Agency Response Letter GRAS Notice No. GRN 000411
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CFSAN/Office of Food Additive Safety
November 20, 2012
Edward A. Steele
EAS Consulting Group, LLC
1940 Duke Street, Suite 200
Alexandria, VA 22314
Re: GRAS Notice No. GRN 000411
Dear Mr. Steele:
The Food and Drug Administration (FDA) is responding to the notice, dated December 9, 2011, that you submitted on behalf of BNLfood Investments SARL (BNLfood) 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 December 19, 2011, filed it on January 4, 2012, and designated it as GRAS Notice No. GRN 000411.1
The subject of the notice is egg yolk-derived phospholipids (EYPL). The notice informs FDA of the view of BNLfood that EYPL is GRAS, through scientific procedures, for use as an ingredient in term and preterm infant formula at levels up to 2 grams phospholipids per liter (g/L) of formula, providing a maximum of 0.28% and 0.18% of total fatty acids as arachidonic acid (AA) and docosahexaenoic acid (DHA), respectively, from EYPL.
EYPL is available in six commercial forms, each containing a stated amount of phospholipids (30, 50, or 85 percent by weight) and varying relative amounts of AA (0.6-6.5% of fatty acids) and DHA (0.8-5.6% of fatty acids); ratios of AA:DHA are either 1:2 or 2:1 depending on the egg yolk starting material. EYPL contains lower AA and DHA levels than other GRAS sources of AA and DHA and BNLfood intends EYPL to be used in combination with other GRAS sources of AA and DHA to provide these fatty acids at levels in finished formulas that are consistent with respective standards and expert recommendations for term and preterm infant formulas. In its discussion of use levels of AA and DHA in finished formulas, BNLfood cites previous GRAS notices where FDA did not questioned the GRAS determinations (i.e., GRNs 000041, 000080, 000094, and 000326).
Our use of “EYPL” 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 (ONLDS) in the Center for Food Safety and Applied Nutrition.
As part of its notice, BNLfood includes the report of a panel of individuals (BNLfood’s GRAS panel) who evaluated the data and information that are the basis for BNLfood’s GRAS determination. BNLfood considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. BNLfood’s GRAS panel reviewed information regarding the identity, composition, allergenicity, safety and toxicity information on egg yolk phospholipids, long-chain omega-3 and omega-6 fatty acids (especially AA and DHA), and published animal and human studies on EYPL. Based on this review, BNLfood’s GRAS panel concluded that EYPL, produced in accordance with good manufacturing practices (GMPs), that meets its established food grade specifications is GRAS under the conditions of its intended use.
BNLfood describes six EYPL commercial forms2 representing three different concentrations of phospholipids (30, 50, and 85 percent by weight) and 2 different ratios of AA:DHA (2:1 and 1:2). For the ratio of AA:DHA included within the abbreviated names: EYPL 85 (2:1), EYPL 85 (1:2), EYPL 50 (2:1), EYPL 50 (1:2), EYPL 30 (2:1), and EYPL 30 (1:2). In this letter, when the ratio of AA:DHA is not relevant to the discussion, the generic terms EYPL 85, EYPL 50, and EYPL 30 are used.
EYPL contains some residual triglyceride, cholesterol, and free fatty acids from extraction (ethanolic) of egg yolk. BNLfood notes that, at the highest levels of use of any of the EYPL commercial products in infant formula, residual cholesterol will be 16 milligrams per 100 milliliters (mg/100 mL) finished formula, a level that is within the range of cholesterol in breast milk according to published literature. The typical compositions of the EYPL commercial products, expressed on a weight percent basis (g component/100 g ingredient (wt %), are as follows:
- EYPL 85: 85% phospholipids, 1% triglycerides, 7% free fatty acids, and 7% cholesterol.
- EYPL 50: 51% phospholipids, 0.6% triglycerides, 4.2% free fatty acids, 4.2% cholesterol, and 40% potato maltodextrin (added).
- EYPL 30: 30% phospholipids, 9% triglycerides, 2.5% free fatty acids, 3.5% cholesterol, and 55% high oleic sunflower oil (added).
