HPLC-ICP-MS联用检测转基因大豆中的硒形态

Determination of Selenium Species in Genetically Modified Soybean by High-Performance Liquid Chromatography with Inductively Coupled Plasma Mass Spectrometry

本文研究建立了高效液相色谱-电感耦合等离子体质谱（HPLC-ICP-MS）联用检测转基因大豆中的硒酸盐（Se VI）、亚硒酸盐（Se IV）、硒代蛋氨酸（selenomethionine,Se Met）、硒代胱氨酸（selenocystine,Se Cys2）和硒代乙硫氨酸（selenoethionine,Se Et）的方法。探讨了色谱柱、流动相及其酸度对分离效果的影响,使用10 mmol/L的柠檬酸溶液（p H值4.5）作流动相,Hamilton PRP X-100色谱柱分离,碰撞反应池技术消除^40Ar^40Ar^＋和^40Ar^2H^2＋等多原子离子干扰,ICP-MS检测^82Se同位素,在21 min内可完全分离检测5种硒形态。探讨了不同提取方法的提取效果,优化了提取条件,采用蛋白酶提取,针对美国进口的转基因大豆样品进行加标回收实验,结果表明采用蛋白酶提取,Se IV和Se VI的回收率在100%左右,Se Met的回收率在92.6%-109.3%之间,Se Cys2和Se Et的回收率为81.2%-95.9%。该方法可完全满足转基因大豆中的硒形态定量分析鉴定。

An analytical method to detect selenium species such as selenate [Se（IV）], selenite [Se（VI）], selenomethionine（Se Met）, selenocystine（Se Cys2）, and selenoethionine（Se Et） in genetically modified soybean was established using high-performance liquid chromatography coupled with inductively coupled plasma mass spectrometry（HPLC-ICP-MS）. The effects of the chromatographic column, mobile phase, and acidity on separation were evaluated. Hamilton PRP X-100 reversed phase anion exchange column with 10 mmol/L citric acid solution as the mobile phase was used for separation. Collision cell technology was adopted to eliminate the interference of polyatomic ions such as ^40Ar^40Ar^＋ and ^40Ar^2H^2＋. Isotope 82 Se was detected by ICP-MS. The five species were completely separated within 21 min. The extraction procedure was discussed and optimized. A spiked recovery experiment was carried out with proteinase extration, using genetically modified soybean imported from the USA. The result showed that the recoveries were around 100% for Se（IV） and Se（VI）, was in the range 92.6%–109.3% for Se Met, and they were in the range 81.2%–95.9% for Se Cys2 and Se Et. This method can be used for the quantitative determination of selenium species in genetically modified soybeans.