磁性分子印迹聚合物提取-超高效液相色谱-串联质谱法测定乳及乳制品中的4种伪蛋白
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摘要
建立磁性分子印迹聚合物(magnetic molecular imprinted polymers,MMIPs)提取结合超高效液相色谱-串联质谱法测定乳及乳制品中4种伪蛋白(三聚氰胺、环丙氨嗪、双氰胺和缩二脲)的方法。以缩二脲-~(13)C_2和环丙氨嗪-D_4为印记分子,Fe_3O_4为磁性组分,制备具有特异识别性的MMIPs提取样品中的目标化合物,超高效液相色谱-串联质谱法检测。结果表明,4种化合物在5～200 ng/mL范围内具有良好的线性关系,线性相关系数大于0.999。三聚氰胺、环丙氨嗪、双氰胺和缩二脲在牛奶样品中的检出限分别为0.10、0.02、0.05、0.50μg/kg,在奶粉样品中的检出限分别为0.25、0.05、0.13、1.25μg/kg。回收率为80.5%～96.1%,相对标准偏差为1.1%～9.2%。利用本方法对实际样品中4种化合物进行测定,分析结果显示该方法所得到的数值准确、可靠。
A simple and rapid method for the determination of 4 pseudo proteins including melamine, cyromazine, dicyandiamide and biuret in milk and dairy products has been developed by using magnetic molecularly imprinted polymers (MMIPs) extraction combined with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The MMIPs used for the extraction of 4 pseudo proteins were synthesized by polymerization using biuret-~(13)C_2 and cyromazine-D_4 as templates and Fe_3O_4 magnetite as magnetic component. Under the optimized conditions, good linearity was observed for all target analytes in the concentration range of 5–200 ng/mL with correlation coefficient above 0.999. The limits of detection (LODs) for melamine, cyromazine, dicyandiamide and biure were 0.10, 0.02, 0.05 and 0.50 μg/kg in milk samples and 0.25, 0.05, 0.13 and 1.25 μg/kg in powered milk samples, respectively. The recoveries were between 80.5%and 96.1% with relative standard deviations (RSDs) between 1.1% and 9.2%. The developed method was applied with good precision and accuracy to determine real dairy samples.
A simple and rapid method for the determination of 4 pseudo proteins including melamine, cyromazine, dicyandiamide and biuret in milk and dairy products has been developed by using magnetic molecularly imprinted polymers (MMIPs) extraction combined with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The MMIPs used for the extraction of 4 pseudo proteins were synthesized by polymerization using biuret-~(13)C_2 and cyromazine-D_4 as templates and Fe_3O_4 magnetite as magnetic component. Under the optimized conditions, good linearity was observed for all target analytes in the concentration range of 5–200 ng/mL with correlation coefficient above 0.999. The limits of detection (LODs) for melamine, cyromazine, dicyandiamide and biure were 0.10, 0.02, 0.05 and 0.50 μg/kg in milk samples and 0.25, 0.05, 0.13 and 1.25 μg/kg in powered milk samples, respectively. The recoveries were between 80.5%and 96.1% with relative standard deviations (RSDs) between 1.1% and 9.2%. The developed method was applied with good precision and accuracy to determine real dairy samples.
引文
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[20]程莎,李建兵,王晖,等.高效液相法测定化肥中双氰胺和三聚氰胺的方法研究[J].现代化工,2014,34(3):166-168.
[21]张超,赵风年,洪思慧,等.三聚氰胺磁性分子印迹的合成与应用[J].分析仪器,2016(B11):69-74.
[22]黄超囡,段西健,麻海鹏,等.磁性分子印迹聚合物萃取/高效液相色谱检测蔬菜中的灭多威[J].分析测试学报,2013,32(3):275-280.DOI:10.3969/j.issn.1004-4957.2013.03.002.
[23]BADRUDDOZA A Z,TAY A S,TAN P Y,et al.Carboxymethyl-β-cyclodextrin conjugated magnetic nanoparticles as nano-adsorbents for removal of copper ions:synthesis and adsorption studies[J].Journal of Hazardous Materials,2011,185(2):1177-1186.DOI:10.1016/j.jhazmat.2010.10.029.
