期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 63, 期 35, 页码 7855-7861出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.5b02513
关键词
azaspiracid; AZA-1; ELISA; immunoassay; antibody; polyclonal; shellfish toxin; mussel
资金
- Norwegian Research Council [139593/140]
- BIOTOX project (European Commission) [514074]
- European Union [311820]
- Norwegian Veterinary Institute
- New Zealand Foundation for Research, Science and Technology (International Investment Opportunities Fund) [C10X0406]
- National Institute of Environmental Health Sciences (NIEHS), NIH, USA [ES10615]
- Marine Institute
- Marine Research Sub-Programme of the National Development Plan [PBA/AF/08/001(01)]
- European Regional Development Fund
Azaspiracids (AZAs) are a group of biotoxins that cause food poisoning in humans. These toxins are produced by small marine dinoflagellates such as Azadinium spinosum and accumulate in shellfish. Ovine polyclonal antibodies were produced and used to develop an ELISA for quantitating AZAs in shellfish, algal cells, and culture supernatants. Immunizing antigens were prepared from synthetic fragments of the constant region of AZAs, while plate coating antigen was prepared from AZA-1. The ELISA provides a sensitive and rapid analytical method for screening large numbers of samples. It has a working range of 0.45-8.6 ng/mL and a limit of quantitation for total AZAs in whole shellfish at 57 mu g/kg, well below the maximum permitted level set by the European Commission. The ELISA has good cross-reactivity to AZA-1-10, -33, and -34 and 37-epi-AZA-1. Naturally contaminated Irish mussels gave similar results whether they were cooked or uncooked, indicating that the ELISA also detects 22-carboxy-AZA metabolites (e.g., AZA-17 and AZA-19). ELISA results showed excellent correlation with LC-MS/MS analysis, both for mussel extract spiked with AZA-1 and for naturally contaminated Irish mussels. The assay is therefore well suited to screening for AZAs in shellfish samples intended for human consumption, as well as for studies on AZA metabolism.
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