高效液相色谱-电感耦合等离子体质谱法检测富硒苹果中5种硒形态

Determination of 5 kinds of selenium species in selenium-enriched apples by high performance liquid chromatography-inductively coupled plasma mass spectrometry

目的建立高效液相色谱-电感耦合等离子体质谱联用技术(high performance liquid chromatographyinductively coupled plasma mass spectrometry,HPLC-ICP-MS)测定富硒苹果中5种硒形态的方法。方法对样品提取方法、流动相的类型、浓度、p H值等条件进行考察及确定,采用柠檬酸超声提取的方式处理样品,流动相为浓度为5 mmol/L、p H=5.0的柠檬酸溶液。选用Hamiltion PRP-X100阴离子分析柱,色谱进样量为100μL,质谱采用碰撞池模式进行测定。结果本方法在15 min内可以完全分离5种硒形态,硒代胱氨酸(Se Cys2)、甲基硒代半胱氨酸(Me Se Cys)、亚硒酸根(Se(Ⅳ))、硒代蛋氨酸(Se Met)、硒酸根(Se(Ⅵ))的检出限分别为0.6、0.7、1、0.9、1μg/L,样品加标回收率范围为82.1%~98.3%,相对标准偏差为1.6%~3.8%。结论本方法可以简单、快速、准确地测定富硒苹果中硒的5种形态。

Objective To establish a method for determination of 5 kinds of selenium species in selenium-enriched apples by high performance liquid chromatography-inductively coupled plasma mass spectrometry（HPLC-ICP-MS）. Methods The sample extraction method, the type, concentration and p H value of mobile phase and other conditions were investigated and determined. The samples were extracted by citric acid with ultrasonic extraction method, and the citric acid solution with the concentration of 5 mmol/L, p H=5.0 was used as the mobile phase. A Hamiltion PRP-X100 anion analytical column was selected, the chromatographic injection volume was 100 μL, and collision cell mode of the mass spectrometry was used to proceed the determination. Results The 5 kinds of selenium species could be completely separated within 15 min by this method. The limits of detection of Se Cys2, Me Se Cys, Se（Ⅳ）, Se Met and Se（Ⅵ） were 0.6, 0.7, 1, 0.9, 1 μg/L, respectively. The spiked recoveries ranged from 82.1% to 98.3%, and the relative standard deviations were 1.6%~3.8%. Conclusion This method can simply, quickly and accurately determine the 5 kinds of selenium species in selenium-enriched apples.