Bovine Bone Sourcing Practices and Bone Gelatin Manufacturing Processes in the United States FDA TSE ADVISORY COMMITTEE MEETING July 17, 2003 J. Michael Dunn, Ph.D Vice President, Regulatory Affairs, GELITA North America Chairman, Regulatory Committee Gelatin Manufacturers Institute of America (GMIA)

US BONE GELATIN MANUFACTURERS Eastman Gelatine Corp. Peabody, MA GELITA USA Inc. Sioux City, IA US manufacturers produce primarily limed bone gelatin (Type B) and to some extent acid bone gelatin (Type A) The sourcing practices and processing conditions discussed today will be representative of both US manufacturers

BOVINE BONE SOURCING PRACTICES AND BONE GELATIN MANUFACTURING PROCESSES IN THE US Overall Objectives Adequately describe the bovine bone sourcing practices and processing conditions for the manufacturing of bone gelatin in the US Confirm that the bone gelatin processing conditions employed in the US are: Virtually the same as those used in the EU Meet or exceed all of the minimum processing requirements as specified in the GME TSE Inactivation Study Protocol

BOVINE BONE SOURCING PRACTICES Degreased gelatin bone is sourced exclusively from USDA inspected beef processing facilities in the US This raw material is derived solely from healthy cattle that have been deemed fit for human consumption based upon both ante and postmortem inspections

BOVINE BONE SOURCING PRACTICES SRM’s All US gelatin bone suppliers have been removing Specified Risk Materials (SRM’s) with the exception of vertebrae since as early as 1998 Limited quantities of vertebrae-free gelatin bone have been available since the Fall of 2002 Currently, there are no FDA or USDA requirements for the removal of SRM’s in the US

GELATIN MANUFACTURING PROCESS OVERVIEW Daily Bone Gelatin Production Degreased Gel Bone HCl Acid Water Labor Gelatin Process Dicalcium Phosphate Gelatin 6

