高效液相色谱串联质谱法检测蚯蚓体内硒代氨基酸
|
摘要
建立了高效液相色谱-电喷雾质谱联用技术(HPLC-ESI-MS)检测蚯蚓体内硒代氨基酸的分析方法.采用振荡超声辅助酶解法提取蚯蚓(赤子爱胜蚓(Eisenia fetida))体内的硒代蛋氨酸(SeMet)和甲基硒代半胱氨酸(MeSeCys).用HPLC-ESI-MS正离子多反应离子监测模式(MRM)对这两种硒代氨基酸进行分离检测,外标法定量.SeMet和MeSeCys在4—600 ng·mL~(-1)有良好的线性关系,线性相关系数R~2均大于0.9999.SeMet和MeSeCys的方法检出限分别为0.49μg·g~(-1)和0.67μg·g~(-1).样品加标回收率在81.1%—112.4%之间,并且相对标准偏差RSD均小于3.1%.本方法不需要柱前衍生化,增加反向气等繁琐操作,简单快速的实现对两种硒代氨基酸的分离检测.
A high performance liquid chromatography-electrospray ionization-mass spectrometry(HPLC-ESI-MS) method was developed for the determination of the seleno amino acids in earthworms. Selenomethione(SeMet) and methylselenocysteine(MeSeCyS) were extracted from earthworms by oscillating and ultrasound assisted enzymatic hydrolysis. These two seleno amino acids were isolated and determined by HPLC-ESI-MS using external standard method. Then the positive ion and multiple reaction monitoring(MRM) was applied and operated. The results indicated that both SeMet and MeSeCys had a good calibration curve in a range of 4—600 ng·mL~(-1), with R~2 > 0.9999. The method detection limits of SeMet and MeSeCys were 0.49 μg·g~(-1) and 0.67 μg·g~(-1), respectively. The recoveries of standard addition were 81.1%—112.4%, with RSD values less than 3.1%. This detection method was preferred over others as it had eliminated the need for more cumbersome procedures, such as pre-column derivatization and increased the opposite direction gas.
A high performance liquid chromatography-electrospray ionization-mass spectrometry(HPLC-ESI-MS) method was developed for the determination of the seleno amino acids in earthworms. Selenomethione(SeMet) and methylselenocysteine(MeSeCyS) were extracted from earthworms by oscillating and ultrasound assisted enzymatic hydrolysis. These two seleno amino acids were isolated and determined by HPLC-ESI-MS using external standard method. Then the positive ion and multiple reaction monitoring(MRM) was applied and operated. The results indicated that both SeMet and MeSeCys had a good calibration curve in a range of 4—600 ng·mL~(-1), with R~2 > 0.9999. The method detection limits of SeMet and MeSeCys were 0.49 μg·g~(-1) and 0.67 μg·g~(-1), respectively. The recoveries of standard addition were 81.1%—112.4%, with RSD values less than 3.1%. This detection method was preferred over others as it had eliminated the need for more cumbersome procedures, such as pre-column derivatization and increased the opposite direction gas.
引文
[1] 张择洲,朱元元,李梦,等.生物样品中硒的形态分析方法研究进展[J].生物技术进展,2017,7(5):409-414.ZHANG Z Z,ZHU Y Y,LI M,et al.Progress on selenium speciation analysis in biological samples [J].Current Biotechnology,2017,7(5):409-414(in Chinese).
[2] 魏宁果,何冲,范芳芳.食品中硒形态的研究进展[J].农产品加工,2018(11):71-73,76.WEI N G,HE C,FAN F F.Research progress of selenium speciation in foods [J].Farm Products Processing,2018(11):71-73,76(in Chinese).
[3] 何彩梅,吴桂容,陈春岚,等.食品中硒与硒形态分析方法研究进展[J].食品工业,2017,38(9):218-222.HE C M,WU G R,CHEN C L,et al.Research progress in determination method and speciation analysis of selenium in food [J].The Food Industry,2017,38(9):218-222(in Chinese).
