Advanced multi-mycotoxin analysis with Agilent 6460c LC-MS/MS

OBJECTIVE  

This abstract presents a multi-mycotoxin sample preparation and analysis method for EU-regulated and emerging mycotoxins with Agilent 6460c UHPLC-MS/MS.

MATERIALS & METHODS  

The method is based on “dilute and shoot” principle.

Sample preparation involves:

• 1. Extraction with an organic solvent.
• 2. Centrifugation of the extracts.
• 3. To compensate the matrix effects in electrospray ionization, the extracts are mixed with isotopically labelled internal standards (9) for each group of mycotoxins.

⇰ The addition of the internal standard is performed in the needle of the autosampler, before injection into UHPLC-MS/MS.

Method validation in terms of linearity, selectivity, sensitivity, accuracy, and precision was performed in 8 complex feed matrixes (corn, compound feed, wheat, barley, soya meal, wheat bran, sunflower meal, total mixed ration) to detect 34 mycotoxins:

• ⇰ Aflatoxins B1, B2, G1, and G2
• ⇰ α – zearalenol, β – zearalenol, zearalanone, and zearalenone
• ⇰ Diacetoxyscirpenol
• ⇰ HT-2 toxin and T-2 toxin
• ⇰ Neosolaniol
• ⇰ 3-acetyl deoxynivalenol, 15- acetyl deoxynivalenol, deoxynivalenol, and deoxynivalenol-3-glucoside
• ⇰ Nivalenol
• ⇰ Fumonisins B1, B2, and B3
• ⇰ Fusaric acid
• ⇰ Moniliformin

• ⇰ Fusarenon X
• ⇰ Ochratoxin A
• ⇰ Beauvericin
• ⇰ Enniatins A, A1, B, and B1
• ⇰ Citrinin
• ⇰ Patulin
• ⇰ Ergosine
• ⇰ Ergocryptinine
• ⇰ Alternariol

The limits of quantitation (LOQs) were from 0.05 ngmL-1 to 4 ngmL-1.

The recoveries of the 34 mycotoxins spiked in the 8 feed categories were in the range of 65.5%–125.7% after internal standard correction, with relative standard deviations (RSDs) below 15.1%.

Results showed the robustness of this method for analysis of 34 mycotoxins using LC-MS/MS.

The implementation of effective monitoring programs and stringent legal regulations can significantly contribute to the reduction and carry over of these mycotoxins in food chain.

Autores