超高效液相色谱-串联质谱法快速检测动物饲料中5种霉菌毒素
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摘要
目的建立一种同步、快速的测定饲料中5种主要霉菌毒素的超高效液相色谱-串联质谱(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)检测方法。方法样品经84%乙腈水(含0.1%甲酸)提取液提取, MLJ-1杂质吸附型固相萃取柱净化。采用0.1mmol/L乙酸铵/0.1%甲酸-水(A)和0.1%甲酸-甲醇(B)作为流动相进行梯度洗脱,质谱采用多反应监测模式(multiplereactionmonitoring,MRM)对霉菌毒素及其同位素内标的定量离子和定性离子进行监测。结果本方法在1min内完成样品净化处理,15 min内完成5种目标化合物的分离分析。5种霉菌毒素在高、中和低浓度添加水平的回收率为83.9%～113.8%,相对标准偏差为0.6%～15.0%(n=6),方法定量限为0.3～4.0μg/kg。结论该方法操作简单、快速,适合饲料中5种主要霉菌毒素同步确证检测。
Objective To establish a rapid method for simultaneous determination of 5 major mycotoxins in animal feed samples by ultra performance liquid chromatography-tandem mass spectrometry(UPLC–MS/MS).Methods The 5 analytes were extracted from the feed samples by acetonitrile-water-formic acid, and purified by a commercial MLJ-1 pass-through solid-phase extraction(SPE) clean-up cartridge, and then analyzed by UPLC-MS/MS. The 5 analytes were then separated on a reversed phase column using a gradient elution program of0.1 mmol/L ammonium acetate/0.1% formic acid aqueous solution and 0.1% formic acid methanol solution. The quantitative and qualitative ions of mycotoxin and their internal isotopes were detected with multiple reaction monitoring(MRM) mode. Results This method could purify the samples within 1 min and separate and analyze 5 target compounds within 15 min. The recoveries of 3 spiked levels were in range of 83.9%-113.8%, with the relative standard deviations(RSDs) of 0.6%-15.0%(n=6). The limits of quantification(LOQ) for all analytes were between0.3 and 4.0 μg/kg. Conclusion The proposed method is simple, rapid and accurate, which is suitable for detecting 5 major mycotoxins in animal feed samples.
Objective To establish a rapid method for simultaneous determination of 5 major mycotoxins in animal feed samples by ultra performance liquid chromatography-tandem mass spectrometry(UPLC–MS/MS).Methods The 5 analytes were extracted from the feed samples by acetonitrile-water-formic acid, and purified by a commercial MLJ-1 pass-through solid-phase extraction(SPE) clean-up cartridge, and then analyzed by UPLC-MS/MS. The 5 analytes were then separated on a reversed phase column using a gradient elution program of0.1 mmol/L ammonium acetate/0.1% formic acid aqueous solution and 0.1% formic acid methanol solution. The quantitative and qualitative ions of mycotoxin and their internal isotopes were detected with multiple reaction monitoring(MRM) mode. Results This method could purify the samples within 1 min and separate and analyze 5 target compounds within 15 min. The recoveries of 3 spiked levels were in range of 83.9%-113.8%, with the relative standard deviations(RSDs) of 0.6%-15.0%(n=6). The limits of quantification(LOQ) for all analytes were between0.3 and 4.0 μg/kg. Conclusion The proposed method is simple, rapid and accurate, which is suitable for detecting 5 major mycotoxins in animal feed samples.
引文
[1]Cao X,Wu S,Yue Y,et al.A high-throughput method for the simultaneous determination of multiple mycotoxins in human and laboratory animal biological fluids and tissues by PLE and HPLC-MS/MS[J].J Chromatogr B,2013,942-943:113-125.
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[3]Ahmad A,Jae-Hyuk Y.Occurrence,toxicity,and analysis of major mycotoxins in food[J].Int J Environ Res Public Health,2017,14(6):632-652.
[4]Marin S,Ramos AJ,Cano-Sancho G,et al.Mycotoxins:Occurrence,toxicology,and exposure assessment[J].Food Chem Toxicol,2013,60:218-237.
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[7]World Health Organization.WHO global strategy for food safety:safer food for better health[M].Switzerland:WHO,2003.
[8]GB 2761-2011食品安全国家标准食品中真菌毒素限量[S].GB 2761-2011 National food safety standard-Maximum levels of mycotoxins in foods[S].
