流动相对西罗莫司异构体在反相液相色谱上分离效果影响的研究

Effect of Mobile Phases on the Isomeric Separation of Sirolimus in Reversed-phase Liquid Chromatography

研究不同流动相对西罗莫司异构体在反相液相色谱上分离效果的影响,为准确测定西罗莫司的含量提供依据。色谱柱采用YMC J’sphere ODS-H80,流动相分别采用不同体积比例的水/乙腈(40/60,35/65,30/70,v/v)或水/甲醇(20/80,15/85,10/90,v/v)。结果表明:以水/乙腈或水/甲醇二元溶剂为流动相时,随着有机溶剂比例的增加,西罗莫司异构体(β峰与γ峰)的保留时间都变短,峰宽都变狭窄,分离度也相应降低。选择流动相水/乙腈(35/65)或水/甲醇(15/85),西罗莫司的异构体(β峰与γ峰)均能完全分离。与流动相水/乙腈(35/65)相比,以水/甲醇(15/85)作流动相,西罗莫司β异构体色谱峰的保留时间更短,峰宽更狭窄,是水/甲醇与水/乙腈流动相体系中的最佳条件。

The complete chromatographic separation of sirolimus isomers （β-peak and γ-peak） is helpful for accurately determining the content of sirolimus.The main purpose of this study was to examine the effect of mobile phases on the isomeric separation of sirolimus in reversed-phase liquid chromatography.Detection of sirolimus was performed using HPLC System, equipped with a YMC J＇sphere ODS-H80 column.The mobile phases used were binary solvent mixtures of water/acetonitrile （40/60, 35/65, 30/70, v/v） and water/methanol （20/80, 15/85, 10/90, v/v）.The experimental data showed that the retention time, peak width and resolution of sirolimus isomers （β-peak and γ-peak） all decreased significantly with the increasing organic content in the mobile phases.By adjusting the ratio of water/acetonitrile （35/65）, or water/methanol （15/85）, β-peak and γ-peak of sirolimus were separated to achieve good resolution.Furthermore, β-peak was observed to have a shorter retention time and narrower peak width in water/methanol （15/85, v/v） mixture than in water/acetonitrile （35/65, v/v） mixture.Consequently, these results show that rapid and high-resolution separation of sirolimus isomers is obtained in reversed-phase liquid chromatography with the mobile phase of water/methanol （15/85, v/v）.