高效液相色谱法测定烧烤制品中5种杂环胺的含量
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摘要
目的建立高效液相色谱法检测烧烤制品中5种杂环胺(heterocyclic aromatic amines,HAAs)含量的方法。方法样品经乙酸乙酯、三乙胺、氨水提取,丙基磺酸(propanesulfonic acid,PRS)和C_(18)固相萃取小柱净化,采用TSK-gel ODS-80_(TM)色谱柱分析,以乙腈和0.05 mol/L醋酸-醋酸铵缓冲液(pH=3.4)为流动相进行梯度洗脱,紫外-荧光检测器的串联方式检测杂环胺含量。结果 2-氨基-1-甲基-6-苯基-咪唑并[4,5-b]吡啶(2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine,PhIP)、9H-吡啶并[4,3-b]吲哚(9H-pyrido[4,3-b]indole,Norharman)、1-甲基-9H-吡啶并[4,3-b]吲哚(1-methyl-9H-pyrido[4,3-b]indol,Harman)、2-氨基-9H-吡啶并[2,3-b]吲哚(2-amine-9H-pyrido[2,3-b]indol,AaC)在0.1～500μg/L范围呈良好的线性关系,2-氨基-3-甲基咪唑并[4,5-f]喹啉(2-amino-3-methylimidazo(4,5-f)quinoline,IQ)在0.1～1000μg/L范围呈良好的线性关系,相关系数r~2均大于99.3%,加标回收率为74.06%～93.77%(n=6),相对标准偏差(relative standard deviation,RSD)为3.41%～5.93%(n=6)。结论该方法简便快速、结果准确、灵敏度高,适合于肉制品中的杂环胺含量的检测。通过检测结果可知不同原料中5种杂环胺含量不同,相同原料烧烤条件不同,产生的杂环胺含量也不相同。
Objective To establish a method for determination of 5 kinds of heterocyclic aromatic amines(HAAs) in the grilled meat. Methods The sample was purified by ethyl acetate, triethylamine, aqueous ammonia, propanesulfonic acid(PRS) and C_(18) solid phase extraction cartridge, and analyzed by TSK-gel ODS-80 TM column. The gradient phase elution was carried out with acetonitrile and 0.05 mol/L acetic acid-ammonium acetate buffer(pH=3.4) as the mobile phase, and the heterocyclic amine content was detected by the ultraviolet-fluorescence detector in series. Results 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine(PhIP), 9 H-pyrido[4,3-b]indole(Norharman), 1-methyl-9 H-pyrido[4,3-b]Indol(Harman), and 2-amine-9 H-pyrido [2,3-b]indol(AaC) showed good linear relationships in the range of 0.1-500 μg/L, 2-amino-3-methylimidazo(4,5-f)quinoline(IQ) showed a good linear relationship in the range of 0.1-1000 μg/L, the correlation coefficients(r~2) were greater than 99.3%, the recoveries were 74.06%-93.77%(n=6), and the relative standard deviations(RSDs) were 3.41% to 5.93%(n=6). Conclusion The method is simple, rapid, accurate and sensitive, which is suitable for detecting the content of heterocyclic amines in meat products. The content of HAAs is different in the different raw materials. The same raw material barbecue conditions are different, and the content of heterocyclic amines produced is also different.
Objective To establish a method for determination of 5 kinds of heterocyclic aromatic amines(HAAs) in the grilled meat. Methods The sample was purified by ethyl acetate, triethylamine, aqueous ammonia, propanesulfonic acid(PRS) and C_(18) solid phase extraction cartridge, and analyzed by TSK-gel ODS-80 TM column. The gradient phase elution was carried out with acetonitrile and 0.05 mol/L acetic acid-ammonium acetate buffer(pH=3.4) as the mobile phase, and the heterocyclic amine content was detected by the ultraviolet-fluorescence detector in series. Results 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine(PhIP), 9 H-pyrido[4,3-b]indole(Norharman), 1-methyl-9 H-pyrido[4,3-b]Indol(Harman), and 2-amine-9 H-pyrido [2,3-b]indol(AaC) showed good linear relationships in the range of 0.1-500 μg/L, 2-amino-3-methylimidazo(4,5-f)quinoline(IQ) showed a good linear relationship in the range of 0.1-1000 μg/L, the correlation coefficients(r~2) were greater than 99.3%, the recoveries were 74.06%-93.77%(n=6), and the relative standard deviations(RSDs) were 3.41% to 5.93%(n=6). Conclusion The method is simple, rapid, accurate and sensitive, which is suitable for detecting the content of heterocyclic amines in meat products. The content of HAAs is different in the different raw materials. The same raw material barbecue conditions are different, and the content of heterocyclic amines produced is also different.
