SPR技术检测奶粉中生物素

Detection of biotin in milk powder basing on surface plasmon resonance

中文关键词:  表面等离子共振  生物芯片  生物素  微生物法

英文关键词:surface plasmon resonance  biochip  biotin  microbiological method

中文摘要:

      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0.1 μg/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。

英文摘要:

      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 competitive 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 0.1 μg/100 g. Biotin contents below the required threshold level were observed in none of ten milk samples. Less than four hours was consumed for the determination of biotin by the developed method. Conclusion This method may offer a simple and efficient approach for biotin quantification.