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Reference materials for agricultural, food and consumer products


Specific Objectives:
To provide agricultural, food and consumer products CRMs to users world-wide as required to support the implementation of European Commission policies and legislation, consumer protection and to prepare and/or certify new CRMs according to developing needs with emphasis on GMOs, TSE test evaluation and standardisation, certification of mycotoxin reference materials and production of DNA reference materials for GMO and pathogen detection.
Planned Deliverables:

Specific Deliverables to DGs:
- Support CRLs in their task of NRL inter-comparisons by preparing and distributing CRMs or PT samples and by data handling and evaluating the results from inter-comparison measurements, in particular organisation of a proficiency testing programme heavy metals in tuna fish with CRL/ISS Roma;
- Production and certification of one genomic DNA CRM from pathogenic bacteria;
- Development of six new CRMs for the detection of GMOs;
- Completion of short term stability and homogeneity study for mycotoxin M1 in milk powder CRMs (certification 2003);
- Further support to BSE test evaluation and standardisation.

Actual requirements depend on developments which are currently not under IRMMs control:
- Deliverables as a result of the research;
- Clone for one GMO available for plasmid CRM production;
- Clone for one food pathogen available for plasmid CRM production;
- Establishment of concepts for stability monitoring of DNA reference materials.

Summary of deliverables made by: 31/12/2001
Detection of food-borne pathogens (FOOD-PCR)
EU expert laboratories, JRC-IHCP, DG SANCO
PCR-based methods need to be established for food safety and this task involves the extraction, stability control and amplification of DNA from various food borne pathogens (e.g. E Coli, Salmonella).
Freeze drying of bacteria DNA in propylene vials was carried out and proved to be best method for long term storage and reconstitution. DNA from Listeria, E.coli Salmonella and Yersinia were received from 5 expert laboratories. About 200 units of each of the purified DNA materials each were prepared by freeze-drying. The materials proved sufficiently stable (transport at ambient temperature possible), the DNA quality remained intact after reconstitution. This technique can also be implemented for other types of DNA reference materials (clinical, GMO).
In principle this task has been finalised from IRMM's point of view, but IRMM will stay in contact for possible future reference material production and/or a possible follow up project.

Definition of performance criteria for testing gender determination of beef including PT round.
On request of DG AGRI - Regulations EC 2456/1993 and EC 2772/2000 (amending EC 1964/1982)
EU export and import of beef carries a subsidy difference of some 130 EURO per 100 kg of meat depending on whether the beef is declared as male (considered to be higher quality) or female. This subsidy difference requires strict legislation to ensure the correct gender declaration of beef at the EU level.
The feasibility study was planned to include 7 customs laboratories. 430 proficiency samples were prepared by March 2001 and the samples were shipped. 2 PCR methods were set up at IRMM and proficiency testing was completed and the evaluation of results from the 8 laboratories was also completed. The internal report and summary report including the technical requirements as requested by DG AGRI is presently in preparation.

Preparation and certification of CRM 801 (triglycerides in cocoa butter).

Directive 2000/36/EC DG AGRI:
The addition of vegetable fat other than cocoa butter may not exceed 5 %, so detection and quantification by triglyceride analysis is required.
Homogeneity, short-term stability and mid-term stability studies and the certification procedure were carried out. The calculation of assigned values and combined uncertainties was completed and the long-term stability study is now in progress. The certification report is being drafted.

Preparation and certification of a CRM for PCB in krill ISS, Roma, DG SANCO.
The production of a CRM for the low-level residue analysis of PCBs in arctic krill is required for food safety and consumer protection.
The production of 2892 units containing 3 g plus 42 units containing 6 g was completed. PCB-analysis of bottled material was carried out. Different extraction methods and clean up methods were investigated. Part of the batch produced was transferred to ISS for further distribution to the laboratories taking part in the certification.

Preparation and certification of two honey CRMs (IRMM-802 and -803);
- Directive 96/23/EC ISS Roma, DG SANCO;%- LCRMs are needed to measure trace metal elements in honey - an important parameter for food safety and consumer protection.
The honey material was prepared, bottled and sterilised. CRL-ISS samples were sent to ISS for the NRL inter-comparison. Electronic forms for the collection of results for the inter-comparison and the certification exercise were developed. The NRL inter-comparison was completed and evaluated by IRMM. Tenders for the IRMM certification exercise were requested and received and the participants were selected.

Certification of Rice Flour (IRMM-804 and IMEP 19):
Heavy metal loads in irrigation water in Asia has led to concerns about the safety of rice especially with regard to possible contamination with cadmium.
Rice material grown on Cd contaminated water in Japan was obtained from NICM, Japan. Based on preliminary measurements of micro-homogeneity (6% for Cd, 10% for Zn) and particle size (large but unimodal <500 µm), the decision was taken to produce a CRM. Homogeneity, stability, monitoring and characterisation measurements were planned for the certification of 4 elements (Cd, Zn, Cu and Pb). 1126 samples of 15g were prepared and labelled as IRMM-804. The homogeneity measurements of all 4 elements were performed.