[9]GB 13078-2017饲料卫生标准[S].GB 13078-2017 Hygienical standard for feeds[S].
[10]赵颖,刘洪美,卢静华,等.基于适配体生物传感器检测黄曲霉毒素B1的研究进展[J].食品安全质量检测学报,2018,9(22):5806-5815.Zhao Y,Liu HM,Lu JH,et al.Research progress on detection of aflatoxin B1 based on aptamer biosensor[J].J Food Saf Qual,2018,9(22):5806-5815.
[11]Qian M,Yang H,Li Z,et al.Detection of 13 mycotoxins in feed using modified QuEChERS with dispersive magnetic materials and UHPLC-MS/MS[J].J Sep Sci,2018,41(3):756-764.
[12]孙雪,郗存显,唐柏彬,等.复合免疫亲和柱净化-液相色谱-串联质谱法测定动物源食品中6种黄曲霉毒素和6种玉米赤霉醇类真菌毒素残留量[J].分析化学,2016,44(6):970-978.Su X,Xi CX,Tang BB,et al.Determination of aflatoxins and zeranols in animal-originated foods by high performance liquid chromatography-tandem mass spectrometry combined with immunoaffinity clean-up column[J].Chin J Anal Chem,2016,44(6):970-978.
[13]马俊美,刘东,张雷雷,等.在线固相萃取-液相色谱-串联质谱法快速测定玉米粉和面粉中伏马毒素[J].食品科学,2017,(20):307-312.Ma JM,Liu D,Zhang LL,et al.Liquid chromatography-tandem mass spectrometry method for determination of fumonisins in corn and flour samples based on-line solid phase extraction purification[J].Food Sci.2017,(20):307-312.
[14]王少敏,黄晓静,毛丹,等.QuEChERS-超高效液相色谱串联质谱法同时测定中药瓜蒌皮中22种真菌毒素[J].食品安全质量检测学报,2018,9(22):5843-5855.Wang SM,Huang XJ,Mao D,et al.Simultaneous determination of 22kinds of mycotoxins in trichosanthis pericarpium by ultra performance liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2018,9(22):5843-5855.
[15]雷方,李承龙,张新,等.免疫磁珠亲和纯化-超高效液相色谱串联质谱法快速检测小麦中多种真菌毒素[J].食品安全质量检测学报,2016,7(3):1252-1260.Lei F,Li CL,Zhang X,et al.Rapid determination of mycotoxins in wheat by immune magnetic beads affinity purification with ultra performance liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2016,7(3):1252-1260.
[16]林涛,邵金良,陈兴连,等.固相萃取-超高效液相色谱-串联质谱法快速测定农产食品中14种真菌类生物毒素[J].食品安全质量检测学报,2018,9(13):3237-3244.Lin T,Shao JL,Chen XL,et al.Rapid determination of 14 kinds of fungal biotoxins in agricultural products and foods by solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2018,9(13):3237-3244.
[17]王瑞国,苏晓鸥,程芳芳,等.液相色谱-串联质谱法测定饲料原料中26种霉菌毒素[J].分析化学,2015,43(2):264-270.Wang RG,Su XO,Cheng FF,et al.Determination of 26 mycotoxins in feedstuffs by multifunctional cleanup column and liquid chromatography-tandem mass spectrometry[J].Chin J Anal Chem,2015,43(2):264-270.
[18]张大伟,高和杨,周旌,等.超高效液相色谱-串联质谱法同时检测饲料原料、饲料成品中18种真菌毒素含量[J].食品安全质量检测学报,2018,9(22):5867-5876.Zhang DW,Gao HY,Zhou J,et al.Simultaneous determination of 18kinds of mycotoxins in feed ingredients and feed products by ultra performance liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2018,9(22):5867-5876.
[2]Bryden WL.Mycotoxin contamination of the feed supply chain:Implications for animal productivity and feed security[J].Anim Feed Sci Technol,2012,173:134-158.
[3]Ahmad A,Jae-Hyuk Y.Occurrence,toxicity,and analysis of major mycotoxins in food[J].Int J Environ Res Public Health,2017,14(6):632-652.
[4]Marin S,Ramos AJ,Cano-Sancho G,et al.Mycotoxins:Occurrence,toxicology,and exposure assessment[J].Food Chem Toxicol,2013,60:218-237.
