食品中真菌毒素高分辨质谱测定与定量方法的建立

Tu Vu Ngoc, Huong Hoang Lan, Tien Bui Cao, Son Tran Cao
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摘要

高分辨率液相色谱-质谱法(LC-HRMS)是同时检测和定量三种真菌毒素Alternariol (AOH)、Alternariol Monomethyl Ete (AME)、Tenuazonic acid (TeA)的一种高灵敏度、特异性和准确性的方法。色谱条件为:柱反相C18,流动相为梯度模式,水和甲醇两通道,含0.1%甲酸和10 mM甲酸铵。采用负电子电离(ESI-)源HRMS高分辨质谱检测器。使用Oasis HLB固相萃取柱对样品进行清洗和富集。该方法AOH和AME的定量限为1.0µg/kg, TeA的定量限为1.0µg/kg,回收率为80.0 ~ 114.8 %,重复性为0.07 ~ 9.9%,符合AOAC和欧洲标准的要求。应用该方法对河内市采集的80份食品样品中的互孢菌毒素进行了检测和定量。04/20蔬菜样品(3.3 ~ 178µg/kg)、03/20水果样品(5.6 ~ 7.3µg/kg)、06/20谷物样品(13.4 ~ 17.4µg/kg)、03/20油籽样品(13.2 ~ 25.2µg/kg)中均检测到AOH;01/20蔬菜样品(7.5µg/kg)、02/20水果样品(< 3µg/kg)、01/20谷物样品(9.5µg/kg)、02/20油籽样品(< 3µg/kg)检测出AME;在02/20蔬菜样品(9.0µg/kg)、01/20谷物样品(< 3µg/kg)、02/20油籽样品(< 3µg/kg)中检测到TeA,水果样品中未检测到TeA。
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Development of method for determination and quantification of Alternaria mycotoxins in food by high resolution mass spectrometry
High-resolution liquid chromatography-mass spectrometry (LC-HRMS) is a highly sensitive, specific and accurate method for the simultaneous detection and quantification of three mycotoxins Alternaria (Alternariol (AOH), Alternariol Monomethyl Ete) (AME), Tenuazonic acid (TeA)). Chromatographic conditions include: column reversed phase C18, mobile phases in gradient mode with two channels of H2O and MeOH containing 0.1 % formic acid and 10 mM ammonium formate. HRMS high resolution mass spectrometer detector with negative electron ionization (ESI-) source is used. Samples were cleaned and enriched using an Oasis HLB solid phase extraction column. The method has a limit of quantitation (LOQ) for AOH and AME of 1.0 µg/kg and TeA of 1.0 µg/kg with recovery from 80.0 to 114.8 %, repeatability from 0.07 - 9.9 %, meeting the requirements of AOAC and European regulations. The method was applied to detect and quantify Alternaria mycotoxins in 80 food samples collected in Hanoi city. AOH was detected in 04/20 vegetable samples (3.3 - 178 µg/kg), 03/20 fruit samples (5.6 - 7.3 µg/kg), 06/20 cereal samples (13.4 - 17.4 µg/kg), 03/20 oil seed samples (13.2 - 25.2 µg/kg); detected AME in 01/20 vegetable samples (7.5 µg/kg), 02/20 fruit samples (< 3 µg/kg), 01/20 cereal samples (9.5 µg/kg), 02/20 oil seed samples (< 3 µg/kg); detected TeA in 02/20 vegetable samples (9.0 µg/kg), 01/20 cereal samples (< 3 µg/kg), 02/20 oil seed samples (< 3 µg/kg), not detected in fruit samples.
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