应用液相色谱-原子荧光联用仪测定富硒大米中的5种硒形态
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摘要
目的建立了富硒大米中硒酸根、亚硒酸根、硒代蛋氨酸、硒代胱氨酸和甲基硒代半胱氨酸5种硒形态的高效液相色谱-氢化物发生原子荧光光谱(HPLC-HG-AFS)分析方法。方法样品采用链酶蛋白酶E水解超声提取方式,30mmol/L磷酸氢二铵+0.5mmol/L四丁基溴化铵+2%甲醇为流动相,用甲酸调节pH至5.8～6.0,经C18(5μm,100.4.6×150mm)色谱柱分离后HG-AFS测定。对流动相浓度、pH值以及流速进行了选择优化。结果10min内可以完全分离5种硒形态,各种硒形态标准曲线的线性关系良好(r2≥0.995),方法检出限为2.27～3.89μg/L,平均回收率为85.4%～105.5%,相对标准偏差<5.8%。结论本方法前处理简单、灵敏度高、设备便宜、运行费用低廉,适用于富硒大米中硒的形态分析。
The determined seleniums species were Se(Ⅳ),Se(Ⅵ),selenomethionine(SeMet),selenocystine(SeCys2)and Se-methylselenocysteine(MeSeCys).The Pronase E hydrolysis with ultrasonic extraction was taken as the sample pretreatment method.The target compounds were separated on a C18 column(5μm,100.4.6×150 mm)and analyzed by HG-AFS.The mobile phase was 30 mmol/L(NH_4)H_2PO_4+0.5 mmol/L teltrabutylammonium bromide+2% methanol in the pH range of 5.8—6.0 adjusted with formic acid.Good linearity was obtained for all the selenium species,with the linear correlation coefficients higher than 0.995.The LODs of the four species were between 2.27 and 3.89μg/L.Average recoveries for the five analytes were in the range of 85.4%—105.5%,with the RSD lower than 5.8%.The method is simple,sensitive and of low-cost.
The determined seleniums species were Se(Ⅳ),Se(Ⅵ),selenomethionine(SeMet),selenocystine(SeCys2)and Se-methylselenocysteine(MeSeCys).The Pronase E hydrolysis with ultrasonic extraction was taken as the sample pretreatment method.The target compounds were separated on a C18 column(5μm,100.4.6×150 mm)and analyzed by HG-AFS.The mobile phase was 30 mmol/L(NH_4)H_2PO_4+0.5 mmol/L teltrabutylammonium bromide+2% methanol in the pH range of 5.8—6.0 adjusted with formic acid.Good linearity was obtained for all the selenium species,with the linear correlation coefficients higher than 0.995.The LODs of the four species were between 2.27 and 3.89μg/L.Average recoveries for the five analytes were in the range of 85.4%—105.5%,with the RSD lower than 5.8%.The method is simple,sensitive and of low-cost.
引文
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[15]赵秋香,冯超,莫书伟,等.形态的硒的研究过程中硒代胱氨酸的稳定性[J].光谱实验室,2011,28(4):2074-2078.
[2]周云,刘忠荣.硒与人体健康[J].微量元素与健康研究,2002,19(4):79-80.
[3]Jorge M P,Antonio M P,Vanessa R H,et al.In vitro bioavailability of total selenium and selenium species from seafood[J].Food Chem,2013,139:872-877.
[4]Jiri S,Miroslav P,Branislav R N,et al.Effect of selenium in organic and inorganic form on liver,kidney,brain and muscle of Wistar rats[J].Cent Eur J Chem,2012,10(5):1442-1451.
[5]程建中,杨萍,桂仁意.植物硒形态分析的研究综述[J].浙江农林大学报,2012,(29):288-295.
[6]张涛.富硒大米及富硒酵母中的硒形态分析[D].内蒙古:内蒙古科技大学包头医学院,2007.
[7]张永志,王钢军,叶雪珠.联用技术测定富硒农产品中硒的形态研究进展[J].广东微量元素科学,2012,19(8):1-5.
[8]李红卫,王开萍,吴娱,等.柱前衍生-反相高效液相色谱法研究富硒米曲霉中有机硒形态[J].食品科学,2015,36(22):137-141.
[9]杨修斌,王丙涛,俞坤,等.HPLC-ICP-MS联用检测转基因大豆中的硒形态[J].现代食品科技,2015,31(2):280-284.
[10]Zembrzuska J,Matusiewicz H,Polkowska M H,et al.Simultaneous quantitation and identification of organic and inorganic selenium in diet supplements by liquid chromatography with tandem mass spectrometry[J].Food Chem,2014,142:178-187.
[11]Ni Z L,Liu Y H,Qu M H.Determination of 5natural selenium species in selenium-enriched bamboo shoots using LC-ICP-MS[J].Food Sci Biotechnol,2014,23:1049-1053.
[12]Galinha C,Sánchez-Martínez M,Pacheco A M G,et al.Characterization of selenium-enriched wheat by agronomic biofortification[J].J Food Sci Technol,2015,52:4236-4245.
[13]尚德荣,秦德元,赵艳芳,等.高压液相色谱-氢化物发生-原子荧光光谱(HPLC-HG-AFS)联用技术检测水产品中硒的赋存形态[J].食品安全质量检测学报,2013,4(6):1847-1852.
[14]李红卫,王开萍,吴娱,等.柱前衍生-反相高效液相色谱法研究富硒米曲霉中有机硒形态[J].食品科学,2015,36(22):137-141.
[15]赵秋香,冯超,莫书伟,等.形态的硒的研究过程中硒代胱氨酸的稳定性[J].光谱实验室,2011,28(4):2074-2078.