Agency Response Letter GRAS Notice No. GRN 000375
CFSAN/Office of Food Additive Safety
September 2, 2011
Ashley Roberts, Ph.D.
Cantox Health Science International
An Intertek Company
2233 Argentia Road, Suite 308
Mississauga, Ontario L5N 2X7
Re: GRAS Notice No. GRN 000375
Dear Dr. Roberts:
The Food and Drug Administration (FDA) is responding to the notice, dated February 23, 2011, that you submitted on behalf of Toyo Sugar Refining Co., Ltd and Nippon Paper Chemicals Co., Ltd (Toyo Sugar and Nippon Paper) 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 this notice on March 2, 2011, filed it on March 9, 2011, and designated it as GRN No. 000375.
The subject of the notice is enzyme-modified steviol glycosides (EMSG). EMSG is obtained by enzyme treatment of steviol glycosides (SGs) purified from the leaves of Stevia rebaudiana (Bertoni) Bertoni. The notice informs FDA of the view of Toyo Sugar and Nippon Paper that EMSG is GRAS, through scientific procedures, for use as a general-purpose sweetener in foods, excluding meat and poultry products, at levels determined by good manufacturing practices. Toyo Sugar and Nippon Paper note that purified SGs have recently been the subject of several GRAS notices and that FDA responded to these GRAS notices informing the notifiers that, at the time of its response, the agency had no questions regarding their conclusions that the purified SGs that are the subjects of their respective notices are GRAS for their intended use as a sweetener in food.
Our use of "EMSG," "enzyme-modified steviol glycosides," "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, Labeling, and Dietary Supplements in the Center for Food Safety and Applied Nutrition.
As part of its notice, Toyo Sugar and Nippon Paper include the report of a panel of individuals (Toyo Sugar and Nippon Paper's GRAS panel) who evaluated the data and information that are the basis for Toyo Sugar and Nippon Paper's GRAS determination. Toyo Sugar and Nippon Paper consider the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. Toyo Sugar and Nippon Paper's GRAS panel evaluated the identity, method of manufacture, product specifications, and the potential exposure resulting from the intended uses of EMSG as well as published and unpublished studies related to the safety evaluation of EMSG. Based on this review, Toyo Sugar and Nippon Paper's GRAS panel concluded that EMSG, produced consistent with good manufacturing practice and meeting appropriate purity and food grade specifications, is GRAS under the conditions of its intended use.
Toyo Sugar and Nippon Paper provide information about the identity and composition of EMSG. Toyo Sugar and Nippon Paper describe EMSG as a white to slightly off-white powder composed of >95% SGs, including both the group of structurally-related sweet-tasting compounds that are natural constituents of the stevia leaf, and their 1,4-α-D-glucosylated forms. The α-D-glucosylated SGs constituents are generated by treatment of SGs extract preparations with a food-grade α-glycosyltransferase enzyme in the presence of dextrin. The α-D-glucosylated SGs have glucose side chains that have been extended, generally with 1 to 3 additional glucose moieties, as compared to the naturally occurring SGs. These additional glucose residues are attached by stereo- and regio-specific 1,4-α-D-glycosidic bonds, whereas the glucose in naturally occurring SGs is attached by β-glycosidic bonds.
Toyo Sugar and Nippon Paper provide information about the manufacturing process and specifications for EMSG. The starting material for the manufacturing process is high-purity (≥95%) SGs obtained from the leaves of S. rebaudiana (Bertoni) Bertoni. Toyo Sugar and Nippon Paper state that the SGs starting material meets the specifications for SGs established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) at its 69th meeting in June 2009. In the first step of the manufacturing process, the starting material is dissolved in hot water with a food-grade α-glycosyltransferase enzyme in the presence of dextrin as a glucose source. At this step, a glycosylation reaction takes place adding glucose moieties to the SGs, resulting in the formation of 1,4-α-D-glucosylated SGs constituents in the preparation. The mixture is then subjected to filtration, adsorption and ion-exchange resin (strong acidic and strong basic resins) with ethanol or methanol. The resulting mixture then undergoes concentration, sterilization and spray-drying to yield the final EMSG product. Toyo Sugar and Nippon Paper provide specifications for its EMSG that include the content of total SGs and α-D-glucosylated SGs (>95% by weight (w/w)) and limits for lead (≤1 milligram per kilogram (mg/kg)), arsenic (≤1 mg/kg), residual ethanol (≤5000 mg/kg), residual methanol (≤200 mg/kg), total ash (≤1%) and microbial contaminants (within specified limits).
Toyo Sugar and Nippon Paper estimate the intake of EMSG resulting from its intended use in foods. Toyo Sugar and Nippon Paper utilize the method of estimation reported by Renwick that predicted dietary exposures for the intense sweetener rebaudioside A (Ref. 1). In this study, published data on intake of intense sweeteners were collected from a large number of countries and were converted to sucrose equivalents using estimates of sweetness relative to sucrose. Based on a panel evaluation, Toyo Sugar and Nippon Paper determined the relative sweetness for EMSG to be 137 times that of sucrose. Based on this sweetness intensity factor, the maximum estimated use of EMSG would yield an estimated daily intake (expressed as steviol equivalents) of 2.12 mg/kg body weight per day (mg/kg bw/day).Toyo Sugar and Nippon Paper state that the estimated intake of EMSG in food is below the ADI of 4 mg/kg bw/day, expressed as steviol equivalents, defined by JECFA for steviol glycosides (JECFA, 2009). Toyo Sugar and Nippon Paper state that the use of EMSG in food is self-limiting due to its organoleptic properties.