[24]OZALP V C,BAYRAMOGLU G,ARICA M Y,et al.Design of a core-shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples[J].Applied Microbiology&Biotechnology,2013,97(21):9541-9551.DOI:10.1007/s00253-013-5231-0.
[25]YUE Q,ZHANG Y,JIANG Y,et al.Nanoengineering of core-shell magnetic mesoporous microspheres with tunable surface roughness[J].Journal of the American Chemical Society,2017,139(13):4954-4961.DOI:10.1021/jacs.7b01464.
[26]SONG F,ZHANG H,WANG D,et al.Imine-linked porous organic polymers showing mesoporous microspheres architectures with tunable surface roughness[J].Journal of Polymer Science Part APolymer Chemistry,2017,56(3):319-327.DOI:10.1002/pola.28902.
[27]DU L,WU Y,ZHANG X,et al.Preparation of magnetic molecularly imprinted polymers for the rapid and selective separation and enrichment of perfluorooctane sulfonate[J].Journal of Separation Science,2017,40(13):2819-2826.DOI:10.1002/jssc.201700157.
[28]LAHCEN A A,BALEG A A,BAKER P,et al.Synthesis and electrochemical characterization of nanostructured magnetic molecularly imprinted polymers for 17-β-estradiol determination[J].Sensors&Actuators B Chemical,2017,241:698-705.DOI:10.1016/j.snb.2016.10.132.
[29]ANSARI S,KARIMI M.Recent configurations and progressive uses of magnetic molecularly imprinted polymers for drug analysis[J].Talanta,2017,167:470-485.DOI:10.1016/j.talanta.2017.02.049.
[30]ITO T,SUGAI H,WATANBE M.Process for producing metal nanoparticle and metal nanoparticle produced by the process:8167972B2[P].2012-01-05.
[31]WANG S,LI Y,DING M,et al.Self-assembly molecularly imprinted polymers of 17β-estradiol on the surface of magnetic nanoparticles for selective separation and detection of estrogenic hormones in feeds[J].Journal of Chromatography B Analytical Technologies in the Biomedical&Life Sciences,2011,879(25):2595-2600.DOI:10.1016/j.jchromb.2011.07.017.
[32]PATEL A K,SHARMA P S,PRASAD B B.Electrochemical sensor for uric acid based on a molecularly imprinted polymer brush grafted to tetraethoxysilane derived sol-gel thin film graphite electrode[J].Materials Science and Engineering,2009,29(5):1545-1553.DOI:10.1016/j.msec.2008.12.008.
[33]YANG H,ZHOU W,GUO X,et al.Molecularly imprinted polymer as SPE sorbent for selective extraction of melamine in dairy products[J].Talanta,2009,80(2):821-825.DOI:10.1016/j.talanta.2009.07.067.
[34]涂志江,张宝林,冯凌云,等.聚乙二醇/聚乙烯吡咯烷酮修饰的纳米Fe_3O_4粒子的制备与表征[J].化工学报,2012,63(12):132-137.DOI:10.3969/j.issn.0438-1157.2012.12.050.
[35]ELLWANGER A,BERGGREN C,BAYOUDH S,et al.Evaluation of methods aimed at complete removal of template from molecularly imprinted polymers[J].Analyst,2001,126(6):784-792.
[36]The Commission of the European Communities.Corporate social responsibility:a business contribution to sustainable development[R].Brussels:European Community European Communities,2002.
[2]AKIYAM H,MORIMOTO S,HAYATSU M,et al.Nitrification,ammonia-oxidizing communities,and N_2O and CH_4,fluxes in an imperfectly drained agricultural field fertilized with coated urea with and without dicyandiamide[J].Biology&Fertility of Soils,2013,49(2):213-223.DOI:10.1007/s00374-012-0713-2.
[3]SMITH I,SCHALLENBERG M.Occurrence of the agricultural nitrification inhibitor,dicyandiamide,in surface waters and its effects on nitrogen dynamics in an experimental aquatic system[J].Agriculture Ecosystems&Environment,2013,164(4):23-31.DOI:10.1016/j.agee.2012.09.002.
