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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.