Simultaneous broad-spectrum detection of over 50 mycotoxins in maize silage via liquid chromatography-tandem mass spectrometry: Method development and application

Abstract

For a comprehensive assessment of mycotoxin contamination in maize silage, a novel method using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) that can simultaneously detect 53 mycotoxins in maize silage was developed. The method streamlines the extraction and purification processes with a simplified dilute-and-shoot sample pretreatment step. After extraction with acetonitrile/water/formic acid (70:30:0.1, v/v/v) followed by the LC-MS/MS detection. Furthermore, isotope internal standards were employed to eliminate matrix effects. The method was validated by linearity, matrix effects, selectivity, recovery, repeatability, limits of detection (LOD), limits of quantification (LOQ) and measurement uncertainties. The LODs ranged from 0.3 to 162.2 μg/kg and the LOQs from 0.9 to 535.4 μg/kg. The method developed enables the effective quantitative analysis of 51 out of 53 mycotoxins, with mean recoveries for 51 qualitatively determinable analytes applicable to this method ranging from 33.20% to 119.50% and relative standard deviations of less than 13.33%; all 53 mycotoxins could be qualitatively analyzed, and the method was successfully applied for analyzing 63 maize silage samples from China. Most of the mycotoxins detected were attributed to the Fusarium species. highlighting the prevalence of this genus in maize silage contamination. The developed LC-MS/MS method provides an excellent analytical performance for detecting multiple mycotoxins, marking it as the first report of such broad-spectrum detection in silage, and offers an effective tool for the accurate assessment of the true mycotoxin content in maize silage and the precise evaluation of feed quality.