QuEChERS-UPLC-MS/MS测定焙烤食品中4种链格孢菌毒素
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摘要
建立了超高效液相色谱-串联质谱法测定焙烤食品中4种链格孢菌毒素(腾毒素、链格孢菌酚、交链胞酚单甲醚、细交链胞菌酮酸)快速检测方法。样品经乙腈-0.1 mol/L盐酸溶液超声萃取,萃取液经QuEChERS净化处理,在RP18色谱柱(2.1 mm×100mm,1.7μm)上以乙腈和1 mmol/L碳酸氢铵为流动相梯度洗脱,采用电喷雾离子源、正负离子多反应监测模式检测,外标法定量。结果表明,4种待测物在相应的浓度范围内线性良好,相关系数均大于0.999;方法定量限(S/N≥10)为0.03～0.3μg/kg;进行3个水平的添加回收实验,平均回收率(n=6)为83.2%～105.3%;相对标准偏差(RSD,n=6)为1.2%～6.4%。应用本方法检测45个焙烤食品样品,4种真菌毒素均有检出。其中24份样品检出AME,含量为0.1～4.6μg/kg;6份样品检出AOH,含量为0.47～1.5μg/kg;27份样品检出Te A,含量为0.6～3.7μg/kg;9份样品检出TEN,含量为0.3～2.1μg/kg。
An analysis method using ultra-high performance liquid chromatography-tandem mass spectrometru(UPLC-MS/MS) was established for the determination of four alternaria toxins [Tenuazonic acid(TeA), Alternariol(AOH), Alternariol Monomethyl Ether(AME),Tentoxin(TEN)] in baked food. The samples were extracted with acetonitrile and 0.1 mol/L HCl, and the extracted solutions were purified using the Qu ECh ERS approach. The target analytes were then separated on a RP18 column(2.1 mm×100 mm, 1.7 μm) using acetonitrile and 1 mmol/L NH4 HCO3 as the mobile phase and detected by MS/MS under multiple reaction monitoring(MRM) mode. The mass spectrometry measurement was conducted using positive(ESI+) and negative(ESI-) electrospray ionization. Under the optimized conditions, the linearity of the four analytes were good in the corresponding concentration range with the correlation coefficients more than 0.999. The MLOQ was0.03～0.3 μg/kg and the average recoveries(n=6) ranged from 83.2% to 105.3%, and relative standard deviation(RSD, n=6) in a range of1.2%～6.4% when three levels of standard were spiked. This developed method was successfully applied for the detection of 45 baking food samples, and mycotoxins were detected in several samples. AME was detected in 24 samples with the concentrations ranged from 0.1 to 4.6μg/kg; AOH was detected in 6 samples with the concentrations ranged from 0.47 to 1.5 μg/kg; Te A was detected in 27 samples with the concentrations ranged from 0.6 to 3.7 μg/kg; and TEN was detected in 9 samples with the concentrations ranged from 0.3 to 2.1 μg/kg.
An analysis method using ultra-high performance liquid chromatography-tandem mass spectrometru(UPLC-MS/MS) was established for the determination of four alternaria toxins [Tenuazonic acid(TeA), Alternariol(AOH), Alternariol Monomethyl Ether(AME),Tentoxin(TEN)] in baked food. The samples were extracted with acetonitrile and 0.1 mol/L HCl, and the extracted solutions were purified using the Qu ECh ERS approach. The target analytes were then separated on a RP18 column(2.1 mm×100 mm, 1.7 μm) using acetonitrile and 1 mmol/L NH4 HCO3 as the mobile phase and detected by MS/MS under multiple reaction monitoring(MRM) mode. The mass spectrometry measurement was conducted using positive(ESI+) and negative(ESI-) electrospray ionization. Under the optimized conditions, the linearity of the four analytes were good in the corresponding concentration range with the correlation coefficients more than 0.999. The MLOQ was0.03～0.3 μg/kg and the average recoveries(n=6) ranged from 83.2% to 105.3%, and relative standard deviation(RSD, n=6) in a range of1.2%～6.4% when three levels of standard were spiked. This developed method was successfully applied for the detection of 45 baking food samples, and mycotoxins were detected in several samples. AME was detected in 24 samples with the concentrations ranged from 0.1 to 4.6μg/kg; AOH was detected in 6 samples with the concentrations ranged from 0.47 to 1.5 μg/kg; Te A was detected in 27 samples with the concentrations ranged from 0.6 to 3.7 μg/kg; and TEN was detected in 9 samples with the concentrations ranged from 0.3 to 2.1 μg/kg.
