克仑特罗食物中毒的酶联免疫法筛选与气-质联机确证研究

本文采用酶联免疫法筛选和气-质确证对克仑特罗食物中毒病因学诊断技术进行研究。组织样品经匀浆初提,液液萃取;生物材料经离心适当稀释,上酶标板显色进行筛选测定。克仑特罗浓度的自然对数与吸光度比值呈线性关系,线性方程Y=-22.078ln(X)+199.24,相关系数为0.9922。检出限为0.1μg/kg。在对猪肝、鸡肉和尿液样品基质的不同加标水平的回收率在50.5%～92.0%之间,相对标准偏差在12.3%～18.1%之间。中毒者食用中毒食品(酱猪肝)及血液和尿液的测定结果与气质联机法确证结果基本吻合,而且生物体液(特别是尿液在中毒7d内仍可以作为克仑特罗食物中毒的辅助病因学诊断材料。

Application in Food Clenbuterol Poisoning of ELISA Screening and Confirmation by GC-MS

An enzyme-linked immunosorbant assay (ELISA) method and validation by GC-MS for clenbuterol residue in liver,kidney and muscle samples and biological samples was set up. The method was developed to enable rapid, specific, sensitivedetermination of clenbuterol residues in tissue samples and biological samples. Tissue samples were comminuted and extractedby perchloric acid solution, after liquid-liquid partition by mixture of isopropanol and ethyl acetate (40:60), the extracts wereapplied into the plate. Clenbuterol standard was spiked into liver and chicken samples by different levels respectively. Therecovery rate was from 50.5 to 92.0%, RSD was from 12.3 to 18.1%. Cooked toxic liver and blood and urine samples of toxicsubjects were determined. Positive samples from ELISA method were confirmed by gas chromatography-mass spectrometry.The ELISA results were consistent with the results of GC-MS.