间接竞争化学发光酶联免疫分析法检测番茄酱和果汁中异细交链孢菌酮酸

针对番茄酱和果汁中链格孢霉菌毒素污染问题,建立异细交链孢菌酮酸(iso-tenuazonic acid,ITe A)间接竞争化学发光酶联免疫分析法。通过棋盘法和单因素试验确定最佳包被原质量浓度为15.6 ng/m L、抗体质量浓度为稀释96 000倍、缓冲体系pH 7.4、竞争反应时间和二抗反应时间分别为40 min和50 min。方法IC50为2.13 ng/m L,线性范围为0.50~9.12 ng/m L,检测限为0.10 ng/m L,与结构功能类似物交叉反应率均小于0.1%,样品添加回收率在78.60%~110.83%之间,变异系数均小于15%。本方法适用于样品中ITe A污染的特异性快速筛查。

Development of Indirect Competitive Chemiluminescene Enzyme Immunoassay (icCLEIA) Method for Determination of Iso-Tenuazonic Acid in Fruit Juice and Tomato Sauce

An indirect competitive chemiluminescene enzyme immunoassay (icCLEIA) for the rapid determination of isotenuazonic acid (ITeA) in fruit juice and tomato sauce was developed. The optimized experimental parameters were obtained as follows: the coating antigen concentration was 15.6 ng/mL, the antibody was diluted 96 000 folds, the buffer pH was 7.4, and the time of competition and anti-mouse IgG reaction were 40 and 50 min, respectively. The half maximal inhibitory concentration (IC50) of the proposed method was 2.13 ng/mL with limit of detection (LOD) of 0.10 ng/mL in the linear range from 0.50 to 9.12 ng/mL. The cross-reactivity with ITeA analogues was lower than 0.1%. The recoveries of ITeA from spiked samples ranged from 78.60% to 110.83%. The coefficient of variation was lower than 15%. This proposed icCLEIA provided an efficient method for rapid detection of ITeA in real samples.