BNLfood describes the phospholipid profile of EYPL as predominantly glycerophospholipids with minor amounts of sphingophospholipids. BNLfood provides the relative percentages of phospholipids, which are similar for all EYPL products regardless of the AA:DHA ratio: 70-75% phosphatidylcholine, 15-20% phosphatidylethanolamine, 3-4% lysophosphatidylcholine, 2-3% sphingomyelin, 1-2% lysophosphatidylethanolamine, and 2-3% phosphatidylinositol.
BNLfood describes the method of manufacture of EYPL. Source eggs from hens (Gallus domesticus) are produced in accordance with good agricultural practices and 21 CFR Part 118.3 The starting material is pasteurized egg yolk powder that is processed in a United States facility, in accordance with GMPs and must test negative for Salmonella. Egg yolk powder and phospholipid products are regularly tested for environmental contaminants, animal drugs, and pesticides and comply with all applicable standards and regulations outlined in 21 CFR 109.30, 21 CFR Part 556, and 40 CFR Part 180. BNLfood cites publications that discuss dietary strategies to modify lipid composition of hen eggs. To produce the 2:1 AA:DHA egg yolk, the hen is fed a diet rich in corn, soy, and sunflower. To produce the 1:2 AA:DHA egg yolk, the hen is fed a diet rich in wheat, peas, alfalfa, and flax.
BNLfood describes the extraction and formulation of EYPL 30, EYPL 50, and EYPL 85 from egg yolk. BNLfood states that all materials used in the method of manufacture and packaging of EYPL are food grade. For EYPL 30, phospholipids are extracted from egg yolk powder using ethanol, and separated by centrifugation. Residual aggregated protein is removed by filtration. The resultant lipid/ethanol phase is then mixed with high oleic sunflower oil. Ethanol is removed via distillation and evaporation; the product is a viscous, orange/brown oil that is heated to ensure microbiological safety, then packaged for storage up to 12 months. For EYPL50 and EYPL85, phospholipids are extracted from egg yolk powder using ethanol and water, then separated by centrifugation. Phospholipids are further purified by removal of triglycerides by crystallization (induced by cooling) and decantation; phospholipids (as a percent of dry matter) are increased from approximately 72% to 85% at this stage. Residual aggregated protein is then removed by filtration, and remaining ethanol is removed by distillation and evaporation. For EYPL 85, the phospholipid extract is lyophilized, yielding orange/brown flakes (about 0.5 to 1 centimeter in length) (12-month storage life). To produce EYPL 50, the phospholipid extract is combined with potato maltodextrin, then homogenized and heated twice. The product is spray dried and sieved, producing a yellow powder that is stable for 18 months. General specifications provided by BNLfood are listed in Table 1. In addition to Chronobacter sakazakii, BNLfood provides microbial specifications for total bacterial count < 100 colony forming units (cfu)/g; Salmonella absent/25g; moulds/yeasts < 10 cfu/g.
|Parameter||EYPL 85||EYPL 50||EYPL 30|
|Total lipid||98.5% min.||55.0% min.||95.0% min.|
|Moisture||1.50% max.||5.0% max.||1.50% max.|
|Phospholipids||85.0 ± 2.5%||50.0 ± 1.5%||30.0 ± 2.5%|
|Nitrogen||2.0 ± 0.5%||1.2 ± 0.2%||0.5 ± 0.5%|
|Cholesterol||7.0 ± 0.5%||4.0 ± 0.5%||3.5 ± 0.5%|
|Free fatty acid||7.0 ± 0.5%||4.0 ± 0.5%||2.5 ± 0.5%|
|Lead||0.1 mg/kilograms (kg) max.||0.1 mg/kg max.||0.1 mg/kg max.|
|Peroxide||1.0 mg/kg max.||1.0 mg/kg max.||1.0 mg/kg max.|
|Protein||< 100 mg/kg||< 100 mg/kg||< 100 mg/kg|
|C. sakazakii||0 cfu/10g||0 cfu/10g||0 cfu/10g|
|Shelf life||12 mos.||18 mos.||12 mos.|
The maximum levels of AA and DHA are similar for the EYPL 85 and EYPL 50 ingredients, which vary from EYPL 30. Triglycerides are removed (by decantation) during manufacture of EYPL 85 and EYPL 50, but not EYPL 30; hence, EYPL 30 contains more triglycerides from egg yolk. EPA is less than 0.5% of total fatty acids for all EYPL ingredients; therefore, EPA levels do not exceed those of DHA for any ingredient. Specifications for AA, DHA, and EPA are shown in Table 2.
