HPLC-ECD法测定大观霉素的含量

Determination of Spectinomycin by HPLC-ECD

建立了HPLC-ECD法测定大观霉素含量的分析方法。采用高效液相色谱-电化学检测器,Waters XBridge C18色谱柱(4.6 mm×250 mm,5μm),以pH 3.2的草酸盐缓冲溶液-乙腈(1000∶105,V∶V)为流动相,流速1.0 mL/min;柱后衍生溶液为21 g/L的NaOH溶液,流速0.5mL/min;ECD系统参数:极性为阳性,检波+0.12 V,氧化电位+0.70 V,还原电位-0.60 V,范围5μA。结果表明,大观霉素及4R-二氢大观霉素在40～320μg/mL范围内呈良好的线性关系(大观霉素r=0.9999,4R-二氢大观霉素r=0.9992),平均回收率为100.0%(n=9),RSD为0.2%。该方法解决了微生物效价测定大观霉素含量偏高的难题,且操作简便、结果准确,可用于大观霉素及相关制剂产品的质量控制。

The quantitative method for determination of spectinomycin by HPLC - ECD was established. The column used was C18 Waters XBridge packed with 5 μm particles（4.6 mm ×250mm,5μm）. The mobile phase was a mixture of pH 3.2 oxalate buffer and acetonitrile in ratio of 1000 ： 105 （ V ： V） with the flow rate of 1.0 mL/min. The post- column derivatization solution was sodium hydroxide solution at the concentration of 21 g/L with flow rate of 0.5 mL/min. ECD system parameters read as follows ： positive polarity, detection of 0. 12 V, oxidation of 0.70 V, reduction of - 0. 60 V, range of 5 μA. The results indicated that the spectinomycin dihydrochloride and （4R） -dihydrospectinomycin dihydrochloride showed linearity in the range of 40 -320 μg/mL,the linear coefficient of speetinomycin dihydrochloride was 0. 9999, the linear coefficient of （4R） - dihydrospectinomycin dihydrochloride was 0. 9992, and the average recovery rate was 100.0% （ n = 9 ） , the relative standard deviation was 0.2%. This method was easy, precise and could be used for quality control of spectinomycin and its related preparation products.