高效液相色谱法测定猪血清中的依维菌素

目的　建立猪血清中依维菌素 (iverm ectin)的高效液相色谱测定方法。方法　以 0 .0 5 %偏磷酸 -甲醇(7∶ 3)沉淀猪血清蛋白后用乙酸乙酯提取依维菌素 ,离心后取上清液用旋转真空蒸发器挥干溶剂 ,用甲醇溶解残渣后进样分析。色谱柱为反相 C18柱 (5μm ,2 5 0 m m× 4 .6 m m) ,流动相为甲醇 -水 (90∶ 10 ) ,流速 1ml/ m in,紫外检测波长为 2 4 5 nm。结果　方法检出下限为 0 .0 10 mg/ L ,线性范围为 0 .0 10～ 2 0 m g/ L ,相对标准差为 0 .78%～ 3.82 % ,加标回收率为 94 .0 %～ 10 0 .0 %。结论　本法操作简便 ,结果准确可靠 ,适用于猪血清中微量依维菌素的测定

Determination of Ivermectin in Pig Serum by High Performance Liquid Chromatography

OBJECTIVE: To establish a high performance liquid chromatographic method for the determination of micro-amount of ivermectin in pig serum. METHODS: Ivermectin in pig serum was extracted with ethyl acetate after the serum protein was precipitated with 0.05% metaphosphoric acid-methanol in the ratio of 7:3 (V/V). Then the sample was centrifuged at 4000 r/min for 5 min and the supernate was evaporated to dryness with rotary vacuum evaporator. The residue was dissolved with 0.20 ml of methanol as the sample solution for HPLC analysis. HPLC column used was a phenomenex C18 (5 microm, 250 mm x 4.6 mm) with a same type of guard column. Mobile phase consisted of methanol and water in the ratio of 90:10 (V/V) and the flow rate was 1.0 ml/min. The detection wavelength was 245 nm. RESULTS: The linear range of the method was found to be 0.010-20 mg/L and its detection limit was 0.010 mg/L. The relative standard deviation of the method was 0.78%-3.82% and the recoveries varied from 94.0% to 100.0%, with an average recovery of 97.3%. CONCLUSION: This method is simple, reproducible, accurate and suitable to the determination of micro-amount of ivermectin in pig serum.