Feasibility study of micro kinetic-chromogenic method for bacterial endotoxin detection
10.19778/j.chp.2024.06.013
- VernacularTitle:微量动态显色法用于细菌内毒素检测的可行性研究
- Author:
Suzhen CHEN
1
;
Qi CHEN
1
;
Qiumin ZENG
1
;
Yanhong WU
1
Author Information
1. 广东省药品检验所,广州 510180
- Publication Type:Journal Article
- Keywords:
micro kinetic-chromogenic method;
bacterial endotoxins;
methodological validation;
variety adaptabil-ity research;
kinetic-chromogenic method
- From:
Drug Standards of China
2024;25(6):607-611
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a micro kinetic-chromogenic method and verify its feasibility for bacterial endo-toxin testing.Methods:Establish an addition system for micro kinetic-chromogenic lysate reagent with 25 μL and 25 μL sample,respectively.Methodological validation was conducted by examining accuracy and precision,linear-ity 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(0.997 7±0.002 3).When the minimum sensitivity concentration of lysate reagent is 0.01 EU·mL-1,the reaction time is(3 896.0±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.
