高效液相色谱–电感耦合等离子体质谱联用测定稻米中5种硒形态  

Determination of Five Selenium Species in Rice by High Performance Liquid Chromatography with Inductively Coupled Plasma Mass Spectrometry

作　　者：曹赵云[1] 李洪艳 柴爽爽 许萍[1] 陈铭学[1] CAO Zhao-yun;LI Hong-yan;CHAI Shuang-shuang;XU Ping;CHEN Ming-xue(Rice Product Quality Supervision and Inspection Center,Ministry of Agriculture and Rural Affairs,China National Rice Research Institute,Hangzhou,Zhejiang 310006,China;Guizhou Lianda Testing Technology Co.,LTD,Guiyang,Guizhou 550025,China)

机构地区：[1]中国水稻研究所农业农村部稻米及制品质量监督检验测试中心,浙江杭州310006 [2]贵州省联大检测技术有限公司,贵州贵阳550025

出　　处：《粮油食品科技》2023年第6期20-27,共8页Science and Technology of Cereals,Oils and Foods

基　　金：国家水稻产业技术体系(CARS-01-47);中央级公益性科研院所基本科研业务费专项(CPSIBRF-CNRRI-202207)。

摘　　要：利用高效液相色谱–电感耦合等离子体质谱联用技术建立了富硒大米中5种硒形态的检测方法。采用反向C18色谱柱,对流动相离子对浓度、洗脱盐浓度、pH值进行了优化,最终确定流动相组成为15 mmol/L乙酸铵溶液,pH5.5和0.2 mmol/L四丁基氢氧化铵(TBAH),硒–甲基硒代半胱氨酸(SeMeCys)、硒代半胱氨酸(SeCys)、硒代蛋氨酸(SeMet)、四价硒Se(Ⅵ)和六价硒Se(Ⅳ)5种硒形态获得基线分离。大米样品用酶提法提取,通过对酶的种类、酶的用量及酶解时间等对提取效率的研究,确定样品与蛋白酶(protease)XIV酶量比为20∶1为最佳,且最佳酶解时间为20 h。结果表明本方法分离效率高,方法线性良好,灵敏度高,回收率为77.1%~107.3%,硒–甲基硒代半胱氨酸(SeMeCys)、硒代半胱氨酸(SeCys)、硒代蛋氨酸(SeMet)的检出限和定量限分别为0.0015mg/kg和0.005 mg/kg,四价硒Se(Ⅵ)和六价硒Se(Ⅳ)检出限和定量限分别为0.0025 mg/kg和0.009 mg/kg。为全面评价富硒大米质量和后续开展富硒大米膳食暴露评估提供技术支撑。A detection method for five selenium species in Se-enriched rice was established using HPLC-ICP MS/MS.The ion pair concentration of mobile phase,elution salt concentration,pH value were optimized by using a reverse C18 chromatographic column,and the final mobile phase composition was determined to consist of 15 mmol/L ammonium acetate solution,pH5.5 and 0.2 mmol/L TBAH,the baseline separation of SeMeCys,SeCys,SeMet,Se(Ⅵ) and Se(Ⅳ) was obtained.Selenium species in rice were extracted by enzyme extraction method.The effects of enzyme selection,enzyme dosage and extraction time on extraction efficiency were investigated.The optimal ratio of sample to protein XIV enzyme content was determined to be 20∶1,and the optimal enzymatic hydrolysis time was 20 h.The method has high separation efficiency,good linearity,good precision,and high sensitivity,and the recovery was 77.1%~107.3%.The limits of detection and the limits of quantitation of SeMeCys(methylselenocysteine),SeCys(selenocystine),and SeMet(selenomethionine)were 0.001 5 mg/kg and 0.005 mg/kg,respectively,while the limits of detection and the limits of quantitation of Se(Ⅵ) and Se(Ⅳ) were 0.002 5 mg/kg and 0.009 mg/kg.The method provide technical support for objective and comprehensive evaluation of the quality of Se-enriched rice and subsequent assessment of dietary exposure to Se-enriched rice.

关 键 词：RP-HPLC-ICP MS/MS 富硒大米 硒形态 酶提法 

分 类 号：TS201.4[轻工技术与工程—食品科学] S-3[轻工技术与工程—食品科学与工程]