高效液相色谱法分析交沙霉素及交沙霉素片中的有关物质

Analysis of Related Substances in Josamycin and Josamycin Tablets by HPLC

目的:采用反相高效液相色谱法对交沙霉素原料及交沙霉素片中的有关物质进行分析。方法:采用Ultimate-AQ-C18色谱柱(250 mm×4.6 mm,5μm),流动相A为0.2 mol·L-1四丁基硫酸氢铵-0.2 mol·L-1磷酸氢二钠(pH3.0)-乙腈-水(3∶5∶24∶68),流动相B为0.2 mol·L-1磷酸氢二钠一水合物(pH 3.0)-乙腈-水(5∶50∶45),线性梯度洗脱。流速为1.5 ml·min-1,检测波长为232 nm,柱温为50℃。结果:交沙霉素主峰与杂质A、B、C、D、E相邻各峰间分离度均较好;交沙霉素主峰与其保留时间为1.1倍处杂质峰的分离度大于1.7。交沙霉素最低检出限为1.43 ng。交沙霉素原料和交沙霉素片中杂质A、B含量在1.5%以下,杂质D所占比例在2.0%以下,杂质E所占比例在3.0%以下,交沙霉素其它最大杂质所占比例在1.0%以下,总杂质所占比例在12%以下。交沙霉素原料中杂质C所占比例在1.0%以下,而交沙霉素片中杂质C所占比例在3.0%以下。结论:本方法灵敏度高,重现性好,专属性强,结果准确可靠,可以有效地分析交沙霉素原料及交沙霉素片中有关物质。

Objective： To establish a determination method for the related substances in josamycin and josamycin tablets by HPLC. Methods：High performance liquid chromatography was used. The column was Ultimate-AQ-C18（250 mm ×4.6 mm,5 μm）, the mobile phase respectively was mobile phase A of 0. 2 mol·L^-1 tetrabutylammonium hydrogen sulphate R-0. 2 mol·L^-1 disodium hydrogen phosphate（pH 3. 0）-acetonitrile R-water（3∶5∶24∶68） and mobile phase B of 0. 2 mol·L^-1 disodium hydrogen phosphate （pH 3. 0）-acetonitrile R-water（5∶50∶45）. The flow rate was 1. 5 ml·min^-1 with the detection wavelength of 232nm. The column temperature was 50℃. Results：The main component josamycin had a good separation with the other related substances. The resolution between josamycin and the related substances with the relative retention time of 1. 1-fold of josamycin was above 1. 7. The detection limit of josamycin was 1. 43ng. In josamycin raw materiel and josamycin tablets, the percentage of related substance A and B was less than 1. 5%. The percentage of related substance D was below 2. 0%. The percentage of related substance E was less than 3. 0%. The percentage of the other maximum related substances was less than 1. 0%. The percentage of all related substances was less than 12%. The percentage of related substance C was less than 1. 0% in josamycin raw materiel while 3. 0% in josamycin tablets. Conclusion：The method is accurate, sensitive and reliable in the determination of related substances in josamycin and josamycin tablets.