Development of liquid chromatography-tandem mass spectrometrymeth | 53350
Journal of Chromatography & Separation Techniques

Journal of Chromatography & Separation Techniques
Open Access

ISSN: 2157-7064

Development of liquid chromatography-tandem mass spectrometrymethods for the quantitation of Anisakis simplex proteins in fish

2nd International Conference on Current Trends in Mass Spectrometry

July 20-22, 2016 Chicago, USA

Christiane Kruse F�?¦ste

Norwegian Veterinary Institute, Norway

Scientific Tracks Abstracts: J Chromatogr Sep Tech

Abstract :

Larvae of the parasite Anisakis simplex are present in many marine fish species used for food. They infect mostly the gut and inner organs but have alsobeen shown to penetrate into the fish fillet. Thus, human health can be at risk, either by contracting anisakiasis through the consumption of live parasites in raw or under-cooked fish, or by sensitisation to anisakid proteins inprocessed food. Methods for the detection of anisakid larvae include visual techniques and PCR, and immunological assays for protein determination. Recently, mass spectrometry-based proteomics has been used for the characterisation of A. simplex proteins, preparing for the development of two quantitative liquid chromatography-tandem mass spectrometry methods in the present study. Both, the label-free semi-quantitative nLC-nESI-Orbitrap-MS/MS (MS1) and the heavy peptide-applying absolute-quantitative (AQUA) LC-TripleQ-MS/MS (MS2)use unique reporter peptides derived from anisakid hemoglobin and SXP/RAL-2 protein as analytes. Standard curves in buffer and in salmon matrix showed good linearity and sufficient sensitivity for the intended method use. Preliminary validation included the assessment of specificity, repeatability, reproducibility, and applicability to incurred and naturally-contaminated samplesfor both assays. By further optimisation and full validation the LC��?MS/MS methods could be standardised and used as confirmative techniques for thedetection of A. simplex protein in fish and products.

Biography :

Christiane Kruse Fæste has completed her PhD in biochemistry at the age of 28 from the Leibniz University in Hanover, Germany, and the Max-Planck Institute for Plant Breeding Research in Cologne, Germany. Shehas been working in Preclinical Pharmaceutical Industry and is now expert for xenobiotics metabolism, toxicokinetics, food allergy and proteomics in the Section of Chemistry and Toxicology of the Norwegian Veterinary Institute, Oslo, Norway. She is member of the Norwegian Committee for Food Safety (panel on contaminants), CEN (Food Allergens), AOAC (Food Allergens), EU-Research Executive Agency (Chemistry), ILSI (Allergens) and COST (Allergens). She has published 40 articles on food allergens and mycotoxin metabolism.