反相高效液相色谱法手性分析和制备全反式虾青素3种异构体

Chiral analysis and separation of the three stereoisomers of all-trans-astaxanthin by reversed-phase high performance liquid chromatography

中文关键词:  虾青素  立体选择性  立体异构体  反相高效液相色谱法  制备

英文关键词:astaxanthin  enantioselective  stereoisomer  reversed-phase high performance liquid chromatography  preparation

中文摘要:

      目的 建立反相高效液相色谱法(reversed-phase high performance liquid chromatography, RP-HPLC)检测全反式虾青素三种异构体(3R,3′R、3R,3′S和3S,3′S)的分析方法。方法 采用了固定相填料为纤维素-三(3,5-二甲基苯基氨基甲酸酯)的手性色谱柱(Ultimate Cellu-DR column), 综合考察了流动相和柱温对全反式虾青素3种异构体的分离效果, 并采用半制备液相色谱法获得全反式虾青素的3种异构体。结果 得到最佳的分析色谱条件: 流速为0.80 mL/min, 柱温30 ℃, 流动相为甲醇, 等度洗脱, 检测波长为480 nm。在此条件下, 全反式虾青素的3种异构体达到良好分离, 分离度均大于2。优化后的半制备色谱条件为: 制备柱Ultimate Cellu-DR, 流动相为纯甲醇, 流速为15.00 mL/min。获得的3种异构体纯度均大于97%。结论 该分析方法能有效地区分全反式虾青素的3种异构体, 可用于不同来源虾青素的鉴定。

英文摘要:

      Objective To establish a method for the determination of all-trans-astaxanthin (3R,3′R, 3R,3′S and 3S,3′S) by reversed-phase high performance liquid chromatography (RP-HPLC). Methods The separation was performed on a column packed with cellulose-tris (3,5-dimethylcarbamate) chiral stationary phase (Ultimate Cellu-DR column). The influence of mobile phase, and column temperature on the separation of 3 stereoisomers of all-trans-astaxanthin was studied. While the three stereoisomers of all-trans-astaxanthin were obtained by semi preparative liquid chromatography. Results The optimal separation conditions were as follows: the mobile phase was using a methanol with a isocratic elution profile and flow rate of 0.80 mL/min, column temperature was 30 ℃, the detection wavelength was 480 nm. Under these conditions, three stereoisomers of all-trans-astaxanthin were separated well, and the resolution (Rs) was more than 2. The optimized semi preparative separation condition was carried on preparative Ultimate Cellu-DR column using methanol as mobile phase with the flow of 15.00 mL/min. The purity of the three stereoisomers obtained was over 97%, respectively. Conclusion The analytic method can differentiate different stereoisomers of all-trans-astaxanthin efficiently and it is suitable for the determination of different sources of astaxanthin.