难溶性香菇多糖无菌原料药细菌内毒素定量检测方法建立

Establishment of a Quantitative Determination Method for Bacterial Endotoxin in Insoluble Lentinan Aseptic API

目的建立定量检测难溶性香菇多糖无菌原料药中细菌内毒素的动态浊度法。方法参考2020年版《中国药典(四部)》通则1143细菌内毒素检查法,建立细菌内毒素检查标准曲线,通过干扰试验确定样品的质量浓度范围,并考察研磨法溶解样品的细菌内毒素回收率。结果香菇多糖质量浓度在2.5000μg/mL及以下时不干扰试验,研磨法溶解样品的细菌内毒素回收率在50%~200%范围内,样品中内毒素含量可定量测定。结论动态浊度法可定量检测香菇多糖无菌原料药中的细菌内毒素。研磨法操作方便,对细菌内毒素的生物活性无影响。

Objective To establish a kinetic turbidimetric assay for quantitative determination of bacterial endotoxin in the insoluble lentinan aseptic active pharmaceutical ingredients(API).Methods According to the requirements of bacterial endotoxin test of general rule 1143 in the Chinese Pharmacopoeia(2020 Edition,VolumeⅣ),standard curves for bacterial endotoxin test were established.The mass concentration range of samples was determined through the interference test,and the recovery rate of bacterial endotoxin in samples dissolved by the grinding method was investigated.Results There was no interference when the mass concentration of lentinan was not more than 2.5000μg/mL.The recovery rate of bacterial endotoxin in samples dissolved by the grinding method was in the range of 50%-200%,and the content of bacterial endotoxin in samples could be quantitatively determined.Conclusion The kinetic turbidimetric assay can be used for the quantitative determination of bacterial endotoxin in the lentinan aseptic API.The grinding method is easy to operate and has no effect on the biological activity of bacterial endotoxin.