UPLC法与HPLC法测定加替沙星微球有关物质及微球的光稳定性评价

目的:采用超高效液相色谱法(UPLC)与高效液相色谱法(HPLC)测定加替沙星微球的有关物质,并进行微球的光稳定性评价。方法:UPLC法、HPLC法的色谱柱分别为Waters ACQUITY uplc HSS T3、Diamosil C18(2),流速分别为0.6、1.0mL·min-1,进样量分别为5、50μL,流动相均为三乙胺磷酸溶液(pH4.3)-乙腈(88:12),检测波长均为293nm;另取微球和原料药在温度为25℃、湿度为60%、光照强度为6600lx下放置5、10d后,采用UPLC法考察杂质谱变化情况。结果:UPLC法和HPLC法检测加替沙星的最低检测限分别为0.001、4.1ng,最低定量限分别为0.002、15.3ng,检测时间分别为8、40min左右;微球和原料药经光照后杂质谱相似,但原料药中的部分杂质含量明显增加。结论:在拟定的色谱条件下,与HPLC法比较,UPLC法更快速、灵敏度更高;加替沙星制成微球后对光的稳定性增加。

Evaluation of Light Stability of Gatifloxacin Microsphere and Related Substances by UPLC and HPLC

OBJECTIVE：To detect the related substances of Gatifloxacin microsphere by UPLC and HPLC,and to evaluate the light stability of microspheres.METHODS：UPLC was performed on Waters ACQUITY uplc~HSS T3 column with the flow rate of 0.6 mL·min-1 and the injection volume was 5 μL;HPLC was performed on Diamosil C18（2） column with the flow rate of 1.0 mL·min-1 and the injection volume was 50 μL.The mobile phase was triethylamine-phosphoric acid solution（pH4.3）-acetonitrile（88：12）,and detection wavelength was set at 293 nm.The chromatograms change of impurity was detected by UPLC after Gatifloxacin microsphere and raw material were kept at 25 ℃ with moisture of 60% and light intensity of 6 600 lx for 5 days and 10 days.RESULTS：LOQ of gatifloxacin were 0.001 ng and 4.1 ng,and LOD were 0.002 ng and 15.3 ng for UPLC and HPLC.The determination duration were 8 and 40 min.The impurity chromatograms of microspheres and raw material were similar and the contents of some impurity in raw material increased.CONCLUSIONS：Under fixed chromatogram condition,UPLC method is proved to be more rapid and sensitive than HPLC method.The light stability of gatifloxacin is improved after being made into microspheres.