Quantification of Ochratoxin A in Swine Kidneys by Enzyme-linked Immunosorbent Assay Using a Simplified Sample Preparation Procedure
Abstract:An improved procedure for sample preparation and quantitation of ochratoxin A (OA) in swine kidneys was developed. Kidney samples were homogenized in acidified ethyl acetate, centrifuged, sub-sampled, dried, reconstituted with methanol and directly assayed using an indirect competitive enzyme-linked immunosorbent assay (ELISA). The rabbit antisera used in the development of this assay was found to have a high degree of cross-reaction with ochratoxins A and C but not with ochratoxins B, α, 4-OH-OA and two structurally similar molecules L-phenylalanine and citrinin with the values being 100, 80, 3.33, 10, 1.4, 0 and 0.04%, respectively. Extraction recoveries as determined by high performance liquid chromatography in kidneys spiked with 0.97 to 15.62 ppb OA were determined. The recovery values ranged from 91 to 110% with acceptable inter-assay coefficients of variation (CV) being obtained at the 3.9 ppb spiking concentration or higher. The lowest reproducible OA detection limit for the ELISA in the spiked swine kidney samples was 7.81 ppb with inter-assay CV of 8.85%. The ELISA analysis of the spiked samples correlated highly with conventional high-performance liquid chromatography (HPLC) analysis but was dependent on the conditions of the assay. Standards prepared in methanol or extract prepared from a kidney had correlation coefficients (r) of 0.91 ± 0.09 and 0.94 ± 0.07, respectively. The assay is sensitive, specific, simple and sufficiently accurate for routine analysis of swine kidneys.
Document Type: Research Article
Affiliations: 1: Department of Foods and Nutrition, University of Manitoba, Winnipeg, Manitoba, Canada, R3T 2N2 2: Department of Animal Science in the Faculty of Agriculture and Food Sciences, University of Manitoba, Winnipeg, Manitoba, Canada, R3T 2N2
Publication date: November 1, 1994
- IAFP Members with personal subscriptions to JFP Online: To access full-text JFP or JMFT articles, you must sign-in in the upper-right corner using your Ingenta sign-in details (your IAFP Member Login does not apply to this website). The Journal of Food Protection (JFP) is a refereed monthly publication. Each issue contains scientific research and authoritative review articles reporting on a variety of topics in food science pertaining to food safety and quality. The Journal is internationally recognized as the leading publication in the field of food microbiology with a readership exceeding 11,000 scientists from 70 countries. The Journal of Food Protection is indexed in Index Medicus, Current Contents, BIOSIS, PubMed, Medline, and many others.
Print and online subscriptions are available to IAFP Members and institutional subscribers. IAFP Members with a subscription to JFP Online will have access to all available JFP and JMFT content. Online visitors who are not IAFP Members or journal subscribers will be charged on a pay-per-view basis. Membership and subscription information is available at www.foodprotection.org.
- Information for Authors
- Submit a Paper
- Subscribe to this Title
- Membership Information
- Information for Advertisers
- Ingenta Connect is not responsible for the content or availability of external websites