广谱检测磺胺类药物的酶联免疫分析方法的建立

目的 利用抗磺胺类药物多抗建立酶联免疫分析方法,测定磺胺类药物在动物源食品中的残留.方法 以抗磺胺类药物抗体包被微孔板,辣根过氧化物酶(HRP)标记磺胺类半抗原磺胺噻唑(ST),磺胺间甲氧嘧啶为标准品建立酶联免疫分析方法.结果 本方法的标准曲线测定范围为0.3～ 100ng/mL.方法学鉴定结果表明,灵敏度为0.2ng/mL,批内、批间变异分别为9.2％ ～ 14.4％、10.5％～ 12.7％.牛奶、蜂蜜、水产品及鸡肉、鸡蛋样品的添加回收率分别在79.3％～88.9％、65.5％～136.4、63.0％～83.3％、59.3％～78.1％之间.牛奶稀释实验表明,测定值与稀释度呈线性相关,相关系数r=0.997.结论 本方法可同时快速测定动物源性食品中23种磺胺类药物的残留,对19种磺胺类药物的检测灵敏度高于10ng/mL.

Establishment of Enzyme-linked Immunosorbent Assay for Broad-specificity Detection of Sulfonamides

Objective To develop an indirect competitive enzyme-linked immunoassay(ELISA) for detection of residues of sulfonamides in food.Methods The polyclonal antibody against sulfonamides was coated in microplate,N-sulfanilyl-4-amino-thiazol-acetic acid was labeled with horseradish peroxidase.Sulfamonomethoxine was used as calibrator.Results The range of standard curve was 0.3-100ng/mL.The sensitivity of the assay was 0.2ng/mL.The intra-assay and inter-assay CVs were 9.2-14.4%,and 10.4-12.7% respectively.The recovery of SAs in milk,honey,fish and eggs were 79.3%~88.9%,65.5%~136.4%,63.0%~83.3% and 59.3%~78.1% respectively.The correlation coefficient between measured and expected value was 0.997 after serial dilutions of the samples with high concentration of SAs.Conclusion The method developed in this study would detect 23 kinds of sulfonamides rapidly at the same time.The sensitivity of this assay for 19 kinds of sulfonamides is superior to 10ng/mL.