for the analysis of the concentration of bezafibrate in plasma and also suitable for pharmacokinetic studies of bezafibrate.