1. Raw Materials Sourcing and Bone Gelatin Manufacturing Practices in Europe Presentation to the FDA TSE Advisory Committee July 17, 2003 by Reinhard Schrieber Presentation to the FDA TSE Advisory Committee July 17, 2003 by Reinhard Schrieber

2. TSEAC July 17, 2003 No.2 Sourcing of Bovine Bones (1) GME members have taken voluntary steps to ensure the safety of raw materials sourced from European countries Since long before the emergence of BSE, GME members have used only raw materials from healthy slaughtered animals that are released for human consumption. No bones at all sourced from the UK. 1997: Removal of heads in Europe. 1998: Removal of spinal cord in Europe. 1998: Partial replacement of European bones by imports. 1999: Removal of vertebrae from animals of every age. GME members have taken voluntary steps to ensure the safety of raw materials sourced from European countries Since long before the emergence of BSE, GME members have used only raw materials from healthy slaughtered animals that are released for human consumption. No bones at all sourced from the UK. 1997: Removal of heads in Europe. 1998: Removal of spinal cord in Europe. 1998: Partial replacement of European bones by imports. 1999: Removal of vertebrae from animals of every age.

3. TSEAC July 17, 2003 No.3 Sourcing of Bovine Bones (2) GME members have taken voluntary steps to ensure the safety of raw materials sourced from GBR II countries Only use of raw materials from healthy slaughtered animals, released for human consumption. 1997: Removal of heads. 1998: Removal of spinal cord. 1998: Removal of vertebrae from animals of every age from India, Pakistan and Nigeria. GME members have taken voluntary steps to ensure the safety of raw materials sourced from GBR II countries Only use of raw materials from healthy slaughtered animals, released for human consumption. 1997: Removal of heads. 1998: Removal of spinal cord. 1998: Removal of vertebrae from animals of every age from India, Pakistan and Nigeria.

4. TSEAC July 17, 2003 No.4 Sourcing of Bovine Bones (3) Measures required by European Regulation 1996: No bones at all from the UK. 1999: Implementation of the Edible Gelatin Regulation. 2000: Removal of heads. 2000: Removal of spinal cord. 2001: BSE testing of all cattle older than 30 months. 2001: Removal of vertebrae from animals older than 12 months. Measures required by European Regulation 1996: No bones at all from the UK. 1999: Implementation of the Edible Gelatin Regulation. 2000: Removal of heads. 2000: Removal of spinal cord. 2001: BSE testing of all cattle older than 30 months. 2001: Removal of vertebrae from animals older than 12 months.

5. TSEAC July 17, 2003 No.5 Risk of Bone Contamination (1) Because of the steps taken by the industry there was always a very little chance that BSE infectivity could be present in the raw materials used to produce bovine-origin gelatin. Other more recent controls provide additional safety, for example: –Rapid post mortem BSE testing, –the careful removal of specified risk materials (SRMs) make it almost impossible for highly infective materials to enter the supply chain. Because of the steps taken by the industry there was always a very little chance that BSE infectivity could be present in the raw materials used to produce bovine-origin gelatin. Other more recent controls provide additional safety, for example: –Rapid post mortem BSE testing, –the careful removal of specified risk materials (SRMs) make it almost impossible for highly infective materials to enter the supply chain.

6. TSEAC July 17, 2003 No.6 Risk of Bone Contamination (2) As with any processes and systems, there is a possibility of error. For example –animals with low infectivity may be undetected, –surveillance systems might be non-adequate, –removal of SRMs may not be done perfectly, –infectivity of bone marrow has not been finally clarified. Based on our experience, we believe that those risks are low. As with any processes and systems, there is a possibility of error. For example –animals with low infectivity may be undetected, –surveillance systems might be non-adequate, –removal of SRMs may not be done perfectly, –infectivity of bone marrow has not been finally clarified. Based on our experience, we believe that those risks are low.

7. TSEAC July 17, 2003 No.7 Comparison of actual Raw Material Risks and the GME Study Design To account for the potential risk presented by raw materials, GME has studied the effectiveness of the gelatin manufacturing process in eliminating BSE infectivity by assuming that the raw materials are highly contaminated. The tests have been done with a level of infectivity at least 10,000 times higher than could theoretically have been possible in practice. –The tests have assumed that ALL animals used were clinically infective. –The tests have assumed that the bones from all animals contain the full quantity of infective spinal cord and dorsal root ganglia. To account for the potential risk presented by raw materials, GME has studied the effectiveness of the gelatin manufacturing process in eliminating BSE infectivity by assuming that the raw materials are highly contaminated. The tests have been done with a level of infectivity at least 10,000 times higher than could theoretically have been possible in practice. –The tests have assumed that ALL animals used were clinically infective. –The tests have assumed that the bones from all animals contain the full quantity of infective spinal cord and dorsal root ganglia.