|Fatty acids (wt % TFA)|| EYPL 85 (2:1);
EYPL 50 (2:1)
| EYPL 85 (1:2);
EYPL 50 (1:2)
|EYPL 30 (2:1)||EYPL 30 (1:2)|
|EPA||< 0.5||< 0.5||< 0.5||< 0.5|
BNLfood provides estimates of intake of EYPL 85 (0.35 g per kg of body weight per day (g/kg bw/d)), EYPL 50 (0.6 g/kg bw/d), and EYPL 30 (1 g/kg bw/d), that correspond to a 2 g phospholipid/L formula (or approximately 0.3 g phospholipid/100 kcal formula) target use level for infant formula. These estimates assume that the recommended intake of formula for term and preterm infants is 150 mL formula per kg body weight per day for term and preterm infants and the energy density of formula is approximately 67 kcal/100 mL.4 Assuming AA and DHA are present at levels representing midpoints of specifications for each ingredient, BNLfood estimates corresponding intakes of AA and DHA to be 14 and 7 mg/kg bw/d, respectively, for EYPL ingredients with AA:DHA ratio 2:1; and, 4.7 and 9.4 mg/kg bw/d, respectively, for EYPL ingredients with AA:DHA ratio 1:2. BNLfood estimates corresponding intakes of EPA to be < 0.25 mg/kg bw/d for EYPL ingredients with AA:DHA ratio 2:1 and < 0.75 mg/kg bw/d for EYPL ingredients with AA:DHA ratio 1:2. BNLfoods notes that levels of AA and DHA are 2-3 fold below those in previous GRAS notices (GRNs 000041, 000080, 000094, 000326) which provided for up to 45 mg/kg bw/d AA and up to 30 mg/kg bw/d DHA, with AA:DHA ratios from 1:1 to 2:7:1. Further, EYPL will be used in conjunction with other GRAS sources of AA and DHA to provide stated target levels of these components in finished formulas. As a percent of TFA, BNLfood’s maximum AA levels are 0.28% and 0.09% for EYPL 2:1 and 1:2 respectively, and maximum DHA levels are 0.14% and 0.18%, for EYPL 2:1 and 1:2 ingredients, respectively.
BNLfood notes that EYPL 85 was approved for use in infant formulae and follow-on formulae by the European Commission in 2000.5 Since that time, the maximum use level of EYPL in formulae in Europe has increased to 2 g phospholipids per L, and additional commercial forms of EYPL (i.e., EYPL 30 and EYPL 50), have been introduced into infant formula and toddler foods outside the United States. BNLfood also notes that both the European Society for Pediatric Gastroenterology, Hepatology and Nutrition report, published in 2005 and the 2006 European Commission Directive (2006/141/EC), both state a maximum level of 2 g/L phospholipid in infant formula and follow-on formula.
The notifier’s safety evaluation focused on three areas: the safety of phospholipids which contain AA and DHA, the safety of adding AA and DHA to infant formula, and the issue of allergenicity as it pertains to any residual egg yolk proteins that may be in EYPL.
In discussing the safety of phospholipids, BNLfood describes current sources, stability, average amounts found in human milk at various times (colostrum, transitional, mature milk), as well as basic phospholipid metabolism. In addition, the notifier discusses published animal studies (rat, piglet, and baboon) which examined the effects of adding egg phospholipids to the diet at levels of up to 1.02 g/100 mL with no adverse effects noted. Furthermore, the notifier discusses published clinical trials in both term and pre-term infants who consumed formula with phospholipid levels of up to 0.396 g/100 mL with normal growth and no adverse effects reported. Finally, BNLfood notes that fatty acids from dietary phospholipids are absorbed at least as well as fatty acids from dietary triglycerides.
