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

XU Xiaojie; MENG Xiaoli; REN Lihong; DING Bo

doi:10.13506/j.cnki.jpr.2023.12.008

Abstract：Objective This study aims to establish microbial limit test of clarithromycin products on the basis of enzy-molysis test.Methods The different dose units of macrolide esterase was used in enzymolysis test of clarithromycin.Clar-ithromycin 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 clar-ithromycin tablets and clarithromycin granules.Results After 30 min of macrolide esterase,the equilibrium of enzyme hy-drolysis 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 2 000 mL trypticase soy broth(TSB)including 20 U macrolide esterase.Conclusion The microbial limit test of clarithromycin preparations was estab-lished 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.