胆固醇的细菌内毒素检查方法研究

Study on Bacterial Endotoxin Test of Cholesterol

关键词:  胆固醇  难溶性辅料  细菌内毒素  方法学研究

Key words:  cholesterol  insoluble excipients  bacterial endotoxin  methodological research

摘要:

目的 为难溶性辅料胆固醇(供注射用)建立细菌内毒素检测方法。方法 通过筛选出适合溶解胆固醇的有机溶剂,并经过9种排除干扰的方法筛选,采用凝胶法和动态浊度法进行干扰试验,排除了有机溶剂和胆固醇对内毒素检查法的干扰,最终建立了胆固醇的细菌内毒素质量控制方法。检测条件:取胆固醇用无水乙醇配制成20mg·mL^-1溶液,用0.5mg·mL^-1聚氧乙烯(35)蓖麻油水溶液代替细菌内毒素检查用水稀释100倍,按内毒素检查法动态浊度法检测。结果 3批样品使用2个厂家的动态浊度法鲎试剂的回收率均在50%~200%,并且内毒素检测值均<0.1 EU·mg^-1。如使用凝胶法,该品种只能使用1个厂家最高灵敏度的试剂进行检测。因此,胆固醇不建议使用凝胶法进行检测。结论 建立了胆固醇(供注射用)细菌内毒素动态浊度法检测方法,用于质量控制。

Abstract:

OBJECTIVE To establish a bacterial endotoxin detection and control method for insoluble excipient cholesterol(for injection).METHODS The bacterial endotoxin quality control method for cholesterol was established by screening the organic solvent suitable for cholesterol dissolution and screening through 9 methods to eliminate the interference of organic solvent and cholesterol to the endotoxin test. The detection conditions were as follows: cholesterol was prepared with anhydrous ethanol into 20 mg·mL^-1 solution, diluted 100 times with 0.5 mg·mL^-1 polyoxyethylene(35) castor oil solution instead of bacterial endotoxin test water, and detected by kinetic-turbidimetric assay of endotoxin test.RESULTS The recoveries of the kinetic-turbidimetric assay limulus reagent from two manufacturers were 50%-200%, and the values of endotoxin were all <0.1 EU·mg^-1. At present, this variety could only be tested with the most sensitive reagent from one manufacturer when use gel method. Therefore, cholesterol detection by gel method was not recommended.CONCLUSION A kinetic-turbidimetric assay for the determination of bacterial endotoxin in cholesterol(for injection) is established for quality control.