苏丹红Ⅱ纯物质中有机杂质的定性和定量分析
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摘要
本实验对苏丹红Ⅱ标准物质候选物中的主要有机杂质进行定性、定量分析,采用高效液相色谱-离子阱-飞行时间串联质谱法(HPLC-IT-TOF MS)和液相色谱-三重四极杆串联质谱法(HPLC-MS/MS)对有机杂质进行定性分析,鉴别出杂质1和杂质2分别为苏丹红Ⅰ和溶剂橙2,推导出杂质3和4的可能分子结构为苏丹红Ⅱ的同分异构体。利用液相色谱外标法测定苏丹红Ⅰ和溶剂橙2的含量分别为0.017%、0.258%,相对标准偏差分别为1.6%、2.5%;采用与主成分苏丹红Ⅱ响应因子一致的方法定量分析杂质3和4,其含量分别为0.012%、0.028%,相对标准偏差分别为7.0%、4.9%。本研究可为苏丹红Ⅱ一级标准物质的研制奠定基础,对苏丹红Ⅱ的准确测定具有重要意义。
SudanⅡis one of synthetic organic azo-compounds,which is abundantly used in industry for coloring.A variety of foodstuffs contaminated with Sudan dyes have been detected throughout Europe and Asia.This issue has become one focal point for addressing food safety.Sudan Ⅱ could produce aniline and its derivatives,which could attack the hepatocytes and lead to the toxic hepatic disease.For this reason,accurate and reliable determination of Sudan Ⅱin foodstuff is required for the assurance of consumer's healthiness.But the traceability of SudanⅡ has not been settled yet.Qualita-tive and quantitative analysis of the major organic impurities in pure Sudan Ⅱ was performed in this work.The related organic impurities were qualitatively determined by high performance liquid chromatography-ion trap-time of flight mass spectrometry(HPLC-IT-TOF MS)and high performance liquid chromatography-triple quadrupole tandem mass spectrometry(HPLC-MS/MS).Full scan of LC/MS in positive ion mode was firstly executed to obtain the m/z values of the molecules.Then LC-MS/MS was carried out on target compounds to obtain as much structural information as possible.According to the first and the second mass spectra,retention time of standard materials and published reports,the impurities in Sudan Ⅱ were identified and their fragmentation pathways were concluded.Four unknown related organic impurities were observed in Sudan Ⅱ.From analysis of possible structures and fragmentation pathways of fragments,impurities 1 and 2 were successfully identified as SudanⅠand Solvent Orange 2,respectively.The results were verified by standards.The possible molecular structures of impurity 3 and 4 were derived as isomers of SudanⅡ.High performance liquid chromatography external standard method was used to determine the contents of the identified organic impurities.The results showed that there are 0.017% Sudan Ⅰ with the RSD of 1.6% and 0.258% Solvent Orange 2 with the RSD of 2.5%in pure Sudan Ⅱsubstance.Impurities 3 and 4 were quantified in accordance with the SudanⅡresponse factor under DAD detector.The contents of impurities 3 and 4 are 0.012%and 0.028%with the RSD of 7.0% and 4.9%,respectively.Moreover,this method has instructive significance to the qualitative and quantitative analysis of pure Sudan dyes and other pure organic substances.Also it will assure the accuracy of determination of Sudan Ⅱand lay the foundation for development of a national first grade certified reference material of Sudan Ⅱ.
SudanⅡis one of synthetic organic azo-compounds,which is abundantly used in industry for coloring.A variety of foodstuffs contaminated with Sudan dyes have been detected throughout Europe and Asia.This issue has become one focal point for addressing food safety.Sudan Ⅱ could produce aniline and its derivatives,which could attack the hepatocytes and lead to the toxic hepatic disease.For this reason,accurate and reliable determination of Sudan Ⅱin foodstuff is required for the assurance of consumer's healthiness.But the traceability of SudanⅡ has not been settled yet.Qualita-tive and quantitative analysis of the major organic impurities in pure Sudan Ⅱ was performed in this work.The related organic impurities were qualitatively determined by high performance liquid chromatography-ion trap-time of flight mass spectrometry(HPLC-IT-TOF MS)and high performance liquid chromatography-triple quadrupole tandem mass spectrometry(HPLC-MS/MS).Full scan of LC/MS in positive ion mode was firstly executed to obtain the m/z values of the molecules.Then LC-MS/MS was carried out on target compounds to obtain as much structural information as possible.According to the first and the second mass spectra,retention time of standard materials and published reports,the impurities in Sudan Ⅱ were identified and their fragmentation pathways were concluded.Four unknown related organic impurities were observed in Sudan Ⅱ.From analysis of possible structures and fragmentation pathways of fragments,impurities 1 and 2 were successfully identified as SudanⅠand Solvent Orange 2,respectively.The results were verified by standards.The possible molecular structures of impurity 3 and 4 were derived as isomers of SudanⅡ.High performance liquid chromatography external standard method was used to determine the contents of the identified organic impurities.The results showed that there are 0.017% Sudan Ⅰ with the RSD of 1.6% and 0.258% Solvent Orange 2 with the RSD of 2.5%in pure Sudan Ⅱsubstance.Impurities 3 and 4 were quantified in accordance with the SudanⅡresponse factor under DAD detector.The contents of impurities 3 and 4 are 0.012%and 0.028%with the RSD of 7.0% and 4.9%,respectively.Moreover,this method has instructive significance to the qualitative and quantitative analysis of pure Sudan dyes and other pure organic substances.Also it will assure the accuracy of determination of Sudan Ⅱand lay the foundation for development of a national first grade certified reference material of Sudan Ⅱ.
