高效液相色谱法测定动物性食品中4种β_2-兴奋剂

Determination of four β_2-agonists in animal food by high performance liquid chromatography

目的：建立动物性食品中4种β2-兴奋剂：奥西那林、克伦特罗、沙丁胺醇和非诺特罗的高效液相色谱-二级管阵列检测的方法。方法：动物组织样品用缓冲溶液匀浆后，酶水解，高氯酸沉淀蛋白后，滤液调至碱性，用乙酸乙酯：异丙醇（6：4）提取后，旋转蒸发去除溶剂。缓冲液溶解后，过C18-SCX串联固相萃取小柱，甲醇淋洗后分析，对于含蛋白、脂肪少的动物食品样品，酶解并沉淀蛋白，滤液调至pH3后过C18-SCX串联固相萃取小柱，洗脱液用于分析。结果：对含蛋白、脂肪少的样品，样品加标回收率在60．6％～88．0％，精密度在2．72％～7．86％；含蛋白、脂肪多的样品，加标回收率在60．1％～84．9％，精密度在1．26％～6．20％，方法在0．2～50．0μg／ml有良好线性。样品的最低检出浓度：奥西那林为0．07mg／kg；沙丁胺醇为0．10mg／kg；非诺特罗为0．08mg／kg；克伦特罗为0．04mg／kg。结论：本方法操作简便、准确灵敏、重现性好、分析成本较低。

Objective：In this study, a enzymatic hydrolyzation- C18 -SCX （Strong Cation Ion Exchange） tandem SPE -high performance liquid chromatography - PAD method for four β2 - agonists in animal food was developed and evaluated. Methods ： Homogenized in buffer solution and hydrolyzed by enzyme, sample tissue with much protein was deposited with HClO4, centrifuged, and extracted with additional organic solution at adjusted alkaline pH which was evaporated revolvely later. The residue was dissolved in salt, transited by SCX - C18 tandem SPE and analysed after washed by aminated methanol. On the other hand, after hydrolyzed by enzyme, sample tissue with little protein was centrifuged at pH3.0, followed by SCX - C18 tandem SPE and was analysed after elution by aminated methanol. Results：On samples with little protein, the recoveries for the spiked samples ranged from 60. 6% to 88.0% , the relative standard deviation ranged from 2. 72% to 7.86% ; on samples with much protein, the recoveries for the spiked samples ranged from 56.4% to 84. 9% , the relative standard deviation ranged from 1.26% to 6. 20%. The analysis has a good correlativity from 0. 2 μg/ml to 50 μg/ml. The detection limit are 0. 070 mg/kg for Orciprenaline, 0. 10 mg/kg for Salbutamol, 0. 081 mg/kg for Fenoterol and 0. 038 mg/kg for Clenbuterol. Conclusion：It was concluded that this method was simple, rapid, accurate, sensitive and economic for determining the four β2 -agonists in animal food.