Optimization of the QuEChERS-Based Analytical Method for Investigation of 11 Mycotoxin Residues in Feed Ingredients and Compound Feeds

Abstract

Mycotoxins are toxic substances naturally produced by various fungi, and these compounds not only inflict economic damage, but also pose risks to human and animal health. The goal of the present study was to optimize the QuEChERS-based extraction and liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for the analysis of 11 mycotoxins, such as aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FBs), T-2 toxin, HT-2 toxin, zearalenone (ZEN), and deoxynivalenol (DON), commonly found in feed. The QuEChERS method, characterized by being “quick, easy, cheap, effective, rugged, and safe”, has become one of the most common extractions and clean-up procedures for mycotoxin analyses in food. Therefore, in this experiment, an optimal method for the analysis of 11 mycotoxins in feed was established by modifying the general QuEChERS method. In this process, it was confirmed that even if feed samples of different weights were extracted, the quantitative value of mycotoxins in the feed was not affected. To reduce matrix effects, 13C-labeled compounds and deuterium were used as internal standards. This optimized method was then applied in the determination of 11 mycotoxins in 736 feed ingredients and compound feeds obtained from South Korea. The results showed that the occurrence rates of FBs, ZEN, and DON were 59.4%, 38.0%, and 32.1%, respectively, and OTA, AFs, and T-2 toxin and HT-2 toxin were found in fewer than 1% of the 736 feeds. The mean concentration ranges of FBs, ZEN, and DON were 757–2387, 44–4552, and 248–9680 μg/kg, respectively. Among the samples in which DON and ZEN were detected, 10 and 12 samples exceeded the management recommendation standards presented by the Ministry of Agriculture, Food and Rural Affairs (MAFRA). However, when the detected concentrations of DON and ZEN were compared with guideline levels in foreign countries, such as the US, Japan, China, and the EU, the number of positive samples changed. In addition, the co-occurrence of mycotoxins in the feed was analyzed, and the results showed that 43.8% of the samples were contaminated with two or three mycotoxins, among which the co-occurrence rate of FBs, ZEN, and DON was the highest. In conclusion, these results suggest the need for stricter management standards for FBs, DON, and ZEN in South Korea, and emphasize the importance of the continuous monitoring of feeds.

Simultaneous Determination of 11 Mycotoxins in Maize via Multiple-Impurity Adsorption Combined with Liquid Chromatography–Tandem Mass Spectrometry

“…Multifunction impurity adsorption (MIA) cleaning is a method based on matrix dispersion solid-phase extraction, which is mainly based on the selection of a variety of functionalized adsorbent materials to adsorb the main interfering impurities in a sample. This effectively removes phospholipids, pigments, and other substances that may be present in the matrix while leaving the measured substances in the sample solution and achieving purification and enrichment, which can save time for sample pre-treatment [ 25 ]. According to the adsorption characteristics of different materials such as BONDESIL-SI sorbent, which is based on a bonded reversed-phase (with high-purity irregular silica gel as the matrix and end-group capping treatment), a typical reversed-phase extraction retention mechanism has excellent strength retention properties for non-polar compounds and can remove lipids from samples.…”

“…Even when relying only on LC, it may have low sensitivity and is prone to false positives. LC-MS/MS, with good selectivity and specificity and a low detection limit, has become a popular direction for the study of mycotoxins in recent years, and several methods for detecting multiple mycotoxins in maize have been published [ 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ]. The limits of quantification (LOQ) of various toxins are as follows: AFs (0.05~1.6 μg/kg), T-2 (0.05~0.3 μg/kg), DON (0.94~13.6 μg/kg), ZEN (0.5~0.72 μg/kg), OTA (0.03~0.3 μg)/kg), and FBs (1.0∼8.2 μg/kg).…”

“…In recent years, the technique of purification via multiple-impurity adsorption (MIA) principles has gradually emerged; it mainly involves adsorption of the main interfering impurities in a sample through a variety of functionalized adsorbent materials, effectively removing phospholipids, fats, and some proteins that may be present in the matrix, while leaving the measured substances in the sample solution and achieving purification and enrichment. This has the significant advantages of rapid and simple operation and high detection throughput [ 25 ] and has been successfully applied to the simultaneous detection of various drugs in food [ 26 ]. Therefore, the development of novel multiple-impurity adsorption methods is necessary to monitor or study multiple mycotoxins in maize.…”

Application of response surface methodology in optimizing simultaneous extraction of multiple mycotoxins in maize via QuEChERS

“…QuEChERS extraction dynamics and mechanism have already been extensively discussed (Nakhjavan et al, 2020;Pantano et al, 2021;Seo et al, 2021). However, the QuEChERS procedures developed for multiple mycotoxins extraction have been based on single-factor optimization.…”

Risks for animal health related to the presence of ochratoxin A (OTA) in feed

“…In the last few years much of the literature has focused on multi‐mycotoxin methods especially using QuEChERS and LC–MS/MS in different feed materials (e.g. Nakhjavan et al., 2020 ; Nualkaw et al., 2020 ; Jo et al., 2021 ; Gonzales‐Jartin et al., 2021 ; Konak et al., 2021 ; Seo et al., 2021 ; Bi et al., 2022 ; Mackay et al., 2022 ; Nochetto and Li, 2022 ). However, there has also been focus on other and often faster methods including those with biosensors that often involve specific antibodies, aptamers or molecular imprinting polymers in the analytical work (e.g.…”