Development of a Novel Immunoaffinity Column for the Determination of Deoxynivalenol and Its Acetylated Derivatives in C
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Development of a Novel Immunoaffinity Column for the Determination of Deoxynivalenol and Its Acetylated Derivatives in Cereals Hai Gen Zuo 1 & Jian Xin Zhu 1 & Lei Shi 1 & Chun Rui Zhan 1 & Ping Guo 1 & Ying Wang 2 & Yanming Zhang 2 & Jiapeng Liu 2 Received: 2 January 2018 / Accepted: 20 February 2018 # Springer Science+Business Media, LLC, part of Springer Nature 2018
Abstract 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 15ADON 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 15ADON in cereals, with the advantages of simplicity, rapidness, good selectivity and sensitivity, and environmental friendliness. Keywords Monoclonalantibody . Immunoaffinitycolumn . Deoxynivalenol . 3-acetyl-deoxynivalenol . 15-acetyl-deoxynivalenol . HPLC-MS/MS
Introduction Deoxynivalenol (DON), also known as vomitoxin, is an epoxy-sesquiterpenoid mycotoxin produced mainly by Fusarium species (mainly F. graminearum and F. culmorum) under appropriate conditions of temperature and humidity. DON is produced and commonly found in cereals, especially wheat, barley, maize, sorghum, and cereal-based products (Li et al. 2012; Wu and Wang 2015; Ennouari et al. 2013). The acetylated derivatives of DON, i.e., 3-acetyl-deoxynivalenol Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12161-018-1211-4) contains supplementary material, which is available to authorized users. * Hai Gen Zuo [email protected] 1
Jiangxi Entry-Exit Inspection and Quarantine Bureau, Nanchang 330002, China
2
Hua-an Magnech Biotech Co., Ltd, Beijing 102200, China
(3-ADON) and 15-acetyl-deoxynivalenol (15-ADON), are intermediaries in its biosynthesis; hence, they are often described as fungal precursors or as free mycotoxins (Broekaert et al. 2014). In addition, there may exist conversion between the acetylated derivatives (3-ADON and 15-ADON) and DON dur
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