Determination of Ochratoxin A in Cereals and Feeds by Ultra-performance Liquid Chromatography Coupled to Tandem Mass Spe
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Determination of Ochratoxin A in Cereals and Feeds by Ultra-performance Liquid Chromatography Coupled to Tandem Mass Spectrometry with Immunoaffinity Column Clean-up Hui Meng & Zhanhui Wang & Sarah De Saeger & Ying Wang & Kai Wen & Suxia Zhang & Jianzhong Shen
Received: 5 March 2013 / Accepted: 6 August 2013 / Published online: 6 September 2013 # Springer Science+Business Media New York 2013
Abstract This paper describes the preparation of reusable immunoaffinity columns and the development of an ultraperformance liquid chromatography tandem mass spectrometry method combined with immunoaffinity column clean-up (IAC-UPLC-MS/MS) for the determination of ochratoxin A (OTA) in cereals and feeds. The monoclonal antibody (mAb) was produced from a stable hybridoma cell line (4H10), which belongs to the immunoglobulin G1 (κ-light chain) isotype. A competitive indirect enzyme-linked immunosorbent assay was used to characterize the mAb. The concentrations causing 50 % inhibition of binding of mAb to OTA-ovalbumin by free OTA, ochratoxin B, and ochratoxin C were 1.29, 4.78, and 0.94 ng mL−1, respectively. The IAC-UPLC-MS/MS method offers a limit of quantification (LOQ, S/N >10) ranging from 0.5 to 1.0 μg kg−1 and a limit of detection (LOD, S/N >3) ranging from 0.2 to 0.3 μg kg−1 in cereal and feed samples. The IAC-UPLC-MS/MS method offers a good LOQ and LOD for OTA in cereal and feed samples. The accuracy and precision at this level fall within the EU regulatory limit. This methodology has been validated in four different matrices (millet, maize, soybean, and swine finisher diet) with highly satisfactory results and applied to the analysis of samples collected from the markets. H. Meng : Z. Wang : Y. Wang : K. Wen : S. Zhang : J. Shen (*) Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory For Food Quality and Safety, College of Veterinary Medicine, China Agricultural University, Yuanmingyuan West Road 2, Haidian District, Beijing 10193, People’s Republic of China e-mail: [email protected] S. De Saeger Laboratory of Food Analysis, Faculty of Pharmaceutical Sciences, Ghent University, Harelbekestraat 72, 9000 Ghent, Belgium
Keywords Ochratoxin A . Monoclonal antibody . Reusable immunoaffinity column . UPLC-MS/MS . Cereals . Feeds
Introduction Ochratoxins, a group of highly toxic fungal secondary metabolites, are mainly produced by several species of Aspergillus ochraceus and Penicillium verrucosum molds. According to the Joint FAO/WHO Expert Committee on Food Additives, ochratoxin A (OTA) is a potent nephrotoxin and hepatotoxin with teratogenic, mutagenic, carcinogenic, and immunosuppressive effects even at a trace level (Joint FAO/WHO Expert Committee on Food Additives 2001). OTA is considered to be an important factor in Balkan endemic nephropathy occurring in many European countries. It has been classified in group 2B as a potential human carcinogen by the International Agency for Research on Cancer (IARC 1993). This mycotoxin occurs in a large variety of agricultural commodities
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