Multiple mycotoxins analysis in animal feed with LC‐MS/MS: Comparison of extract dilution and immunoaffinity clean‐up

Abstract

The aim of this study was a performance comparison of two clean‐up procedures (dilutions versus immunoaffinity columns) in the simultaneous determination of eight mycotoxins (aflatoxin B1, deoxynivalenol, fumonisin B1 & B2, ochratoxin A, toxin T‐2 & HT‐2 and zearalenone) in the animal feed. After extraction the analytes were separated on a Kinetex Biphenyl column with a gradient elution using methanol/0.01 M ammonium acetate as a mobile phase and analyzed with the LC‐MS/MS technique. Both of the procedures were validated by analysis of a series of spiked feed samples (n = 6) at three different concentration levels. Better signal to noise ratios were observed for immunoaffinity clean‐up. The recoveries of analyses were in the range 88–110% for the dilution procedure and 78–120% for the immunoaffinity clean‐up. The dilution procedure was more precise (coefficient of variation of the within‐laboratory reproducibility for it was 7.8–22.4% in comparison to 12–35.5% for the immunoaffinity clean‐up. The results show that both procedures fulfilled the requirements for mycotoxin analysis and can be used successfully in multi‐analyte determination. Although the dilution procedure shows better precision and trueness, the immunoaffinity clean‐up procedure can have advantages in more complex feed samples thanks to lower matrix effect and limits of detections.

Simultaneous Determination of Deoxynivalenol, Its Modified Forms, Nivalenol and Fusarenone-X in Feedstuffs by the Liquid Chromatography–Tandem Mass Spectrometry Method

“…The suitability for extraction and clean-up of DONTEST IAC, OASIS HLB and Mycosep 225 was tested by using them according to the manufacturer’s instruction. The QuEChERS technique and SLE were prepared based on our previous experience [ 41 , 42 ]. As is shown in Figure 4 the best results were obtained for Mycosep 225 columns which are dedicated products for Trichothecene analysis.…”

“…In addition to IACs, several other techniques of sample preparation have been tested: SLE, QuEChERS, SPE with OASIS HLB cartridges and Mycosep 225 Trich column (contain a mixture of adsorbent materials). The protocol for sample preparation for SLE and QuEChERS was based on our previously described methods [ 41 , 42 ].…”

Combined biocompatible medium with molecularly imprinted polymers for determination of aflatoxins B1 in real sample

“…Four milliliters (10 μg/mL) AFB1 solution (prepared in methanol-water) was added into the tubes. The mixture was vigorously shaken at 2500 rpm for a series of time (1,2,3,6,9,12,15,20,25,30,35,40, and 60 min). The tubes were centrifuged and the supernatants were measured by HPLC.…”

“…Mycotoxins are notoriously toxic secondary metabolites produced by a variety of fungal species. The most disturbing mycotoxins include aflatoxins (AFs), ochratoxins (OTs), and zearalenone (ZEN), which were classified as carcinogenic substance by the International Agency of Research on Cancer (IARC) [1][2][3]. These mycotoxins often contaminate crops and their by-products during preparing, processing, and storing columns (IACs) and other commercialized SPE columns were preferentially selected as the purification columns of the mycotoxins [11].…”

Fast and efficient immunoaffinity column cleanup and liquid chromatography–tandem mass spectrometry method for the quantitative analysis of aflatoxins in baby food and feeds

“…LC‐MS/MS has been reported and considered as the universal approach for mycotoxin screening due to their high selectivity for analyzing several analyte in one run [17–22]. Recently, a few multi‐mycotoxin analysis methods have been described in varied food commodities, however the validated methods for baby food analysis are rare as high sensitivity is required to satisfy the strict regulation in such types of matrices [7,23].…”