高效液相色谱-电感耦合等离子体质谱法测定富硒食用菌中的4种硒形态

摘要

目的 建立富硒食用菌中硒代胱氨酸(SeCys)、4价硒(Se(IV))、硒代蛋氨酸(SeMet)、6价硒(Se(VI))4种硒形态的高效液相色谱-电感耦合等离子体质谱(high performance liquid chromatography-inductively coupled plasma-mass spectrometry,HPLC-ICP-MS)的分析方法.方法 以Hamilton PRP-X100为色谱分离柱,以40 mmol/L磷酸氢二铵(pH 6.0)为流动相,对富硒食用菌中4种硒形态进行分离,利用电感耦合等离子体质谱法对质量数为77的硒同位素(77Se)进行定量检测,样品提取采用蛋白酶(来源于地衣芽孢杆菌)为提取剂,60℃水浴提取1 h.结果 4种硒形态在0~100μg/L浓度范围内线性良好,相关系数r2大于0.997,检出限在2μg/L以下.结论 该方法适用于富硒食用菌中硒的形态分析.%Objective To establish a method for determination of 4 selenium species (SeCys, Se(IV) , SeMet, Se(VI)) in selenized edible fungi using high performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS). Methods An Hamilton PRP-X100 column was used to separate the 4 compounds using 40 mmol/L ammonium dibasic phosphate (pH 6.0) as the mobile phase, and the contents of isotope with a mass of 77 of these 4 compounds were determined by plasma mass spectrometry. The 4 selenium compounds were effectively extracted using protease (from Bacillus licheniformis) with water bath for 60 min. Results Four kinds of selenium compounds had good linear relationships in the range of 0~100 μg/L, and the correlation coefficients were larger than 0.997. The limits of detection were less than 2 μg/L. Conclusion The method is suitable for the simultaneous determination of selenium compounds in selenized edible fungi.