引文
[1]Ohgaki H,Takayama S,Sugimura T.Carcinogenicities of heterocyclic amines in cooked food[J].Mut Res Genet Toxicol,1991,259(3-4):399-410.
[2]Nagao M,Honda M,Seino Y,et al.Mutagenicities of smoke condensates and the charred surface of fish and meat[J].Canc Lett,1977,2(4):221-226.
[3]Felton JS,Fultz E,Dolbeare FA,et al.Effect of microwave pretreatment on heterocyclic aromatic amine mutagens/carcinogens in fried beef patties[J].Food Chem Toxicol,1994,32(10):897-903.
[4]孟晓霞,彭增起,冯云.煎炸对肉制品中杂环胺及多环芳香烃化合物含量的影响及其控制措施[J].肉类研究,2009,(6):52-55.Meng XX,Peng ZQ,Feng Y.Effect factors of formation of polycyclic aromatic hydrocarbon and heterocyclic amines in fried meat and control measures[J].Meat Res,2009,(6):52-55.
[5]Wakabayashi K,Totsuka Y,Fukutome K,et al.Human exposure to mutagenic/carcinogenic heterocyclic amines and comutagenicβ-carbolines[J].Mut Res Fund Mol Mech Mutagenesis,1997,376(1):253-259.
[6]薛菲,彭英云.食品烹调过程中产生的致突变物-杂环胺[J].科技资讯,2007,(36):221-222.Xue F,Peng YY.Mutagenic products produced during food cookingheterocyclic amines[J].Sci Technol Inf,2007,(36):221-222.
[7]Marchand IL,Hankin JH,Wilkens LR,et al.Combined effects of well-done red meat,smoking and rapid N-Acetylrransferase 2 and CYP1A2 phenotypes in increasing colorectal cancer risk[J].Canc Epidemiol Biomark Prev,2001,10(12):1259-1266.
[8]姚瑶,彭增起,邵斌,等.加工肉制品中杂环胺的研究进展[J].食品科学,2010,31(23):447-453.Yao Y,Peng ZQ,Shao B,et al.Heterocyclic aromatic amines in processed meat products[J].Food Sci,2010,31(23):447-453.
[9]Cheng KW,Wong CC,Chao J,et al.Inhibition of mutagenic Ph IPformation by epigallocatechin gallate via scavenging of phenylacetaldehyde[J].Mol Nutr Food Res,2009,53(6):716-725.
[10]Liao GZ,Wang GY,Xu XL,et al.Effect of cooking methods on the formation of heterocyclic aromatic amines in chicken and duck breast[J].Meat Sci,2010,85(1):149-154.
[11]廖国周.烧烤肉制品中杂环胺形成规律研究[D].南京:南京农业大学,2008.Liao GZ.Study on the formation discipline of heterocyclic aromatic amines in fried meat[D].Nanjing:Nanjing Agricultural University,2008.
[12]Lee J,Dong A,Jung K,et al.Influence of extra virgin olive oil on the formation of heterocyclic amines in roasted beef steak[J].Food Sci Biotechnol,2011,20(1):159-165.
[13]Monti SM,Ambrosino P,Skog K,et al.Characterization of phenolic compounds in virgin olive oil and their effect on the formation of carcinogenic/mutagenic heterocyclic amines in a model system[J].J Agric Food Chem,2001,49(8):3969-3975.
[14]Vttaglione P,Moni SM,Ambrosino P,et al.Carotenoids from tomatoes inhibit heterocyclic amine formation[J].Eur Food Res Technol,2002,215(2):108-113.
[15]Kataoka H,Nishioka S,Kobayashi M,et al.Analysis of mutagenic heterocyclic amines in cooked food samples by gas chromatography with nitrogen-phosphorus detector[J].Bull Environ Contam Toxicol,2002,69(5):682-689.
[16]PaiS P,Moyano E,Puignou L,et al.Liquid chromatography-atmospheric-pressure chemical ionization mass spectrometry as a routine method for the analysis of mutagenic amines in beef extracts[J].J Chrom A,1997,778(1):207-218.
[17]Murkovic M,Weber HJ,Geiszler S,et al.Formation of the food associated carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine(PhIP)in model systems[J].Food Chem,1999,65(2):233-237.
