Statement of Measurement Beef

Statement of measurement 4005

Beef Reference Material LGC7221

Assessed value

Ground raw beef

Date of issue: May 2017 Certificate revised April 2018

Signed: ______Gill Holcombe (Mrs) for the Government Chemist

Material number: LGC7221 Batch number: 004 Page 1 of 4

Material Preparation Beef was purchased from a commercial source as one topside joint, labelled as beef topside joint. The surface of the meat was removed and discarded to minimise the risk of cross-contamination and then the sample was prepared by removing any separable fat, retaining the lean meat. The lean meat was cubed, homogenised in a food processor, combined and then mixed. The resultant paste was transferred, in portions of at least 10 g, to glass pots with tamper-evident lids.

Homogeneity As the material is intended for use as a reference material for meat species identification, and was prepared using meat from one source, a formal homogeneity check by analysis was not carried out.

Stability When stored under the recommended conditions, the material is expected to remain stable. Tests carried out at LGC to check the effect of one additional freeze-thaw cycle in a similar meat species material showed no significant change. However, freeze-thaw cycles should be avoided to prevent changes in the extraction profile.

Assessment and Methods Used Confirmation of the meat as beef was obtained using three approaches. These were DNA sequencing, a PCR based method and an immunoassay method. The material was also checked for absence of cross-contamination using both immunoassay and PCR based techniques. No evidence was found of the presence of pork, horse meat, sheep meat, goat meat, chicken meat or turkey meat. The highest detection limit for these techniques is 1 % of one meat species in the presence of another. Immunoassay Analysis Commercially available meat speciation kits (sandwich Enzyme Linked ImmunoSorbent Assays (ELISA)) were used. Samples of the homogenised meat were extracted according to the manufacturer’s protocol. Two independent meat extracts from each sample were added to a pre- coated microtitre plate supplied with each kit and the assay was carried out in accordance with the manufacturer’s protocol. DNA Analysis Prior to homogenisation, DNA was extracted from a cut of the meat using a proprietary method, and was analysed by Sanger DNA sequencing. Mitochondrial genes MT-RNR1, which encodes RNA 12S, and MT-CYB, which encodes cytochrome b, were used for species specific identification. The meat was identified as the correct species by alignment using validated DNA sequence databases. Confirmation of species identity on the final product was based on amplification of specific genomic targets using PCR for beef, pork, horse meat, sheep meat, goat meat, chicken meat and turkey meat. Only the PCR product specific to beef was detected. The results also confirmed that no cross-contamination had occurred during preparation and packaging.

Intended Use This material is intended for use as a reference material in procedures for the identification of beef.

Instructions for Use The material should be defrosted and allowed to reach room temperature, (20 ± 5) °C, before use. Once defrosted, store refrigerated and use within 5 days.

Material number: LGC7221 Batch number: 004 Page 2 of 4

Storage The unopened pot should be stored frozen, at (-20 ± 5) °C. Freeze-thaw cycles should be avoided to prevent changes in extraction profile.

Shelf Life Provided the samples are stored under the recommended conditions, this statement of measurement will remain valid for 3 months from the date of shipment.

Certificate Revision This certificate was revised in April 2018 to clarify the advice for users regarding opened units

Material number: LGC7221 Batch number: 004 Page 3 of 4

Unit Number Date of Shipment

Legal notice

The information provided in this document is based on the techniques described herein. No warranty or representation, express or implied, is made that the use of the product or any information, material, apparatus, method or process which is the subject of or referred to in this document does not infringe any third party rights. Further, save to the extent: (a) prohibited by law; or (b) caused by a party's negligence; no party shall be liable for the use made of the product, any information, material, apparatus, method or process which is the subject of or referred to in this document. In no event shall the liability of any party exceed whichever is the lower of: (i) the value of the product; or (ii) £500,000; and any liability for loss of profit, loss of business or revenue, loss of anticipated savings, depletion of goodwill, any third-party claims or any indirect or consequential loss or damage in connection herewith is expressly excluded.

------This material was produced by LGC in its role as the UK’s designated National Measurement Institute (NMI) for chemical and bio-measurement.

This document may not be published except in full, unless permission for the publication of an approved extract has been obtained in writing from LGC Limited. It does not of itself impute to the subject of measurement any attributes beyond those shown by the data contained herein .

Material number: LGC7221 Batch number: 004 Page 4 of 4