[4]王祖翔,蒋俊,孙莉,等.高效液相色谱-串联质谱法测定食品中的尿素、缩二脲与双氰胺[J].分析测试学报,2012,31(5):593-595.DOI:10.3969/j.issn.1004-4957.2012.05.016.
[5]MACMAHON S,BEGLEY T H,DIACHENKOG W,et al.A liquid chromatography-tandem mass spectrometry method for the detection of economically motivated adulteration in protein-containing foods[J].Journal of Chromatography A,2012,1220(2):101-107.DOI:10.1016/j.chroma.2011.11.066.
[6]樊苑牧,俞雪钧,顾晓俊,等.高效液相色谱法测定含脂羊毛中灭蝇胺和环虫腈[J].分析化学,2010,38(1):113-116.
[7]谌芳,毛洪祥,张淑娟,等.高效液相色谱法测定尿素中缩二脲[J].磷肥与复肥,2011,26(2):69-70.DOI:10.3969/j.issn.1007-6220.2.11.02.027.
[8]陈梅兰,潘广文,戴琨,等.离子排斥色谱紫外检测法测定奶粉中双氰胺含量[J].分析化学,2013,41(11):1734-1738.DOI:10.3724/SP.J.1096.2013.30289.
[9]曾凯,刘峙嵘,宁雅君,等.气相色谱-质谱联用法同时测定乳及乳制品中的三聚氰胺及肌酐[J].色谱,2013,31(5):477-480.DOI:10.3724/SP.J.1123.2012.11008.
[10]金瓯,任天辉,潘咏梅.反相高效液相色谱DAD法测定尿囊素及其相关物的含量[J].化学工业与工程技术,2013,34(1):59-62.DOI:10.3969/j.issn.1006-7906.2013.01.015.
[11]刘萍.超高效液相色谱紫外法测定水基食品模拟物中双氰胺的特定迁移量[J].食品安全导刊,2016(18):87-87.
[12]王奇.高效液相色谱法测定奶粉中双氰胺[J].化工时刊,2015(5):17-18.DOI:10.3969/j.issn.1002-154X.2015.05.006.
[13]林品言.高效液相色谱法检测生鲜乳中三聚氰胺[J].湖南畜牧兽医,2017(1):44-46.DOI:10.3969/j.issn.1006-4907.2017.01.020.
[14]LEE H M,LEE B J.A novel approach to simultaneous screening and confirmation of regulated pharmaceutical compounds in dietary supplements by LC/MS/MS with an information-dependent acquisition method[J].Food Additives&Contaminants,2011,28(4):396-407.
[15]张宪臣,王勇,张朋杰,等.SA-UPLC-MS-MS联用测定奶粉中双氰胺的残留量[J].食品科学,2013,34(18):231-238.DOI:10.7506/spkx1002-6630-201318047.
[16]张立佳,胡雪,白艳梅,等.液相色谱-串联质谱法检测乳及乳制品中双氰胺的研究进展[J].乳业科学与技术,2015,38(3):34-38.DOI:10.7506/rykxjs1671-5187-201503009.
[17]侯建波,谢文,李杰,等.液相色谱-串联质谱法同时测定牛奶和奶粉中5种非蛋白含氮化合物[J].质谱学报,2016,37(4):336-342.DOI:10.7538/zpxb.youxian.2016.0022.
[18]夏明星,黄均艳,李光满,等.动物源血液中三聚氰胺的液相色谱质谱联用检测方法的建立[J].安徽农业大学学报,2017,44(3):392-397.DOI:10.13610/j.cnki.1672-352x.20170524.011.
[19]曹倩,朱森森.三聚氰胺检测方法简述[J].广州化学,2017,42(2):77-81.DOI:10.16560/j.cnki.gzhx.20170205.
[20]程莎,李建兵,王晖,等.高效液相法测定化肥中双氰胺和三聚氰胺的方法研究[J].现代化工,2014,34(3):166-168.
[21]张超,赵风年,洪思慧,等.三聚氰胺磁性分子印迹的合成与应用[J].分析仪器,2016(B11):69-74.
