大环内酯酶酶解试验在克拉霉素制剂微生物限度检查中的应用  

Application of macrolide esterase enzymolysis test in the microbial limit test of clarithromycin preparations

作　　者：徐晓洁 孟晓丽 任丽宏 丁勃 XU Xiaojie;MENG Xiaoli;REN Lihong;DING Bo(Shandong Research Center of Engineering and Technology for Safety Inspection of Food and Drug,Shandong Institute for Food and Drug Control,Jinan 250101,China;School of Pharmaceutical Sciences,Shandong University,Jinan 250012,China)

机构地区：[1]山东省食品药品检验研究院,山东省食品药品安全检测工程技术研究中心,山东济南250101 [2]山东大学药学院,山东济南250012

出　　处：《药学研究》2023年第12期994-999,共6页Journal of Pharmaceutical Research

基　　金：国家药典委员会国家药品标准提高项目(No.2022Y21、2022Y22)。

摘　　要：目的用克拉霉素酶解试验指导建立克拉霉素制品的微生物限度检查方法。方法采用不同剂量单位的大环内酯酶酶解克拉霉素,高效液相色谱法(HPLC)测定克拉霉素含量以确定酶解时间与克拉霉素降解速度之间的关系,选择最小剂量单位和最佳酶解时间,用于克拉霉素片和克拉霉素颗粒的微生物限度检查。结果大环内酯酶作用20 min后接近最大酶解效果,30 min后达到酶解平衡,克拉霉素含量不再显著变化。20 U和10 U的大环内酯酶对0.02 mg·mL^(-1)克拉霉素的酶解作用没有显著差异。需氧菌总数选择1∶100供试液,采用薄膜过滤法,冲洗量300 mL,并在最后一次冲洗液中加入约10 U的大环内酯酶作用20 min,霉菌和酵母菌总数选择1∶10的供试液采用平皿法(倾注法)。大肠埃希菌检查采用直接接种法,2000 mL胰酪大豆胨液体培养基联合20 U大环内酯酶,可检出所加试验菌。结论大环内酯酶酶解试验指导建立了克拉霉素制剂的微生物限度检查方法,提高了检验效率,开拓了大环内酯类抗生素微生物限度检查的新思路。Objective This study aims to establish microbial limit test of clarithromycin products on the basis of enzymolysis test.Methods The different dose units of macrolide esterase was used in enzymolysis test of clarithromycin.Clarithromycin content was determined by HPLC to determine the relationship between the enzymolysis time and the degradation rate of clarithromycin.The minimum dose unit and the best enzymolysis time were selected for the microbial limit test of clarithromycin tablets and clarithromycin granules.Results After 30 min of macrolide esterase,the equilibrium of enzyme hydrolysis was reached,and the clarithromycin content did not change significantly.There was no significant difference between 20 U and 10 U macrolide esterase on the enzymolysis test of clarithromycin at 0.02 mg·mL^(-1).The total aerobic microbial count was performed using the membrane filtration method with a 1∶100 dilution of the sample product.Filter membrane was washed with 300 mL rinsing fluid,and 10 U or 20 U macrolide esterase was added to the last flush with 20 min action.The plate-count method(pour-plate method)was used for the total combined yeasts and molds count with a 1∶10 dilution of the sample product.Test for E.coli was carried out using the direct inoculation method,with 2000 mL trypticase soy broth(TSB)including 20 U macrolide esterase.Conclusion The microbial limit test of clarithromycin preparations was established by the guidance of Macrolide enzymolysis test.The method improves the efficiency of testing and opens up a new idea of microbial limit test for macrolide antibiotics.

关 键 词：克拉霉素 大环内酯酶 酶解试验 微生物限度检查 薄膜过滤法 

分 类 号：R927.1[医药卫生—药学]