Multimycotoxin Analysis by LC-MS/MS in Cereal Food and Feed: Comparison of Different Approaches for Extraction, Purification, and Calibration

Abstract

Twelve different approaches commonly used for the simultaneous LC tandem MS (MS/MS) determination of mycotoxins (deoxynivalenol, aflatoxins, ochratoxin A, T-2 and HT-2 toxins, fumonisins, and zearalenone) were tested in cereals and feed materials. They comprised different extraction solvents, types of cleanup [solid-phase extraction, QuEChERS, and immunoaffinity (IMA)], and calibration approaches (external or matrix-matched). The percentage of mycotoxins with acceptable recovery, according to Regulation (EC) No. 401/2006, ranged from 9 to 100%. The approach giving the highest percentage of acceptable results was selected and further tested for corn, rice, and feed spiked at three different mycotoxin levels (low, medium, and high). The method is based on extraction with MeOH-water (70 + 30, v/v) and cleanup with two multiantibody IMA columns. For corn and rice spiked at low mycotoxin levels, a significant matrix effect was observed and was compensated by using 13C calibration. At higher mycotoxin levels (medium and high), matrix effects were negligible as no significant differences were observed for the majority of recovery results calculated by 13C calibration and external calibration. Although the proposed method still needs improvement in terms of accuracy and, to a lesser extent, precision, it was successfully tested with four proficiency tests in buckwheat, corn, rice, and feed, giving acceptable z-scores for 97% (34 out of 35) of results.

Multiple Mycotoxins Determination in Food by LC-MS/MS: An International Collaborative Study

“…The matrix matched calibration curve (which is restricted to one or two sample types analyzed per batch with the additional requirement to have blank matrices available for spiking purposes) or standard addition (requiring several extractions for one single analysis) quantifications are indeed time consuming, not user friendly, and not adapted for laboratories dealing with a broad range of food matrices on a daily basis. The use of labelled ISTD also improves method precision and accuracy, as recently highlighted in a study evaluating different approaches (extraction, clean-up, quantification) for mycotoxins determination in cereals [27]. The best performances were achieved by a method making use of 13 C-labeled internal standards for quantification.…”

Multiple mycotoxins analysis in animal feed with LC‐MS/MS: Comparison of extract dilution and immunoaffinity clean‐up

“…In this bracket of retention time (where methanol concentration is high in the LC gradient programme), the matrix effect can be higher and very different for feed samples and in potential effect on immunoaffinity columns. Reduction of the matrix effect can be obtained using labeled internal standards (Figure ) .…”

“…This can be explained by the degradation of those analytes during sample preparation [24,25]. This phenomenon was not observed in the study of Solfizzo et al [20].…”

“…Many authors have published multi-mycotoxin methods [9,[11][12][13][14][15][16] but only a few have used immunoaffinity columns developed for multi-analysis of food [17][18][19] and feed [20]. The main advantage of these columns is high clean-up potential, helpful in work with complex matrixes.…”

Development of an Improved Method of Sample Extraction and Quantitation of Multi-Mycotoxin in Feed by LC-MS/MS

“…Multi-mycotoxin extractions were prepared by using three different sample preparation techniques in corn. Immunoaffinity (method A) [23], solid-phase extraction (method B) [24] and QuEChERS (quick, easy, cheap, effective, rugged and safe) (method C) [25,26] methods applied for this study are shown in Table 1. They were evaluated based on analyte recovery (Rec.)…”