高效液相色谱法测定大鼠血浆中依托贝特及其代谢物烟酸和氯贝酸浓度

Simultaneous determination of etofibrate,its metabolites nicotinic acid and clofibric acid in rat plasma by HPLC

目的:建立HPLC方法同时测定大鼠血浆中依托贝特及其代谢物烟酸和氯贝酸的浓度。方法:采用Hypersil氰基柱(4.6mm×250mm,5μm),柱温30℃,流动相为甲醇-乙腈-0.01%磷酸溶液和0.6mmol.L-1四丁基溴化铵水溶液(33∶12∶55),紫外检测波长λ=265 nm,流速为1.0mL.min-1。结果:依托贝特、烟酸、氯贝酸的保留时间分别为12.9min、3.8min和6.8min;线性范围分别为0.05～50μg.mL-1、0.05～50μg.mL-1和4～200μg.mL-1;工作曲线方程分别为A=0.020 3 C+0.003 2(r=0.999 6)、A=0.060 9 C+0.004 8(r=0.999 9)和A=0.004 2 C+0.011 4(r=0.999 8);平均回收率分别为78.2%、82.6%和90.4%;日内、日间RSD均小于15%。结论:所建立的HPLC方法灵敏、准确,可为血浆样品中同时检测依托贝特及其代谢物提供方法参考。

OBJECTIVE To establish an HPLC method for simultaneous determination of etofibrate（EF） and its metabolites nicotinic acid（NA） and clofibric acid（CA） in rat plasma. METHODS The HPLC analysis was performed on a Hypersil CN column （250mm×4.6mm,5μm）. The column temperature was set at 30 ℃. A mixture of methanol acetonitrile-0. 01 phosphoric acid and 0. 6 mmol.L-1 tetrabutyl ammonium bromide aqueous solution （33：12：55） was used as the mobile phase with the flow rate at 1 mL.min-1. The detection wavelength was set at 265 nm. RESULTS EF, NA and CA showed good linearity at the ranges of 0. 05 - 50. 00 mL.min-1, 0. 05 - 50. 00 mL.min -1 and 4. 00 - 200. 00 mL.min-1, linear regression equation of A = （7. 020 3 C＋ 0. 003 2（r= 0. 999 6） ,A = 0. 060 9C＋ 0. 004 8（r= 0. 999 9） ,A = 0. 004 2C ＋ 0. 011 4（r = 0. 999 8）, re tent ion time of 12. 9 min,3.8 min and 6. 8 min, with average recoveries of 78. 2%,82. 6 % and 90. 4%, rspectively. The intraday and inter-day variations of RSD were less than 15 %. CONCLUSION The method has high sensitivity and good selectivity, and it is suitable for the simultaneous detection of EF and its metabolites for plasma samples.