超声辅助酶提取-高效液相原子荧光测定抗癌类海洋中药中硒的形态

Determination of selenium species in anticancer marine traditional Chinese medicine using ultrasonic assisted enzymatic extraction coupled with high performance liquid chromatography-atomic fluorescence spectrometry

建立了高效液相色谱-氢化物发生原子荧光光谱(HPLP-HG-AFS)联用技术测定抗癌类海洋中药中4种硒形态的方法。样品经超声辅助蛋白酶XIV提取,采用Hamilton PRP-X100阴离子交换柱(250 mm×4. 1 mm,10μm),以5 mmol/L(pH 4. 5)的柠檬酸为流动相,在8 min内可分离4种硒形态:硒代胱氨酸(SeCys2),甲基-硒代半胱氨酸(Me SeCys),亚硒酸盐(Se(Ⅳ)),硒代蛋氨酸(SeMet)。检出限分别为1. 25,1. 03,0. 56,1. 85μg/L,相关系数均大于0. 9990,加标回收率范围为90. 7%~104. 5%。方法可满足海洋中药中硒形态的定量分析。

A high performance liquid chromatography-atomic fluorescence spectrometry(HPLC-AFS)method for the analysis of 4 selenium species in anticancer marine traditional Chinese medicine was proposed. 4 selenium species were selenocystine(Se Cys2),methylselenocysteine(Me Se Cys),selenite(Se(Ⅳ)) and selenomethionine(Se Met). The selenium species in marine traditional Chinese medicine samples were extracted using ultrasonic extraction system with protease XIV. Chromatographic separation was performed using Hamilton PRPX-100 anion exchange column(250 mm × 4. 1 mm,10 μm) with5 mmol/L citric acid solution(pH 4. 5) within 8 min. The limits of detection for Se Cys2,Me Se Cys,Se(Ⅳ) and Se Met were obtained as 1. 25,1. 03,0. 56,1. 85 μg/L,respectively. All the linear correlation coefficients of 4 selenium species were greater than 0. 9990. The recoveries of 4 selenium species were in the range of 90. 7%~ 104. 5%. The developed analytical method was successfully applied for the determination of selenium species in marine traditional Chinese medicines.