Context
SIMBAG-FEED project responds to the specific programme “competitive and Sustainable Growth”, Generic Activity “Measurements & Testing”, Dedicated Call 10/99, Topic II.15, “Screening and identification methods for official control of possible illegal use of banned antibiotics and growth promoters in feedingstuffs”.After several crises affecting feed and food sectors, there is a great need for gaining consumers’s confidence in relation with health and quality of european products.
In order to protect humain health, the autorisation of several antibiotics and growth promoters as feed additives have been banned in animal feed by the European Union (EU) in 1999. The antibiotics were banned due to their possible contribution to the development of bacteria resistance. The growth promoters were banned because of their toxic properties.
By the availability of official and efficient analytical methods, the implementation of official directives and decisions are facilited.
Objectives
The main technical and scientific objective of SIMBAG-FEED, is to develop, improve and validate adequate multi-methods for the detection and determination of the banned antibiotics and the banned growth promoters in feedingstuffs.The second objective of SIMBAG-FEED project is the set-up of a reference standards bank of banned substances with guaranteed stability for the use by the official control laboratories.
The banned antibiotic molecules studied in this project are: avoparcin (AVO), bacitracin-zinc (BAC), spiramycin (SPIR), tylosin (TYL) and virginiamycin (VIRG). In parallel, two growth promoters: carbadox (CAR) and olaquindox (OLA) are studied.
Description of tasks
The methods are implemented in an overall control strategy applying the following techniques :- microbiological inhibition system (MIS),
- high voltage electrophoresis (HVE),
- thin layer chromatography (TLC),
- high performance liquid chromatography (HPLC),
- liquid chromatography-mass spectrometry (LC-MS).
For the project execution, eight workpackages were established. Five workpackages focus on the development, improvement and in-house validation of the five analytical techniques. The methods that are selected as part of overall control strategy are validated by means of collaborative studies (1 workpackage). Two workpackages cover final validations by means of collaborative studies and the establishment of a bank of reference standards.
The proposed control strategy is composed of methods that have been developed and validated with successful results.
For the antibiotics, a three-step controle strategy is proposed: a screening step, a post-screening step and a confirmation step. In the case of growth promoters, two-step control strategy is proposed: a screening and a confirmation step.
For banned antibiotics, methods used are :
- MIS as a screening method,
- HVE and TLC for the post-screening,
- LC-MS for the results confirmation.
For growth promoters, methods used are :
- HPLC (UV) for the screening,
- HPLC (DAD) and LC- MS for the confirmation.
The SIMBAG-FEED project collaborates with an other project named Feedstuffs-RADIUS (Rapid Antibiotic Detection for Illegal and Unlicensed substances in animal feedingstuffs, contract QLK5-CT-2000-00900). The collaboration mainly concerns the LC-MS work of both projects.
Expected results
The detecion limits should be at least 5 times lower than the lowest contents formerly authorised in the Directive 70/524/CEE. Consequently, the detection limit for each of the antibiotics should be 1 mg/kg and for growth promoters, the detection limit should be 3 mg/kg for olaquindox and 4 mg/kg for carbadox.Sufficient amount of reference standards must be stored to provide the official control laboratories in EU with the material for at least 10 years.
Contribution
The CRA-W was invited by the project for its expertise in the antibiotic domain (detection and determination in various matrix). The CRA-W took part in meetings and in a collaborative trial study.1. Meetings
A plenary meeting was organised in Rome at 20 and 21 june 2002. During this meeting the progress of methods development and within-lab validation was discussed.
The CRA-W participed at the mid-term meeting in Rennes (19-21 May 2003). Results of method development and between-lab validation were presented and discussed in details.
CRA-W was among participants of the Lisbon meeting (4-6 february 2004). During this meeting, the progress of method development and within-lab validation was rediscussed. The candidate methods for the control strategy were selected.
2. Collaborative trial study
CRA-W took part in a SIMBAG-FEED collaborative trial study on animal feedingstuffs: simultaneous determination of carbadox and olaquindox content using HPLC method. The workpackage leader is Istituto Superiore di Sanità, Lab. Med.Veterinaria (ISS) of Roma in Italy.
The HPLC method for olaquindox and carbadox was reproduced by the CRA-W laboratory. For carbadox, we detected matrix interferences in specific feeds. The use of HPLC-DAD is a very important factor for the HPLC analysis of carbadox.
The detection limits of the method have been confirmed by the CRA-W to be better than 3 mg/kg and 4 mg/kg respectivery for olaquindox and carbadox (in relation with contains formerly authorised in the Directive 70/524/CEE). The limits of quantification were 0.08 and 0.83 mg/kg respectivery for olaquindox and carbadox.
In control strategy to detect banned growth promoters in feed, The CRA-W suggests that the use of UV detection is suitable as a screening method. On the other hand, the use of DAD detection is suitable as a confirmatory method.
Partners
The SIMBAG-FEED project includes 10 partners (Official laboratories and institutes, Research centres, Universities) from 8 European countries involved in the feedingstuffs control. The project is co-ordinated by the Institute of Food Safety of Wageningen in Netherlands.The CRA-W participe in Workpackage working on HPLC method for the carbadox and the olaquindox determination. In this domain, the main partners are:
- Gianfranco Brambilla, ISS, Roma, Italy,
- Christophe Genouel, DGCCRF, Rennes, France,
- Erik Nordkvist, SVA, Uppsala, Sweden
CRAW off coordinator
De Jong JacobInstitute of food Safety - RIKILT
P.O. Box 230
NL-6700 AE Wageningen
Website http://workplace.wur.nl/simbag-feed
Funding
- European Commission