8. TSEAC July 17, 2003 No.8 The Production Process of Bone Gelatin in Europe Bone transport, inspection, degreasing, drying and storage Demineralization Acid / alkaline pre-treatment Washing and extraction Filtration, demineralization and concentration Sterilization Drying, testing, storage, blending, testing and shipping All plants are ISO 9000 certified and have a HACCP system in place. They have been FDA inspected as well. All relevant production parameters of the processes have been validated in our plants against the GME study conditions by the independent “SGS European Quality Certification Institute” Bone transport, inspection, degreasing, drying and storage Demineralization Acid / alkaline pre-treatment Washing and extraction Filtration, demineralization and concentration Sterilization Drying, testing, storage, blending, testing and shipping All plants are ISO 9000 certified and have a HACCP system in place. They have been FDA inspected as well. All relevant production parameters of the processes have been validated in our plants against the GME study conditions by the independent “SGS European Quality Certification Institute”

9. TSEAC July 17, 2003 No.9 From Raw Bone to Bone Chips (Degreasing in Europe) The fresh bones are collected from the meat processors. The only bones collected are those from healthy slaughtered animals released for human consumption following ante and post mortem inspection. Before processing, the bones are inspected for foreign materials (also SRM) on sorting belts. The bones are crushed (max. 5/8 inch) and in a continuous process degreased by hot water (appr. 185 o F for appr. 20 minutes with high agitation). The solid bone particles are separated, dried with hot air (the surface temperature will remain below 150 o F), sieved to remove fine materials and stored in silos. The fresh bones are collected from the meat processors. The only bones collected are those from healthy slaughtered animals released for human consumption following ante and post mortem inspection. Before processing, the bones are inspected for foreign materials (also SRM) on sorting belts. The bones are crushed (max. 5/8 inch) and in a continuous process degreased by hot water (appr. 185 o F for appr. 20 minutes with high agitation). The solid bone particles are separated, dried with hot air (the surface temperature will remain below 150 o F), sieved to remove fine materials and stored in silos.

10. TSEAC July 17, 2003 No.10 From Bone Chips to Ossein (Demineralization) The bone chips, produced in Europe or imported, are treated with diluted hydrochloric acid (min. 4 %) in a countercurrent system for 4 days minimum. The phosphate of the bones dissolves and only the protein matrix, called ossein, remains. The ossein is washed and then ready for further pre- treatment with either more acid or alkaline (lime or caustic). The procedure is the same as in the US. The bone chips, produced in Europe or imported, are treated with diluted hydrochloric acid (min. 4 %) in a countercurrent system for 4 days minimum. The phosphate of the bones dissolves and only the protein matrix, called ossein, remains. The ossein is washed and then ready for further pre- treatment with either more acid or alkaline (lime or caustic). The procedure is the same as in the US.

11. TSEAC July 17, 2003 No.11 From Demineralization to Extraction (1) (Pre-treatment) Two variations of this process are commonly used 1.The wet ossein is directly treated with sulfuric acid for another 24 hours and then prepared for low pH acid extraction. (This process is applied to 2 - 3% of the production for special gelatin capsules). 2.The wet ossein is treated for at least 20 days with over- saturated lime solution at appr. pH 12,5. It is then washed and prepared for extraction at appr. neutral pH. (This process is the standard for more than 95% of the gelatin). These manufacturing conditions are applied in Europe and the US. Two variations of this process are commonly used 1.The wet ossein is directly treated with sulfuric acid for another 24 hours and then prepared for low pH acid extraction. (This process is applied to 2 - 3% of the production for special gelatin capsules). 2.The wet ossein is treated for at least 20 days with over- saturated lime solution at appr. pH 12,5. It is then washed and prepared for extraction at appr. neutral pH. (This process is the standard for more than 95% of the gelatin). These manufacturing conditions are applied in Europe and the US.

12. TSEAC July 17, 2003 No.12 From Demineralization to Extraction (2) (Pre-treatment) Another process variation under review and not in common use The wet ossein is kept after the addition of 0.3 M NaOH for a minimum of 2 hours at a pH of 13.0 and then washed, again treated with sulfuric acid for 24 hours and prepared for low pH acid extraction. (The interest in and potential applications for this type of product are under review). Another process variation under review and not in common use The wet ossein is kept after the addition of 0.3 M NaOH for a minimum of 2 hours at a pH of 13.0 and then washed, again treated with sulfuric acid for 24 hours and prepared for low pH acid extraction. (The interest in and potential applications for this type of product are under review).

13. TSEAC July 17, 2003 No.13 From Ossein to Gelatin (Washing and Extraction) After pre-treatment the ossein is washed and the pH adjusted to the desired extraction value. Extraction is done stepwise by using 4 to 6 times new hot water with increasing temperature starting from about 125 o F up to 200 o F. Each time the length of stay is about 4 hours. The different extracts have different physical properties The intended use of each extract of one production day might be different (photo, pharma, food). All gelatins comply with the regulatory requirements for food and pharma. Same procedures in Europe and the US. After pre-treatment the ossein is washed and the pH adjusted to the desired extraction value. Extraction is done stepwise by using 4 to 6 times new hot water with increasing temperature starting from about 125 o F up to 200 o F. Each time the length of stay is about 4 hours. The different extracts have different physical properties The intended use of each extract of one production day might be different (photo, pharma, food). All gelatins comply with the regulatory requirements for food and pharma. Same procedures in Europe and the US.

