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LNR NRL NRL - Favv

LNR NRL NRL - Favv

Analysis of

Analysis of glyceroltriheptanoate in animal by-products In the nineties it was shown that certain transferable animal diseases (e.g. TSE: transferable spongiform encephalopathy) can be spread by animal by-products. Moreover, cannibalism (feeding proteins from cadavers of the same species) could entail an additional risk for spreading diseases. To prevent animal by-products that are not destined for human consumption from re-entering the food chain, Regulation 1774/2002 (European Communities, 2002) was drawn up. According to Regulation 1774/2002 animal by-products are divided into three categories: category 1 (high risk), category 2 (average risk) and category 3 (low risk). At the request of the DG Health and Consumer Protection (DG SANCO) the Joint Research Centre Institute for Reference Materials and Measurements of the European Commission (IRMM) has evaluated the characteristics of the different substances that are suitable for use as marker for animal by-products of category 1 and category 2. In 2002 the IRMM proposed to use glyceroltriheptanoate (GTH) as a marker. After all, GTH proved to be the ideal candidate. GTH is a fat that consists of 3 C7-fatty acids (n-heptane acid) that have been esterifised with glycerol. Therefore, GTH does not appear in nature. The structural formula is illustrated in figure 1. The physico-chemical characteristics of GTH make sure it mixes well with fats and products containing fat. Moreover, GTH cannot be removed after it has been added. It is a safe and non-toxic product that is already used in the food industry for other applications, such as the marking of butter. Apart from that, GTH is a very stable substance that can resist heat treatment under steam pressure at 133°C, as it is applied in the fat rendering industry. GTH is commercially available as a clear transparent fluid. The price is low and not a lot needs to be added (legal requirement: 250 mg/ kg fat). After all, there is a sensitive and simple method to analyse GTH. Figure 1: Structural formula of glyceroltriheptanoate 4

The analysis method was developed by the IRMM and is suitable for detecting GTH in animal meal and animal fats. The method consists of three basic steps: (i) an extraction of the fat fraction with petroleum ether (only for animal meal) (ii) purification of the extracted fat (for animal meal) or of the animal fat (for fat samples) and (iii) determination of the GTH concentration by gas chromatography. Two detectors can be used for detection, the Flame Ionisation Detector (FID) or Mass Spectrometer (MS). FID is only suitable for quantification of GTH in the target materials (category 1 and category 2 animal meal and animal fat). With MS, however, the absence of GTH in category 3 animal by-products can also be shown. The in-house validation of the IRMM, in which several performance characteristics (such as robustness, sensitivity, precision (repeatability and reproducibility) and accuracy) were determined, indicated that this method is suitable for the intended purpose. After the analysis method for GTH determination had been fine-tuned, an implementation study was carried out in cooperation with different rendering plants in Europe. The objective of this study was to check whether the application of GTH as marker in category 1 and category 2 animal by-products is practically feasible under realistic circumstances. The study was carried out by the IRMM and CCL Research (Cooperative Central Laboratory) in close cooperation with DG SANCO and EFRA (European Fat Processors and Renderers Association). Every rendering plant showed high GTH levels in the end products (animal meal and fat). GTH levels were well over the demonstrability limit. These results indicate that GTH can be completely mixed with fat, and this for both marking animal meal and fat. A stability study also showed that GTH is stable in animal meal at room temperature during at least 58 weeks. Therefore, the results of the implementation study make it possible to conclude that GTH is a suitable and a practically usable marker for category 1 and category 2 materials. On the basis of this study report, the European Commission approved the use of the GTH marker system on September 11, 2007 (European Commission, 2007). Finally, on December 2007 Regulation 1432/2007 was published (European Union, 2007), which came into force on July 1, 2008. According to this regulation, category 1 and category 2 animal by-products have to be marked permanently with GTH. Marking such materials insures the identification and traceability of products that have to be destroyed and that by no means can end up in the food chain again. The risk of fraude is also eliminated. Adding GTH needs to be done in such a way that the processed products contain a minimum concentration of at least 250 mg per kg of fat. Moreover, GTH needs to be homogeneously distributed in the substance and has to be added after the products have undergone a prior sanitising heat treatment at a core temperature of at least 80°C. After all, experiments at laboratory scale have indicated that GTH is not stable in raw animal by-products. GTH is cleaved by gut enzymes. These enzymes, however, turn inactive as of the moment when the animal by-products reach a temperature of 80°C. Therefore, GTH is not added in slaughterhouses, but in the processing companies. Figure 2 contains an example of a fat rendering process. The moment of GTH addition is indicated with the little pipette. 5

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