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Development and validation of a mass spectrometric method for the detection of forbidden animal proteins in feeding stuff
Project
Project code: BfR-LMS-08-1322-627
Contract period: 01.01.2015
- 31.12.2015
Purpose of research: Applied research
In the framework of the combat against BSE, complex legal rules have been endorsed in the European Union which prohibit the feeding of processed animal protein (PAP) - apart from e.g. egg or milk protein - to farmed animals (Regulation (EC) No. 999/2001). Irrespective of few exemptions, the highest priority is still pending on PAP derived from ruminants as possible carriers of transmissible spongiform encephalopathies (TSE). The traceability of forbidden animal compounds signifies a big challenge since the obligatory heat- and pressure treatment imposed on PAP (133 oC, 300 kPa, 20 min) destroys proteins and nucleic acids as potential analytical targets to a considerable degree. For this reason a pragmatic qualitative method based on light microscopy has been established, which detects smallest animal fragments like bone spicules, gills, feathers a. o. particles. At present this approach is the legally required European standard method (Regulation (EC) No. 152/2001). Following the readmission of non-ruminant PAP in aquaculture (since June 2013) in the EU, a validated real time PCR method for the detection of ruminant DNA was introduced in parallel (Regulation (EU) No. 51/2013). However, both classical PCR as well as light microscopy, exhibit striking analytical gaps. PCR as a DNA-based method will result in false-positive signals with allowed components like milk powder. The microscopist – on the other hand – cannot differentiate animal species merely on the morphology of particles. Immunological approaches so far did not reach the desired sensitivity. As a direct way in protein detection, mass spectroscopic methods could be considered. The state of method development for the scope of investigation is still in the very beginning. It is therefore the aim of the project, to develop and validate a sensitive mass spectrometric method based on the identified heat-stable marker protein myoglobin which shall enable the reliable detection of 0.1 % (w/w) meat and bone meal from beef or sheep origin in feeding stuff.
Section overview
Subjects
- Animal nutrition
- Toxicology
