两性霉素B效价测定方法的改进

摘要

目的 改进两性霉素B效价含量测定方法,提高方法的可靠性与可操作性.方法 管碟法.新制培养基pH6.0～6.2；pH7.0磷酸盐缓冲液(含1％聚山梨醇)；检定菌为啤酒酵母(ATCC9763);培养温度为(36±1)℃；培养时间为(20±2)hr;高、低剂量分别为10和5u/mL.结果 抑菌圈大小符合规定,边缘清晰可以使用仪器进行测量;在1.09～10.90u/mL浓度范围内线性良好(回归方程:y=0.10763x+0.50076,r=0.99965);重现性与重复性好,日内精密度RSD=0.46％(n=6),日间精密度RSD=1.25％(n=12).结论 可以作为两性霉素B的常规质量控制方法.%Objective Improve the potency determination method of amphotericin B for the reliability and operability of the method.Methods The cylinder plate method with new medium(pH6.0～6.2) and pH7.0 phosphate buffer (1％ tween-80) was used.The test organism was Saccharomyces cerevisiae (ATCC9763); Incubation lasted 18～22 hours at the temperature(36±1)C.The high and low doses were 10 and 5 amphotericin B units per mL respectively.Results The size of circular area (zone) of growth inhibition around the cylinder was moderate and the brim of ones was clear.The linearity was achieved over the range of 1.09～10.90u/mL (y=0.10763x+0.50076,r=0.99965).The reproducibility and recovery were acceptable.The average relative standard deviation (RSD) of intraday repeatability was 0.46％ (n=6) and the average RSD of inter-day repeatability was 1.25％ (n=12).Conclusion The method has the potential to become the routine quality control method for amphotericin B in pharmacopoeias.