一种测定蛹虫草中虫草酸含量的酶标仪微量反应方法

A Microplate Reader Micro-Reaction Method to Determine Cordycepic Acid Content in Cordyceps militaris

研究和建立一种基于酶标仪-96孔板高通量测定虫草酸含量的检测方法,并对该方法进行性能评价。以酶标仪为检测仪器,在96孔板内按照设定反应条件微量加入样品和试剂进行显色反应,利用酶标仪测定吸光度值并计算虫草酸含量。通过检测精密度、重复性、回收率,并与分光光度计法进行比较,综合评价该方法的准确度、精确度。结果表明,测定数据具有较高的精密度(样品CM1的RSD值0.829%;样品CM2的RSD值1.772%)、重复性(标准样品B40的RSD值2.061%;样品CM2的RSD值1.599%)、回收率(平均回收率99.24%,RSD值3.666%),测定结果与分光光度法检测结果无显著差异(P>0.05)。结果表明,酶标仪微量法测定准确、重复性好,并可大大减少样品和试剂的用量,该方法方便、快捷、高效,可以替代分光光度法用于虫草酸含量的测定。

A high-throughput determination method of cordycepic acid content based on microplate reader-96-well plate was established and evaluated for its performance.Microplate reader as detection instrument,micro-add samples and reagents in 96-well plate according to the set up reaction conditions to carry out color reaction,and use microplate reader to measure the absorbance value and calculate the content of cordycepic acid.The accuracy and precision of the method were comprehensively evaluated by testing the precision,repeatability and recovery rate,and compared with the spectrophotometric method.As a result,the measurement data has high precision(RSD value of sample CM1 was at 0.829%;RSD value of sample CM2 at 1.772%),repeatability(RSD value of standard sample B40 was 2.061%;RSD value of sample CM2 at 1.599%),recovery(average recovery rate at 99.24%,RSD value at 3.666%),and there was no significant difference(P>0.05)between the determination results and the spectrophotometric results.In conclusion,the micro-reaction method of microplate reader was accurate and repeatable,and could greatly reduce the amount of samples and reagents.This method was convenient,fast and efficient,and could replace the spectrophotometric method for the determination of cordycepic acid content.