[4] 秦玉燕,时鹏涛,王运儒,等.高效液相色谱-氢化物发生-原子荧光光谱法测定富硒食品中5种形态硒的含量[J].理化检验-化学分册,2018,54(5):566-572.QIN Y Y,SHI P T,WANG Y R,et al.HPLC-HG-AFS determination of 5 species of selenium in foodstuffs rich in selenium [J].PTCA (PART B:CHEM,ANAL.),2018,54(5):566-572(in Chinese).
[5] 秦海波,朱建明,李社红,等.环境中硒形态分析方法的研究进展[J].矿物岩石地球化学通报,2008,27(2):180-187.QIN H B,ZHU J M,LI D H,et al.Advances in the speciation analysis of selenium in the environment [J].Bulletin of Mineralogy,Petrology and Geochemistry,2008,27(2):180-187(in Chinese).
[6] 王欣,幸苑娜,陈泽勇,等.高效液相色谱-电感耦合等离子体质谱法检测富硒食品中6种硒形态[J].分析化学,2013,41(11):1669-1674.WANG X,XING Y N,CHEN Z Y,et al.Determination of 6 selenium species in selenium-enriched food by hyphenated technique of high performance liquid chromatography inductively coupled plasma mass spectrometry [J].Chinese Journal of Analytical Chemistry,2013,41(11):1669-1674(in Chinese).
[7] 徐肖雅,丁亮,林少彬.人血液和尿液中硒元素的形态分析研究进展[J].环境卫生学杂志,2012,2(2):91-96.XU X Y,DING L,LIN S B.Speciation analysis of selenium in human blood and urine [J].Journal of Environmental Hygiene Apr,2012,2(2):91-96(in Chinese).
[8] 牟仁祥,陈铭学,朱智伟.硒的形态分析方法综述[J].光谱实验室,2004,21(1):27-35.MOU R X,CHEN M X,ZHU Z W.Review of the analytical methods for selenium speciation [J].Chinese Journal of Spectroscopy Laboratory,2004,21(1):27-35(in Chinese).
[9] 雷绍荣,杨定清,周娅.硒的总量及形态分析综述[J].中国测试,2009,35(5):1-6.LEI S R,YANG D Q,ZHOU Y.Summary on total content determination and speciation analyses of selenium [J].China Measurement & Test,2009,35(5):1-6(in Chinese).
[10] 梁若玉,和娇,史雅娟,等.典型富硒农业基地土壤硒的生物有效性与剖面分布分析[J].环境化学,2017,36(7):1588-1595.Bioavailability and profile distribution of selenium in soils of typical Se-enriched agricultural base [J].Environmental Chemistry,2017,36(7):1588-1595 (in Chinese).
[11] SUN G X,LIU X,WILLIANMS P,et al.Distribution and translocation of selenium from soil to grain and its speciation in paddy rice (Oryzasativa L.) [J].Environmental Science & Technology,2010,44:6706-6711.
[12] MARIA M,MARIA J,ELISA B,et al.Evaluation of different sample extraction strategies for selenium determination in selenium-enriched plants (Allium sativum and Brassica juncea) and Se speciation by HPLC-ICP-MS [J].Talanta,2006,68:1287-1293.
[13] 李瑶佳.高效液相色谱-氢化物原子荧光联用技术分析富硒苦荞中的硒形态[J].山东化工,2018,47(21):89-91,103.LI Y J.Analysis of selenium forms in selenium-enriched buckwheat using HPLC-HG-AFS [J].Shandong Chemical Industry,2018,47(21):89-91,103(in Chinese).
[14] 开建荣,王彩艳,李冬.农产品中硒形态研究进展分析[J].宁夏农林科技,2017,58(3):36-38,51.KAI J R,WANG C Y,LI D.Research progress on speciation analysis of selenium in agricultural products [J].Ningxia Journal of Agriculture and Forestry Science and Technology,2017,58(3):36-38,51(in Chinese).
