超高效液相色谱-串联质谱法测定燕麦粉和小米粉中白僵菌素和恩镰孢菌素

Determination of beauvericin and enniatins in oat flours and millet flours by ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  超高效液相色谱-串联质谱  白僵菌素  恩镰孢菌素  小米粉  燕麦粉

英文关键词:ultra performance liquid chromatography-tandem mass spectrometry  beauvericin  ennitatin  millet flour  oat flour

中文摘要:

      目的 建立一种超高效液相色谱-串联质谱法测定谷物性食品燕麦粉和小米粉中白僵菌素（beauvericin，BEA）、恩镰孢菌素A（enniatin A，ENA）、恩镰孢菌素A1（enniatin A1，ENA1）、恩镰孢菌素B（enniatin B，ENB）、恩镰孢菌素B1（enniatin B1，ENB1）残留的分析方法。方法 样品采用乙腈-水-甲酸（84:15:1，V/V/V）提取、经过Oasis Prime HLB固相萃取柱净化后，Waters BEH C18色谱柱(2.1 mm×100 mm, 1.8 μm)分离，在正离子模式下，以5 mmol/L乙酸铵水溶液-乙腈为流动相进行梯度洗脱，在多反应监测模式下进行定性定量检测分析，基质匹配标准曲线外标法定量。结果 BEA和4种恩镰孢菌素在各自的线性范围内具有良好的线性关系(r2>0.999)，方法检出限为0.02~0.05 μg/kg，定量限为0.05~0.15 μg/kg。按线性范围的最低浓度、中浓度和最高浓度3个水平进行加标回收实验，燕麦粉和小米粉的平均回收率分别是83.6%~105.2%和88.5%~104.2%，相对标准偏差分别为1.3%~8.2%和1.3%~3.2% (n=6)。对北京市采集的10份燕麦粉和10份小米粉样品进行检测，5种化合物均有不同程度检出，其中，ENB和ENB1的检出率为100%。结论 本方法简单快速、准确性好、灵敏度高，可以实现对燕麦粉和小面粉中白僵菌素和恩镰孢菌素进行准确的定性定量。

英文摘要:

      Objective To develop a method for the determination of beauvercin (BEA), enniatin A (ENNA), enniatin A1 (ENNA1), enniatin B (ENNB) and enniatin B1 (ENNB1) in oat flours and millet flours by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The samples were extracted with acetonitrile-water-formic acid, cleaned-up by an Oasis Prime HLB solid phase extraction column, and separated on a Waters BEH C18 column (2.1 mm×100 mm, 1.8 μm) in positive ionization mode with a gradient elution using a 5 mmol/L aqueous ammonium acetate-acetonitrile as the mobile phases, and then analyzed by multiplexed reaction monitoring (MRM) with UPLC-QqQ-MS/MS. The qualitative and quantitative analysis was carried out in UPLC-QqQ-MS/MS multi-reaction monitoring mode, and quantified by matrix-matched standard curve external standard method. Results The results showed good linearity (r2>0.999) in the respective linear ranges of BEA and four kinds of enniatins, with the limits of detection of 0.02-0.05 μg/kg and the limits of quantification of 0.05-0.15 μg/kg. The accuracy of the method was evaluated by conducting a recovery test at three spiked levels: the lowest, middle and highest concentrations of the linear ranges. The average recoveries for oat flours and millet flours were 83.6%~105.2% and 88.5%~104.2%, respectively, with the relative standard deviations of 1.3%~8.2% and 1.3%~3.2% (n=6), and the matrix effects were 78.1%~98.3% and 80.3%~100.1%, respectively. The five compounds were detected in different degrees in 10 samples of oat flour and 10 samples of millet flour collected in Beijing, among which, the detection rates of ENB and ENB1 were 100%. Conclusion The method is quick, easy, accurate and sensitive, which is suitable for the accurate qualitative and quantitative determination of beauvericin and enniatins in oat flour and wheat flour.