BNLfood specifically discusses the safety of the use of AA and DHA in infant formula, noting AA and DHA have been the subjects of several previous GRAS notices and have been discussed in numerous publicly-available scientific reviews. BNL food summarizes data available on AA and DHA, including a discussion of published results from genotoxicity studies on AA and DHA rich oils (Ames assay, mouse lymphoma assay, and chromosomal aberration assay in Chinese hamster ovary cells) that showed no mutagenic or genotoxic potential. The notifier further reviewed multiple published subchronic (90 days) rat studies in which rats received DHA or AA at levels of at least 3000 mg/kg bw/d with no adverse effects. In addition the notifier discusses a published rat developmental toxicity study in which exposures to 1250 mg/kg bw/d DHA and 2500 mg/kg/bw/d AA showed a lack of maternal or fetal toxicity. Finally, BNLfood summarizes numerous published clinical studies involving both term and pre-term infants that resulted in no adverse effects from exposure to infant formula with AA and DHA.
In discussing allergenicity as it relates to any residual egg yolk protein, BNLfood considers that EYPL is a highly purified lipid fraction which has less than 1 mg/kg of egg white protein and less than 5 mg/kg egg yolk protein. The notifier also discusses known egg protein allergens as well as which particular egg proteins have been found in their specific products. Furthermore, the notifier discusses an unpublished skin contact hypersensitivity assay utilizing albino guinea pigs which showed no positive reactions from exposure to their product. Finally, BNLfood states that the ingredient will be labeled as an egg derived product.
Potential Labeling Issues
In describing data and information on clinical studies that BNLfood relies on to conclude that EYPL is GRAS under the conditions of its intended use, BNLfood raises 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 EYPL bear any claims on the label or in labeling, such claims are the purview of ONLDS. The Office of Food Additive Safety 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 EYPL on the label or in labeling.
The Food Allergen Labeling and Consumer Protection Act (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. EYPL derived from egg yolk may require labeling under FALCPA, because it may contain protein derived from eggs. Although issues associated with labeling food are the responsibility of the ONLDS, issues associated with FALCPA exemptions are the responsibility of the Food Allergen Coordinator in the Office of Food Additive Safety.
Intended Use in Infant Formula
Under section 412 of the FD&C Act a manufacturer of a new infant formula must make a submission to FDA, providing required assurances about the formula, at least 90 days before the formula is marketed. BNLfood should be aware that FDA’s response to BNLfood’s GRAS notice does not alleviate the responsibility of any infant formula manufacturer who intends to market an infant formula that contains EYPL to make the submission required by section 412.
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 BNLfood’s notice that EYPL is GRAS for the intended use, FDA did not consider whether section 301(ll) or any of its exemptions apply to foods containing EYPL. Accordingly, this response should not be construed to be a statement that foods that contain EYPL, if introduced or delivered for introduction into interstate commerce, would not violate section 301(ll).
Based on the information provided by BNLfood, and other information available to FDA, the agency has no questions at this time regarding BNLfood’s conclusion that EYPL 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 EYPL. As always, it is the continuing responsibility of BNLfood 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 000411, 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
1Clarifying information, including updated specifications, was provided by the notifier on April 16, 2012, and June 14, 2012.
2In the notice, BNLfood refers to EYPL by trade names. Further, to denote the ratio of AA:DHA, BNLfood uses the terms “AA-rich” to describe EYPL with a 2:1 ratio AA:DHA and “DHA-rich” to describe EYPL with a 1:2 ratio AA:DHA. FDA has not evaluated the terms “AA-rich” or “DHA-rich” within the context of GRN 000411.
3FDA’s 2009 egg rule (74 FR 33030; July 9, 2009; Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation).
4FDA notes that energy density needs and recommendations are different for term and preterm infants, and the assumptions of Danone do not reflect those differences. However since the use of EYPL is not based on energy density, but on volume of formula, the assumptions do not affect BNLfood’s safety determination.
5Commission Decision of 22 February 2000 authorizing the use of phospholipids purified to 85% from egg yolk as a novel food ingredient. Directive 95/2/EC on food additives permitted addition of phospholipids to infant formula at a maximum level of 1 g/L.