手性柱高效液相色谱拆分测定营养强化剂硒-甲基硒代半胱氨酸对映体

Chiral ligand- exchange HPLC anlysis of nutrient supplement se-methylselenocysteine

目的:建立营养强化剂硒-甲基硒代半胱氨酸对映体拆分与定量测定高效液相色谱法,为其质量标准的建立奠定基础。方法:采用配体交换手性柱,以硫酸铜为手性配体流动相,考察了铜离子浓度、D,L-硒-甲基硒代半胱氨酸进样量和柱温等因素对硒-甲基硒代半胱氨酸对映体的拆分效果。结果:优化拆分条件为:检测波长240nm,柱温为35℃,流动相硫酸铜浓度为0.4mmol/L,流速为1.0mL/min。在上述色谱条件下,L-硒-甲基硒代半胱氨酸的进样量在0.95μg^30.28μg范围内与峰面积之间呈现良好线性关系,线性回归方程为Y=725.52X-62.13,R=0.9999,加标回收率为97.67%~99.88%,检测限和定量限分别为0.0528μg和0.132μg。归一化法测得该色谱条件下能准确检出0.6%以上的D型异构体。结论:该方法专属性强,灵敏度较高,重现性好,结果准确,适用于L-硒-甲基硒代半胱氨酸的工艺和质量控制。

Objective：To establish a Chiral Ligand-exchange HPLC method for the separation and analysis of Se-methylselenocysteine enantiomer.Methods：The determination was performed on chiral ligand-exchange column,the mobile phase was composed of copper sulfate as chiral mobile phase ligands.The effect of copper concentration,sample volume and column temperature were investigated.Results：The optimum separation condition of D,L-Se-methylselenocysteine was obtained as the followings ; mobile phase：0.4mmol/L CuSO4-H2O,wavelength：254nm,flow rate：1.0mL/min,column temperature：35℃.This method showed good linearity for L-Se-methylselenocysteine （ranging from 0.95μg to 30.28μg） and the hnear regression equation was Y =725.52X-62.13 （R =0.9999）.The recoveries were in the range of 97.67％ ～99.88％ while the detection limit was 0.0528μg and the limit of quantification was 0.132μg.The result indicated that the detection limit of D-Se-methylselenocysteine was 0.6％.Conclusion：The results demonstrated this method was sensitive,accurate and suitable for the quality control of L-Se-methylselenocysteine.