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Development of a Novel Immunoaffinity Column for the Determination of Deoxynivalenol and Its Acetylated Derivatives in Cereals

Zuo, Hai Gen, Zhu, Jian Xin, Shi, Lei, Zhan, Chun Rui, Guo, Ping, Wang, Ying, Zhang, Yanming, Liu, Jiapeng
Food analytical methods 2018 v.11 no.8 pp. 2252-2260
15-acetyldeoxynivalenol, 3-acetyldeoxynivalenol, bovine serum albumin, cleaning, corn, cross reaction, deoxynivalenol, detection limit, haptens, high performance liquid chromatography, mice, monoclonal antibodies, myeloma, oatmeal, solid phase extraction, splenocytes, statistical analysis, tandem mass spectrometry, wheat
Deoxynivalenol (DON) hapten 3-O-hemisuccinyl-deoxynivalenol (3-HS-DON) was synthesized and conjugated to bovine serum albumin (BSA) to prepare immunogen by active ester method. A novel anti-DON monoclonal antibody (mAb) showing good cross-reactivity against both 3-acetyl-deoxynivalenol (3-ADON) and l5-acetyl-deoxynivalenol (15-ADON) was produced by fusing mouse myeloma cells lines SP 2/0 with spleen cells from Bal b/c mice which were immunized by the immunogen. The mAb was coupled with sepharose-4B, then packed into a solid phase extraction (SPE) cartridge to prepare an immunoaffinity columns (IAC). A method for simultaneous determination of DON, 3-ADON, and 15-ADON in cereals was developed. The cereal samples were extracted by water, cleaned up with IAC, and determined by high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). The results showed that the limits of detection (LODs) for DON, 3-ADON, and 15-ADON were 1, 6, and 3 μg kg⁻¹, respectively. Recoveries of DON, 3-ADON, and 15-ADON from wheat, oatmeal, and maize were 67.5–93.8, 63.8–113.2, and 75.5–106.6% at spiking levels of 20–2000 μg kg⁻¹, and the relative standard deviations (RSDs) were 2.4–13.7, 2.4–12.6, and 7.6–12.2%, respectively. This method was suitable for determination of DON, 3-ADON, and 15-ADON in cereals, with the advantages of simplicity, rapidness, good selectivity and sensitivity, and environmental friendliness.