HPLC法测定5-氨基水杨酸灌肠液的含量及有关物质

目的：建立5-氨基水杨酸灌肠液的含量及其有关物质的测定方法。方法：采用高效液相色谱梯度洗脱法测定,色谱柱为Hypersil,ODS C18（250 mm×4.6 mm,5.0μm）,流动相为流动相A为pH 7.5磷酸缓冲液（PBS）∶四丁基氢氧化铵溶液（TBAH）∶甲醇∶水（600∶50∶50∶300）;流动相B为pH7.5磷酸缓冲液∶TBAH∶甲醇∶水（200∶50∶300∶450）。检测波长为208 nm;柱温40℃,进样量20μl。结果：5-氨基水杨酸与其两种降解产物可完全分离。5-氨基水杨酸在1~250μg·ml-1的浓度范围内,浓度与峰面积具有良好的线性关系,r=0.999 5,平均回收率为99.44%,RSD为0.94%（n=9）。结论：方法简单、灵敏、重复性好,可用于本品的含量和有关物质检查。

Determination of Content and Related Substances in 5-Aminosalicylic Acid Enemas by HPLC

Objective： To establish an HPLC method to determine 5-amino salicylic acid（ 5-ASA） and its related substances in the enemas. Methods： The liquid chromatograph was equipped with a Hypersil ODS C18column（ 250 mm × 4. 6 mm,5. 0 μm） with gradient elution. The mobile phase A was composed of pH 7. 5 phosphate buffer（ PBS）,tetrabutyl ammonium hydroxide solution（ TBAH）,methanol and water（ 600∶50∶50∶ 300）,the mobile phase B was composed of pH 7.5 PBS,TBAH,methanol and water（ 200∶50∶300∶450）. The wavelength was 208 nm,the column temperature was 40 ℃ and the injection volume was 20μl. Results： 5-ASA and its two degradation products could be completely separated. The concentration of 5-ASA and the corresponding peak area had a good linear relationship within the range of 1-250μg·ml- 1（ r =0.999 5）. The average recovery was 99.44%（ RSD =0.94%,n =9）. Conclusion： The method is simple,sensitive and reproducible,and can be used in the determination of the content and related substances in the enemas.