Agency Response Letter GRAS Notice No. GRN 000632
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See also Generally Recognized as Safe (GRAS).
CFSAN/Office of Food Additive Safety
June 24, 2016
James La Marta, Ph.D.
DSM Nutritional Products
45 Waterview Boulevard
Parsippany, NJ 07054
Re: GRAS Notice No. GRN 000632
Dear Dr. La Marta:
The Food and Drug Administration (FDA) is responding to the notice, dated February 16, 2016, that you submitted in accordance with the agency’s proposed regulation, proposed 21 CFR 170.36 (62 FR 18938; April 17, 1997; Substances Generally Recognized as Safe (GRAS); the GRAS proposal). FDA received the notice on February 18, 2016, filed it on March 18, 2016, and designated it as GRAS Notice No. GRN 000632. FDA received correspondence on June 16, 2016 that includes revisions to the estimated dietary exposure presented in GRN 000632.
The subject of the notice is purified rebaudioside A from Yarrowia lipolytica (rebaudioside A). The notice informs FDA of the view of DSM that rebaudioside A is GRAS, through scientific procedures, for use as a general-purpose sweetener in foods, excluding infant formula and meat and poultry products, at levels determined by good manufacturing practices (GMP), as well as use as a table top sweetener.
Our use of “rebaudioside A,” “steviol glycosides,” or “SGs” in this letter should not be considered an endorsement or recommendation of that term as an appropriate common or usual name for the purpose of declaring the substance in the ingredient statement of foods that contain that ingredient. Title 21 CFR 101.4 states that each ingredient must be declared by its common or usual name. In addition, 21 CFR 102.5 outlines general principles to use when establishing common or usual names for nonstandardized foods. Issues associated with labeling and the appropriate common or usual name of a food are the responsibility of the Office of Nutrition and Food Labeling in the Center for Food Safety and Applied Nutrition.
As part of its notice, DSM includes the statement of a panel of individuals (DSM’s GRAS panel) that evaluated the data and information that are the basis for DSM’s GRAS determination. DSM considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. DSM’s GRAS panel evaluated information describing the identity and composition, manufacturing process and specifications, and estimated dietary exposure to rebaudioside A, as well as published and unpublished studies supporting the safety of rebaudioside A. Based on this review, DSM’s GRAS panel concludes that rebaudioside A produced in accordance with GMP and meeting appropriate purity and food grade specifications, is GRAS, by scientific procedures, under the conditions of its intended use.
DSM provides information about the identity and composition of rebaudioside A. Rebaudioside A (CAS Reg. No. 58543-16-1), a glycoside of steviol, is identified as 19-O-β-glucopyranosyl-13-O-(β-glucopyranosyl-(1-2)-β-glucopyranosyl(1-3))-β-glucopyranosyl-13-hydroxykaur-16-en-19-oic acid. Rebaudioside A is one of a group of known steviol glycosides (SGs), which differ from each other by the number of glycoside moieties and bonding order.
DSM provides information about the manufacturing process for rebaudioside A. Rebaudioside A is obtained from Y. lipolytica that is engineered to produce SGs. DSM provides information on the parent Y. lipolytica strain and describes the production strain and the genes used to express the proteins that encompass the production pathway of SGs (e.g., rebaudioside A). DSM states that the production strain is neither toxigenic nor pathogenic and that Y. lipolytica has a history of safe use as a production source for food ingredients. DSM states that the production strain contains no residual antibiotic resistance genes and is susceptible to antifungal agents. DSM uses a controlled, submerged, aerobic fed batch fermentation process with the Y. lipolytica production strain. DSM states that rebaudioside A is biosynthesized and excreted during fermentation. Following fermentation, the production organism is removed by centrifugation and the supernatant is heat treated to inactivate any remaining microorganisms. The supernatant is subsequently clarified by centrifugation or filtration. The supernatant is subjected to adsorption chromatography, and the SGs are eluted with aqueous ethanol. The eluate is decolorized using activated carbon and subjected to ion exchange resins. The resulting solution is concentrated to produce a dry powder and subjected to recrystallization to obtain rebaudioside A, which is then dried to form a powder.
DSM provides specifications for rebaudioside A that include the minimum content of rebaudioside A (≥ 95%) and total SGs (> 95%). Specifications also include limits for moisture (≤ 6%), ash (≤ 1%), lead (≤ 1 milligram per kilogram (mg/kg)), arsenic (≤ 1 mg/kg), ethanol (≤ 0.5%), and limits on microbial contaminants. DSM states that rebaudioside A meets the specifications for SGs established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) at its 73rd meeting in June 2010. DSM provides the results of six batch analyses to demonstrate that rebaudioside A meets specifications.
