Immunoaffinity Cleanup and Isotope Dilution-Based Liquid Chromatography Tandem Mass Spectrometry for the Determination of Six Major Mycotoxins in Feed and Feedstuff

Abstract

In this study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of deoxynivalenol, aflatoxin B1, zearalenone, ochratoxin A, T-2 toxin and fumonisin B1 in feed and feedstuff was established. The sample was extracted with an acetonitrile–water mixture (60:40, v/v), purified by an immunoaffinity column, eluted with a methanol–acetic acid mixture (98:2, v/v), and reconstituted with a methanol–water mixture (50:50, v/v) after drying with nitrogen. Finally, the reconstituted solution was detected by LC-MS/MS and quantified by isotope internal standard method. The six mycotoxins had a good linear relationship in a certain concentration range, the correlation coefficients were all greater than 0.99, the limits of detection were between 0.075 and 1.5 µg·kg−1, and the limits of quantification were between 0.5 and 5 µg·kg−1. The average spike recoveries in the four feed matrices ranged from 84.2% to 117.1% with relative standard deviations less than 11.6%. Thirty-six actual feed samples were analyzed for mycotoxins, and at least one mycotoxin was detected in each sample. The proposed method is reliable and suitable for detecting common mycotoxins in feed samples.

Monitoring Mycotoxin Exposure in Food-Producing Animals (Cattle, Pig, Poultry, and Sheep)

“…In other cases, the extract after SLE may not be clear enough, necessitating a second cleanup step. Several authors have used immunoaffinity columns (IAC) before LC-FLD [70][71][72][73][74][75][76][77][78], LC-MS/MS [54,79,80], or LC coupled with an ultraviolet detector [73,77]. IAC-based clean-up is highly specific since these columns are composed of monoclonal antibodies against the mycotoxins of interest.…”

“…This is advantageous when the aim is to analyze mycotoxins individually. However, the drawbacks of these methods include their high cost, the fact that they are usually designed for the analysis of a single or very few analytes, and the fact that there is a limited variety of columns available [79]. For these reasons, the simultaneous detection of multiple mycotoxins in routine analysis remains a challenge, and more research is needed to obtain new IAC columns capable of retaining multiple mycotoxins.…”

“…For these reasons, the simultaneous detection of multiple mycotoxins in routine analysis remains a challenge, and more research is needed to obtain new IAC columns capable of retaining multiple mycotoxins. For instance, Liu et al (2022) [79] prepared a novel in-house multi-IAC for the extraction of six mycotoxins (DON, AFB1, ZEA, OTA, T-2 toxin, and FB1) in feed.…”

“…Regarding the techniques for detecting and quantifying mycotoxins in animal feed, most authors used LC (on reversed-phase columns) coupled with mass spectrometry [54,60,61,64,[66][67][68][79][80][81][83][84][85][86][87][88], employing electrospray ionization and triple quadrupoles (QqQs) or quadrupole ion traps (QTraps) as mass analyzers. In addition, LC coupled with FLD has been widely used [53,63,68,[70][71][72][73][74][75][76][77][78]82], and, in some cases (for DON quantification), LC has been coupled with an ultraviolet detector [73,77].…”

Advancements in Sample Preparation Methods for the Chromatographic and Mass Spectrometric Determination of Zearalenone and Its Metabolites in Food: An Overview

“…In addition to magnetic nanomaterials as adsorbents, many examples of different adsorbents used in these methods have been reported in the literature, including reduced graphene oxide and gold nanoparticle composites [60], graphitized carbon black [61], chitosan nanofibers [62], immunosorbents [63], and molecularly imprinted polymers (MIPs) [64,65]. Utilizing immunoaffinity columns (IACs) for mycotoxin purification and concentration has garnered significant research attention.…”