葡萄酒中16种真菌毒素的同位素稀释-超高效液相色谱-串联质谱同时测定法

职业与健康 ›› 2023, Vol. 39 ›› Issue (11): 1447-1454.

葡萄酒中16种真菌毒素的同位素稀释-超高效液相色谱-串联质谱同时测定法

杨雪丽, 王猛, 李菁, 李方

新疆维吾尔自治区疾病预防控制中心卫生监测检验中心,新疆乌鲁木齐 830001

收稿日期:2022-09-15 修回日期:2022-11-03 发布日期:2026-03-15
通信作者: 李方,副主任医师,E-mail：115144679@qq.com
作者简介:杨雪丽,女,实验师,主要从事为食品理化检验工作。
基金资助:
新疆维吾尔自治区自然科学基金（2020D01C083）

Simultaneous determination of 16 kinds of mycotoxins in wine by isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry

YANG Xue-li, WANG Meng, LI Jing, LI Fang

Health Monitoring and Inspection Center,Xinjiang Uygur Autonomous Region Center for Disease Prevention and Control,Urumqi Xinjiang 830001,China

Received:2022-09-15 Revised:2022-11-03 Published:2026-03-15
Contact: LI Fang,Associate chief physician,E-mail：115144679@qq.com

摘要/Abstract

摘要： 目的建立稳定同位素内标超高效液相色谱-串联质谱联用法（ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS）检测葡萄酒中16种真菌毒素的分析方法。方法葡萄酒样品经乙腈提取后盐析,上清液用纯水1 ∶ 1稀释后过0.22 μm聚四氟乙烯针式滤膜,加入稳定同位素内标降低基质效应。采用ACQUITY UPLCBETH C18色谱柱（100 mm×2.1 mm,1.7 μm）分离,多反应监测模式（multiple reaction monitoring,MRM）下电喷雾离子源正、负离子模式,内标法定量。结果 16种真菌毒素在一定浓度范围内具有良好的线性关系,相关系数（r）均>0.999,方法检出限（limit of detection,LOD）在0.1~50.0 μg/kg之间,方法定量限（limit of quantification,LOQ）为0.3~165.0 μg/kg。在3个浓度的加标水平下,平均回收率为在62.7%~117.5%之间,相对标准偏差为2.0%~9.5%（n=6）。结论该方法操作简单、高效、成本低,具有较好的准确度和精密度,可用于大批量葡萄酒中16种真菌毒素的快速、准确定量检测。

关键词: 真菌毒素, 同位素稀释, 葡萄酒, 超高效液相色谱-串联质谱

Abstract: Objective To establish a method for the simultaneous determination of 16 kinds of mycotoxins in wine by isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）. Methods The wine samples were extracted by acetonitrile,the extract was salted out and dehydration. The supernatant diluted with water 1 ∶ 1 and then filtrated with 0.22 μm polytetrafluoroethylene millipore filter,the matrix effects were efficiently reduced with the stable isotope. The target substances were separated on ACQUITY UPLCBETH C18（100 mm×2.1 mm,1.7 μm）. The multiple reaction monitoring（MRM） mode was used for detection in electrospray ionization with positive and negative ion modes,and quantified by internal standard method. Results There was a good linearity for 16 kinds of mycotoxins within a certain concentration range,the correlation coefficients（r） were greater than 0.999,the limits of detection（LOD） and limit of quantification（LOQ） were 0.1-50.0 μg/kg and 0.3-165.0 μg/kg,respetively. The average recoveries were 62.7%-117.5% at three different levels,and the relative standard deviations were 2.0%-9.5%（n=6）. Conclusion The method is simple,efficient,low-consumption,and has good accuracy and precision,which is uitable for the rapid and accurate quantitative detection of 16 mycotoxins in large quantities of wine.

