反相离子对色谱法测定2,5-二羟基苯磺酸盐及有关物质

Ion-pair-RP-HPLC determination of dobesilates and their related substances

目的:建立一种反相离子对高效液相色谱法测定2,5-二羟基苯磺酸盐(包括钙盐、铵盐)的含量及其有关物质。方法:选用Kromasil 100-5C18柱(250 mm×4.6 mm,5μm),以乙腈-甲醇-0.2%磷酸二氢钾溶液(含0.2%四丁基溴化铵,用磷酸调pH为3.10±0.05)(10∶25∶65)为流动相,流速1.0 mL.min-1,检测波长260 nm,柱温30℃。结果:IP-HPLC测定2,5-二羟基苯磺酸钙、2,5-二羟基苯磺酸铵的线性范围均为20.0～500μg.mL-1,相关系数分别为0.9999和0.9998;最低检测限(S/N≥3)分别为1.44和1.64 ng;精密度RSD(n=6)分别为0.21%和0.23%。结论:本法准确、可靠,应用范围广泛。

Objective：To establish an ion-pair-RP-HPLC method for the determination of dobesilates（including calcium dobesilate and ammonium dobesilate） and their related substances.Methods：The separation was performed on a Kromasil 100-5C18 column（250 mm×4.6 mm,5 μm）.The mobile phase was composed of acetonitrile-methyl alcohol-0.2% tetrabutyl ammonium bromide in 0.2% potassium dihydrogen phosphate solution（pH 3.10±0.05）,（10∶25∶65）.The detection was carried out with a UV detector and the detective wavelength was set at 260 nm.The flow rate was 10 mL·mL-1 and the column temperature was 30 ℃.Results：The calibration curves of calcium dobesilate and ammonium dobesilate were both linear between 20.0-500 μg·mL-1;The correlation coefficient were 0.9999,0.9998,separately.The corresponding least detection limits were 1.44 ng and 1.64 ng,separately.The precisions were good（RSD=0.21%,0.23%）.Conclusion：The method is credible and the results of the determination are accurate.