Agency Response Letter GRAS Notice No. GRN 000399
July 31, 2012
Re: GRAS Notice No. GRN 000399
Dear Dr. Endres:
The Food and Drug Administration (FDA) is responding to the notice, dated August 11, 2011, that you submitted on behalf of Ganeden Biotech, Inc. (Ganeden) 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 August 19, 2011, filed it on August 23, 2011, and designated it as GRAS Notice No. GRN 000399.
The subject of the notice is a preparation of Bacillus coagulans strain GBI-30, 6086 spores (B. coagulans spore preparation). The notice informs FDA of the view of Ganeden that B. coagulans spore preparation is GRAS, through scientific procedures for use as an ingredient in baked goods and baking mixes; beverages and beverage bases; breakfast cereals; chewing gum; coffee and tea; condiments and relishes; confections and frostings; dairy product analogs; fruit juices; frozen dairy desserts and mixes; fruit and water ices; gelatins, puddings, and fillings; grain products and pastas; hard candy; herbs, seeds, spices, seasonings, blends, extracts, and flavorings; jams and jellies; milk; milk products; nuts and nut products; plant protein products; processed fruits; processed vegetables and vegetable juices; snack foods; soft candy; soups and soup mixes; at a maximum level of approximately 2 x 109 colony forming units per serving (cfu/serv). sugar; and sweet sauces, toppings, and syrups.
In its notice, Ganeden includes a statement from a panel of individuals (Ganeden’s GRAS panel) who evaluated the data and information that are the basis for Ganeden’s GRAS determination. Ganeden considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. Ganeden’s GRAS panel evaluated the method of manufacture, specifications, exposure, and safety data and information for B. coagulans spore preparation. Based on this review, Ganeden’s GRAS panel concluded that B. coagulans spore preparation produced in accordance with current good manufacturing practices and meeting established specifications is GRAS under the conditions of its intended use.
Ganeden discusses the history of B. coagulans and the identification and characteristic properties of B. coagulans strain GBI-30, 6086 as follows: B. coagulans was first described in 1915 at the Iowa Agricultural Experiment Station associated with the coagulation of evaporated milk. B. coagulans strain GBI-30, 6086 was identified by genetic comparison with a known B. coagulans strain. Published information describes B. coagulans strain GBI-30, 6086 as a Gram-positive, sporeforming, L+ lactic acid producing bacterium.
Ganeden describes the manufacture of B. coagulans spore preparation.All ingredients, fermentation tanks, and culture media are heat sterilized prior to inoculation with B. coagulans GBI-30, 6086 vegetative cells. The fermentation medium contains soy- and milk-derived ingredients. The strain is fermented under pH- and temperature-controlled, aseptic conditions. The pH is controlled by adding ammonium hydroxide or sodium hydroxide. The flow of nutrients to the vegetative cells is discontinued during fermentation to induce spore formation. After fermentation, the spores undergo centrifugation and freeze- or spray-drying before packaging. Ganeden states that all materials used in the manufacturing process are food-grade and meet appropriate regulations.
Ganeden provides specifications for appearance and moisture for B. coagulans spore preparation. Limits for microbiological contaminants include yeast, total coliforms, Escherichia coli, Staphylococci, Salmonella, and Pseudomonas aeruginosa. Also included are limits for heavy metal contaminants: arsenic (≤ 2 milligrams per kilogram (mg/kg)), cadmium (≤ 2 mg/kg), lead (≤ 5 mg/kg), and mercury (≤ 2 mg/kg).
Ganeden states that B. coagulans spore preparation is comprised of nearly 100% spores, which survive harsher conditions than vegetative cells. A study shows that B. coagulans spore preparation is stable for three months under storage at 40°C and 75% relative humidity. Ganeden states that the relative proportions of spores and vegetative cells are consistent among batches.
Ganeden states that B. coagulans spore preparation is currently added to the intended foods at levels of 108 to 2 x 109 cfu/serv. A publication from The United States Department of Agriculture Center for Nutrition Policy and Promotion (October 2000) states that males aged 51 and older consume the largest number of servings of food a day at 18.2 servings per day. Using this estimate of the number of servings per day at a level of 2 x 109 cfu/serv, Ganeden estimates the daily intake of B. coagulans spores at 36.4 x 109 cfu per day (cfu/day).
