高效液相色谱-电感耦合等离子体质谱法测定海产品中5种硒形态的含量  

Determination of5 Selenium Speciesin Seafood Products by High Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry

作　　者：朱羽庄 梅光明[2,3] 杨盈悦 顾捷 孟春英[2,3] ZHU Yuzhuang;MEI Guangming;YANG Yingyue;GU Jie;MENG Chunying(School of Food Science and Pharmaceutics of Zhejiang Ocean University,Zhoushan 316022,China;Laboratory of Aquatic Product Processing and Quality Safety,Marine Fisheries Research Institute of Zhejiang Province,Zhoushan 316021,China;Zhejiang Key Laboratory of Marine Aquaculture,Zhoushan 316021,China)

机构地区：[1]浙江海洋大学食品与药学学院,舟山316022 [2]浙江省海洋水产研究所水产品加工与质量安全研究室,舟山316021 [3]浙江省海水增养殖重点实验室,舟山316021

出　　处：《理化检验（化学分册）》2022年第3期291-298,共8页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

基　　金：浙江省科技计划项目(LGN20C200015)。

摘　　要：制备好的冻干海产品样品0.2 g(精确至0.0001 g)在pH 8.0的水溶液体系下,经30 mg木瓜蛋白酶(800 U·mg-1)在50℃、180 r·min^(-1)的条件下酶解12 h,涡旋3 min,离心10 min.所得的上清液经0.22μm滤膜过滤,以Agilent ZORBAX SB-AQ色谱柱(250 mm×4.6 mm,5μm)为固定相,10 mmol·L^(-1)的柠檬酸溶液(pH 4.75)为流动相,于0.6 mL·min^(-1)下等度洗脱,可在11 min内实现对5种硒形态的分离,再采用电感耦合等离子体质谱法测定硒酸盐(Se^(6+))、亚硒酸盐(Se^(4+))、甲基硒代半胱氨酸(Me-Se-Cys)、硒代蛋氨酸(Se-Met)和硒代乙硫氨酸(Se-Et)的含量.结果表明,5种硒形态标准曲线的线性范围均在500μg·L^(-1)内,检出限(3S/N)为1.231~2.423μg·L^(-1).按标准加入法对海产品样品进行3个浓度水平的回收试验,回收率为81.3%~118%,测定值的日内相对标准偏差(n=6)和日间相对标准偏差(n=6)均小于9.0%.方法用于市售海产品分析,结果表明小黄鱼和带鱼中硒代蛋氨酸的测定结果比文献报道的大西洋鲑鱼和大西洋鳕鱼的测定值都要高,说明在我国食用小黄鱼和带鱼能够为居民补充较高水平的硒代蛋氨酸.此外,口虾蛄也是补充硒代蛋氨酸的重要来源.0.2 g(accurateto 0.0001 g)ofthe preparedfreeze-dried seafood product sample was hydrolyzed with30 mg papain(800 U·mg^(-1))inanaqueous solutionsystem with pH 8.0 at50℃for12 h withthe speed of 180 r·min^(-1).After vortexfor3 min and centrifugationfor 10 min,the obtained supernatant was passedthrough 0.22μmfilter membrane,and 5 selenium species in the filtrate could be separated in 11 min,using Agilent ZORBAX SB-AQcolumn(250 mm×4.6 mm,5μm)asthestationaryphaseand10 mmol·L^(-1)citricacidsolution(pH 4.75)asthe mobile phaseforisocratic elutionat0.6 mL·min^(-1).Then selenate(Se^(6+)),selenite(Se^(4+)),L-Se-methylselenocysteine(Me-Se-Cys),selenomethionine(Se-Met)and selenoethionine(Se-Et)in seafood product samplesolution were determined byinductively coupled plasma mass spectrometry.As shown by the results,the samelinearranges of the standard curves of 5 selenium species were found within 500μg·L^(-1),with detection limits(3S/N)intherange of1.231-2.423μg·L^(-1).Testsforrecovery were made on seafood product samples at three concentrationlevels bystandardaddition method,givingresultsintherangeof81.3%-118%,withintra-day RSDs(n=6)andinter-day RSDs(n=6)ofthedeterminedvalueslessthan9.0%.This method wasappliedforthe analysis ofcommercialseafood productsample,andit wasshownthatthedeterminationresultsofselenomethionine insmall yellow croaker and hairtail were higher than those of Atlantic salmon and Atlantic cod reported in the literature,indicating that eating small yellow croaker and hairtail in China could provide a higher level of selenomethioninefor people.Inaddition,squilla was also animportantsource ofselenomethioninesupplement.

关 键 词：高效液相色谱-电感耦合等离子体质谱法 硒形态 海产品 

分 类 号：O657.63[理学—分析化学]