1. Substance Structure, production process and indented use Is there adequate published safety data and exposure data for this substance to meet current GRAS standards? Insufficient Data to Proceed NO YES Food Substance Decision Tree for Independent GRAS Determinations Assumptions Animal & Human Studies Conducted and Published Identify deficiencies Conduct Studies & Evaluate Data Make data publically available New to U.S. Food Market History of dietary exposure in other countries Assumptions: Substance has existing safety data in public literature Dietary exposure without previous FDA review/approval Approach: May require additional safety testing and publication of study results in support of GRAS status of proposed use (see FDA Redbook for testing guidelines) Must account for cumulative exposure of intended uses in safety assessment due to increase in dietary exposure Clinical studies may be required Safety concerns to consider are dependant on category of independent GRAS determination I Forgot what wording to put here…… Modified Production Manufacturing modification for currently FDA-approved or GRAS substances Assumptions: Substance has current FDA approval or GRAS status for use in food Substance has existing and adequate safety data in the public literature Approach: May require additional safety testing and publication of study results in support of GRAS status of proposed use (see FDA Redbook for testing guidelines) Extension of Food Use Extension of food uses for currently FDA-approved or GRAS substances Assumptions: Substance has current FDA approval or GRAS status for use in food Substance has existing and adequate safety data in the public literature Intended food uses may change with no increase in dietary exposure Approach: Perform/revise exposure estimates

2. ProteinsEnzyme Preparations Has the source of the enzyme or other proteins been well characterized and described? Potential for allergic response? Identities and levels of diluents, stabilizers, preservatives, and any other substances used in the enzyme/protein formulation Information on secondary enzymes/proteins derived from the production organism(s) that may be present in the enzyme/protein preparation. The content of total organic solids (TOS) for both the commercial enzyme/protein preparation and the enzyme/protein batch used in toxicology studies Does the enzyme/protein preparation conform to specifications for purity provided in the most recent relevant edition of the Food Chemicals Codex? ChemicalsLipids Extracts In the case that additional testing is required in order to achieve GRAS status the following considerations may be addressed General Testing Considerations ●Single Dose-animals & human ●Comparative biodisposition ● Repeated doses ● Subchronic dosing ● Genetic Toxicity ●Reproductive /developmental toxicity studies● Adequate for ADI * May refer to FDA Redbook for testing guidelines Carbohydrates Determine Natural Occurrence and Presence in the Human Diet Define Any History of Use and Consumption in U.S. Population Determine Current Regulatory Status of the Purified Carbohydrate in the USA and Identify Any Prior GRAS determinations for Its Use in Food in the Publicly Available Literature Develop Detailed Specifications/Description of the Purified Carbohydrate Prepare Full Chemical Characterization of the Purified Carbohydrate Provide Complete Manufacturing Process Description (Demonstration of Consistency/Reproducibility in Manufacturing Process)