复方酮康唑乳膏中酮康唑、丙酸氯倍他索和硫酸新霉素的含量测定

Contents determination of ketoconazole,clobetasol propionat and neomycin sulfate in compound ketoconazole cream

目的:建立HPLC法、抗生素微生物检定法测定复方酮康唑乳膏中酮康唑、丙酸氯倍他索和硫酸新霉素的含量。方法:用HPLC法测定酮康唑和丙酸氯倍他索含量,色谱条件如下:Kromasil 100-5C18柱(150mm×4.6mm,5μm);流动相为甲醇-0.02mol/L磷酸二氢钠溶液(内含0.15%三乙胺)=68∶32;流速1.0ml/min;柱温为室温;检测波长为239nm;进样量20μl。以金黄色葡萄球菌作为实验菌株,采用抗生素微生物检定法中管碟法中的二剂量法测定硫酸新霉素的效价。结果:酮康唑在25~800μg/ml范围内线性关系良好(r=0.999 9),平均回收率为98.52%,RSD为1.33%(n=6)。丙酸氯倍他索在0.625~20μg/ml范围内线性关系良好(r=0.999 9),平均回收率为97.60%,RSD为1.44%(n=6)。硫酸新霉素的回归非常显著(P<0.01),偏离平行不显著(P>0.05),可靠性测验结果成立,可信限率(FL%)良好,效价符合要求。结论:两种测定方法准确、灵敏,专属性强,重现性好,对复方酮康唑乳膏质量控制标准的提高具有参考意义。

Objective：To establish HPLC method and microbiological assay method for contents determination of ketoconazole,clobetasol propionat and neomycin sulfate in compound ketoconazole cream.Methods：The contents of ketoconazole and clobetasol propionat were determined by HPLC method.The separation column of Kromasil 100-5 C18（150 mm×4.6 mm,5μm）was used.Methanol-0.02 mol/L sodium dihydrogen phosphate（containing 0.15%triethylamine,68∶32）was applied as the mobile phase.The flow rate was 1.0 ml/min and the column temperature was set at room temperature.The detection wavelength was 239 nm and the sample size was 20μl.The two-dose assay of incylinder-plate method of microbiological assay was used to determine the potency of neomycin sulfate with staphylococcus aureus as the tester strain.Results：Ketoconazole and clobetasol propionat displayed good linearity within the range of 25-800μg/ml（r=0.999 9）and 0.625-20μg/ml（r=0.999 9）,respectively.The average recoveries were 98.52%（RSD 1.33%,n=6）and 97.60%（RSD 1.44%,n=6）,respectively.The regression of neomycin sulfate was very significant（P〈0.01）.The parallel deviate was not significant（P〉0.05）.The reliability test results were established.The confidence-limit rate（FL%）of the experiment results was good.Conclusion：The two methods are accurate,sensitive,specific and reproducible,which could provide the reference for heightening the quality standards of compound ketoconazole cream.