[18]Vanderlaan M,Watkins BE,Hwang M,et al.Monoclonal antibodies for the immunoassay of mutagenic compounds produced by cooking beef[J].Carcinogenesis,1988,9(1):153-160.
[19]李可,韩雪,谢美娟,等.HPLC法检测市售传统肉制品中的杂环胺含量[J].现代食品科技,2017,(10):294-301.Li K,Han X,Xie MJ,et al.Analysis of heterocyclic amines in the traditional meat productsdetermined by HPLC[J].Mod Food Sci Technol,2017,(10):294-301.
[20]张峰,吕泉福,储晓刚,等.固相萃取-超高效液相色谱-电喷雾串联质谱法同时测定烤鱼中15种杂环胺含量[J].分析化学,2011,39(4):501-505.Zhang F,Lv CF,Chu XG,et al.simultaneous determination of 15heterocyclic amines in grilled fish using solid phase extraction and ultra performance liquid chromatography with electrospray ionization tandem mass spectrometry[J].Chin J Anal Chem,2011,39(4):501-505.
[21]王盼,洪燕婷,朱雨辰,等.烹调食品中杂环胺检测的分析方法[J].食品工业科技,2013,34(22):343-347.Wang P,Hong YT,Zhu YC,et al.Analytical methods of heterocyclic aromatic amines in foods[J].Sci Technol Food Ind,2013,34(22):343-347.
[22]李可,韩雪,谢美娟,等.SPE-UPLC-MS/MS法测定酱卤鸡腿老汤中10种杂环胺[J].食品工业,2017,(8):306-310.Li K,Han X,Xie MJ,et al.Determination of 10 heterocyclic aromatic amines in halogen soup ofdrumstick products by ultra-high performance liquid chromatographytandem mass spectrometry[J].Food Ind,2017,(8):306-310.
[23]Yao Y,Peng ZQ,Shao B,et al.Effects of frying and boiling on the formation of heterocyclic amines in braised chicken[J].Poultry Sci,2013,92(11):3017-3025.
[24]曾茂茂,李洋,何志勇,等.液相色谱-质谱联用法结合主成分分析考察食品中前体物质对杂环胺生成的影响[J].分析化学,2014,42(1):71-76.Zeng MM,Li Y,He ZY,et al.Effects of different precursors on formation of heterocyclic amines of processed food by ultra-high performance liquid chromatography tandem mass spectrometry combined with principal component analysis[J].Chin J Anal Chem,2014,42(1):71-76.
[2]Nagao M,Honda M,Seino Y,et al.Mutagenicities of smoke condensates and the charred surface of fish and meat[J].Canc Lett,1977,2(4):221-226.
[3]Felton JS,Fultz E,Dolbeare FA,et al.Effect of microwave pretreatment on heterocyclic aromatic amine mutagens/carcinogens in fried beef patties[J].Food Chem Toxicol,1994,32(10):897-903.
[4]孟晓霞,彭增起,冯云.煎炸对肉制品中杂环胺及多环芳香烃化合物含量的影响及其控制措施[J].肉类研究,2009,(6):52-55.Meng XX,Peng ZQ,Feng Y.Effect factors of formation of polycyclic aromatic hydrocarbon and heterocyclic amines in fried meat and control measures[J].Meat Res,2009,(6):52-55.
[5]Wakabayashi K,Totsuka Y,Fukutome K,et al.Human exposure to mutagenic/carcinogenic heterocyclic amines and comutagenicβ-carbolines[J].Mut Res Fund Mol Mech Mutagenesis,1997,376(1):253-259.
[6]薛菲,彭英云.食品烹调过程中产生的致突变物-杂环胺[J].科技资讯,2007,(36):221-222.Xue F,Peng YY.Mutagenic products produced during food cookingheterocyclic amines[J].Sci Technol Inf,2007,(36):221-222.
[7]Marchand IL,Hankin JH,Wilkens LR,et al.Combined effects of well-done red meat,smoking and rapid N-Acetylrransferase 2 and CYP1A2 phenotypes in increasing colorectal cancer risk[J].Canc Epidemiol Biomark Prev,2001,10(12):1259-1266.
[8]姚瑶,彭增起,邵斌,等.加工肉制品中杂环胺的研究进展[J].食品科学,2010,31(23):447-453.Yao Y,Peng ZQ,Shao B,et al.Heterocyclic aromatic amines in processed meat products[J].Food Sci,2010,31(23):447-453.