14. TSEAC July 17, 2003 No.14 The Purification of Gelatin (Filtration, Demineralization, Concentration) The diluted (3 - 5 %) gelatin solution is filtered by different types of centrifuges, cellulose and/or diatomaceous earth filters. Mineral salts, anion and cation, remaining from the pre- treatments of the ossein are removed by ion-exchange columns. Afterwards the gelatin solution is concentrated by ultrafiltration and/or vacuum evaporation. Same procedures and equipment as in the US. The diluted (3 - 5 %) gelatin solution is filtered by different types of centrifuges, cellulose and/or diatomaceous earth filters. Mineral salts, anion and cation, remaining from the pre- treatments of the ossein are removed by ion-exchange columns. Afterwards the gelatin solution is concentrated by ultrafiltration and/or vacuum evaporation. Same procedures and equipment as in the US.

15. TSEAC July 17, 2003 No.15 The Sterilization of Gelatin (UHT Treatment) The concentrated gelatin solution (25 to 45 % gelatin) is sterilized by direct steam injection. The temperature under pressure in the liquid phase is min. 280 o F for at least 4 seconds. After expansion the temperature drops back to about 130 o F. Same procedures and equipment as in the US. The concentrated gelatin solution (25 to 45 % gelatin) is sterilized by direct steam injection. The temperature under pressure in the liquid phase is min. 280 o F for at least 4 seconds. After expansion the temperature drops back to about 130 o F. Same procedures and equipment as in the US.

16. TSEAC July 17, 2003 No.16 Final Procedures (Drying, Testing, Blending, Testing, Shipping) Finally the sterilized gelatin solution is chilled to set and then dried with purified and conditioned air on belt dryers. Each production batch (single extract) is tested for physical, chemical and bacteriological properties. Different production batches with different properties are dry blended according to customer specification. The final blends are again tested for physical, chemical and bacteriological compliance with regulatory and customer requirements. After testing, the released gelatin is shipped. Same procedures and equipment as in the US. Finally the sterilized gelatin solution is chilled to set and then dried with purified and conditioned air on belt dryers. Each production batch (single extract) is tested for physical, chemical and bacteriological properties. Different production batches with different properties are dry blended according to customer specification. The final blends are again tested for physical, chemical and bacteriological compliance with regulatory and customer requirements. After testing, the released gelatin is shipped. Same procedures and equipment as in the US.

17. TSEAC July 17, 2003 No.17 Heat and Pressure Process ( a Special Case for Low Gelling Gelatin) Done by only one company in Europe Degreasing like standard gelatin. 1. Autoclaving the bone chips (>=270 o F, >3 bar, >=20 minutes). Extraction with hot water. 2. - 7. Autoclaving at lower temperature and shorter time. Collection of different extracts. Flocculation, ion-exchange, evaporation. Drying, testing, blending, testing, shipping. Done by only one company in Europe Degreasing like standard gelatin. 1. Autoclaving the bone chips (>=270 o F, >3 bar, >=20 minutes). Extraction with hot water. 2. - 7. Autoclaving at lower temperature and shorter time. Collection of different extracts. Flocculation, ion-exchange, evaporation. Drying, testing, blending, testing, shipping.

18. TSEAC July 17, 2003 No.18 Conclusions (1) This review of the gelatin manufacturing process is intended to show the commercial minimum manufacturing conditions reflected by the GME study. The study design has been validated against the actual process parameters used at GME plants. The study demonstrates the ability of the gelatin manufacturing process to remove and inactivate infectivity - even under conditions in which the raw materials contain unrealistically high infectivity levels. This review of the gelatin manufacturing process is intended to show the commercial minimum manufacturing conditions reflected by the GME study. The study design has been validated against the actual process parameters used at GME plants. The study demonstrates the ability of the gelatin manufacturing process to remove and inactivate infectivity - even under conditions in which the raw materials contain unrealistically high infectivity levels.

19. TSEAC July 17, 2003 No.19 Conclusions (2) The safety of European bovine bone gelatin is established on two principles: –The safety of the raw materials (as required by GME practices and EU law) and, –the safety of the manufacturing process (as demonstrated by the GME study). The Scientific Steering Committee of the European Union has concluded, based on these principles, in its “Opinion on the safety of gelatin” that the “risk is close to zero”. The safety of European bovine bone gelatin is established on two principles: –The safety of the raw materials (as required by GME practices and EU law) and, –the safety of the manufacturing process (as demonstrated by the GME study). The Scientific Steering Committee of the European Union has concluded, based on these principles, in its “Opinion on the safety of gelatin” that the “risk is close to zero”.