Agency Response Letter GRAS Notice No. GRN 000355
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CFSAN/Office of Food Additive Safety
March 7, 2011
Matthias Liebergesell, Ph.D.
DuPont Applied Biosciences
Route 141 & Powder Mill Road
Wilmington, DE 19803
Re: GRAS Notice No. GRN 000355
Dear Dr. Liebergesell:
The Food and Drug Administration (FDA) is responding to the notice, dated August 30, 2010, 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 September 1, 2010, filed it on September 8, 2010, and designated it as GRAS Notice No. GRN 000355.
The subject of the notice is eicosapentaenoic acid (EPA)-rich triglyceride oil from Yarrowia lipolytica (EPA-rich oil). The notice informs FDA of the view of DuPont Applied BioSciences (DuPont) that EPA-rich oil is GRAS, through scientific procedures, for use as an ingredient in food categories as listed in 21 CFR 184.1472(a)(3) at a maximum intake of 3.0 grams per person per day (g/p/d) EPA and not to be combined or augmented with any other food ingredient containing EPA and/or another omega-3 fatty acid, docosahexaenoic acid.
Our use of “EPA-rich oil” 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 all ingredients must be declared by their 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, DuPont includes the report of a panel of individuals (DuPont’s GRAS panel) who evaluated the data and information that are the basis for DuPont’s GRAS determination. DuPont considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. DuPont’s GRAS panel evaluated the manufacturing process, including the safety of the production organism, specifications, current regulatory status, genotoxicity studies as well as animal and human studies. Based on this review, DuPont’s GRAS panel concluded that EPA-rich oil produced in accordance with current good manufacturing practices (cGMPs) and meeting appropriate food grade specifications is GRAS under the conditions of intended use.
DuPont states that EPA-rich oil is produced by fermentation of a genetically engineered strain of Y. lipolytica, which is a non-toxigenic and non-pathogenic yeast found in nature. The EPA-rich oil may be produced with an EPA content ranging from 35 to 55 percent through use of strains with different copy numbers and promoters. DuPont describes the fermentation of Y. lipolytica as a 2-stage process; the first stage promotes rapid growth of the yeast, while in the second stage, the yeast are deprived of nitrogen and fed glucose to promote oil accumulation. Following fermentation, the yeast biomass is dewatered, washed to remove residual salts and medium, and dried. Isohexane is used to extract the EPA-rich triglyceride oil from the biomass. The isohexane is then removed via evaporation to yield crude oil. The crude oil is refined and degummed to remove free fatty acids, phospholipids, and trace metals. DuPont provides specifications for EPA-rich oil including a limit for lead (less than 0.01 milligrams/kilogram) and microbiological specifications (yeasts and molds, less than 100 colony forming units (CFU)/milliliter (mL); Staphylococcous aureus, less than 5 CFU/mL; and coliforms, less than 5 CFU/mL).
The production organism Y. lipolytica produces and accumulates oil as an energy source in the presence of excess glucose. DuPont has integrated into the Y. lipolytica genome single or multiple copies of a variety of well-characterized genes involved in efficient fatty acid synthesis.
DuPont states that the linear DNA is integrated into the Y. lipolytica genome by homologous or non-homologous recombination, and cells that contain the introduced DNA are identified through the use of selectable markers. DuPont states that Y. lipolytica is inoculated, and then fermented in culture medium containing glucose under controlled conditions to ensure consistent process performance and final EPA-rich oil quality. DuPont states that the EPA content of EPA-rich oil ranges from 35–55 percent. In order to maintain an exposure less than 3.0 g/p/d to EPA, DuPont has proposed intended use levels appropriate for each end of the range. For the 35 percent EPA-rich oil an intake of 8.57 g of oil would result in an exposure of 3.0 g to EPA. For the 55 percent EPA-rich oil, 5.45 g of oil would also result in an exposure of 3.0 g to EPA.