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Overall Objectives Hydrolyze collagen Hydrolyze sufficient cross-links and peptide bonds within the collagen matrix to render the hydrolyzed collagen (gelatin) highly soluble in water Purify the gelatin Employ appropriate processing steps to ensure its physical, chemical and microbiological purity Concentrate the finished product Use additional processes to remove water from the gelatin liquor to produce a dry, stable product Acidification Washing Extraction Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Incoming Gel Bone Acidification Washing Extraction Initial Filtration Typical Characteristics Fat content = 1 to 2.5% Size range = 1/8 to 5/8” Mineral/protein ratio = 1.9 - 2.1 Moisture content = 6 to 9% In contrast, EU gelatin producers have their own degreasing facilities Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Acidulation Acidification Washing Extraction Objectives / Conditions Produce ossein by removing the mineral content of the bone Initiate the hydrolysis of collagen Remove non-collagen impurities HCL concentration: 4-6% Acidulation time: 5-7 days (counter-current process) Acidulation temperature: 50° to 65° F Washing: Rinse up to 24 hours to remove acid salts, fat and other impurities Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Liming / Type B Acidification Washing Objectives / Conditions Continue to hydrolyze collagen Continue to remove non-collagen impurities Extraction Initial Filtration Ion Exchange Initial Concentration Final Filtration Convert asparagine and glutamine to their respective acids (pI decreases) Lime slurry concentration: Over saturated Lime slurry pH: Approximately 12.5 Liming time: 25-70 days Liming temperature: 50-70° F Agitation: Daily Lime slurry changes: Weekly pH Adjustment Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing / Acidification Washing Acid Treatment Acidification Washing Objectives / Conditions Remove and neutralize excess lime Remove non-collagen impurities Extraction Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Adjust pH of the ossein slurry Wash time: 24 to 48 hours Agitation: Vigorous Water temperature: 45° to 70° F Neutralizing acids: Hydrochloric & sulfuric Target pH: 5 to 7 Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Acid Treatment / Type A Acidification Washing Extraction Objective / Conditions Condition ossein for extraction at low pH Traditional Treatment: Sulfuric acid at pH = 1.0 - 2.0 Treatment time: = 6 hours Washing: Rinse up to 24 hours to a pH of 2.8 - 3.2 Alkaline pre-treatment option (under review) pH is maintained at 13 or greater with NaOH for 3 hours (prior to acid treatment) Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration > Sterilization Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Gelatin Extraction Acidification Washing Objective / Conditions Solubilize hydrolyzed collagen (gelatin) from the ossein Extraction Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Water purity: Demineralized Procedure: Series of extractions with increasing water temperatures Water temperatures: 120° to 200° F Conditioning time: 1 to 6 hours / extraction Number of extractions: 4 to 6 Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Initial Filtration Acidification Washing Extraction Objective / Conditions Clarify dilute gelatin solution Protect ion exchange system Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Vertical leaf pressure filter Pre-coated with diatomaceous earth and cellulose Gelatin concentration: 4-6% Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Ion Exchange Acidification Washing Extraction Initial Filtration Objective / Conditions Deionize the thin gelatin liquor Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Cations removed: Ca+2, Mg+2 and Fe+2 Anions removed: Cl- or S04= Ash (product): 0.1 to 1% Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Initial Concentration Acidification Washing Extraction Initial Filtration Objective / Conditions Concentrate the thin gelatin liquor Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Evaporator type: Multiple effect rising film or plate and frame Concentration target: 15-25% Output temperature: 115° F to 150° F Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Final Filtration Acidification Washing Extraction Objectives / Conditions Clarify concentrated gelatin solution Remove additional coagulated protein and particulate Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Chilled Extrusion Plate and flame pressure filters Cellulose / diatomaceous earth filter pads Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment pH Adjustment Acidification Washing Extraction Initial Filtration Objective / Conditions Fine adjustment of the finished product pH Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Chilled Extrusion Final pH targets: 5-7 Caustic adjustment: NaOH Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Final Concentration Acidification Washing Extraction Initial Filtration Objective / Conditions Concentrate the thick gelatin liquor Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Evaporator type: Thin film / wiped surface or plate and frame multiple effect Output temperature: 115° to 125° F Concentration target: 25 to 50% (viscosity dependent) Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Sterilization Acidification Washing Extraction Initial Filtration Objective / Conditions Ensure microbiological purity / product hygiene Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Equipment: Direct steam injection / HTST Temperature: 138° to 149° C Time: 8 to 16 seconds Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Chilled Extrusion Acidification Washing Extraction Initial Filtration Objective / Conditions Increase the surface area of the gel mass Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Glycol - cooled heat exchanger / extruder Temperature: Approx. 70° F Noodle dimensions: 1’ to 2’ long / 1/8” thick Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Drying Acidification Washing Extraction Objective / Conditions Produce a dry, stable product Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Air quality: Heated, dehumidified and filtered Number of zones: 10 to 12 Temperature gradient: 80° to 160° F Time: 1 - 3 hours Moisture content (gelatin): 10 to 12% Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Milling and Screening Acidification Washing Extraction Objective / Conditions Grind gelatin to a uniform size Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Equipment: Cage mill Particle size: 8 to 30 mesh Chilled Extrusion Drying Milling/Screening Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Washing Acid Treatment Blending Acidification Washing Extraction Blender Initial Filtration Drumming and weighing Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening QC/QA Analysis Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option High Capacity Blender Washing Acid Treatment Formulation Acidification Washing Extraction Component Selection Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening QC/QA Analysis Blending Formulation Final Packaging

GELATIN MANUFACTURING PROCESS Initial Concentration Bone /Type B Bone/Type A GELATIN MANUFACTURING PROCESS Acidulation Acidulation Lime Treatment NaOH Option Final Packaging Washing Acid Treatment FIBCs Acidification Washing Drums Extraction Initial Filtration Ion Exchange Initial Concentration Final Filtration pH Adjustment Final Concentration Sterilization Chilled Extrusion Drying Milling/Screening Small Bags Blending Formulation Final Packaging