Determination of deoxynivalenol sulphonates in cereal samples: method development, validation and application

The first short and simple RP-UHPLC-MS/MS based method for co-determination of the Fusarium mycotoxin deoxynivalenol (DON) and its reaction products upon treatment with sulphur reagents, the DON sulphonates (DONS) 1, 2 and 3, in cereals was developed and validated. Recoveries of extraction from maize, wheat and barley were between 88 and 107%. Matrix effects ranging between 93 and 234% in concentrated extracts and between 99 and 134% in diluted extracts were compensated by quantitation against matrix matched standards. Measurement of concentrated and diluted extracts enabled determination of DONS-1, -2, -3 and DON in cereals in a concentration range from 0.015 to 120 mg/kg. The method was then applied to investigate the influence of reagent concentration, type of sulphur reagent, moisture content, presence of propionic acid, storage time and thermal treatment in the presence of sodium metabisulphite (SBS) and monomethylamine (MMA) on DON reduction and DONS formation. DON reduction greater than 80% was obtained in several experiments and required storage at moisture contents between 25 and 30% (at reagent concentration of 0.5%) for less than one week, storage with 0.5% SBS at 14% moisture for 6 weeks or thermal treatment in the presence of SBS and MMA. The efficiency of sodium sulphite for DON reduction was systematically compared with that of SBS and found to be lower at 14% moisture content, but similar at 30% moisture. Under most storage conditions, DONS-3 was the main reaction product, followed by DONS-2. Prolonged storage for more than 6-8 weeks shifted the pattern of formed DON sulphonates towards DONS-2. Likewise, thermal treatment in the presence of SBS and MMA caused exclusive formation of DONS-1 and -2. Due to partial degradation of DONS-3 to DON under physiological conditions, predominant conversion of DON into DONS-1 and DONS-2 is desirable.