title: 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 ultra-performance 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.
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, 430062, China.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, 430062, China.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, 430062, China.|Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, 430062, China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Wuhan Institute of Physics and Mathematics, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Donghu Rd, Wuhan, 430072, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Hu
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, 430062, China. zwzhang@whu.edu.cn.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, 430062, China. zwzhang@whu.edu.cn.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, 430062, China. zwzhang@whu.edu.cn.
authors.forename: Zhaowei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, 430062, China. peiwuli@oilcrops.cn.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, 430062, China. peiwuli@oilcrops.cn.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, 430062, China. peiwuli@oilcrops.cn.|Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, 430062, China. peiwuli@oilcrops.cn.
authors.forename: Peiwu
authors.initials: P
authors.lastname: Li
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, 430062, China.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, 430062, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Wuhan Institute of Physics and Mathematics, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Donghu Rd, Wuhan, 430072, China.
authors.forename: Min
authors.initials: M
authors.lastname: Wang
mesh_terms: D000821:Animal Feed / Q000032:analysis; D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057230:Limit of Detection; D008807:Mice, Inbred BALB C; D009183:Mycotoxins / Q000032:analysis*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D009183:Mycotoxins
keywords: Feed; Multiple immunoaffinity columns; Multiple mycotoxins; Simultaneous determination; UPLC–MS/MS