基于UPLC-MS-MS技术定量确证小麦中10种真菌毒素
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摘要
建立了小麦中黄曲霉毒素B_1、黄曲霉毒素B_2、黄曲霉毒素G_1、黄曲霉毒素G_2、脱氧雪腐镰刀菌烯醇、雪腐镰刀菌烯醇、3-乙酰脱氧雪腐镰刀菌烯醇、脱氧雪腐镰刀菌烯醇-3-葡萄糖苷、15-乙酰脱氧雪腐镰刀菌烯醇和玉米赤霉烯酮等10种真菌毒素的超高液相色谱-三重四级杆串联质谱法检测的方法。小麦样品经乙腈-水-乙酸溶液震荡超声提取后,采用多功能净化柱净化。待测物经Kinetex SB-C_(18)色谱柱分离,甲醇-0.1%甲酸为流动相,梯度洗脱,正负模式同时扫描,三重串联四级杆质谱多反应离子监测(MRM)方式检测,基质标准校正,外标法定量。结果表明,在一定的浓度范围内,10种真菌毒素线性关系良好,相关系数均大于0.999,方法的检出限为0.015~1μg/kg,空白样品中添加高中低不同浓度的真菌毒素的回收率为65.12%~89.16%,RSD%(n=6)在4.26%~9.46%之间。该方法分析速度快、重复性好、灵敏度高,适合小麦中多种真菌毒素的高灵敏度快速测定。
A method for the quantitative determination of 10 mycotoxins, such as aflatoxin B_1, aflatoxin B_2, aflatoxin G_1,aflatoxin G_2, deoxynivalenol, 3-deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol and zearalenone in wheat by ultra-high liquid chromatography triple quadrupole tandem mass spectrometry was developed. Wheat samples were extracted by ultrasonic with acetonitrile-water-acetic acid solution, and purified by multi-functional purification column. Chromatographic analysis was carried out in a Kinetex SB-C_(18) column, the binary mobile phase consisted of methanol and 0.1% formic acid, and the system was run with a gradient program. Mass spectrometric detection was carried out on Xevo TQ mass spectrometer with negative and positive ionization mode. Quantification analysis was performed in multiple reaction monitoring(MRM), matrix standard calibration, and external standard method quantitative. The results demonstrated that the 10 mycotoxins had good linear relationship and the correlation coefficients were more than 0.999. The detection limit(LOD) of the method was 0.015-1 μg/kg, the average recoveries in the blank matrix ranged from 65.12% to 89.16% with relative standard deviation(RSD, n=6) of 4.26%-9.46%. The method in this study was fast analysis, good reproducibility and high sensitivity, it is suitable for rapid determination of various fungal toxins in wheat.
A method for the quantitative determination of 10 mycotoxins, such as aflatoxin B_1, aflatoxin B_2, aflatoxin G_1,aflatoxin G_2, deoxynivalenol, 3-deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol and zearalenone in wheat by ultra-high liquid chromatography triple quadrupole tandem mass spectrometry was developed. Wheat samples were extracted by ultrasonic with acetonitrile-water-acetic acid solution, and purified by multi-functional purification column. Chromatographic analysis was carried out in a Kinetex SB-C_(18) column, the binary mobile phase consisted of methanol and 0.1% formic acid, and the system was run with a gradient program. Mass spectrometric detection was carried out on Xevo TQ mass spectrometer with negative and positive ionization mode. Quantification analysis was performed in multiple reaction monitoring(MRM), matrix standard calibration, and external standard method quantitative. The results demonstrated that the 10 mycotoxins had good linear relationship and the correlation coefficients were more than 0.999. The detection limit(LOD) of the method was 0.015-1 μg/kg, the average recoveries in the blank matrix ranged from 65.12% to 89.16% with relative standard deviation(RSD, n=6) of 4.26%-9.46%. The method in this study was fast analysis, good reproducibility and high sensitivity, it is suitable for rapid determination of various fungal toxins in wheat.