引文
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[13]王孜一,戴新华,张伟,等.皮质醇纯物质中杂质分析与定值[J].质谱学报,2014,35(5):413-419.WANG Ziyi,DAI Xinhua,ZHANG Wei,et al.Impurities analysis and purity determination of pure hydrocortisone[J].Journal of Chinese Mass Spectrometry Society,2014,35(5):413-419(in Chinese).
[2]江晨舟,孙洁敏,张驰中,等.食品中苏丹红检测方法的比较和优化[J].上海计量测试,2016,43(3):18-24.JIANG Chenzhou,SUN Jiemin,ZHANG Chizhong,et al.Comparison and optimization of detection methods for Sudan red in foods[J].Shanghai Measurement and Testing,2016,43(3):18-24(in Chinese).
[3]SRICHAROENA P,LIMCHOOWONGA N,SRIPAKDEE T,et al.Electrolyte-assisted microemulsion breaking in vortex-agitated solidified floating organic drop microextraction for preconcentration and analysis of Sudan dyes in Chili products[J].Anal Methods,2017,9(25):3 810-3 818.
[4]宁啸骏,王林,虞成华,等.UPLC-MS/MS同位素内标法测定食品中对位红、苏丹红Ⅰ~Ⅳ的研究[J].质谱学报,2009,30(1):41-46.NING Xiaojun,WANG Lin,YU Chenghua,et al.UPLC-MS/MS isotopic internal standard determination of parared and sudanⅠ-Ⅳin food[J].Journal of Chinese Mass Spectrometry Society,2009,30(1):41-46(in Chinese).
[5]CHAILAPAKUL O,WONSAWAT W,SIANGPROH W,et al.Analysis of SudanⅠ,SudanⅡ,SudanⅢ,and SudanⅣin food by HPLC with electrochemical detection:comparison of glassy carbon electrode with carbon nanotubeionic liquid gel modified electrode[J].Food Chem,2008,109(4):876-882.
[6]喻凌寒,宋之光,苏流坤,等.高效液相色谱-离子阱质谱联用测定食品中7种苏丹红类染料[J].质谱学报,2006,27(4):221-225.YU Linghan,SONG Zhiguang,SU Liukun,et al.Determination of 7kinds of sudan dye in food by HPLC-ESI-MS/MS[J].Journal of Chinese Mass Spectrometry Society,2006,27(4):221-225(in Chinese).
[7]ZHANG C J,LI G K,ZHANG Z M,et al.A hydrazone covalent organic polymer based microsolid phase extraction for online analysis of trace Sudan dyes in food samples[J].J Chromatogr A,2015,(1 419):1-9.
[8]范文锐,吴青,劳扬,等.高效液相色谱法同时测定食品中7种非食用色素[J].分析化学,2012,40(2):292-297.FAN Wenrui,WU Qing,LAO Yang,et al.Determination of seven inedible pigments in food using high performance liquid chromatography[J].Chinese Journal of Analytical Chemistry,2012,40(2):292-297(in Chinese).
[9]XIE X Y,CHEN L,PAN X Y,et al.Synthesis of magnetic molecularly imprinted polymers by reversible addition fragmentation chain transfer strategy and its application in the Sudan dyes residue analysis[J].J Chromatogr A,2015,(1 405):32-39.
[10]WESTWOOD S,CHOTEAU T,DAIREAUX A,et al.Mass balance method for the SI value assignment of the purity of organic compounds[J].Anal Chem,2013,85(6):3 118-3 126.
[11]暴海霞,戴新华,张伟,等.17α-羟孕酮纯度及其杂质分析方法的研究[J].化学试剂,2014,36(3):226-230.BAO Haixia,DAI Xinhua,ZHANG Wei,et al.Analysis of the purity and impurities in 17α-hydroxyprogesterone[J].Chemical Reagents,2014,36(3):226-230(in Chinese).
[12]赵贵平,孟云,仇雯荔,等.苏丹红Ⅰ的LC-MS/MS分析[J].分析测试学报,2005,24(6):290-291.ZHAO Guiping,MENG Yun,QIU Wenli,et al.Analysis method of Sudan I by liquid chromatography-tandem mass spectrometry[J].Journal of Instrumental Analysis,2005,24(6):290-291(in Chinese).
[13]王孜一,戴新华,张伟,等.皮质醇纯物质中杂质分析与定值[J].质谱学报,2014,35(5):413-419.WANG Ziyi,DAI Xinhua,ZHANG Wei,et al.Impurities analysis and purity determination of pure hydrocortisone[J].Journal of Chinese Mass Spectrometry Society,2014,35(5):413-419(in Chinese).