超高效液相色谱-质谱联用法检测青贮玉米中9种真菌毒素
|
摘要
为了建立超高效液相色谱-串联质谱检测青贮玉米中黄曲霉毒素B_1(AFB_1)、B_2(AFB_2)、G1(AFG1)、G2(AFG2)、T-2毒素、HT-2毒素、脱氧雪腐镰刀烯醇(DON)、玉米赤霉烯酮(ZEN)、赭曲霉毒素A(OTA)的分析方法,样品采用乙腈提取,C18为分散净化剂的QuEChERS净化柱净化,超高效液相色谱分离,多反应离子监测模式检测,外标法定量。结果显示:青贮玉米中9种真菌毒素的线性关系良好,9种毒素的平均添加回收率为71.86%~110.00%之间,相对标准偏差为0.09%~8.94%。方法检出限(LOD)在0.05~7.5μg/kg之间,定量限(LOQ)在0.18~25μg/kg之间。该方法快速灵敏,准确可靠,适用于青贮玉米中9种真菌毒素的检测。
A method was established to determine aflatoxin B_1(AFB_1),aflatoxin B_2(AFB_2),aflatoxin G1(AFG1),aflatoxin G2(AFG2),T-2 toxin,HT-2 toxin,Deoxynivalenol(DON),Zearalmone(ZEN)and ochratoxins A(OTA)in corn silage by UPLC-MS/MS.Homogenized samples were extracted with acetonitrile,cleaned up by QuEChERS cartridge with C18 as sorbent for cleaning up.Chromatographic separation was achieved by UPLC and the determination was performed by tandem mass spectrometry under the mode of multiple reaction monitoring,the quantification analysis of external standard calibration curves was used.There were good linear correlations between the concentrations of the nine mycotoxins in corn silage.The recoveries of 9 kinds of toxins were in range of 71.86%—110.00% with the relative standard deviations(RSDs)of 0.09%—8.94%.The detection limit(LOD)and the limit of quantification(LOQ)were in range of 0.05—7.5μg/kg and 0.18~25μg/L,respectively.This method is rapid,sensitive,accurate and reliable,and it is suitable for the detection of 9 kinds of toxins in corn silage.
A method was established to determine aflatoxin B_1(AFB_1),aflatoxin B_2(AFB_2),aflatoxin G1(AFG1),aflatoxin G2(AFG2),T-2 toxin,HT-2 toxin,Deoxynivalenol(DON),Zearalmone(ZEN)and ochratoxins A(OTA)in corn silage by UPLC-MS/MS.Homogenized samples were extracted with acetonitrile,cleaned up by QuEChERS cartridge with C18 as sorbent for cleaning up.Chromatographic separation was achieved by UPLC and the determination was performed by tandem mass spectrometry under the mode of multiple reaction monitoring,the quantification analysis of external standard calibration curves was used.There were good linear correlations between the concentrations of the nine mycotoxins in corn silage.The recoveries of 9 kinds of toxins were in range of 71.86%—110.00% with the relative standard deviations(RSDs)of 0.09%—8.94%.The detection limit(LOD)and the limit of quantification(LOQ)were in range of 0.05—7.5μg/kg and 0.18~25μg/L,respectively.This method is rapid,sensitive,accurate and reliable,and it is suitable for the detection of 9 kinds of toxins in corn silage.
引文
[1]Ferraretto L F,Shaver R D,Luck B D.Silage review:Recent advances and future technologies for wholeplant and fractionated corn silage harvesting[J].Journal of Dairy Science,2018,101(5):3937-3951.
[2]李剑南,杨武,冯进华,等.玉米类型和生育时期对其青贮饲料品质的影响[J].河北农业大学学报,2015,38(5):17-2167.
[3]Garon D,Richard E,Sage L,et al.Mycoflora and multimycotoxin detection in corn silage:experimental study[J].Journal of Agricultural&Food Chemistry,2006,54(9):3479-3484.