引文
[1]Council for Agricultural Science and Technology(CAST)[S].Mycotoxins:Risks in plant,animal and human systems.Task force report,No.139,CAST,Ames,Iowa,2003.
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[5]陈慧菲,朱天仪,陈凤香,等.QuEChERS-超高效液相色谱串联质谱法测定谷物中的8种真菌毒素[J].粮食与油脂,2016(05):67-70.
[6]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(06):206-210.
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[8]李可,梁肇海,曾胜波,等.免疫亲和柱层析-超高效液相色谱法测定动物性食品中6种黄曲霉毒素[J].中国食品卫生杂志,2016(03):310-313.
[9]胡晓科,孙丹红,文君.柱前衍生-高效液相色谱法检测食品中的黄曲霉毒素B1、B2、G1、G2[J].食品工业科技,2015(12):49-52,56.
[10]王叶群,杨增玲,张绍英,等.用于污染黄曲霉毒素花生分选的荧光信号研究[J].农业工程学报,2016(01):187-192.
[11]涂春蓉,董晓娟,马良,等.分散液液微萃取/荧光分光光度法快速测定乳制品中的黄曲霉毒素M1[J].分析测试学报,2014(09):1062-1067.
[12]管笛,潘灿平,王文,等.磁微粒酶联免疫吸附法测定玉米中的伏马毒素B1[J].食品科学,2014(08):208-211.
[13]熊勇华,陈雪岚,许杨.检测赭曲霉毒素A(OTA)的酶联免疫吸附法(ELISA)体系的建立[J].食品科学,2006(05):30-35.
[14]YIBADATIHAN S,JINAP S,MAHYUDIN NA.Simultaneous determination of multi-mycotoxins in palm kernel cake(PKC)using liquid chromatography/tandem mass spectrometry(LC-MS/MS)[J].Food Addit Contam,2014,31(12):2071-2079.
[15]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(06):970-978.
[16]朱建国.粮油中真菌毒素和农药残留多组分检测技术研究[D].北京:中国农业科学院,2016.
[17]侯建波,谢文,李杰,等.液相色谱-串联质谱法测定牛奶中11种真菌毒素残留量[J].理化检验(化学分册),2015(05):600-604.
[18]应永飞,朱聪英,韦敏珏,等.液相色谱-串联质谱法测定饲料中14种霉菌毒素及其类似物[J].分析化学,2010(12):1759-1764.
[19]朱聪英,应永飞,韦敏珏,等.液相色谱-串联质谱法测定饲料中黄曲霉毒素的研究[J].质谱学报,2010(04):240-246.
[20]孙娟,李为喜,张妍,等.用超高效液相色谱串联质谱法同时测定谷物中12种真菌毒素[J].作物学报,2014(04):691-701.
[21]宫小明,任一平,董静,等.超高效液相色谱串联质谱法测定花生、粮油中18种真菌毒素[J].分析测试学报,2011(01):6-12.
[22]陈勇,陈重均,李劲,等.超高效液相色谱串联质谱法检测三七药材中10种真菌毒素[J].药学学报,2015(01):81-85.
[23]SULYOK M,FRANZ B,RUDOLF K,et al.Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize[J].Rapid Commun Mass Spectrom,2006,20(18):2646-2659.
[24]Commission Decision of 12 August 2002 Implementing Council Diretive 96/23/EC,Concerning the Performance of Analytical Methods and the Interpretation of Resuits(2002/657/EC)[R].2002-8-12.
[25]勾新磊,赵新颖,高峡,等.超高效液相色谱-串联质谱法测定保健食品中10种磷酸二酯酶-5抑制剂[J].质谱学报,2016(06):554-561.
[2]MOSS M O.Risk assessment for aflatoxins in foodstuffs[J].International Biodeterioration and Iodegradation,2002(50):137-142.
