Title: The certification report of the genetic identification of a Hippoglossus hippoglossus (Atlantic halibut) fish powder EURM-020
Authors: KORTEKAAS ANNA MARIALEYS HANNESEGHERS JOHNTRAPMANN STEFANIE
Publisher: Publications Office of the European Union
Publication Year: 2020
JRC N°: JRC119890
ISBN: 978-92-76-16230-8 (online)
ISSN: 1831-9424 (online)
Other Identifiers: EUR 30097 EN
OP KJ-NA-30097-EN-N (online)
URI: https://publications.jrc.ec.europa.eu/repository/handle/JRC119890
DOI: 10.2760/170544
Type: EUR - Scientific and Technical Research Reports
Abstract: This report describes the production of EURM-020, a fish tissue material certified for the identification of the fish species Hippoglossus hippoglossus (Atlantic halibut). This material was produced following ISO 17034:2016 and is certified in accordance with ISO Guide 35:2017. The material was produced from one fillet originating from a single fish which was genetically identified as Hippoglossus hippoglossus (Atlantic halibut). In facilitating storage, handling and shipment, the fish fillet was cut into small pieces, slowly dried using a freeze drying process and cryogenically milled to obtain a fine powder. The powder was bottled, and the bottles were closed under an argon atmosphere to ensure the conservation of the material. In storing the material, the bottles were placed at 4 °C. The fish species present in EURM-020 was determined to be Hippoglossus hippoglossus (Atlantic halibut) via genetic identification based upon the sequencing results of two specific regions of mitochondrial DNA (also called DNA barcodes): the cytochrome b gene (cytb) and the cytochrome c oxidase subunit I gene (COI). Between-unit homogeneity was established by evaluating the quality and quantity of the extracted DNA from multiple bottles. In addition, the two DNA barcodes were amplified by polymerase chain reaction (PCR) and analysed by bi-directional Sanger sequencing for each bottle sampled. The stability during dispatch and storage was assessed through quantification of extracted DNA by UV spectrophotometry, and gel-electrophoresis of the extracted DNA and PCR products of the two DNA barcodes. The process was conducted following ISO Guide 35:2017. The material was characterised based on a comparison of measurements produced by two laboratories with demonstrated competencies and adhering to ISO/IEC 17025. Data analysis and species identification was completed at the JRC. The material is only certified for its identity which was verified with two different mitochondrial DNA based methods. It is unlikely that there is an alternative species that shares both the sequences of the cytb and COI gene barcode regions with the species in EURM-020, therefore, the identity cannot be doubted. Consequently, uncertainties of the certified value relating to inhomogeneity, instability and characterisation are negligible. The material is intended for the quality control or assessment of method performance. As with any reference material, it can be used for validation studies. EURM-020 is available in glass vials containing at least 200 mg of dried Hippoglossus hippoglossus (Atlantic halibut) powder. The minimum amount of sample to be used is 7 mg.
JRC Directorate:Health, Consumers and Reference Materials

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