Development of an Improved Method of Sample Extraction and Quantitation of Multi-Mycotoxin in Feed by LC-MS/MS

Toxins (Basel). 2020 Jul 19;12(7):462. doi: 10.3390/toxins12070462.


A multi-mycotoxin chromatographic method was developed and validated for the simultaneous quantitation of aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), zearalenone (ZON), deoxynivalenol (DON), nivalenol (NIV), diacetoxyscirpenol (DAS), fumonisins (FB1, FB2 and FB3), T-2 toxin (T-2) and HT-2 toxin (HT-2) in feed. The three most popular sample preparation techniques for determination of mycotoxins have been evaluated, and the method with highest recoveries was selected and optimized. This modified QuEChERS (quick, easy, cheap, effective, rugged and safe) approach was based on the extraction with acetonitrile, salting-out and cleanup with lipid removal. A reconstitution process in methanol/water was used to improve the MS responses and then the extracts were analyzed by LC-MS/MS. In this method, the recovery range is 70-100% for DON, DAS, FB1, FB2, FB3, HT-2, T-2, OTA, ZON, AFG1, AFG2, AFB1 and AFB2 and 55% for NIV in the spike range of 2-80 µg/kg. Method robustness was determined with acceptable z-scores in proficiency tests and validation experiments.

Keywords: LC-MS/MS; feed; modified QuEChERS; mycotoxins.

Publication types

  • Validation Study

MeSH terms

  • Animal Feed / microbiology*
  • Chromatography, Affinity*
  • Chromatography, High Pressure Liquid*
  • Food Microbiology*
  • Fungi / metabolism*
  • Mycotoxins / analysis*
  • Reproducibility of Results
  • Solid Phase Extraction*
  • Spectrometry, Mass, Electrospray Ionization*
  • Tandem Mass Spectrometry*


  • Mycotoxins