Agency Response Letter GRAS Notice No. GRN 000440
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CFSAN/Office of Food Additive Safety
February 12, 2013
Christopher Wahl, M.D.
141 Avenue du President Kennedy
5th Floor, UQAM-Biological Sciences Bld.
Montreal, Quebec, H2X 3Y7
Re: GRAS Notice No. GRN 000440
Dear Dr. Wahl:
The Food and Drug Administration (FDA) is responding to the notice, dated July 27, 2012, 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 this notice on August 16, 2012, filed it on August 16, 2012, and designated it as GRN No. 000440.
The subject of the notice is Lactobacillus reuteri strain NCIMB 30242. The notice informs FDA of the view of Micropharma Ltd. (Micropharma) that L. reuteri strain NCIMB 30242 is GRAS, through scientific procedures, for use as an ingredient in beverages and beverage bases, breakfast cereals, cheeses, dairy product analogs, fats and oils, frozen dairy desserts, grain products and pastas, milk products, processed fruits and fruit juices, and sugar substitutes at levels ranging from 3.3 X 108 to 1010 colony forming units per serving (CFU/serving).
As part of its notice, Micropharma includes the report of a panel of individuals (Micropharma’s GRAS panel) that evaluated the data and information that are the basis for Micropharma’s GRAS determination. Micropharma considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. Micropharma’s GRAS panel evaluated information describing the identity, method of manufacture and specifications, and estimated dietary exposure for L. reuteri strain NCIMB 30242, as well as published and unpublished safety studies. Based on this review, Micropharma’s GRAS panel concluded that L. reuteri strain NCIMB 30242 produced in accordance with current good manufacturing practices (cGMP) and meeting the appropriate food-grade specifications is GRAS under the conditions of its intended use.
Micropharma discusses the identity and characterization of L. reuteri strain NCIMB 30242. Lactobacilli are Gram-positive, nonsporeforming, rod- or coccobacillus-shaped bacteria. The microorganisms can be found in dairy products, fermented meat, sourdough, vegetables, fruits, and beverages. L. reuteri strains are commonly found as commensal organisms in the respiratory, gastrointestinal, and genital tracts of humans and animals. The published literature suggests that L. reuteri strain NCIMB 30242 may have been isolated from a porcine source. The strain was deposited in the National Collection of Industrial and Food Bacteria and its identity was confirmed using phenotypic and genotypic analyses.
Micropharma describes the production of L. reuteri strain NCIMB 30242 by fermentation. Micropharma notes that milk-based ingredients are used during fermentation. The fermentation medium is sterilized and cooled before inoculation with a pure culture of L. reuteri strain NCIMB 30242. The fermentation is conducted under controlled conditions, and upon completion, the pH of the culture is adjusted, and the resulting biomass is concentrated via centrifugation. Food-grade cryoprotectants are added to the concentrated slurry, which is then freeze-dried, packaged, and stored under cool and dry conditions. Micropharma states that the freeze-dried product is formulated into a frozen slurry, lyophilized powder, or encapsulated powder depending on the specific food use for the ingredient. Reformulation for encapsulation or coating is conducted under controlled conditions using permitted food-grade ingredients. The manufacturing process is conducted under cGMP using suitable food-grade raw materials that are either GRAS or permitted for use in the United States.
Micropharma provides specifications for each ingredient formulation including identity and levels of formulation components, sodium alginate and epsilon-polylysine. Micropharma provides limits for heavy metal contaminants (e.g., lead, < 0.05 milligram/kilogram) and microbial contaminants (e.g., Listeria monocytogenes, absent in 25 grams). Micropharma provides data showing that L. reuteri strain NCIMB 30242 is stable in the lyophilized powder at 2–8° C for up to 12 months and in the frozen pellet formulations at -80° C for up to 13 months.
Micropharma used data from the National Health and Nutrition Examination Survey 2003–06 to estimate dietary exposure to L. reuteri strain NCIMB 30242 from its use in the food products specified above. Based on these data, Micropharma estimates the mean exposure to be 1.8 X 1010 CFU per person per day (p/d) and the 90th percentile exposure to be 3.5 X 1010 CFU/p/d based on the intended uses.
Micropharma states that L. reuteri strains have a history of safe use by the food industry as fermentation starters in the manufacture of sourdough and other breads. The history of safe food use of L. reuteri strains is recognized by the European Food Safety Authority and L. reuteri strains meet their criteria for “Qualified Presumption of Safety” status. FDA evaluated two GRAS notices for the uses of another L. reuteri strain (L. reuteri strain DSM 17938) and responded with “no questions” letters to both GRN 000254 and GRN 000410, which determined that L. reuteri strain DSM 17938 is GRAS for use in certain foods and term infant formula, respectively.
