应用高效液相色谱-电感耦合等离子体质谱联用技术分析研究畜肉中硒形态

Determination of selenium species in livestock meat using HPLC-ICP/MS

目的应用高效液相色谱-电感耦合等离子体质谱联用技术(HPLC-ICP/MS)分析研究畜肉中Se(Ⅵ)、Se(Ⅳ)、SeCys、MeSeCys和SeMet 5种硒形态。方法采用超声辅助蛋白酶ⅩⅣ和胰脂肪酶酶解提取畜肉中硒形态,以C18反相色谱柱为分析柱,10 mmol/L柠檬酸和5 mmol/L己烷磺酸钠(pH=4.0,含1%甲醇)为流动相,等度洗脱,HPLC-ICP/MS分析。结果5种硒形态可在7 min内有效分离,在线性范围内相关系数均>0.999,线性良好;Se(Ⅵ)、SeCys、MeSeCys和SeMet 4种硒形态检出限为0.0003~0.002 mg/kg,不同浓度水平的加标回收率为80.0%~103.3%,相对标准偏差均小于6.8%,有证标准物质ERM■BC210a测定值在标准值范围内。结论该方法适用畜肉中Se(Ⅵ)、SeCys、MeSeCys和SeMet的分析测定。畜肉中的硒形态主要为SeMet,同时含有少量的MeSeCys和SeCys。

Objective To apply high performance liquid chromatography-inductively coupled plasma mass spectrometry(HPLC-ICP/MS)to the analysis of five forms of selenium[Se(Ⅵ),Se(Ⅳ),SeCys,MeSeCys,and SeMet]in livestock meat.Methods The selenium species were extracted using the ultrasonic-assisted enzyme method,with C18 reversedphase column(250 mm×4.6 mm,5μm)as the analytical column,10 mmol/L citric acid and 5 mmol/L sodium hexane sulfonate(pH=4.0,containing 1%methanol)as the mobile phase,and an injection volume of 20μL.Results Four selenium forms were separated in 7 min,with correlation coefficients greater than 0.999,indicating good linearity.The detection limits of Se(Ⅵ),SeCys,MeSeCys,and SeMet ranged from 0.0003-0.002 mg/kg.The spiked recoveries of selenium-enriched pork at three concentrations(low,medium,and high)ranged from 80.0%to 103.3%,with a relative standard deviation of less than 6.8%.Certified Reference Material ERM■ BC210 revealed values within the standard range.Conclusion The developed method was suitable for determining Se(Ⅵ)SeCys,MeSeCys,and SeMet in livestock meat.The meat was found to mainly contain SeMet and small amounts of MeSeCys and SeCys.