Toyo Sugar and Nippon Paper discuss the metabolic fate and safety of EMSG. In its discussion, Toyo Sugar and Nippon Paper describe several published studies on the metabolism of SGs. Based on these studies, Toyo Sugar and Nippon Paper conclude the final metabolic fate of EMSG is the same as that of naturally occurring SGs, whose metabolism has been well studied. Toyo Sugar and Nippon Paper state that given this expected metabolic fate, the data and information to support the safety evaluation of SGs is also applicable to the safety evaluation of EMSG. As such, Toyo Sugar and Nippon Paper discuss published and unpublished studies pertaining to the safety evaluation of SGs, including studies on rebaudioside A, stevioside, steviol, and crude stevia extracts. Among the published studies considered were acute toxicity studies in rats, mice, and hamsters, subchronic toxicity studies in rats, chronic toxicity/carcinogenicity studies in rats, and reproductive/developmental toxicity studies in rats and hamsters. Toyo Sugar and Nippon Paper also consider published clinical studies and published and unpublished absorption, distribution, metabolism and excretion studies in animals and humans and published in vitro and in vivo mutagenicity/genotoxicity studies. Toyo Sugar and Nippon Paper discuss additional unpublished studies on the toxicity and genotoxicity (in vivo and in vitro) of EMSG using mice and rats. Based on its consideration of all SGs studies and the studies on EMSG, Toyo Sugar and Nippon Paper conclude that EMSG is safe for its intended use in foods.
Toyo Sugar and Nippon Paper note that JECFA, the Food Standards Australia New Zealand (FSANZ) and the European Food Safety Authority (EFSA) have recently made decisions on the safety of SGs for use in food as sweeteners. Toyo Sugar and Nippon Paper further note that JECFA established an acceptable daily intake (ADI) for SGs of 0 - 4 mg/kg bw/d (expressed as steviol) and FSANZ and EFSA established an ADI for SGs of 4 mg/kg bw/d (expressed as steviol ).
Standards of Identity
In the notice, Toyo Sugar and Nippon Paper state its intention to use EMSG 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)
The Food and Drug Administration Amendments Act of 2007, which was signed into law on September 27, 2007, amends the FD&C Act to, among other things, add section 301(ll). 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 Toyo Sugar and Nippon Paper' notice that EMSG is GRAS for the intended use, FDA did not consider whether section 301(ll) or any of its exemptions apply to foods containing EMSG. Accordingly, this response should not be construed to be a statement that foods that contain EMSG, if introduced or delivered for introduction into interstate commerce, would not violate section 301(ll).
Based on the information provided by Toyo Sugar and Nippon Paper, as well as other information available to FDA, the agency has no questions at this time regarding Toyo Sugar and Nippon Paper's conclusion that EMSG is GRAS under the intended conditions of use. The agency has not, however, made its own determination regarding the GRAS status of the subject use of EMSG. As always, it is the continuing responsibility of Toyo Sugar and Nippon Paper to ensure that food ingredients 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 000375, 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.
Mitchell A. Cheeseman, 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.
2. European Food Safety Authority. 2011. Revised exposure assessment for steviol glycosides for the proposed uses as a food additive. EFSA Journal: 9(1):1972.
 FDA notes that EFSA had expressed concern that the ADI would likely be exceeded at the maximum use levels proposed by the petitioners whose submission was reviewed by EFSA. The European Commission then requested EFSA to conduct a revised exposure assessment. Following this request, EFSA carried out a revised exposure assessment for SGs in children and adults from its use as a food additive (Ref. 2). In the revised assessment, the exposure estimate calculation uses the same methodology that was used by EFSA in its prior opinion on SGs but with revised use levels and food consumption data for children and the adult population. The mean dietary exposure to SGs (as steviol equivalents) in European children (1 to 14 years of age) ranged from 0.4 to 6.4 mg/kg bw/day, and from 1.7 to 16.3 mg/kg bw/day at the 95th percentile. Due to observed overestimations of consumption levels of non-alcoholic flavored drinks (soft drinks) by children at the 95th percentile within certain datasets, EFSA concluded that a correction factor was necessary. The corrected exposure estimate at the 95th percentile ranges from 1.0 to 12.7 mg/kg bw/day for children. For adults (UK population) mean dietary exposure to SGs (as steviol equivalents) were calculated to be 1.9 to 2.3 mg/kg bw/day and from 5.6 to 6.8 mg/kg bw/day for high level consumers (97.5th percentile). EFSA concluded that the main contributors to the total anticipated exposure to SGs are non-alcoholic flavored drinks (soft drinks) for both populations.
FDA further notes that studies conducted in the U.S., Canada, Australia, New Zealand, and Europe estimated the intakes of aspartame and other high intensity sweeteners using post-market surveillance data. Intake modeling methods are commonly used to estimate the consumption of a particular food or food ingredient. However, a realistic, but conservative approach is to estimate the intake of a sweetener based on the data reported in published studies. Estimates of intake for different sweeteners can be calculated based on adjustments for relative sweetness intensity. In the published study by Renwick (Ref. 1), the estimated intake of rebaudioside A, one of the primary SGs, was determined assuming a relative sweetness of 200 times that of sucrose. The consumption data were primarily from studies that used specifically designed food diaries combined with actual use levels or approved levels in different foods and beverages. These data were aggregated to provide an estimate of potential consumption of rebaudioside A.