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Review of common methods for the analysis of allergens in food and the validation of ELISA method for egg and soy proteins
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
2012 (English)Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Food allergy is an abnormal immune response triggered by foods harmless to the body. A food company must follow legislation regarding production and labeling of a product, since the allergen content should not pose a risk for allergy sufferers. Allergen testing of the products is common and the outcome of the testing must be reliable. The food industry demands that laboratories validate their methods. In this study, a market survey was conducted to select the most suitable method for the company ALcontrol, when analyzing presence of egg, soy and wheat allergen in meat product and spice matrices. The method selected was sandwich enzyme-linked immunosorbent assay (sandwich ELISA). A set of analysis data was created to be used in an accreditation application to the Swedish board of accreditation and conformity assessment (SWEDAC). Data was interpreted via two parameters; limit of quantification (LOQ) and recovery. The LOQs of the egg assay were determined to 0.01 mg/kg in regular extraction buffer and 0.15 mg/kg in tannin extraction buffer. In a validation of the kit, performed by the supplier ELISA Systems (ELISA Systems 2010) the limit of detection (LOD), calculated from the standard deviation (STD) x 3, was determined to 0.091, hence giving a LOQ of 0.273. Compared to the validation, the result in this study showed the potential of even lower LOQ’s. The recoveries of egg protein in the meat product matrix in tannin extraction buffer were determined to 102%, 98% and 118% for 1.048 mg/kg, 2.096 mg/kg and 5.24 mg/kg respectively. The recoveries of egg protein in the meat product matrix in regular extraction buffer were determined to 116%, 116% and 93 % for 1.048 mg/kg, 2.096 mg/kg and 5.24 mg/kg respectively. The recoveries of egg protein in the spice matrix in tannin extraction buffer were determined to 90%, 88% and 69% for 1.048 mg/kg, 2.096 mg/kg and 5.24 mg/kg respectively. The recoveries of egg protein in the spice matrix in regular extraction buffer were determined to 89%, 83% and 70% for 1.048 mg/kg, 2.096 mg/kg and 5.24 mg/kg respectively. The LOQs of the soy assay were determined to 0.133 mg/kg in regular extraction buffer and 0.07 mg/kg in tannin extraction buffer. The recoveries of soy protein in the meat product matrix in regular extraction buffer were determined to 42%, 25% and 16% for 1.78 mg/kg, 3.56 mg/kg and 7.12 mg/kg, respectively. The recoveries of soy protein in the spice matrix in tannin extraction buffer were determined to 38%, 31% and 25% for 1.78 mg/kg, 3.56 mg/kg and 7.12 mg/kg, respectively. In a validation of the kit, performed by the supplier ELISA Systems, the recovery was tested in several matrices. None of the matrices are fully comparable to the matrices of sausage and spice. If compared, the results of this study showed significantly lower recoveries than the ELISA Systems’ validation. An evaluation was conducted and reviewed by the Quality Manager of ALcontrol. The ELISA methods for both egg and soy were considered reliable and approved by the Quality Manager of ALcontrol, within the concept of allergen testing. The methods will be used by ALcontrol in their analyses, and the results from this report will be used when applying to SWEDAC for accreditation for the ELISA method of egg and soy allergen.

Place, publisher, year, edition, pages
2012. , 45 p.
Keyword [en]
allergen, analysis, ELISA, accreditation, validation, egg, soy
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:lnu:diva-22625OAI: oai:DiVA.org:lnu-22625DiVA: diva2:573317
External cooperation
ALcontrol Laboratories, Linköping
Subject / course
Food Science
Uppsok
Physics, Chemistry, Mathematics
Supervisors
Examiners
Available from: 2013-01-07 Created: 2012-11-26 Last updated: 2013-01-07Bibliographically approved

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CiteExportLink to record
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Citation style
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