[15] 何梦洁,张双庆,黄振武.人乳硒形态分析的研究进展[J].中国药事,2016,30(8):772-777.HE M J,ZHANG S Q,HUANG Z W.Advances in the analysis of selenium speciation in human milk [J].Chinese Pharmaceutical Affairs,2016,30(8):772-777(in Chinese).
[16] 任江姿,郑平,吴景,等.硒代氨基酸提取与检测技术研究进展[J].湖南农业科技,2018(8):123-127.REN J Z,ZHENG P,WU J,et al.Review on extraction and analysis methods of seleno amino acids [J].Hunan Agricultural Sciences,2018(8):123-127(in Chinese).
[17] 田强兵.分析化学中检出限和测定下限的探讨[J].分析化学计量,2007,16(3):72-73.TIAN Q B.Discussion of detection limit and the lower limit of detection in analytical chemistry [J].Chemical Analysis and Meterage,2007,16(3):72-73(in Chinese).
[18] 铁梅,方禹之,孙铁彪,等.HPLC-ICP-MS联用技术在富硒金针菇硒的形态分析中的应用[J].高等学校化学学报,2007,28(4):635-639.TIE M,FANG Y Z,SUN T B,et al.Application of HPLC-ICP-MS in speciation analysis of selenium in selenized Flammulina velutipes [J].Chemical Journal of Chinese Universities,2007,28(4):635-639(in Chinese).
[19] 熊珺,覃毅磊,龚亮,等.超声辅助酶法提取-高效液相色谱-电感耦合等离子体质谱联用分析食品中无机硒和硒氨基酸6种硒形态[J].食品科技,2016,41(12):266-272.XIONG J,QIN Y L,GONG L,et al.Simultaneous quantification of the speciation of inorganic selenium and Se amino acids in food by ultrasonic assisted enzymatic extraction combined with high performance liquid chromatography-inductively coupled plasma mass spectrometry [J].Food Science and Technology,2016,41(12):266-272(in Chinese).
[20] OHTA Y,KOBAYASHI Y,KONISHI S,et al.Speciation analysis of selenium metabolites in urine and breath by HPLC- and GC-inductively coupled plasma-MS after administration of selenomethionine and methylselenocysteine to rats [J].Chemical Research in Toxicology,2009,22:1795-1801.
[21] BENTE G,KIM G,JESPER B,et al.Separation,purification and identification of the major selenium metabolite from human urine by multi-dimensional HPLC-ICP-MS and APCI-MS [J].Journal of Analytical Atomic Spectrometry,2003,18:65-70.
[22] 倪张林,汤富彬,张玮,等.HPLC-DRC-ICP-MS测定富硒蔬菜中的硒形态[J].分析试验室,2013,32(2):39-43.NI Z L,TANG F B,ZHANG W,et al.Determination of selenium species in se-enriched vegetables by HPLC-DRC-ICP-MS [J].Chinese Journal of Analysis Laboratory,2013,32(2):39-43(in Chinese).
[23] MASEKO T,CALLAHAN D L,DUNSHEA F R,et al.Chemical characterization and speciation of organic selenium in cultivated selenium-enriched Agaricus bisporus [J].Food Chemistry,2013,141:3681-3687.
[24] YUE S Z,ZHANG H Q,ZHEN H Y,et al.Selenium accumulation,speciation and bioaccessibility in seleniumenriched earthworm (Eisenia fetida) [J].Microchemical Journal,2019,145:1-8.
[25] 孙小斐,乔玉辉,孙振钧,等.富硒蚯蚓的培养及硒富集作用研究[J].农业资源与环境学报,2014,31(6):570-574.SUN X F,QIAO Y H,SUN Z J,et al.The cultivation and selenium enrichment of selenium enriched earthworm [J].Journal of Agricultural Resources and Environment,2014,31(6):570-574(in Chinese).
[2] 魏宁果,何冲,范芳芳.食品中硒形态的研究进展[J].农产品加工,2018(11):71-73,76.WEI N G,HE C,FAN F F.Research progress of selenium speciation in foods [J].Farm Products Processing,2018(11):71-73,76(in Chinese).
