Agri-Food and Biosciences Institute, Veterinary Sciences Division, Stoney Road, Belfast BT4 3SD, UK.
Anal Chem. 2011 Jun 15;83(12):5012-6. doi: 10.1021/ac200926e. Epub 2011 May 23.
The illegal adulteration of milk with melamine in 2008 in China led to adverse kidney and urinary tract effects in hundreds of thousands of children and the reported deaths of six. The milk had been deliberately adulterated to elevate the apparent protein content, and subsequently melamine was detected in many milk-related products which had been exported. This led to the banning of imports of milk and milk products from China intended for the nutritional use of children and to the implementation of analytical methods to test products containing milk products. An optical biosensor inhibition immunoassay has been developed as a rapid and robust method for the analysis of infant formula and infant liquid milk samples. A compound with a chemical structure similar to that of melamine was employed as a hapten to raise a polyclonal antibody and as the immobilized antigen on the surface of a biosensor chip. The sensitivity of the assay, given as an IC(50), was calculated to be 67.9 ng mL(-1) in buffer. The antibody did not cross-react with any of the byproducts of melamine manufacture; however, significant cross-reactivity was observed with the insecticide cyromazine of which melamine is a metabolite. When sample matrix was applied to the assay, a limit of detection of <0.5 μg mL(-1) was determined in both infant formula and infant liquid milk. The development of the immunoassay and validation data for the detection of melamine is presented together with the results obtained following the analysis of melamine-contaminated milk powder.
2008 年中国发生的三聚氰胺非法掺入牛奶事件导致数十万儿童出现肾脏和尿道不良影响，并有报道称有 6 名儿童死亡。牛奶被故意掺入以提高表观蛋白质含量，随后在许多奶制品中检测到三聚氰胺，这些奶制品已被出口。这导致禁止进口中国用于儿童营养的牛奶和奶制品，并实施了分析方法来测试含有奶制品的产品。已经开发出一种光学生物传感器抑制免疫测定法，作为一种快速而强大的方法来分析婴儿配方奶粉和婴儿液态奶样品。一种具有与三聚氰胺化学结构相似的化合物被用作半抗原来提高多克隆抗体，并作为生物传感器芯片表面的固定化抗原。该测定法的灵敏度（以 IC50 表示）在缓冲液中计算为 67.9ng/mL。该抗体与三聚氰胺制造的任何副产物均无交叉反应；然而，与杀虫剂虫螨威（三聚氰胺是其代谢物）观察到显著的交叉反应。当将样品基质应用于测定法时，在婴儿配方奶粉和婴儿液态奶中均确定检测限<0.5μg/mL。本文介绍了免疫测定法的开发和用于检测三聚氰胺的验证数据，以及对三聚氰胺污染奶粉进行分析后获得的结果。
