HPLC法检测大鼠血浆中吡非尼酮的浓度及其药动学研究

HPLC determination of pirfenidone in rat plasma and its pharmacokinetics

目的:建立大鼠血浆吡非尼酮检测的高效液相色谱方法,研究吡非尼酮的药代动力学。方法:血浆经乙酸乙酯,采用ZORBAX SB-C18色谱柱;流动相为乙腈-水-0.1%TFA(三氟乙酸),流速为1.0 mL·min-1;检测波长为318(0～8 min)、246nm(8～10 min)。6只雄性大鼠单剂量灌胃给予15 mg.kg-1吡非尼酮,分别在给药后多点尾静脉采血;用该方法检测血浆中吡非尼酮的浓度。用DAS(数据采集系统)计算药代动力学参数。结果:吡非尼酮浓度在0.025～4.00 mg·L-1范围内线性关系良好(r=0.999 9);定量下限为0.025 mg·L-1;低、中、高3个浓度的相对回收率分别为(100.40±4.66)%、(102.17±4.02)%和(101.12±2.89)%;日内RSD分别为4.6%,3.9%,2.9%,日间RSD分别为5.5%,4.8%,3.7%。大鼠吡非尼酮灌胃后,血浆吡非尼酮在0.39 h达到1.69 mg·L-1的峰值浓度,血浆半衰期为2.21 h。吡非尼酮在大鼠体内符合一级吸收的二室模型。结论:本方法准确可靠、简便快速,适用于大鼠血浆吡非尼酮浓度的测定及其药代动力学研究。

Objective：To develop an HPLC method for the determination of pirfenidone in rat plasma and study its pharmacokinetics. Methods：The plasma was extracted by ethyl acetate. The analytical column was packed with ZORBAX SB -C18. The mobile phase were acetonitrile -water- 0. 1% trifluoroacetic acid at the flow rate of 1.0 mL· min-1. The UV detection wavelength was 318 nm （0 -8 min） ,246 nm （8 - 10 rain）. Six Wistar male rats were given a single 20 mg·kg^-1 oral dose of pirfenidone. Blood samples were collected from the tail vein at different time points after oral administration. The concentration of pirfenidone in plasma were detected by this HPLC methods. The pharmacokinetic parameters were analyzed by DAS program. Results： Excellent liner relationship was obtained from the range of 0. 025 mg·kg^-1 to 4. 00 mg·kg^-1（ r = 0. 999 9 ）, the limit determination of pirfenidone was 0. 025 mg·kg^-1 The relative recoveries were （ 100.4± 4.7 ） % , （ 102.2 ±4.0） % and （ 101.1± 2.9） % respectively at three concentrations, the intra -day RSDs were 4. 6% ,3.9% and 2. 8% and inter -day RSDs were 5.5 % ,4. 8 % and 3.7 % respectively. After oral administration of pirfenidone, the maximum plasma concentration of pirfenidone was 1.69 mg·kg^-1 and the time to reach this value was 0. 39 h. the halflife was 2.21 h. Pirfenidone was fitted the two -compartment model with the first absorption. Conclusion：The method is accurate, simple, rapid and can be used to determine the pirfenidone concentration in rat plasma and for study of its pharmacokinetics.