Development of a multiple immunoaffinity column for simultaneous determination of multiple mycotoxins in feeds using UPLC–MS/MS

Abstract

A sensitive and specific immunoaffinity column to clean up and isolate multiple mycotoxins was developed along with a rapid one-step sample preparation procedure for ultraperformance liquid chromatography-tandem mass spectrometry analysis. Monoclonal antibodies against aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, zearalenone, ochratoxin A, sterigmatocystin, and T-2 toxin were coupled to microbeads for mycotoxin purification. We optimized a homogenization and extraction procedure as well as column loading and elution conditions to maximize recoveries from complex feed matrices. This method allowed rapid, simple, and simultaneous determination of mycotoxins in feeds with a single chromatographic run. Detection limits for these toxins ranged from 0.006 to 0.12 ng mL −1 , and quantitation limits ranged from 0.06 to 0.75 ng mL −1 . Concentration curves were linear from 0.12 to 40 μg kg −1 with correlation coefficients of R 2 > 0.99. Intra-assay and inter-assay comparisons indicated excellent repeatability and reproducibility of the multiple immunoaffinity columns. As a proof of principle, 80 feed samples were tested and several contained multiple mycotoxins. This method is sensitive, rapid, and durable enough for multiple mycotoxin determinations that fulfill European Union and Chinese testing criteria.

Occurrence, Toxicity, and Analysis of Major Mycotoxins in Food

“…A variety of cleanup methods have been implemented including liquid–liquid partitioning, solid phase extraction (SPE), immune-affinity columns (IAC), column chromatography, ion-exchange columns, and multifunctional cleanup columns such as Mycosep™ [7,75]. The most commonly used methods for mycotoxins clean-up are SPE and IAC, as these are rapid, efficient, reproducible, and safe, with a wide range of selectivity [79,80]. SPE is a technique based on the specific partitioning of the analyte dissolved in the extract (mobile phase) and the stationary phase (cartridge), which is composed of a solid adsorbent where the mycotoxins are absorbed and then eluted with an organic solvent [75].…”

Eu3+‐labeled IgG‐based time‐resolved fluoroimmunoassay for highly sensitive detection of aflatoxin B1 in feed

“…Generally, analysis of AFB 1 has been achieved by instrumental analysis, [14][15][16] enzyme-linked immunosorbent assay (ELISA) 17,18 and immunochromatographic assay. [19][20][21][22] Immunoassays have been shown to be simple, highly sensitive, cost effective and suitable for high-throughput screening analysis.…”

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

“…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.…”