注射用多西他赛聚合物胶束细菌内毒素定量检测法的建立

Establishment of Quantitative Detection Method of Bacterial Endotoxin in Docetaxel Polymer Micelles for Injection

目的建立注射用多西他赛聚合物胶束中细菌内毒素的定量检测方法。方法采用电镜观察和激光粒度仪检测粒径,筛选破坏胶束的适宜条件(分别以pH9,10,11的氢氧化钠溶液破坏);在样品中加入内毒素,再以pH10的氢氧化钠溶液破坏并稀释,检测内毒素浓度,并计算加标回收率;确定胶束破坏后的内毒素检测方法,并验证方法的准确性和精密度。结果pH10的氢氧化钠溶液可破坏胶束;胶束破坏后加入内毒素并稀释至0.5 mg/mL(内毒素为0.5 EU/mL)时内毒素的加标回收率较高(91.81%),且方法的准确性和精密度均较好,当加入内毒素浓度为1.00,0.10,0.01 EU/mL时,平均回收率在96%~108%之间,变异系数<5%,均满足检测要求;3批样品中均未检出内毒素(<0.02 EU/mg)。结论建立的细菌内毒素定量检测方法可用于注射用多西他赛聚合物胶束细菌内毒素的定量检测。

Objective To establish a quantitative detection method of bacterial endotoxin in Docetaxel Polymer Micelles for Injection.Methods The particle size were observed by electron microscope and laser particle size analyzer to screen the suitable conditions for destroying micelles(destroyed by sodium hydroxide solution with pH9,10 and 11,respectively).Endotoxin was added into the sample,and then destroyed and diluted with pH10 sodium hydroxide solution to detect the concentration of endotoxin,and the recovery rate was calculated.The endotoxin detection method after micelle destruction was determined,and the accuracy and precision of the method were verified.Results pH 10 sodium hydroxide solution could destroy the micelles.When endotoxin was added after micelle destruction and diluted to 0.5 mg/mL(the concentration of endotoxin was 0.5 EU/mL),the recovery rate of endotoxin was higher(91.81%),and the accuracy and precision of the method were good.When the concentration of endotoxin was 1.00,0.10 and 0.01 EU/mL,the average recovery rate was in the range of 96%-108%,and the coefficient of variation was lower than 5%,which met the detection requirements.No endotoxin(<0.02 EU/mg)was detected in the three batches of samples.Conclusion The established quantitative detection method could be used for the quantitative detection of bacterial endotoxin in Docetaxel Polymer Micelles for Injection.