摘要
目的：利用表面等离子共振(surface plasmon resonance, SPR)技术,建立快速定量测定牛奶中生物素的方法。方法将生物素共价偶联到表面等离子共振芯片CM5表面,并对竞争结合的生物素结合蛋白的结合浓度及芯片的再生条件进行优化,检测芯片的稳定性。在无抗生素牛奶中添加系列质量浓度的生物素,利用免疫竞争抑制原理构建标准曲线,并对市售10个奶粉样品进行检测。结果制备的芯片稳定,50个循环相对标准偏差(relative standard deviation, RSD)小于10%。日间批内同一样品差异为8.75%,该方法的检测限为0.1μg/100 g,回收率为80.4%~91.2%。10个牛奶产品中生物素含量全部在固定的允许范围内。所建立的方法可以在4 h内完成样品的前处理和检测。结论该方法是一种简便、快捷的定量检测方法。
Objective To build a method of fast-testing biotin in milk by using the technique of surface plasmon resonance (SPR). Methods Biotin was covalently coupled to laboratory prepared CM5 chip to obtain vitamin chip. The optimal antibody working concentration and chip regeneration conditions were investigated. Meanwhile, the stability of vitamin chip was measured. A series of different concentrations of biotin was added to blank milk samples to construct a standard curve for biotin quantification according to the principle of com-petitive immune inhibition assay. Ten commercial milk samples were determined by the developed method. Results Vitamin chip displayed good stability, and the relative standard deviation(RSD)for 50 cycles was less than 10%.The limit of detection (LOD) of biotin was 0.1μg/100 g. Biotin contents below the required thre-shold level were observed in none of ten milk samples. Less than four hours was consumed for the determina-tion of biotin by the developed method. Conclusion This method may offer a simple and efficient approach for biotin quantification.
出处
《食品安全质量检测学报》
CAS
2014年第6期1723-1727,共5页
Journal of Food Safety and Quality
