手性色谱法测定L-硒-甲基硒代半胱氨酸含量的研究

Determination of L-Se-methylselenocysteine Content by Chiral Chromatography

L-硒-甲基硒代半胱氨酸是一种新型硒源类的食品营养强化剂,而D-硒-甲基硒代半胱氨酸的安全性尚没有相关研究,目前对于L-硒-甲基硒代半胱氨酸含量的测定方法多集中在测定硒-甲基硒代半胱氨酸含量,无法检测到其中L-硒-甲基硒代半胱氨酸的含量。本文建立了通过手性色谱柱分离、标准曲线法测定L-硒-甲基硒代半胱氨酸的高效液相色谱法(HPLC)。实验使用EC250/4NUCLEOSIL Chiral-1手性色谱柱(4 mm×250 mm,5μm),流动相为0.4mmol/L Cu SO4·5H2O水溶液,柱温为35℃,流速为1.0 m L/min,进样量为20μL,检测波长为240 nm;在该条件下L-硒-甲基硒代半胱氨酸的检测线性范围为0.541~1.352 mg/ml,线性相关系数R达0.9999,在线性范围内精密度和稳定性良好,平均回收率为97.73%。该方法操作简单,结果准确,可用于L-硒-甲基硒代半胱氨酸产品及其制品质量的监控。

L-Se-methylselenocysteine is a new selenium-containing food supplement; however, no studies regarding the safety of D-Se-methylselenocysteine have been reported. Current methods for the determination of L-Se-methylselenocysteine content mainly focus on the measurement of total Se-methylselenocysteine content and cannot differentiate it from the content of L-Se-methylselenocysteine. High-pressure liquid chromatography(HPLC) was used in this study to estimate L-Se-methylselenocysteine content using chiral separation, and a standard curve was established. Reliable chromatographic separation was achieved on an EC250/4 NUCLEOSIL Chiral-1 column(4 mm × 250 mm, 5 μm) when the mobile phase was 0.4 mmol/L Cu SO4·5H2O, column temperature was 35 ℃, flow rate was 1.0 m L/min, sample size was 20 μL, and detection wavelength was 240 nm. Under these conditions, the calibration curves established showed good linearity across the concentration range of 0.541~1.352 mg/ml, with a correlation coefficient of 0.9999 and an average recovery of 97.73%. The method is simple, accurate, and can be used to monitor the quality of L-Se-methylselenocysteine and its related products.