[22]黄超囡,段西健,麻海鹏,等.磁性分子印迹聚合物萃取/高效液相色谱检测蔬菜中的灭多威[J].分析测试学报,2013,32(3):275-280.DOI:10.3969/j.issn.1004-4957.2013.03.002.
[23]BADRUDDOZA A Z,TAY A S,TAN P Y,et al.Carboxymethyl-β-cyclodextrin conjugated magnetic nanoparticles as nano-adsorbents for removal of copper ions:synthesis and adsorption studies[J].Journal of Hazardous Materials,2011,185(2):1177-1186.DOI:10.1016/j.jhazmat.2010.10.029.
[24]OZALP V C,BAYRAMOGLU G,ARICA M Y,et al.Design of a core-shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples[J].Applied Microbiology&Biotechnology,2013,97(21):9541-9551.DOI:10.1007/s00253-013-5231-0.
[25]YUE Q,ZHANG Y,JIANG Y,et al.Nanoengineering of core-shell magnetic mesoporous microspheres with tunable surface roughness[J].Journal of the American Chemical Society,2017,139(13):4954-4961.DOI:10.1021/jacs.7b01464.
[26]SONG F,ZHANG H,WANG D,et al.Imine-linked porous organic polymers showing mesoporous microspheres architectures with tunable surface roughness[J].Journal of Polymer Science Part APolymer Chemistry,2017,56(3):319-327.DOI:10.1002/pola.28902.
[27]DU L,WU Y,ZHANG X,et al.Preparation of magnetic molecularly imprinted polymers for the rapid and selective separation and enrichment of perfluorooctane sulfonate[J].Journal of Separation Science,2017,40(13):2819-2826.DOI:10.1002/jssc.201700157.
[28]LAHCEN A A,BALEG A A,BAKER P,et al.Synthesis and electrochemical characterization of nanostructured magnetic molecularly imprinted polymers for 17-β-estradiol determination[J].Sensors&Actuators B Chemical,2017,241:698-705.DOI:10.1016/j.snb.2016.10.132.
[29]ANSARI S,KARIMI M.Recent configurations and progressive uses of magnetic molecularly imprinted polymers for drug analysis[J].Talanta,2017,167:470-485.DOI:10.1016/j.talanta.2017.02.049.
[30]ITO T,SUGAI H,WATANBE M.Process for producing metal nanoparticle and metal nanoparticle produced by the process:8167972B2[P].2012-01-05.
[31]WANG S,LI Y,DING M,et al.Self-assembly molecularly imprinted polymers of 17β-estradiol on the surface of magnetic nanoparticles for selective separation and detection of estrogenic hormones in feeds[J].Journal of Chromatography B Analytical Technologies in the Biomedical&Life Sciences,2011,879(25):2595-2600.DOI:10.1016/j.jchromb.2011.07.017.
[32]PATEL A K,SHARMA P S,PRASAD B B.Electrochemical sensor for uric acid based on a molecularly imprinted polymer brush grafted to tetraethoxysilane derived sol-gel thin film graphite electrode[J].Materials Science and Engineering,2009,29(5):1545-1553.DOI:10.1016/j.msec.2008.12.008.
[33]YANG H,ZHOU W,GUO X,et al.Molecularly imprinted polymer as SPE sorbent for selective extraction of melamine in dairy products[J].Talanta,2009,80(2):821-825.DOI:10.1016/j.talanta.2009.07.067.
[34]涂志江,张宝林,冯凌云,等.聚乙二醇/聚乙烯吡咯烷酮修饰的纳米Fe_3O_4粒子的制备与表征[J].化工学报,2012,63(12):132-137.DOI:10.3969/j.issn.0438-1157.2012.12.050.
[35]ELLWANGER A,BERGGREN C,BAYOUDH S,et al.Evaluation of methods aimed at complete removal of template from molecularly imprinted polymers[J].Analyst,2001,126(6):784-792.
[36]The Commission of the European Communities.Corporate social responsibility:a business contribution to sustainable development[R].Brussels:European Community European Communities,2002.