[5]Turner NW,Bramhmbhatt H,Szabovezse M,et al.Analytical methods for determination of mycotoxins:An update(2009-2014)[J].Anal Chim Acta,2015,901(2):12-33.
[6]Van FHJ,Paulien A,Ans P,et al.Data analyses and modelling for risk based monitoring of mycotoxins in animal feed[J].Toxins,2018,10(2):54-69.
[7]World Health Organization.WHO global strategy for food safety:safer food for better health[M].Switzerland:WHO,2003.
[8]GB 2761-2011食品安全国家标准食品中真菌毒素限量[S].GB 2761-2011 National food safety standard-Maximum levels of mycotoxins in foods[S].
[9]GB 13078-2017饲料卫生标准[S].GB 13078-2017 Hygienical standard for feeds[S].
[10]赵颖,刘洪美,卢静华,等.基于适配体生物传感器检测黄曲霉毒素B1的研究进展[J].食品安全质量检测学报,2018,9(22):5806-5815.Zhao Y,Liu HM,Lu JH,et al.Research progress on detection of aflatoxin B1 based on aptamer biosensor[J].J Food Saf Qual,2018,9(22):5806-5815.
[11]Qian M,Yang H,Li Z,et al.Detection of 13 mycotoxins in feed using modified QuEChERS with dispersive magnetic materials and UHPLC-MS/MS[J].J Sep Sci,2018,41(3):756-764.
[12]孙雪,郗存显,唐柏彬,等.复合免疫亲和柱净化-液相色谱-串联质谱法测定动物源食品中6种黄曲霉毒素和6种玉米赤霉醇类真菌毒素残留量[J].分析化学,2016,44(6):970-978.Su X,Xi CX,Tang BB,et al.Determination of aflatoxins and zeranols in animal-originated foods by high performance liquid chromatography-tandem mass spectrometry combined with immunoaffinity clean-up column[J].Chin J Anal Chem,2016,44(6):970-978.
[13]马俊美,刘东,张雷雷,等.在线固相萃取-液相色谱-串联质谱法快速测定玉米粉和面粉中伏马毒素[J].食品科学,2017,(20):307-312.Ma JM,Liu D,Zhang LL,et al.Liquid chromatography-tandem mass spectrometry method for determination of fumonisins in corn and flour samples based on-line solid phase extraction purification[J].Food Sci.2017,(20):307-312.
[14]王少敏,黄晓静,毛丹,等.QuEChERS-超高效液相色谱串联质谱法同时测定中药瓜蒌皮中22种真菌毒素[J].食品安全质量检测学报,2018,9(22):5843-5855.Wang SM,Huang XJ,Mao D,et al.Simultaneous determination of 22kinds of mycotoxins in trichosanthis pericarpium by ultra performance liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2018,9(22):5843-5855.
[15]雷方,李承龙,张新,等.免疫磁珠亲和纯化-超高效液相色谱串联质谱法快速检测小麦中多种真菌毒素[J].食品安全质量检测学报,2016,7(3):1252-1260.Lei F,Li CL,Zhang X,et al.Rapid determination of mycotoxins in wheat by immune magnetic beads affinity purification with ultra performance liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2016,7(3):1252-1260.
[16]林涛,邵金良,陈兴连,等.固相萃取-超高效液相色谱-串联质谱法快速测定农产食品中14种真菌类生物毒素[J].食品安全质量检测学报,2018,9(13):3237-3244.Lin T,Shao JL,Chen XL,et al.Rapid determination of 14 kinds of fungal biotoxins in agricultural products and foods by solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2018,9(13):3237-3244.
[17]王瑞国,苏晓鸥,程芳芳,等.液相色谱-串联质谱法测定饲料原料中26种霉菌毒素[J].分析化学,2015,43(2):264-270.Wang RG,Su XO,Cheng FF,et al.Determination of 26 mycotoxins in feedstuffs by multifunctional cleanup column and liquid chromatography-tandem mass spectrometry[J].Chin J Anal Chem,2015,43(2):264-270.
[18]张大伟,高和杨,周旌,等.超高效液相色谱-串联质谱法同时检测饲料原料、饲料成品中18种真菌毒素含量[J].食品安全质量检测学报,2018,9(22):5867-5876.Zhang DW,Gao HY,Zhou J,et al.Simultaneous determination of 18kinds of mycotoxins in feed ingredients and feed products by ultra performance liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2018,9(22):5867-5876.