[3] 何彩梅,吴桂容,陈春岚,等.食品中硒与硒形态分析方法研究进展[J].食品工业,2017,38(9):218-222.HE C M,WU G R,CHEN C L,et al.Research progress in determination method and speciation analysis of selenium in food [J].The Food Industry,2017,38(9):218-222(in Chinese).
[4] 秦玉燕,时鹏涛,王运儒,等.高效液相色谱-氢化物发生-原子荧光光谱法测定富硒食品中5种形态硒的含量[J].理化检验-化学分册,2018,54(5):566-572.QIN Y Y,SHI P T,WANG Y R,et al.HPLC-HG-AFS determination of 5 species of selenium in foodstuffs rich in selenium [J].PTCA (PART B:CHEM,ANAL.),2018,54(5):566-572(in Chinese).
[5] 秦海波,朱建明,李社红,等.环境中硒形态分析方法的研究进展[J].矿物岩石地球化学通报,2008,27(2):180-187.QIN H B,ZHU J M,LI D H,et al.Advances in the speciation analysis of selenium in the environment [J].Bulletin of Mineralogy,Petrology and Geochemistry,2008,27(2):180-187(in Chinese).
[6] 王欣,幸苑娜,陈泽勇,等.高效液相色谱-电感耦合等离子体质谱法检测富硒食品中6种硒形态[J].分析化学,2013,41(11):1669-1674.WANG X,XING Y N,CHEN Z Y,et al.Determination of 6 selenium species in selenium-enriched food by hyphenated technique of high performance liquid chromatography inductively coupled plasma mass spectrometry [J].Chinese Journal of Analytical Chemistry,2013,41(11):1669-1674(in Chinese).
[7] 徐肖雅,丁亮,林少彬.人血液和尿液中硒元素的形态分析研究进展[J].环境卫生学杂志,2012,2(2):91-96.XU X Y,DING L,LIN S B.Speciation analysis of selenium in human blood and urine [J].Journal of Environmental Hygiene Apr,2012,2(2):91-96(in Chinese).
[8] 牟仁祥,陈铭学,朱智伟.硒的形态分析方法综述[J].光谱实验室,2004,21(1):27-35.MOU R X,CHEN M X,ZHU Z W.Review of the analytical methods for selenium speciation [J].Chinese Journal of Spectroscopy Laboratory,2004,21(1):27-35(in Chinese).
[9] 雷绍荣,杨定清,周娅.硒的总量及形态分析综述[J].中国测试,2009,35(5):1-6.LEI S R,YANG D Q,ZHOU Y.Summary on total content determination and speciation analyses of selenium [J].China Measurement & Test,2009,35(5):1-6(in Chinese).
[10] 梁若玉,和娇,史雅娟,等.典型富硒农业基地土壤硒的生物有效性与剖面分布分析[J].环境化学,2017,36(7):1588-1595.Bioavailability and profile distribution of selenium in soils of typical Se-enriched agricultural base [J].Environmental Chemistry,2017,36(7):1588-1595 (in Chinese).
[11] SUN G X,LIU X,WILLIANMS P,et al.Distribution and translocation of selenium from soil to grain and its speciation in paddy rice (Oryzasativa L.) [J].Environmental Science & Technology,2010,44:6706-6711.
[12] MARIA M,MARIA J,ELISA B,et al.Evaluation of different sample extraction strategies for selenium determination in selenium-enriched plants (Allium sativum and Brassica juncea) and Se speciation by HPLC-ICP-MS [J].Talanta,2006,68:1287-1293.
[13] 李瑶佳.高效液相色谱-氢化物原子荧光联用技术分析富硒苦荞中的硒形态[J].山东化工,2018,47(21):89-91,103.LI Y J.Analysis of selenium forms in selenium-enriched buckwheat using HPLC-HG-AFS [J].Shandong Chemical Industry,2018,47(21):89-91,103(in Chinese).
[14] 开建荣,王彩艳,李冬.农产品中硒形态研究进展分析[J].宁夏农林科技,2017,58(3):36-38,51.KAI J R,WANG C Y,LI D.Research progress on speciation analysis of selenium in agricultural products [J].Ningxia Journal of Agriculture and Forestry Science and Technology,2017,58(3):36-38,51(in Chinese).