Ganeden discusses published and unpublished information supporting the safety of B. coagulans strain GBI-30, 6086. Published information describes B. coagulans strain GBI-30, 6086 as a nonpathogenic and nontoxigenic bacterium. In addition, an unpublished in vitro test shows that B. coagulans strain GBI-30, 6086 is sensitive to many clinically-used antibiotics, and that there is no evidence that the strain transmits antibiotic resistance to other bacterial species. Unpublished genetic sequencing and bioinformatic analyses show that B. coagulans strain GBI-30, 6086 does not contain genes homologous to those encoding known protein toxins.
Ganeden also discusses a published in vitro study, published studies conducted in animals, and published studies conducted in humans to support the safety of B. coagulans spore preparation. An in vitro study conducted using a continuous culture fermentation system designed to mimic conditions in the human ascending colon provides a basis to conclude that B. coagulans strain GBI-30, 6086 spores germinate and persist for at least 72 hours after discontinuing administration of spores into the system. The study authors state that B. coagulans strain GBI-30, 6086 spores did not alter the human fecal microbiota present in the fermentation system. An in vivo study in piglets conducted using biomass comprised of about 50% spores of B. coagulans strain CNCM I-1061 shows that the strain is transient, persisting for up to seven days in the piglet gastrointestinal tract. Ganeden concludes that taken together, these data show that B. coagulans strain GBI-30, 6086 spores are transient in the human gastrointestinal tract.
Ganeden discusses published studies conducted to assess the potential toxicity of B. coagulans spore preparation. In vitro studies show that B. coagulans spore preparation is not mutagenic or genotoxic. Studies conducted using rats show that B. coagulans spore preparation does not induce acute, subchronic, chronic, or reproductive toxicity following consumption of up to 2000 mg/kg body weight per day (equivalent to approximately 2.6 x 1013 cfu per person per day) spores.
Ganeden cites published human studies designed to investigate effects of B. coagulans spore preparation on patients with certain health conditions. In these studies, B. coagulans spore preparation was administered as a medical food or dietary supplement. Parameters assessed in these studies included arthritis activity, erythrocyte sedimentation rate, C-reactive protein levels, blood levels of cytokines and activated T lymphocytes, irritable bowel syndrome symptoms, gastrointestinal symptoms, and severity of dyspepsia. No adverse events were observed in these studies after the consumption of up to 2 x 109 cfu/day spores.Ganeden also notes that more than one billion servings of B. coagulans spore preparation have been consumed since 2003 at levels up to 2 x 109 cfu/serving, and that there have been no reports of adverse events.
Based on: 1) published and unpublished information demonstrating the safety of B. coagulans strain GBI-30, 6086; 2) published animal studies supporting the safety of B. coagulans spore preparation; 3) published human studies in which no adverse events were observed following consumption of B. coagulans spore preparation, and 4) the history of safe use ofB. coagulans spore preparation, Ganeden concludes that B. coagulans spore preparation is GRAS for the intended uses in foods.
Standards of Identity
In the notice, Ganeden states its intention to use B. coagulans spore preparation 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.
Potential Labeling Issues
In describing the human studies conducted using B. coagulans spore preparation, Ganeden includes studies that describe 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 B. coagulans spore preparation 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 neither consulted with ONLDS on this labeling issue nor evaluated the information in the notice to determine whether it would support any claims made about B. coagulans spore preparation 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. Ganeden states that soy- and milk-derived ingredients used during the manufacturing process for B. coagulans spore preparation will be appropriately labeled in the finished products. Issues associated with labeling food are the responsibility of ONLDS. However, issues associated with FALCPA should be directed to the Food Allergen Coordinator in FDA’s Center for Food Safety and Applied Nutrition (Food Allergen Coordinator, Office of Food Additive Safety, HFS-200, 5100 Paint Branch Parkway, College Park, Maryland 20740).
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 Ganeden’s notice that B. coagulans spore preparation is GRAS for the intended uses, FDA did not consider whether section 301(ll) or any of its exemptions apply to foods containing B. coagulans spore preparation. Accordingly, this response should not be construed to be a statement that foods that contain B. coagulans spore preparation, if introduced or delivered for introduction into interstate commerce, would not violate section 301(ll).
Based on the information provided by Ganeden, as well as other information available to FDA, the agency has no questions at this time regarding Ganeden’s conclusion that B. coagulans spore preparation 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 B. coagulans spore preparation. As always, it is the continuing responsibility of Ganeden 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 000399, 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
B. coagulans spore preparation is not intended for use in any product that would require additional review by the United States Department of Agriculture.
 Ganeden states that B. coagulans spores survive manufacturing conditions and the acidic conditions of the stomach.