Micropharma discusses published in vitro tests and bioinformatic analyses, to support the safety of L. reuteri strain NCIMB 30242. The results of in vitro tests show that L. reuteri strain NCIMB 30242 does not produce biogenic amines or antimicrobial substances and is susceptible to several antibiotics. Bioinformatic analyses revealed that the strain contains antibiotic resistance elements common to other lactobacilli. Micropharma concludes these elements are unlikely to be transmitted to other bacteria. The bioinformatic analyses also showed that L. reuteri strain NCIMB 30242 does not contain genes encoding known protein toxins or proteins known to confer pathogenicity. Furthermore, the notifier’s search of several publicly-available databases did not produce literature documenting incidences of L. reuteri infections.
Micropharma also reports on two published human studies, in which male and female subjects consumed microencapsulated(1) L. reuteri strain NCIMB 30242 at a minimum of 1010 CFU/serving twice a day for six weeks(2) or two capsules per day of L. reuteri strain NCIMB 30242 at 2.9 X 109 CFU/capsule for nine weeks. Micropharma notes that L. reuteri strain NCIMB 30242 was well tolerated in both studies.
Micropharma reported on two unpublished animal studies. In the first study, the bacteria were administered by gavage to hamsters at 109 CFU per day of alginate polylysine encapsulated L. reuteri strain NCIMB 30242 for six weeks. Micropharma considers the findings in the hamster study to be unremarkable. In the second study, 1010 CFU per day of L. reuteri strain NCIMB 30242 was administered by gavage to male rats. L. reuteri strain NCIMB 30242 was not detected in the blood or liver and there was no increase in plasma D-lactic acid compared to the control. Based on the totality of the information in the notice, Micropharma concludes that L. reuteri strain NCIMB 30242 is GRAS for the intended uses in foods.
Standards of Identity
In the notice, Micropharma states its intention to use L. reuteri strain NCIMB 30242 in multiple food and beverage 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.
Potential Labeling Issues
In describing the human studies conducted using L. reuteri strain NCIMB 30242, Micropharma includes studies that mention potential health benefits. These studies raise a potential issue under the labeling provisions of the Federal Food, Drug, and Cosmetic Act (FD&C Act). Under section 403(a) of the FD&C Act, a food is misbranded if its labeling is false or misleading in any particular. Section 403(r) of the FD&C Act lays out the statutory framework for the use of labeling claims that characterize the level of a nutrient in a food or that characterize the relationship of a nutrient to a disease or health-related condition. If products that contain L. reuteri strain NCIMB 30242 bear any claims on the label or in labeling, such claims are the purview of the Office of Nutrition, Labeling, and Dietary Supplements (ONLDS) in the Center for Food Safety and Applied Nutrition. The Office of Food Additive Safety (OFAS) neither consulted with ONLDS on this labeling issue nor evaluated the information in your notice to determine whether it would support any claims made about L. reuteri strain NCIMB 30242 on the label or in labeling.
The Food Allergen Labeling and Consumer Protection Act of 2004 (FALCPA) amends the FD&C Act to require that the label of a food that is or contains an ingredient that bears or contains a “major food allergen” declare the presence of the allergen (section 403(w)). FALCPA defines a “major food allergen” as one of eight foods or food groups (i.e., milk, eggs, fish, Crustacean shellfish, tree nuts, peanuts, wheat, and soybeans) or a food ingredient that contains protein derived from one of those foods. L. reuteri strain NCIMB 30242 grown in a fermentation media that contains milk based ingredients, may require labeling under FALCPA, because it may contain protein derived from milk. Although issues associated with labeling food are the responsibility of ONLDS, issues associated with FALCPA exemptions are the responsibility of the Food Allergen Coordinator in OFAS.
Section 301(ll) of the 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 Micropharma’s notice that L. reuteri strain NCIMB 30242 is GRAS for the intended use, FDA did not consider whether section 301(ll) or any of its exemptions apply to foods containing L. reuteri strain NCIMB 30242. Accordingly, this response should not be construed to be a statement that foods that contain L. reuteri strain NCIMB 30242, if introduced or delivered for introduction into interstate commerce, would not violate section 301(ll).
Based on the information provided by Micropharma, as well as other information available to FDA, the agency has no questions at this time regarding Micropharma’s conclusion that L. reuteri strain NCIMB 30242 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 L. reuteri strain NCIMB 30242. As always, it is the continuing responsibility of Micropharma 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 000440, 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)Micropharma states that the alginate-polylysine coating (used to improve viability) is considered representative of the commercial product.
(2)Micropharma reports that L. reuteri strain NCIMB 30242 is added to the yogurt initially at 5 X 1010 CFU/serving. However, due to reduced viability over time, the organism is consumed at a minimum of 1010 CFU/serving with the concentration more likely to be 2.5 X 1010 CFU/serving consumed twice a day.