[4]田吉鹏.全株玉米青贮饲料中黄曲霉毒素积累规律及调控技术研究[D].北京:中国农业大学,2017.
[5]Shephard G S,Berthiller,Burdaspal,et al.Developments in mycotoxin analysis:An update for 2010-2011[J].World Mycotoxin Journal,2012,3(1):3-23.
[6]Dagnac T,Latorre A,Fern n L B,et al.Validation and application of a liquid chromatography-tandem mass spectrometry based method for the assessment of the co-occurrence of mycotoxins in maize silages from dairy farms in NW Spain[J].Food Addit Contam Part A Chem Anal Control Expo Risk Assess,2016,33(12):1850-1863.
[7]王莎,孔维军,杨美华.同位素内标—超高效液相色谱串联质谱法检测麦芽中11种真菌毒素[J].药学学报,2016,51(1):110-115.
[8]张晓男,聂继云,闫震,等.UPLC-MS/MS同时检测苹果及其制品中的7种真菌毒素[J].分析测试学报,2017,36(5):588-594.
[9]Commission Regulation(EU)No 519/2014,Amending Regulation(EC)No 401/2006as regards methods of sam-pling of large lots,spices and food supplements,perform-ance criteria for T-2,HT-2toxin and citrinin and screening methods of analysis[R].OJ,2016:337-365.
[10]GB 13078-2017饲料卫生标准[S].中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会,2017.
[11]中华人民共和国农业部.NY/T 2071-2011饲料中黄曲霉毒素、玉米赤霉烯酮和T-2毒素的测定液相色谱串联质谱法[S].北京:全国饲料工业标准化技术委员会,2011.
[12]魏云计,冯民,朱臻怡,等.高效液相色谱—串联质谱法同时测定饲料中11种霉菌毒素[J].色谱,2017,35(8):891-896.
[2]李剑南,杨武,冯进华,等.玉米类型和生育时期对其青贮饲料品质的影响[J].河北农业大学学报,2015,38(5):17-2167.
[3]Garon D,Richard E,Sage L,et al.Mycoflora and multimycotoxin detection in corn silage:experimental study[J].Journal of Agricultural&Food Chemistry,2006,54(9):3479-3484.
[4]田吉鹏.全株玉米青贮饲料中黄曲霉毒素积累规律及调控技术研究[D].北京:中国农业大学,2017.
[5]Shephard G S,Berthiller,Burdaspal,et al.Developments in mycotoxin analysis:An update for 2010-2011[J].World Mycotoxin Journal,2012,3(1):3-23.
[6]Dagnac T,Latorre A,Fern n L B,et al.Validation and application of a liquid chromatography-tandem mass spectrometry based method for the assessment of the co-occurrence of mycotoxins in maize silages from dairy farms in NW Spain[J].Food Addit Contam Part A Chem Anal Control Expo Risk Assess,2016,33(12):1850-1863.
[7]王莎,孔维军,杨美华.同位素内标—超高效液相色谱串联质谱法检测麦芽中11种真菌毒素[J].药学学报,2016,51(1):110-115.
[8]张晓男,聂继云,闫震,等.UPLC-MS/MS同时检测苹果及其制品中的7种真菌毒素[J].分析测试学报,2017,36(5):588-594.
[9]Commission Regulation(EU)No 519/2014,Amending Regulation(EC)No 401/2006as regards methods of sam-pling of large lots,spices and food supplements,perform-ance criteria for T-2,HT-2toxin and citrinin and screening methods of analysis[R].OJ,2016:337-365.
[10]GB 13078-2017饲料卫生标准[S].中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会,2017.
[11]中华人民共和国农业部.NY/T 2071-2011饲料中黄曲霉毒素、玉米赤霉烯酮和T-2毒素的测定液相色谱串联质谱法[S].北京:全国饲料工业标准化技术委员会,2011.
[12]魏云计,冯民,朱臻怡,等.高效液相色谱—串联质谱法同时测定饲料中11种霉菌毒素[J].色谱,2017,35(8):891-896.