[3]柳其芳.酶联免疫吸附法和薄层色谱法联合分析黄曲霉毒素B1的研究[J].中国热带医学,2006(02):246-248.
[4]张鹏,赵卫东,张艺兵.高效薄层色谱法测定黄曲霉毒素B1、B2、G1、G2[J].分析化学,2000(03):392.
[5]陈慧菲,朱天仪,陈凤香,等.QuEChERS-超高效液相色谱串联质谱法测定谷物中的8种真菌毒素[J].粮食与油脂,2016(05):67-70.
[6]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(06):206-210.
[7]谢刚,王松雪,张艳.超高效液相色谱法快速检测粮食中黄曲霉毒素的含量[J].分析化学,2013(02):223-228.
[8]李可,梁肇海,曾胜波,等.免疫亲和柱层析-超高效液相色谱法测定动物性食品中6种黄曲霉毒素[J].中国食品卫生杂志,2016(03):310-313.
[9]胡晓科,孙丹红,文君.柱前衍生-高效液相色谱法检测食品中的黄曲霉毒素B1、B2、G1、G2[J].食品工业科技,2015(12):49-52,56.
[10]王叶群,杨增玲,张绍英,等.用于污染黄曲霉毒素花生分选的荧光信号研究[J].农业工程学报,2016(01):187-192.
[11]涂春蓉,董晓娟,马良,等.分散液液微萃取/荧光分光光度法快速测定乳制品中的黄曲霉毒素M1[J].分析测试学报,2014(09):1062-1067.
[12]管笛,潘灿平,王文,等.磁微粒酶联免疫吸附法测定玉米中的伏马毒素B1[J].食品科学,2014(08):208-211.
[13]熊勇华,陈雪岚,许杨.检测赭曲霉毒素A(OTA)的酶联免疫吸附法(ELISA)体系的建立[J].食品科学,2006(05):30-35.
[14]YIBADATIHAN S,JINAP S,MAHYUDIN NA.Simultaneous determination of multi-mycotoxins in palm kernel cake(PKC)using liquid chromatography/tandem mass spectrometry(LC-MS/MS)[J].Food Addit Contam,2014,31(12):2071-2079.
[15]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(06):970-978.
[16]朱建国.粮油中真菌毒素和农药残留多组分检测技术研究[D].北京:中国农业科学院,2016.
[17]侯建波,谢文,李杰,等.液相色谱-串联质谱法测定牛奶中11种真菌毒素残留量[J].理化检验(化学分册),2015(05):600-604.
[18]应永飞,朱聪英,韦敏珏,等.液相色谱-串联质谱法测定饲料中14种霉菌毒素及其类似物[J].分析化学,2010(12):1759-1764.
[19]朱聪英,应永飞,韦敏珏,等.液相色谱-串联质谱法测定饲料中黄曲霉毒素的研究[J].质谱学报,2010(04):240-246.
[20]孙娟,李为喜,张妍,等.用超高效液相色谱串联质谱法同时测定谷物中12种真菌毒素[J].作物学报,2014(04):691-701.
[21]宫小明,任一平,董静,等.超高效液相色谱串联质谱法测定花生、粮油中18种真菌毒素[J].分析测试学报,2011(01):6-12.
[22]陈勇,陈重均,李劲,等.超高效液相色谱串联质谱法检测三七药材中10种真菌毒素[J].药学学报,2015(01):81-85.
[23]SULYOK M,FRANZ B,RUDOLF K,et al.Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize[J].Rapid Commun Mass Spectrom,2006,20(18):2646-2659.
[24]Commission Decision of 12 August 2002 Implementing Council Diretive 96/23/EC,Concerning the Performance of Analytical Methods and the Interpretation of Resuits(2002/657/EC)[R].2002-8-12.
[25]勾新磊,赵新颖,高峡,等.超高效液相色谱-串联质谱法测定保健食品中10种磷酸二酯酶-5抑制剂[J].质谱学报,2016(06):554-561.