The notifier discusses the biochemical pathways and physiologic role of EPA-rich oil in the body, as well as previous concerns raised by FDA (menhaden oil final rule; 62 FR 30751; June 5, 1997) including possible elevated low-density lipoprotein cholesterol levels, prolonged bleeding time, and effects on glycemic control. The notifier reviews published genotoxicity studies including an Ames assay, mouse lymphoma assay, and mouse micronucleus assay on the notified EPA-rich oil and notes that all studies were negative when compared to positive controls. In addition, the notifier discusses published subchronic toxicity studies of 28 and 90 days in rats that received the notified EPA-rich oil by gavage, with no adverse effects on mean body weight, mean body weight gains, nutritional parameters, mortality, clinical signs, and neurobehavioral or ophthalmological observations in male or female rats. Also, the notifier analyzes an unpublished developmental study in rats that also received the notified EPA-rich oil by gavage; noting that there were no effects on reproduction or normal development in the study.
The notifier discusses information pertaining to the safety of the production microorganism and states that Y. lipolytica is a dimorphic, strictly aerobic, ascomycetous yeast that is adaptable to a wide range of physical and chemical conditions. The notifier states that Y. lipolytica is readily isolated from many food products and has traditionally been used in the processing of cheese and other dairy products. The notifier further states that Y. lipolytica is non-pathogenic and non-toxigenic and it is deposited in the American Type Culture Collection with identification number of ATCC 20362. In addition, the notifier discusses the results of published rodent studies showing that Y. lipolytica is unlikely to invade, colonize, and/or produce disease.
Standards of Identity
In the notice, DuPont states its intention to use EPA-rich oil 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.
Use in Products Under USDA Jurisdiction
During its evaluation of GRN 000355, FDA consulted with the Risk and Innovations Management Division of the Food Safety and Inspection Service (FSIS) of the United States Department of Agriculture. Under the Federal Meat Inspection Act, the Poultry Products Inspection Act, and the Egg Products Inspection Act, FSIS is responsible for determining the efficacy and suitability of food ingredients in meat, poultry, and egg products as well as prescribing safe conditions of use. Suitability relates to the effectiveness of the ingredient in performing the intended purpose of use and the assurance that the conditions of use will not result in an adulterated product, or one that misleads consumers.
FSIS requested that FDA advise DuPont to seek regulatory guidance from FSIS about the use of EPA-rich oil in meat, poultry, and egg products. DuPont should direct such an inquiry to Dr. William K. Shaw, Jr., Risk, Innovations, and Management Division, Food Safety and Inspection Service, George Washington Carver Center (GWCC), 5601 Sunnyside Avenue, Mailstop 5272, Beltsville, MD 20705-5271. The telephone number for that office is (301)504-0889.
Section 301(ll) of the FFDCA
The Food and Drug Administration Amendments Act of 2007, which was signed into law on September 27, 2007, amends the FFDCA to, among other things, add section 301(ll). Section 301(ll) of the FFDCA prohibits the introduction or delivery for introduction into interstate commerce of any food that contains a drug approved under section 505 of the FFDCA, 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 DuPont’s notice that EPA-rich oil is GRAS for the intended uses, FDA did not consider whether section 301(ll) or any of its exemptions apply to foods containing EPA-rich oil. Accordingly, this response should not be construed to be a statement that foods that contain EPA-rich oil, if introduced or delivered for introduction into interstate commerce, would not violate section 30l(ll).
Based on the information provided by DuPont, as well as other information available to FDA, the agency has no questions at this time regarding DuPont’s conclusion that EPA-rich oil 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 EPA-rich oil. As always, it is the continuing responsibility of DuPont 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 000355, 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
cc: William K. Shaw, Jr., Ph.D., Director
Risk Innovations, and Management Division
Food Safety and Inspection Service
George Washington Carver Center (GWCC)
5601 Sunnyside Avenue, Mail STOP 5272
Beltsville, MD 20705-5272