一种检测乳过氧化物酶方法的建立

Development of a method to detect lactoperoxidase(LP) activity in milk

【目的】建立一种简便、快速、精确的乳过氧化物酶(LP)检测方法。【方法】以ABTS为底物,利用动力学方法,建立了检测LP含量的ABTS法,并以优化后的体系检验ABTS法的精确度。【结果】以ABTS法检测LP含量的最佳反应条件为:吸收波长416nm,作用时间100s,pH5.5,ABTS浓度2mmol/L,H2O2浓度5mmol/L,工作温度25℃(室温)。经检验,该法的批内变异系数为2.39%,批间变异系数为3.42%,相关系数(R2)为0.9953,准确误差均未超过5.00%。【结论】以ABTS法检测LP精密度和准确度均较好,且操作步骤比较简便、快速,实现了对LP的简便、快速、精确检测。

[Objective]The present experiment was conducted to develop a simple, rapid and accurate method for detection of lactoperoxidase （LP） activity in milk samples. [Method ]Using 2,2＇-azino-bis （3-ethylbenzthiazoline-6-sulphonic acid） （ABTS） as the substrate, the method to detect LP activity in milk was established by determining the reaction kinetics. The reaction conditions were optimized by observing effects of different incubation periods, pH values, substrate and H2O2 concentrations and incubation temperatures. [Result]The optimized conditions for reaction were as follows： 2 mmol/L of substrate con- centration, 5 mmol/L of H202, pH 5.5, and 100 sec of response time, followed by the measurement at 416 nm wavelength. The coefficient of variation in the same group was found to be 2.39% and that of the different group was 3.42%. The correlation coefficient was 0.9953, and the accuracy rating did not exceeded beyond 5%. [Conclusion]Besides being the simple and rapid, the developed ABTS method was found to be highly accurate and precise on detection of LP compared to the other methods.