引文
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[13]赵榕,邵兵,赵婕,等.液相色谱-电喷雾质谱/质谱法测定高温烹制的淀粉类食品中的丙烯酰胺[J].色谱,2005,23(3):289-291ZHAO Rong,SHAO Bing,ZHAO Jie,et al.Determination of acrylamide in heated starchy food by liquid chromatography-electrospray ionization tandem mass spectrometry[J].Chinese Journal of Chromatography,2005,23(3):289-291
[14]Gaspar E M S M,Lopes J F.Simple gas chromatographic method for furfural analysis[J].Journal of Chromatography A,2009,1216(14):2762-2767
[15]Tekel J,Farkas P,Kovác M.Determination of acrylamide in sugar by capillary GLC with alkali flame-ionization detection[J].Food Additives and Contaminants,1989,6(3):377-381
[16]Rosi P,Borsari A,Lasi G,et al.Aflatoxin M1 in milk:Reliability of the immunoenzymatic assay[J].Int.Dairy J.,2007,17(5):429-435
[17]刘辉,张燕.Qu ECh ERS-酶联免疫快速检测法测定茶叶中黄曲霉毒B1[J].食品安全质量检测学报,2015,6(4):1307-1313LIU Hui,ZHANG Yan.Simultaneous determination for aflatoxin B1 in tea leaf by Qu ECh ERS-enzyme-linked immunosorbent assay[J].Journal of Food Safety and Quality,2015,6(4):1307-1313
[18]谭志熹,江俊生,陈若恒,等.免疫亲和柱净化-酶联免疫吸附法检测普洱茶中黄曲霉毒素B1[J].预防医学论坛,2014,20(3):413-415TAN Zhi-xi,JIANG Jun-sheng,CHEN Ruo-heng,et al.Detection of aflatoxin B1 in the Puer tea using affinity column purification-enzyme linked immunosorbent assay[J].Prev.Med.Trib.,2014,20(3):413-415
[19]范妙璇,董娇娇,王京辉,等.Qu ECh ERS-超高效液相-三重四极杆串联质谱测定白茅根中16种真菌毒素[J].中国中药杂志,2017,42(19):3770-3775FAN Miao-xuan,DONG Jiao-jiao,WANG Jing-hui,et al.Simultaneous determination of sixteen mycotoxins contaminants in cogon rootstalk by QuEChERS-UPLC-QqQmass spectrometry[J].China Journal of Chinese Materia Medica,2017,42(19):3770-3775
[2]孟繁磊,牛红红,何智勇,等.基于UPLC-MS-MS技术定量确证小麦中10种真菌毒素[J].食品工业,2018,39(1):149-153MENG Fan-lei,NIU Hong-hong,HE Zhi-yong,et al.Quantitative determination of 10 kinds of mycotoxins in wheat by UPLC-MS-MS[J].The Food Industry,2018,39(1):149-153
[3]姜冬梅,王瑶,姜楠,等.农产品及其制品中交链孢酚和交链孢酚单甲醚研究进展[J].食品科学,2017,38(21):287-293JIANG Dong-mei,WANG Yao,JIANG Nan,et al.Review on alternariol and alternariol monomethyl ether in agro-products[J].Food Science,2017,38(21):287-293
[4]满燕,梁刚,李安,等.链格孢霉毒素检测方法研究进展[J].食品安全质量检测学报,2016,7(2):453-458MAN Yan,LIANG Gang,LI An,et al.Research progress of detecting methods for Alternaria toxins[J].Journal of Food Safety and Quality,2016,7(2):453-458
[5]李志霞,聂继云,闫震,等.果品主要真菌毒素污染检测、风险评估与控制研究进展[J].中国农业科学,2017,50(2):332-347LI Zhi-xia,NIE Ji-yun,YAN Zhen,et al.Progress in research of detection,risk assessment and control of the mycotoxins in fruits and fruit products[J].Scientia Agricultura Sinica,2017,50(2):332-347
[6]蔡玮红,冼燕萍,罗海英,等.UPLC-MS/MS法同时测定焙烤食品中丙烯酰胺、4-甲基咪唑与5-羟甲基糠醛[J].现代食品科技,2014,30(2):249-254CAI Wei-hong,XIAN Yan-ping,LUO Hai-ying,et al.Determination of acrylamide,4-methylimidazole and5-hydroxymethylfurfural in baked food using ultra-high performance liquid chromatography-tandem mass spectrometry[J].Modern Food Science and Technology,2014,30(2):249-254
