HPLC-ECD法测定盐酸大观霉素有关物质

目的:建立HPLC-ECD法测定盐酸大观霉素有关物质。方法:应用高效液相色谱-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)为流动相,流速为1.0 mL.min-1;柱后溶液为21 g.L-1的氢氧化钠溶液,流速0.5 mL.min-1;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20μA。结果:盐酸大观霉素有关物质在2.25～45μg.mL-1范围内呈良好的线性关系,r=0.9999;Y轴截距在100%浓度响应值的7.3%处,在100%浓度的25%～500%的范围内,对任一点的响应值在此点理论值的97.7%至103.6%的范围内;平均回收率(n=9)为98.9%,RSD为2.9%。结论:该方法简便易行,准确度高,重复性好,可用于盐酸大观霉素及相关制剂产品的质量控制。

Determination of spectinomycin hydrochloride related substances by HPLC-ECD

Objective:To establish a quantitative method for spectinomycin hydrochloride related substances by HPLC-ECD.Methods:The detection was performed on a Waters XBridge C18 column(4.6 mm×250 mm,5 μm).The mobile phase was a mixture of pH 3.2 oxalate buffer and acetonitrile in a ratio of 1000∶ 105,with a flow rate of 1.0 mL·min-1;The post-column derivatization solution was 21 g·L-1 sodium hydroxide solution with a flow rate of 0.5 mL·min-1.ECD system parameters:Polarity:Positive;Detector:+0.12 V;Oxidation potential:+0.70 V;Reduction potential:-0.60 V;Range:20 μA.Results:The linear range of the spectinomycin hydrochloride related substances was 2.25-45 μg·mL-1,the correlation coefficient was 0.9999.Y axial intercept in 100% concentration response valve is 7.3%,in 100% the concentration of 25%-500% range,the response value of any point ranged from 97.7% to 103.6% of the theoretical value.The average recovery rate(n=9)was 98.9%,RSD was 2.9%.Conclusion:The method is simple,reliable,accurate and reproducible,which can be used for the quality control of spectinomycin hydrochloride and related finished products.