[15] 何梦洁,张双庆,黄振武.人乳硒形态分析的研究进展[J].中国药事,2016,30(8):772-777.HE M J,ZHANG S Q,HUANG Z W.Advances in the analysis of selenium speciation in human milk [J].Chinese Pharmaceutical Affairs,2016,30(8):772-777(in Chinese).
[16] 任江姿,郑平,吴景,等.硒代氨基酸提取与检测技术研究进展[J].湖南农业科技,2018(8):123-127.REN J Z,ZHENG P,WU J,et al.Review on extraction and analysis methods of seleno amino acids [J].Hunan Agricultural Sciences,2018(8):123-127(in Chinese).
[17] 田强兵.分析化学中检出限和测定下限的探讨[J].分析化学计量,2007,16(3):72-73.TIAN Q B.Discussion of detection limit and the lower limit of detection in analytical chemistry [J].Chemical Analysis and Meterage,2007,16(3):72-73(in Chinese).
[18] 铁梅,方禹之,孙铁彪,等.HPLC-ICP-MS联用技术在富硒金针菇硒的形态分析中的应用[J].高等学校化学学报,2007,28(4):635-639.TIE M,FANG Y Z,SUN T B,et al.Application of HPLC-ICP-MS in speciation analysis of selenium in selenized Flammulina velutipes [J].Chemical Journal of Chinese Universities,2007,28(4):635-639(in Chinese).
[19] 熊珺,覃毅磊,龚亮,等.超声辅助酶法提取-高效液相色谱-电感耦合等离子体质谱联用分析食品中无机硒和硒氨基酸6种硒形态[J].食品科技,2016,41(12):266-272.XIONG J,QIN Y L,GONG L,et al.Simultaneous quantification of the speciation of inorganic selenium and Se amino acids in food by ultrasonic assisted enzymatic extraction combined with high performance liquid chromatography-inductively coupled plasma mass spectrometry [J].Food Science and Technology,2016,41(12):266-272(in Chinese).
[20] OHTA Y,KOBAYASHI Y,KONISHI S,et al.Speciation analysis of selenium metabolites in urine and breath by HPLC- and GC-inductively coupled plasma-MS after administration of selenomethionine and methylselenocysteine to rats [J].Chemical Research in Toxicology,2009,22:1795-1801.
[21] BENTE G,KIM G,JESPER B,et al.Separation,purification and identification of the major selenium metabolite from human urine by multi-dimensional HPLC-ICP-MS and APCI-MS [J].Journal of Analytical Atomic Spectrometry,2003,18:65-70.
[22] 倪张林,汤富彬,张玮,等.HPLC-DRC-ICP-MS测定富硒蔬菜中的硒形态[J].分析试验室,2013,32(2):39-43.NI Z L,TANG F B,ZHANG W,et al.Determination of selenium species in se-enriched vegetables by HPLC-DRC-ICP-MS [J].Chinese Journal of Analysis Laboratory,2013,32(2):39-43(in Chinese).
[23] MASEKO T,CALLAHAN D L,DUNSHEA F R,et al.Chemical characterization and speciation of organic selenium in cultivated selenium-enriched Agaricus bisporus [J].Food Chemistry,2013,141:3681-3687.
[24] YUE S Z,ZHANG H Q,ZHEN H Y,et al.Selenium accumulation,speciation and bioaccessibility in seleniumenriched earthworm (Eisenia fetida) [J].Microchemical Journal,2019,145:1-8.
[25] 孙小斐,乔玉辉,孙振钧,等.富硒蚯蚓的培养及硒富集作用研究[J].农业资源与环境学报,2014,31(6):570-574.SUN X F,QIAO Y H,SUN Z J,et al.The cultivation and selenium enrichment of selenium enriched earthworm [J].Journal of Agricultural Resources and Environment,2014,31(6):570-574(in Chinese).
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |