Agency Response Letter
GRAS Notice No. GRN 000131

Daniel R. Dwyer
Kleinfeld, Kaplan and Becker, LLP
1140 19th Street, NW
Washington, DC 20036

Re: GRAS Notice No. GRN 000131

Dear Mr. Dwyer:

The Food and Drug Administration (FDA) is responding to the notice, dated May 28, 2003, that you submitted on behalf of Loders Croklaan B.V. (Loders Croklaan) 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 May 29, 2003, filed it on June 9, 2003, and designated it as GRAS Notice No. GRN 000131.

The subject of the notice is high 2-palmitic vegetable oil (vegetable oil, enzyme-modified to increase the content of palmitic acid at the C-2 hydroxyl of the glycerol backbone). The notice informs FDA of the view of Loders Croklaan that high 2-palmitic vegetable oil is GRAS, through scientific procedures, for use in infant formula for both term and preterm infants at a level of up to 80 percent of total fat.

Loders Croklaan provides generally available information about the composition of high 2-palmitic vegetable oil. It is a mixture of triglycerides, predominantly 1,3-dioleoyl 2-palmitoyl triglyceride and 1,2-dipalmitoyl 3-oleoyl triglyceride. It is rich in palmitic acid (C16:0; 30-55 percent by weight), with 45-80 percent of the palmitic acid esterified at the sn-2 (middle) position of the glycerol backbone. The sn-1 and sn-3 positions predominantly contain unsaturated fatty acids, mostly oleic acid (C18:1). The fatty acids present in high 2-palmitic vegetable oil are the same as those in edible oils and fats, including human milk.

Loders Croklaan states that high 2-palmitic vegetable oil is manufactured by the enzymatic esterification of food grade vegetable oils (except canola oil or derivatives of canola oil) with unsaturated fatty acids derived from food grade vegetable oils. Palm stearine and oleic acid are the main starting materials, and the processes are primarily those found in traditional oil processing. The enzyme that Loders Croklaan currently uses for interesterification is an immobilized lipase enzyme preparation from Aspergillus oryzae expressing a gene encoding a Rhizomucor miehei lipase. Loders Croklaan provides specifications for high 2-palmitic vegetable oil.

High 2-palmitic vegetable oil is intended to be blended with other vegetable oils to provide an appropriate balance of fatty acids and triglycerides suitable for infant formula. Typically, high 2-palmitic vegetable oil will be 40-80 percent of the vegetable oil blend. Loders Croklaan estimates that intake of high 2-palmitic vegetable oil would not exceed 28 grams per day (g/d)⁽¹⁾.

Loders Croklaan describes generally available information about the digestion, absorption, and metabolism of fat. After ingestion, fat emulsifies, and enzymes hydrolyze the triglycerides to release free fatty acids from the sn-1 and sn-3 positions of the glycerol backbone. The free fatty acids and the sn-2 monoglycerides are then absorbed from the small intestine. Longer chain, saturated fatty acids (C12:0-C18:0) are not absorbed as easily as medium chain (C6:0 to C10:0) or unsaturated fatty acids and form insoluble fatty acid-calcium soaps at the pH of the intestine.

Loders Croklaan discusses published studies on the digestion, absorption, and metabolism of high 2-palmitic vegetable oil in animals, including:

• A study on fat absorption in adult rats fed high 2-palmitic vegetable oil; both palmitic acid and overall fat absorption were increased relative to the control diet (palmitic acid at the sn-1 and sn-3 positions).
• A metabolic study of 1- and 2-palmitoyl triglycerides in the rat; no apparent difference was found in the rate of metabolism and distribution between 1- and 2-palmitoyl triglycerides in the male weanling rat or in the suckling rat.
• Studies conducted with piglets; piglets fed formula containing high 2-palmitic vegetable oil had plasma lipid profiles closer to those of piglets fed sow's milk than to those of piglets fed control formula.

Loders Croklaan discusses published human studies comparing digestion, absorption, and metabolism in preterm and term infants fed formula containing high 2-palmitic vegetable oil relative to infants fed control formula. Loders Croklaan draws the following conclusions from these studies:

• Term and preterm infants fed formula containing high 2-palmitic vegetable oil showed significantly greater absorption of palmitic acid and stearic acid than infants fed control formula.
• Term and preterm infants fed formula containing high 2-palmitic vegetable oil had plasma lipid profiles closer to those of infants fed breast milk than to those of infants fed control formula.
• Some, but not all, of these studies found significantly greater total fat absorption in term infants fed formula containing high 2-palmitic vegetable oil than in term infants fed control formula; this difference was not observed in preterm infants.
• The stools of infants fed formula containing high 2-palmitic vegetable oil contain less calcium and fewer insoluble calcium soaps, and are generally softer, than the stools of infants fed control formula.
• In one study, term infants fed formula containing high 2-palmitic vegetable oil had a higher whole-body bone mineral content than infants fed control formula.

Loders Croklaan describes one generation and two generation feeding studies in rats involving high 2-palmitic vegetable oil at a level of up to 15 percent by weight in the diet. Both studies have been accepted for publication. Loders Croklaan concludes that high 2-palmitic vegetable oil supports the growth and general reproductive performance of the rat without adverse effects and that the no-observed-effect-level (NOEL) was 15 percent of the diet, the highest level used.

As part of its notice, Loders Croklaan includes the findings of a panel of individuals (Loders Croklaan's GRAS panel) who evaluated the data and information that are the basis for Loders Croklaan's GRAS determination. Loders Croklaan considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. Loders Croklaan's GRAS panel discusses the identity and composition, manufacture, specifications, exposure, metabolism, and toxicity studies of high 2-palmitic vegetable oil. Loders Croklaan's GRAS panel concludes that high 2-palmitic vegetable oil is safe for use in infant formula for both term and preterm infants at levels up to 80 percent total fat.

Based on the information provided by Loders Croklaan in GRN 000131 and other information available to FDA, the agency has no questions at this time regarding Loders Croklaan's conclusion that high 2-palmitic vegetable 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 high 2-palmitic vegetable oil. As always, it is the continuing responsibility of Loders Croklaan to ensure that food ingredients that the firm markets are safe, and are otherwise in compliance with all applicable legal and regulatory requirements.

Under section 412 of the Federal Food, Drug, and Cosmetic Act (FFDCA), 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. Loders Croklaan should be aware that FDA's response to Loders Croklaan's GRAS notice does not alleviate the responsibility of any infant formula manufacturer who intends to market an infant formula that contains high 2-palmitic vegetable oil to make the submission required by section 412.

For your information, FDA recently contracted with the National Academy of Sciences to address broad issues related to the addition of ingredients to infant formula. We expect to make significant findings of any final report issued under that contract available to the public

In accordance with proposed 21 CFR 170.36(f), a copy of the text of this letter, as well as a copy of the information in your notice that conforms to the information in proposed 21 CFR 170.36(c)(1), is available for public review and copying on the homepage of the Office of Food Additive Safety (on the Internet at http://www.cfsan.fda.gov/~lrd/foodadd.html).

Sincerely,    /s/ Laura M. Tarantino, Ph.D. Acting Director Office of Food Additive Safety Center for Food Safety      and Applied Nutrition

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⁽¹⁾FDA independently estimated that intake of high 2-palmitic vegetable oil would be 22 g/d at the mean and 35 g/d at the 90th percentile.