[9]Cheng KW,Wong CC,Chao J,et al.Inhibition of mutagenic Ph IPformation by epigallocatechin gallate via scavenging of phenylacetaldehyde[J].Mol Nutr Food Res,2009,53(6):716-725.
[10]Liao GZ,Wang GY,Xu XL,et al.Effect of cooking methods on the formation of heterocyclic aromatic amines in chicken and duck breast[J].Meat Sci,2010,85(1):149-154.
[11]廖国周.烧烤肉制品中杂环胺形成规律研究[D].南京:南京农业大学,2008.Liao GZ.Study on the formation discipline of heterocyclic aromatic amines in fried meat[D].Nanjing:Nanjing Agricultural University,2008.
[12]Lee J,Dong A,Jung K,et al.Influence of extra virgin olive oil on the formation of heterocyclic amines in roasted beef steak[J].Food Sci Biotechnol,2011,20(1):159-165.
[13]Monti SM,Ambrosino P,Skog K,et al.Characterization of phenolic compounds in virgin olive oil and their effect on the formation of carcinogenic/mutagenic heterocyclic amines in a model system[J].J Agric Food Chem,2001,49(8):3969-3975.
[14]Vttaglione P,Moni SM,Ambrosino P,et al.Carotenoids from tomatoes inhibit heterocyclic amine formation[J].Eur Food Res Technol,2002,215(2):108-113.
[15]Kataoka H,Nishioka S,Kobayashi M,et al.Analysis of mutagenic heterocyclic amines in cooked food samples by gas chromatography with nitrogen-phosphorus detector[J].Bull Environ Contam Toxicol,2002,69(5):682-689.
[16]PaiS P,Moyano E,Puignou L,et al.Liquid chromatography-atmospheric-pressure chemical ionization mass spectrometry as a routine method for the analysis of mutagenic amines in beef extracts[J].J Chrom A,1997,778(1):207-218.
[17]Murkovic M,Weber HJ,Geiszler S,et al.Formation of the food associated carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine(PhIP)in model systems[J].Food Chem,1999,65(2):233-237.
[18]Vanderlaan M,Watkins BE,Hwang M,et al.Monoclonal antibodies for the immunoassay of mutagenic compounds produced by cooking beef[J].Carcinogenesis,1988,9(1):153-160.
[19]李可,韩雪,谢美娟,等.HPLC法检测市售传统肉制品中的杂环胺含量[J].现代食品科技,2017,(10):294-301.Li K,Han X,Xie MJ,et al.Analysis of heterocyclic amines in the traditional meat productsdetermined by HPLC[J].Mod Food Sci Technol,2017,(10):294-301.
[20]张峰,吕泉福,储晓刚,等.固相萃取-超高效液相色谱-电喷雾串联质谱法同时测定烤鱼中15种杂环胺含量[J].分析化学,2011,39(4):501-505.Zhang F,Lv CF,Chu XG,et al.simultaneous determination of 15heterocyclic amines in grilled fish using solid phase extraction and ultra performance liquid chromatography with electrospray ionization tandem mass spectrometry[J].Chin J Anal Chem,2011,39(4):501-505.
[21]王盼,洪燕婷,朱雨辰,等.烹调食品中杂环胺检测的分析方法[J].食品工业科技,2013,34(22):343-347.Wang P,Hong YT,Zhu YC,et al.Analytical methods of heterocyclic aromatic amines in foods[J].Sci Technol Food Ind,2013,34(22):343-347.
[22]李可,韩雪,谢美娟,等.SPE-UPLC-MS/MS法测定酱卤鸡腿老汤中10种杂环胺[J].食品工业,2017,(8):306-310.Li K,Han X,Xie MJ,et al.Determination of 10 heterocyclic aromatic amines in halogen soup ofdrumstick products by ultra-high performance liquid chromatographytandem mass spectrometry[J].Food Ind,2017,(8):306-310.
[23]Yao Y,Peng ZQ,Shao B,et al.Effects of frying and boiling on the formation of heterocyclic amines in braised chicken[J].Poultry Sci,2013,92(11):3017-3025.
[24]曾茂茂,李洋,何志勇,等.液相色谱-质谱联用法结合主成分分析考察食品中前体物质对杂环胺生成的影响[J].分析化学,2014,42(1):71-76.Zeng MM,Li Y,He ZY,et al.Effects of different precursors on formation of heterocyclic amines of processed food by ultra-high performance liquid chromatography tandem mass spectrometry combined with principal component analysis[J].Chin J Anal Chem,2014,42(1):71-76.