微量动态显色法用于细菌内毒素检测的可行性研究  

Feasibility study of micro kinetic-chromogenic method for bacterial endotoxin detection

作　　者：陈素珍[1] 陈琪[1] 曾秋敏[1] 吴燕虹 CHEN Suzhen;CHEN Qi;ZENG Qiumin;WU Yanhong(Institute of Drug Control in Guangdong Province,Guangzhou 510180,China)

机构地区：[1]广东省药品检验所,广州510180

出　　处：《中国药品标准》2024年第6期607-611,共5页Drug Standards of China

摘　　要：目的:建立微量动态显色法,并验证其用于细菌内毒素检查的可行性。方法:建立鲎试剂、样品分别为25、25μL的加样体系,通过考察准确度和精密度、线性和范围、最低检测限、试剂耐用度进行方法学验证,并采用动态显色法和微量动态显色法两种方法比较对39个不同品种进行方法适用性研究。结果:0.03、0.3、3.0 EU·mL^(-1)3个浓度6次重复实验的回收率分别为(96.19±16.68)%、(92.53±10.17)%、(89.54±25.84)%;6条标准曲线的相关系数(r)的绝对值为(0.9977±0.0023);鲎试剂最低灵敏度浓度0.01 EU·mL^(-1)时,反应时间为(3896.0±279.6)s,均小于阴性对照的反应时间;2个不同厂家鲎试剂的耐用度检查结果符合规定;品种适应性研究结果显示,39个品种的回收率在50%~200%之间,两种方法的有效性相同。结论:微量动态显色法的准确度和精密度、线性和范围、最低检测限、试剂耐用度均符合方法验证要求,品种适用性与传统的动态显色法相等,该方法可用于常规样品的细菌内毒素检测。Objective:To establish a micro kinetic-chromogenic method and verify its feasibility for bacterial endotoxin testing.Methods:Establish an addition system for micro kinetic-chromogenic lysate reagent with 25μL and 25μL sample,respectively.Methodological validation was conducted by examining accuracy and precision,linearity and range,minimum detection limit,and reagent durability.Two methods,kinetic-chromogenic method and micro kinetic-chromogenic method,were used to compare the applicability of the method to 39 different varieties.Results:The recovery rates of three doses of 0.03,0.3,and 3.0 EU·mL^(-1) in six repeated experiments were(96.19±16.68)%,(92.53±10.17)%,and(89.54±25.84)%,respectively.The absolute value of the correlation coefficient(r)for the six standard curves is(0.9977±0.0023).When the minimum sensitivity concentration of lysate reagent is 0.01 EU·mL^(-1),the reaction time is(3896.0±279.6)s,which is lower than the reaction time of the negative control.The durability test results meet the requirements by two micro kinetic-chromogenic lysate reagents from different manufacturers.The research results on variety adaptability show that the recovery rates of 39 varieties are between 50%and 200%,the effectiveness of the two methods is the same.Conclusion:The accuracy and precision,linearity and range,minimum detection limit,and reagent durability of micro kinetic-chromogenic method all meet the requirements of method validation.The variety applicability is equivalent to that of the traditional kinetic-chromogenic method,so this method can be used for bacterial endotoxin detection of routine samples.

关 键 词：微量动态显色法 细菌内毒素 方法学验证 品种适应性研究 动态显色法 

分 类 号：R921.2[医药卫生—药学]