| Summary: | Standard solutions of mycotoxins prepared in RP HPLC solvents from neat standards are usually used for analytical method development. Multi-mycotoxin HPLC-MS/MS methods necessitate stability estimation for the wide spectrum of fungal metabolites. The stability of individual diluted stock standard solutions of mycotoxins in RP-HPLC solvents and multi-analyte HPLC-MS/MS calibrants was evaluated under standard storage and analysis conditions. Individual stock standard solutions of aflatoxins, sterigmatocystin, A- and B-trichothecenes, zearalenone and its analogues, ochratoxin A, fumonisins, <i>Alternaria</i> toxins, enniatins and beauvericin, moniliformin, citrinin, mycophenolic, cyclopiazonic acids and citreoviridin were prepared in RP-HPLC solvents and stored at −18 °C for 14 months. UV-spectroscopy was utilized to monitor the stability of analytes, excluding fumonisins. The gradual degradation of α-, β-zearalenol and α-, β-zearalanol in acetonitrile was detected. Aflatoxins and sterigmatocystin, zearalenone, <i>Alternaria</i> toxins, enniatins and beauvericin, citrinin, mycophenolic, cyclopiazonic acids and citreoviridin can be referred to as stable. The concentration of the majority of trichothecenes should be monitored. Diluted multi-mycotoxin standard in water/methanol (50/50 <i>v/v</i>) solutions acidified with 0.1% formic acid proved to be stable in silanized glass at 23 °C exposed to light for at least 75 h (CV ≤ 10%). An unexpected manifestation of MS/MS signal suppression/enhancement was discovered in the course of multi-mycotoxin standard solution stability evaluation.
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