Key words: Mycotoxins, Isotope dilution, Wine, Ultra-performance liquid chromatogr-aphy tandem mass spectrometry

中图分类号:

R115

杨雪丽, 王猛, 李菁, 李方. 葡萄酒中16种真菌毒素的同位素稀释-超高效液相色谱-串联质谱同时测定法. [J]职业与健康, 2023, 39(11): 1447-1454.

YANG Xue-li, WANG Meng, LI Jing, LI Fang. Simultaneous determination of 16 kinds of mycotoxins in wine by isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry. [J]OCCUPATION AND HEALTH, 2023, 39(11): 1447-1454.

参考文献

[1] 宋月,杨丽君,吴平谷,等.复合柱净化-超高效液相色谱-串联质谱法测定粮油中黄曲霉毒素和玉米赤霉烯酮类毒素[J].中国食品卫生杂志,2020,32(1):45-50.
[2] 陈鑫璐,邱月,张建友,等.国内外谷物中多种真菌毒素限量和同步检测标准及方法研究进展[J].中国粮油学报,2021,36(12):194-202.
[3] ZHANG B,CHEN X,HAN S Y,et al.Simultaneous analysis of 20 mycotoxins in grapes and wines from hexi corridor region (china):Based on a quechers-uhplc-ms/ms method[J].Molecules,2018,23(8):1926-1942.
[4] KOS J,HAJNAL E J,MALACHOVA A,et al.Mycotoxins in maize harvested in Republic of Serbia in the period 2012-2015.Part 1:Regulated mycotoxins and its derivatives[J].Food Chemistry,2020,312:126034.
[5] 许娇娇,黄百芬,周健,等.直接稀释-超高效液相色谱-串联质谱法快速测定谷物及其制品中16种真菌毒素[J].中国食品卫生杂志,2017,29(6):708-715.
[6] AL-TAHER F,BANASZEWSKI K,JACKSON L,et al.Rapid method for the determination of multiple mycotoxins in wines and beers by lc-ms/ms using a stable isotope dilution assay[J].J Agric Food Chem,2012,61(10):2378-2384.
[7] ADEBO O A,MOLELEKOA,MAKHUVELE R,et al.A review on novel non-thermal food processing techniques for mycotoxin reduction[J].Int J Food Sci Technol,2020,56(1):13-27.
[8] 王刘庆,焦健,王蒙.葡萄及其制品中赭曲霉毒素A的污染与控制研究进展[J].食品安全质量检测学报,2022,13(2):612-619.
[9] 张秋,范光森,李秀婷.我国白酒质量安全现状浅析[J].中国酿造,2016,35(11):15-20.
[10] 王琳,刘睿,孙德鹏,等.葡萄酒酿造过程中风险因素的分析研究[J].酿酒科技,2020(8):122-129.
[11] 柳采秀,陈丽萍,吴长兴,等.葡萄质量安全隐患与生产对策[J].浙江农业科学,2019,60(11):2089-2093.
[12] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.食品安全国家标准食品中真菌毒素限量:GB 2761—2017[S].北京:中国标准出版社,2017.
[13] GORYACHEVA I Y,SAEGER S D,EREMIN S A,et al.Immunochemical methods for rapid mycotoxin detection:Evolution from single to multiple analyte screening:A review[J].Food Addit Contam,2007,24(10):1169-1183.
[14] DIPANWITA B,RAJIB B,TARUN K D.A novel signal amplification technology for ELISA based on catalyzed reporter deposition.Demonstration of its applicability for measuring aflatoxin B₁[J].J Immunol Methods,1999,230(1/2):71-86.
[15] 张思思,陆继伟,王少敏,等.国内外真菌毒素检测方法研究现状及进展[J].食品安全质量检测学报,2016,7(7):2575-2586.
[16] REN Y,ZHANG Y,SHAO S,et al.Simultaneous determination of multi-component mycotoxin contaminants in foods and feeds by ultra-performance liquid chromatography tandem mass spectrometry[J].J Chromatogr A,2007,1143(1/2):48-64.