DSM provides estimates of dietary exposure to rebaudioside A resulting from its intended use in food. DSM discusses dietary exposure data for sweetener use from a published study (Ref. 1) in which the author estimated dietary exposure to rebaudioside A in different subpopulations based on its relative sweetness intensity compared to sucrose. DSM states that rebaudioside A has a sweetness that is 200-300 times that of sucrose. DSM reports that the mean dietary exposure to rebaudioside A, on a steviol equivalents basis, ranges from 0.7 to 1.1 mg/kg body weight (bw)/day for children and 0.4 to 0.5 mg/kg bw/day for adults. DSM reports that the upper percentile (> 90th percentile) dietary exposure, on a steviol equivalents basis, ranges from 1.5 to 1.7 mg/kg bw/day for children and 1.1 to 1.5 mg/kg bw/day for adults.
DSM summarizes published studies pertaining to the metabolic fate of rebaudioside A and other SGs. DSM points to previously reviewed in vitro and in vivo mutagenicity/genotoxicity studies, as well as recently completed unpublished genotoxicity studies on their own rebaudioside A product. DSM also summarizes published acute, subchronic, and chronic toxicity studies, in addition to recently completed unpublished subchronic studies on their own rebaudioside A product. The recently completed unpublished studies on their own rebaudioside A product demonstrated the exact same outcome as previously published studies. DSM also discusses published reproductive and developmental toxicology studies in relation to their safety determination for rebaudioside A. To further support its view that rebaudioside A is GRAS for the intended use, DSM summarizes recent decisions by JECFA and other governmental bodies on the safety of rebaudioside A and other SGs for use in food as a sweetener. DSM notes that JECFA has established an acceptable daily intake (ADI) for SGs of 4 mg/kg bw/d (expressed as steviol). This ADI was based on a no observed adverse effect level of 970 mg/kg bw/d (383 mg/kg bw/d, as steviol) from a two year rat study, and the application of a one hundred-fold safety factor for intra- and inter-species differences. Based on all the available scientific information, DSM concludes that rebaudioside A is GRAS for its intended use in foods.
Standards of Identity
In the notice, DSM states its intention to use rebaudioside A in several food categories, including foods for which standards of identity exist, located in Title 21 of the Code of Federal Regulations. We note that an ingredient that is lawfully added to food products may be used in a standardized food only if it is permitted by the applicable standard of identity.
Section 301(ll) of the Federal Food, Drug, and Cosmetic Act (FD&C Act)
Section 301(ll) of the FD&C Act prohibits the introduction or delivery for introduction into interstate commerce of any food that contains a drug approved under section 505 of the FD&C Act, a biological product licensed under section 351 of the Public Health Service Act, or a drug or a biological product for which substantial clinical investigations have been instituted and their existence made public, unless one of the exemptions in section 301(ll)(1)-(4) applies. In its review of DSM’s notice that rebaudioside A is GRAS for the intended use, FDA did not consider whether section 301(ll) or any of its exemptions apply to foods containing rebaudioside A. Accordingly, this response should not be construed to be a statement that foods that contain rebaudioside A, if introduced or delivered for introduction into interstate commerce, would not violate section 301(ll).
Based on the information provided by DSM, as well as other information available to FDA, the agency has no questions at this time regarding DSM’s conclusion that rebaudioside A is GRAS under the intended conditions of uses. The agency has not, however, made its own determination regarding the GRAS status of the subject uses of rebaudioside A. As always, it is the continuing responsibility of DSM to ensure that food ingredients that the firm markets are safe, and are otherwise in compliance with all applicable legal and regulatory requirements.
In accordance with proposed 21 CFR 170.36(f), a copy of the text of this letter responding to GRN 00 0632, as well as a copy of the information in this notice that conforms to the information in the GRAS exemption claim (proposed 21 CFR 170.36(c)(1)), is available for public review and copying at www.fda.gov/grasnoticeinventory.
Dennis M. Keefe, Ph.D.
Office of Food Additive Safety
Center for Food Safety and Applied Nutrition
1. Renwick, A.G. 2008. The use of a sweetener substitution method to predict dietary exposures for the intense sweetener rebaudioside A. Food and Chemical Toxicology 46:S61–S69.
 DSM uses de novo synthesized genes of the steviol glycoside metabolic pathway from Stevia rebaudiana (Bertoni) Bertoni for the production of steviol glycosides. These genes have been codon optimized to Y. lipolytica preferences.