绵羊血浆阿维菌素荧光高效液相色谱检测法的建立

Determination of avermectin level in plasma of sheep by HPLC with fluorescence detection

建立了一种绵羊血浆中阿维菌素 (AVM )荧光高效液相色谱 (HPLC)检测法。用甲醇提取绵羊血浆中的AVM ,并用ODS C18固相萃取法进行纯化 ,纯化后的AVM经干燥处理后 ,用三氟乙酸酐和N 甲基咪唑对其进行荧光衍生化 ,用荧光HPLC进行检测。本方法的最低检测限为 0 .1ng/mL ,在血浆AVM含量 2 .5～ 5 0 .0ng/mL范围内 ,标准曲线方程为Y =2 72 12x -10 412 (r =0 .9995 ) ,样品的平均回收率为 92 .0 2 %± 6.3 3 %。批内变异系数 1.98%～ 6.2 2 % ,批间变异系数 2 .3 1%～ 8.94%。检测结果显示 ,本方法灵敏度高 ,干扰少 ,重复性好。

A novel method by high performance liquid chromatography (HPLC) with fluorescence detection was developed to determinate avermectin level in plasma of sheep. Samples were extracted with methanol and cleaned up by solid phase extraction (SPE) on a C 18 column, followed by derivation with trifluoroacetic anhydride and N methylimidazole . Then the samples were injected into the HPLC column. Avermectin level was detected by fluorescence detector set at 364 nm excitation and 470 nm emission wavelength. The limit of quantification was 0.1 ng/mL avermectin in plasma of sheep. Mean recovery rate was 90.02% , with a SD 6.33%. The procedure provides a rapid, reliable and sensitive method for determination avermectin in plasma or serum, which is the key to study pharmacokinetics of avermectins and their formulation.