高效液相色谱法测定鸡血浆中帕托珠利和托曲珠利含量研究

Determination of Patozuril and Toltrazuril in Chicken Plasma by High Performance Liquid Chromatography

为建立一种测定鸡血浆中帕托珠利和托曲珠利含量的高效液相色谱法,试验采用C18色谱柱,分别以0.2%乙酸水-乙腈(47∶53)和0.02 mol/L磷酸二氢钾缓冲液(pH 3.0)-乙腈(40∶60)为流动相,柱温分别为35℃和30℃,流速为1 mL/min,紫外检测器检测,检测波长分别为250 nm、240 nm,外标法定量建立检测方法。结果显示,帕托珠利和托曲珠利的浓度与色谱峰面积在0.10~20.00μg/mL浓度范围内线性关系良好(R2>0.999),帕托珠利平均加样回收率为89.65%~96.12%,托曲珠利平均加样回收率90.84%~94.27%,批内和批间变异系数均小于5%。结果表明,建立的高效液相色谱法简便、有效、重现性良好,可用于鸡血浆中帕托珠利和托曲珠利含量的测定。

To establish a high performance liquid chromatography method for determination of patozuril and toltrazuril in chicken plasma,a C18chromatographic column was used for separation,and the mobile phases were 0.2%acetic acid water-acetonitrile(47∶53)and 0.02 mol/L potassium dihydrogen phosphate buffer(pH 3.0)-acetonitrile(40∶60).The column temperatures were 35℃ and 30℃ respectively,the flow rate was 1 mL/min,and the UV detection wavelength was 250 nm and 240 nm respectively,the external standard method was used to quantitatively establish the detection method.The results showed that the plasma concentrations of patozuril and toltrazuril showed a good linear relationship in the concentration range of 0.10 to 20.00μg/mL(R~2>0.999),the average recovery rate of patozuril was 89.65% to 96.12%,the average recovery rate of toltrazuril was 90.84% to 94.27%,the intra-and inter-assay coefficients of variation were both less than 5%.It's suggesed that the established high performance liquid chromatography was simple,effective and reproducible,and could be used for the determination of patozuril and toltrazuril in chicken plasma.