[7]GB 2761-2017,食品安全国家标准食品中真菌毒素限量[S]GB 2761-2017,National food safety standard-Limit of mycotoxins in food[S]
[8]Rizzo I,Vedoya G,Maurutto S,et al.Assessment of toxigenic fungion Argentinean medicinal herbs[J].Microbiol.Res.,2004,159(2):113
[9]Carvalho K L,Gon?alves G A A,Lopes A L,et al.Modeling uncertainty estimation determination of aflatoxin M1 in milk by visual and densitometric thin-layer chromatography with immunoaffinity column clean-up[J].Food Addit.Contam.,2012,29(4):679-693
[10]Campone L,Piccinelli A L,Celano R,et al.Application of dispersive liquid-liquid microextraction for the determination of aflatoxins B1,B2,G1 and G2 in cereal products[J].J.Chromatogr.A,2011,1218(42):7648-7654
[11]付朝晖,黄雪祥,闵顺耕.超高效液相色谱法快速测定发酵茶叶中的黄曲霉毒素[J].分析试验室,2009,28(6):112-115FU Zhao-hui,HUANG Xue-xiang,MIN Shun-geng.Rapid determination of aflatoxins B1,B2,G1 and G2 in fermented tea by ultra performance liquid chromatography[J].Chinese Journal of Analysis Laboratory,2009,28(6):112-115
[12]黎睿,谢刚,王松雪.高效液相色谱法同时检测粮食中常见8种真菌毒素的含量[J].食品科学,2015,36(6):206-210LI Rui,XIE Gang,WANG Song-xue.Simultaneous analysis of 8 mycotoxins in grains by high performance liquid chromatography[J].Food Science,2015,36(6):206-210
[13]赵榕,邵兵,赵婕,等.液相色谱-电喷雾质谱/质谱法测定高温烹制的淀粉类食品中的丙烯酰胺[J].色谱,2005,23(3):289-291ZHAO Rong,SHAO Bing,ZHAO Jie,et al.Determination of acrylamide in heated starchy food by liquid chromatography-electrospray ionization tandem mass spectrometry[J].Chinese Journal of Chromatography,2005,23(3):289-291
[14]Gaspar E M S M,Lopes J F.Simple gas chromatographic method for furfural analysis[J].Journal of Chromatography A,2009,1216(14):2762-2767
[15]Tekel J,Farkas P,Kovác M.Determination of acrylamide in sugar by capillary GLC with alkali flame-ionization detection[J].Food Additives and Contaminants,1989,6(3):377-381
[16]Rosi P,Borsari A,Lasi G,et al.Aflatoxin M1 in milk:Reliability of the immunoenzymatic assay[J].Int.Dairy J.,2007,17(5):429-435
[17]刘辉,张燕.Qu ECh ERS-酶联免疫快速检测法测定茶叶中黄曲霉毒B1[J].食品安全质量检测学报,2015,6(4):1307-1313LIU Hui,ZHANG Yan.Simultaneous determination for aflatoxin B1 in tea leaf by Qu ECh ERS-enzyme-linked immunosorbent assay[J].Journal of Food Safety and Quality,2015,6(4):1307-1313
[18]谭志熹,江俊生,陈若恒,等.免疫亲和柱净化-酶联免疫吸附法检测普洱茶中黄曲霉毒素B1[J].预防医学论坛,2014,20(3):413-415TAN Zhi-xi,JIANG Jun-sheng,CHEN Ruo-heng,et al.Detection of aflatoxin B1 in the Puer tea using affinity column purification-enzyme linked immunosorbent assay[J].Prev.Med.Trib.,2014,20(3):413-415
[19]范妙璇,董娇娇,王京辉,等.Qu ECh ERS-超高效液相-三重四极杆串联质谱测定白茅根中16种真菌毒素[J].中国中药杂志,2017,42(19):3770-3775FAN Miao-xuan,DONG Jiao-jiao,WANG Jing-hui,et al.Simultaneous determination of sixteen mycotoxins contaminants in cogon rootstalk by QuEChERS-UPLC-QqQmass spectrometry[J].China Journal of Chinese Materia Medica,2017,42(19):3770-3775