增强化学发光酶免疫法对猪肉中盐酸克伦特罗的检测

建立了猪肉中克伦特罗(CLB)的直接竞争化学发光酶免疫检测(dc-CLEIA)方法。通过优化离子强度、pH值确立了化学发光酶免疫法的标准曲线,优化后dc-CLEIA的检出限可达0.02μg/L。猪肉中的盐酸克伦特罗用高氯酸提取、SPE净化后绘制基质曲线,基质曲线与标准曲线吻合,说明基质影响基本消除。测定0.10、1.0、5.0μg/kg 3个水平的添加回收率,平均回收率为83%~98%,批内与批间的相对标准偏差均小于15%。进一步研究了dc-CLEIA与HPLC两种方法测定猪肉样品的相关性,结果显示两者测定结果相关性良好,r=0.964 7,说明所建立的直接化学发光酶免疫方法可用于实际样品的检测,结果准确可靠。

Determination of Clenbuterol Residues in Pork by Enhanced Chemiluminescent Enzyme Immunoassay

A direct competitive enhanced chemiluminescent enzyme immunoassay(dc-CLEIA) method for determination of clenbuterol(CLB) residues in pork was developed.The optimal concentration of phosphate buffer solution and its pH value were investigated.The optimal concentration of phosphate buffer solution was 0.2 mol/L and its pH was 7.4.Under the optimized condition,the limit of detection(LOD) of the assay was 0.02 μg/L,with IC_(50) value of the assay was 0.25 μg/L.Matrix interferences with the sensitivity of the assay was systematically studied.The result indicated that the matrix effects could be eliminated when pork samples were extracted with perchloric acid solution,and the extract was centrifuged at 5 000 r/min for 30 min at 4 ℃ followed by clean-up by solid phase extraction (SPE).The mean recoveries of CLB from pork samples at three spiked concentration levels of 0.10,1.0 and 5.0 μg/kg were between 83% and 98%,and the relative standard deviations(RSDs) of intra- and inter-assay were less than 15%.In addition,the developed assay was validated by high performance liquid chromatography(HPLC) and the result obtained by two methods showed better relativity with correlation coefficient of 0.964 7.Therefore,the proposed method was simple,accurate and was suitable for the rapid determination of CLB in real sample.