Abstract
Cereals can be contaminated by several mycotoxins, whose co-presence may represent an undervalued risk for humans and animals. Maize and wheat are the most contaminated cereals and in temperate areas could be affected in field conditions by several Fusarium and Aspergillus infections. To date, only B-fumonisins (FBs), aflatoxins (AFs), zearalenone (ZEA), deoxynivalenol (DON) and T-2 and HT-2 toxins have been regulated in cereals in European Union. The other fungal metabolites, are commonly referred to as “emerging” and “masked” mycotoxins, and more information on their occurrence in combination with the regulated mycotoxins, are needed to design combined toxicological and exposure assessments.This research intends to develop and compare two multiresidue HPLC-ESI-TQ-MS/MS methods for the simultaneous determination of the main regulated, emerging and masked mycotoxins in maize and wheat, among which: FB1, FB2, DON, ZEA, AFB1, AFB2, AFG1, AFG2, moniliformin (MON), deoxynivalenol-3-glucoside (DON-3-G), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV), enniatins A, A1, B, B1 (ENNA, ENNA1, ENNB, ENNB1). The extraction was performed for both methods using a mixture of CH3CN/H2O/CH3COOH (79/20/1, v/v/v), while the dilution/purification was carried out through two different procedures: (1) by the “dilute-and-shoot” technique diluting 1:2 the filtered extract with CH3CN/H2O/CH3COOH (20/79/1, v/v/v) to reduce the matrix effect; (2) using the Oasis® PRiME HLB clean-up columns. The analysis was carried out using CH3OH and H2O both acidified with 0.1% of CH3COOH as eluents. The injection volume was 20 μL and the flow rate 200 μL min-1. The analysis of two reference material (maize and wheat), was performed to evaluate the trueness and precision of the two methods by matrix-matched calibration curves. For all the regulated mycotoxins analyzed by both methods, the range of recovery percentage established by the Regulation (EC) No. 401/2006 was respected, except for ZEA by using the Oasis® PRiME HLB clean-up columns. Nevertheless, the results suggest that the Oasis® PRiME HLB clean-up columns, could be a valid alternative to the dilute-and-shoot method, although an additional cost for the clean-up has to be considered. In conclusion, both two analytical methods considerably reduce the analytical time and costs and therefore result to be promising and applicable for high-throughput routine multi-mycotoxins analysis by the use of a TQ.
Highlights
Preliminary tests were performed using MycoSpinTM 400 clean-up columns, but these tests were not continued because the MON signal in the spiked samples disappeared after the clean-up step, in spite of satisfactory results having been recorded for all the other analyzed mycotoxins
A better ionization yield was recorded using electrospray ionization (ESI)+ and it was selected for aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), 15-ADON, enniatin A (ENNA), enniatin A1 (ENNA1), enniatin B (ENNB), enniatin B1 (ENNB1), fumonisin B1 (FB1) and fumonisin B2 (FB2), whereas the use of ESI− was favored for 3-ADON, DON, DON-3-G, MON, NIV and ZEA
The formation of particular adducts was taken into consideration for both the precursor and product ions during the optimization step for each analyte and these adduct ions were selected if the intensity of the signal associated with each selected reaction monitoring (SRM) improved
Summary
Mycotoxins are secondary metabolites that are produced by microfungi which are capable of causing disease and death in humans and other animals (Bennett and Klich, 2003). The applicability of all the previously described clean-up procedures, except for the dilute-and-shoot approaches, to the simultaneous analysis of ENNs and MON has yet to be proven, and is probably not feasible (Sulyok et al, 2006) In this scenario, the current research is aimed at investigating the applicability of the developed multiresidue analytical methods to a high-throughput routine analysis for the screening and confirmation of the main EU-regulated and most relevant emerging mycotoxins to which EFSA has paid attention through the use of a TQ. Two fast, reliable and repeatable multiresidue HPLC-ESI-MS/MS methods were developed and compared for the quantitative determination of: AFB1, AFB2, AFG1, AFG2, 3-acetyldeoxynivalenol (3-ADON), 15-ADON, DON, DON-3-G, enniatins A, A1, B, B1 (ENNA, ENNA1, ENNB, ENNB1), FB1, FB2, MON, NIV, and ZEA in maize and wheat
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
