Abstract
An immunoaffinity magnetic beads (IMBs) based automatic pretreatment method was developed for the quantitative analysis of deoxynivalenol (DON) by ultra-performance liquid chromatography and ultraviolet detector (UPLC-UV). First, N-hydroxysuccinimide-terminated magnetic beads (NHS-MBs) with good magnetic responsivity and dispersibility were synthesized and characterized by optical microscopy, scanning electron microscopy (SEM), and laser diffraction-based particle size analyzer. Then, the amino groups of anti-DON monoclonal antibody (mAb) and the NHS groups of NHS-MBs were linked by covalent bonds to prepare IMB, without any activation reagent. The essential factors affecting the binding and elution of DON were meticulously tuned. Under optimal conditions, DON could be extracted from a real sample and eluted from IMB by water, enabling environmentally friendly and green analysis. Hence, there was no need for dilution or evaporation prior to UPLC-UV analysis. DON in 20 samples could be purified and concentrated within 30 min by the mycotoxin automated purification instrument (MAPI), allowing for automated, green, high-throughput and simple clean-up. Recoveries at four distinct spiking levels in corn and wheat ranged from 92.0% to 109.5% with good relative standard deviations (RSD, 2.1–7.0%). Comparing the test results of IAC and IMB in commercial samples demonstrated the reliability and superiority of IMB for quantitatively analyzing massive samples.
Highlights
The primary representative of the type B trichothecenes, deoxynivalenol (DON), commonly called vomitoxin, is generated by some Fusarium species [1]
immunoaffinity magnetic beads (IMBs) loaded with ng of DON, to optimize the lowest concentration of fore, we firstly investigated the elution ability of different concentrations of methanolmethanol in in the eluent solvent
DON in 20 corn and wheat samples were analyzed in the range of 100 μg/kg to 2680 μg/kg by mycotoxin automated purification instrument (MAPI)
Summary
The primary representative of the type B trichothecenes, deoxynivalenol (DON), commonly called vomitoxin, is generated by some Fusarium species [1]. DON is one of the most prevalent mycotoxins in various cereals crops and processed grains, such as corn, wheat, bread, beer, and malt [2,3,4,5], with a detection rate of 57% in 11,444 tested samples [6]. The consumption of grains or food products contaminated with DON could result in various adverse effects to humans and animals, including feed refusal, gastroenteritis, nutrient malabsorption, diarrhea, vomiting, teratogenicity, immunotoxicity and cardiotoxicity [7,8,9]. The International Agency for Research on Cancer has categorized DON in the group 3 as not classifiable regarding their carcinogenicity to humans since 1993 [8]. Many countries have established the maximum level (ML) for DON in cereals and its derived products. The ML of DON in Commission Regulation (EC) No 1881/2006 ranged from
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