Certification 282-4R (aflatoxin M1 in milk powder)
Commission Directive 98/53/EC, Commission Regulations EC 1566/1999 and EC 1525/98` DG RTD:
Food safety and consumer protection requires the residue analysis of metabolised aflatoxins in whole milk powder.
Production and labelling of the CRMs was finished and stability and homogeneity studies started with two laboratories. First in-house measurements of the three candidate materials indicate that the target values have been reached very well.

Production of BSE positive and negative CRMs for BSE test calibration.
Food safety and consumer protection within the scope of the BSE problem (production of CRMs containing either BSE positive and BSE negative samples).
The agreement was given by New Zealand authorities and the import permit for 60g of titrated BSE infected brain homogenate was requested from the UK. The titrated pool arrived and three different sets suitable for the three approved BSE tests (Enfer, Prionics, Biorad) were produced. Sets (each containing 80 blinded samples) were sent to 14 of the 15 EU States and the Candidate Countries were contacted (email and fax) for extension of the proficiency testing campaign.

Evaluation of newly developed post-mortem BSE tests.
Food safety and consumer protection within the scope of the BSE problem (evaluation of recently developed post-mortem BSE determining methods).
BSE testing under supervision of IRMM were carried out with homogenates in NL (ID-Lelystad) and In UK (VLA, Addlestone) and the reports were sent to DG SANCO for further evaluation. The BSE test was carried out with homogenates in the USA (UCSF, San Francisco) under supervision of IRMM. Samples were shipped to 4 test developers and the test of Prionics and PE (Wallac) were evaluated. Samples were shipped to the Imperial College, London and a series of new diluted homogenates was prepared because of freeze drying effects observed at UCSF. The evaluation was finalised under supervision of one IRMM representative with the exception from Imperial College, UK and a preliminary report is in preparation.

Third generation CRMS for GMOs Novel Food Reg. (EC) 258/97 DG SANCO
Food safety and consumer protection within the scope of genetically modified food.
IRMM further improved the production process using liquid nitrogen during grinding. For RR-Soya (IRMM-410S): two additional deliveries of raw material were received and checked for being pure non-GMO batches (35S PCR). The production was finalised as well as the bottling.
Contracts were negotiated with Novartis for Bt-176 maize and Bt-11 maize. For T25 maize (new GMO CRM), the raw materials were grown and IRMM is waiting to receive the material from Aventis. Finally the certificate and certification report for the MON810 maize CRMs was completed and this material is now for sale.

Output Indicators and Impact

The Reference Materials unit at IRMM is presently being extended to include special laboratories for the sterile and clean production of biological CRMs for the determination of GMOs, for the production of clinical CRMs and for the detection of food borne pathogens (new type of DNA CRMs). These new production facilities will be operational until end 2002 and will serve IRMM to cope with standardisation issues in the field of new technologies (biotechnology, molecular biology, genome projects). The laboratory facilities will be further optimised to allow better in house characterisation of raw materials and process control. The focus will be clearly on new generations of CRMs like DNA reference materials for pathogen detection, GMO reference materials and implementation of necessary techniques like cloned pathogen and GMO DNA for plasmid production. Since so far all GMO measurements are calibrated with IRMM CRMs, the development of new and optimised CRMs and the co-operation with the network of GMO laboratories will have an impact on the implementation of legislation on GMOs and control measures to be taken.
Summary of the project:

Certified Reference Materials (CRMs) have become a prerequisite to ensure the reliability and comparability of measurements required for agricultural, food and consumer products. The project will be demand driven and target the following:
- Stability control of microbiological CRMs;
- Certification of new CRMs for the control of dairy products;
- Initiation of new CRM-projects and provision of proficiency testing samples for the control of food- and feedstuffs;
- Studies on e.g. the validation of water determination methods for dairy and sugar products through the organisation of laboratory inter-comparisons;
- Bovine Spongiform Encephalopathies (BSE) related studies: production of proficiency testing samples for the detection of bone meal in animal feed and for the evaluation of the efficiency of heat treatments; provision of BSE-infected and non-infected brain and spinal cord samples for the assessment of the performance of candidate official EC post-mortem BSE tests; statistical analysis and scientific-technical evaluation of the results;
- Genetically Modified Organisms (GMO) related studies: production of proficiency testing samples for the development of screening methods of GMO in processed food and for the validation of quantitative methods; development of new CRMs for the detection of modified GMOs in products for human consumption.

Certified Reference Materials (CRM) are required to achieve comparability of measurements for the assessment of the quality of agricultural, food and consumer products. Correct assessment of the composition and quality of the aforementioned products is of primary importance to promote fair international trade, to detect fraud and to provide correct information to consumers. IRMM has developed unique high technology equipment and expertise for the highly efficient production of a large variety of highest quality biological and food reference materials. The Reference Materials laboratory is considered best in the world and hosts a unique multi-functional and flexible production laboratory, clean chambers, cryo-grinding, freeze drying, high purity milling, ultra fine classification and levitation melting. This highly flexible laboratory can quickly adapt it's production facilities to cater for urgent requests that stem from the policy makers and has a modern adjoining analytical laboratory fully equipped to monitor and control the production process.

Call for proposal

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Institute for Reference Materials and Measurements
B-2440 Geel

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