多功能柱净化-超高效液相色谱串联质谱法同时测定水产品中11种真菌毒素

Simultaneous Detection of 11 Mycotoxins in Aquatic Products using Multifunctional Column Clean-up and Ultrahigh-performance Liquid Chromatography-tandem Mass Spectrometry

摘要:

建立了同步测定水产品中黄曲霉毒素B1（AFB1）、黄曲霉毒素B2（AFB2）、黄曲霉毒素G1（AFG1）、黄曲霉毒素G2（AFG2）、黄曲霉毒素M1（AFM1）、赭曲霉毒素A (OTA)、玉米赤霉烯酮（ZEN）、脱氧雪腐镰刀菌烯醇（DON）、3-乙酰基脱氧雪腐镰刀菌烯醇（3-AcDON）、15-乙酰基脱氧雪腐镰刀菌烯醇（15-AcDON）、镰刀菌烯酮（Fus-X）等11种真菌毒素的超高效液相色谱-串联质谱分析方法。试样样品以乙腈/水溶液（84+16）提取，MycoSpin 400多功能柱净化，Waters ACQUITY UPLCTM BEH C18柱分离，多反应监测(MRM)模式进行定量与定性分析，外标法定量。结果表明，11种目标物在各自的线性响应范围内线性关系良好，相关系数(r)≥0.9918，定量限（LOQ）为0.1~20.0 μg/kg，加标平均回收率（n=3）为62.8%~115.3%，相对标准偏差（RSD）为2.6%~19.0%。该方法前处理简单、净化效果好、灵敏度高、重现性好，适合于水产品中痕量真菌毒素的多残留快速检测。

Abstract:

A novel method to simultaneously detect 11 kinds of mycotoxins (aflatoxin B1 [AFB1], aflatoxin B2 [AFB2], aflatoxin G1 [AFG1], aflatoxin G2 [AFG2], aflatoxin M1 [AFM1], ochratoxin A [OTA], zearalenone [ZEN], deoxynivalenol [DON], 3-acetyldeoxynivalenol [3-AcDON], 15-AcDON, and fusarenon-X [Fus-X]) in aquatic products was developed, using ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) with multiple reaction monitoring (MRM). The samples were extracted with acetonitrile-water (84:16, V/V), purified using a multifunctional clean-up column (MycoSpin 400), and separated by a chromatographic column (Waters Acquity UPLCTM BEH C18; 100 × 2.1 mm i.d., 1.7 μm) using (0.01% formic acid-0.05% ammonia-H2O)/methanol as mobile phase. The quantitative and qualitative analyses were conducted in MRM mode with a mass spectrometer and an external standard. The results indicated that 11 mycotoxins showed good linear relationship within their respective linear response ranges, the correlation coefficients were > 0.9918, the limit of quantitation (LOQ) was 0.1 to 20.0 μg/kg, and the average recoveries (n = 3) were from 62.8% to 115.3%, with values of relative standard deviation (RSD) ranging between 2.6% and 19.0%. The developed method with the facile pretreatment, good purification effect, high sensitivity and reproducibility can be used for rapid detection of trace levels of multiple mycotoxins in aquatic products.