[17] JIN P G,HAN Z,CAI Z X,et al.Simultaneous determination of 10 mycotoxins in grain by ultra-high-performance liquid chromatography-tandem mass spectrometry using c-deoxynivalenol as internal standard[J].Food Addit Contam,2010,27(12):1701-1713.
[18] ELARIDI J,YAMANI O,MATARI A A,et al.Determination of Ochratoxin A(OTA),Ochratoxin B(OTB),T-2,and HT-2 Toxins in Wheat Grains,Wheat Flour,and Bread in Lebanon by LC-MS/MS[J].Toxins,2019,11(8):1-12.
[19] PEREIRA V L,FERNANDES J O,CUNHA S C.Comparative assessment of three cleanup procedures after QuEChERS extraction for determination of trichothecenes(type A and type B)in processed cereal-based baby foods by GC-MS[J].Food Chemistry,2015,182(1):143-149.
[20] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.食品安全国家标准食品中赭曲霉毒素A的测定:GB 5009.96—2016[S].北京:中国标准出版社,2016.
[21] 何卓霖,穆蕾,王涛,等.超高效液相色谱-四极杆/静电场轨道阱高分辨质谱快速测定乳及乳制品中21种真菌毒素[J].食品工业科技,2022,43(6):302-310.
[22] 谢慧英,袁娅,于晖,等.同位素稀释-超高效液相色谱-串联质谱法同时测定婴幼儿谷类辅助食品中的15种真菌毒素和6种农药残留[J].食品安全质量检测学报,2021,12(13):5306-5313.
[23] RASMUSSEN R R,STORM I,RASMUSSEN P H,et al.Multi-mycotoxin analysis of maize silage by LC-MS/MS[J].Anal Bioanal Chem,2010,397(2):765-776.
[24] FRENICH A G,ROBERTO R G,MARIA L G P,et al.Multi-mycotoxin analysis in eggs using a QuEChERS-based extraction procedure and ultra-high-pressure liquid chromatography coupled to triple quadrupole mass spectrometry[J].J Chromatogr A,2011,1218(28):4349-4356.
[25] YELKO R C,HOUDA B,GUILLERMINA F,et al.Multi-mycotoxin analysis in wheat semolina using an acetonitrile-based extraction procedure and gas chromatography-tandem mass spectrometry[J].J Chromatogr A,2012,1270(1):28-40.
[26] 韩深,刘萤,王珮玥,等.超高效液相色谱串联四极杆质谱法快速测定葡萄酒中8种真菌毒素[J].环境化学,2013,32(7):1417-1421.
[27] JONATAN V D,MARIA D G P,GRACA P N,et al.Simultaneous determination of pesticides and mycotoxins in wine by direct injection and liquid chromatography-tandem mass spectrometry analysis[J].Food Chemistry,2019,293(2019):83-91.
[28] 栾玉静,黄健,王瑞花,等.同位素内标-高效液相色谱-串联质谱法检测常见食品样品中15种真菌毒素[J].食品安全质量检测学报,2021,12(10):3970-3983.
[29] TREBSTEIN A,LAUBER U,HUMPF H U.Analysis of fusarium toxins via hplc-ms/ms multimethods:Matrix effects and strategies for compensation[J].Mycotoxin Res,2009,25(4):201-213.
[30] CUNHA S C,FERNANDES J O.Development and validation of a method based on a quechers procedure and heart-cutting gc-ms for determination of five mycotoxins in cereal products[J].J Sep Sci,2015,33(4-5):600-609.
[31] 郑翠梅,张艳,王松雪,等.液相色谱-飞行时间质谱同时测定粮食中13种真菌毒素[J].分析测试学报,2012,31(4):383-389.
[32] MICHELANGELO A,LEHOTAY S J,STAJNBAHER D,et al.Fast and easy multiresidue method employing acetonitrile extraction/partitioning and "dispersive solid-phase extraction" for the determination of pesticide residues in produce[J].J AOAC Int,2003,86(2):412-431.