Agency Response Letter
GRAS Notice No. GRN 000206

Ms. Iris Meiri-Bendek
Regulatory Affairs Manager
Enzymotec Ltd.
P.O. Box 6
Migdal HaEmeq
ISRAEL, 23106

Re: GRAS Notice No. GRN 000206

Dear Ms. Meiri-Bendek:

The Food and Drug Administration (FDA) is responding to the notice, dated June 6, 2006, 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 June 13, 2006, filed it on June 14, 2006, and designated it as GRAS Notice No. GRN 000206.

The subject of the notice is the ingredient phytosterol esters and diglycerides resulting from transesterification of vegetable oils/fats with phytosterols (PSEDGV).⁽¹⁾ The notice informs FDA of the view of Enzymotec Ltd. (Enzymotec) that PSEDGV is GRAS, through scientific procedures, for use as an ingredient in baked goods and baking mixes; fats and oils; frozen dairy desserts and mixes; gelatins, puddings, and fillings; grain products and pastas; gravies and sauces; hard candy; milk; milk products; soft candy; soups and soup mixes; and snack foods at a level of 0.65 gram (g) phytosterol esters per reference amount customarily consumed (RACC).

Our use of "phytosterol esters and diglycerides resulting from transesterification of vegetable oils/fats with phytosterols" or "PSEDGV" 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. 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 Nutritional Products, Labeling, and Dietary Supplements in the Center for Food Safety and Applied Nutrition.

As part of its notice, Enzymotec includes the report of a panel of individuals (Enzymotec's GRAS panel) who evaluated the data and information that are the basis for Enzymotec's GRAS determination. Enzymotec considers the members of its GRAS panel to be qualified by scientific training and experience to evaluate the safety of substances added to food. Enzymotec's GRAS panel evaluated use levels in specified food products, estimates of dietary exposure, method of production, and product specifications as well as published and unpublished studies of phytosterols and diglycerides (also referred to as diacylglycerol oil or DAG oil). Based on this review, Enzymotec's GRAS panel concluded that PSEDGV, meeting the specifications described in the notice, is GRAS for the intended use.

Enzymotec describes PSEDGV as a family of structurally related edible oils containing between 20 and 60% phytosterol esters and lesser amounts of diglycerides (5 - 20%). PSEDGV is made by transesterification of phytosterols with commercial vegetable oils or solid vegetable fats (the fatty acid sources). Fatty acid sources are commercial vegetable oils (such as canola, soybean, sunflower, olive, and palm oils) and fats (such as cocoa butter). Phytosterols are generally obtained as byproducts of refining of conventional vegetable oils (soy, corn, sunflower, and rapeseed). The main sterol components of PSEDGV are ß-sitosterol, campesterol, stigmasterol, and brassicasterol; the sterols exist predominantly esterified to fatty acids. These fatty acids reflect the fatty acid composition of the source oil.

Enzymotec produces two grades of PSEDGV (regular and high). These grades reflect differences in the levels of component oil (triglycerides), byproducts of transesterification (diglycerides, monoglycerides, free fatty acids), and phytosterol esters. The regular-grade PSEDGV contains lower levels of phytosterol esters and higher levels of the component oil and its byproducts as compared to the high-grade PSEDGV.

Enzymotec manufactures PSEDGV by two separate methods (enzymatic and chemical). In the enzymatic method, an immobilized lipase enzyme preparation from Candida rugosa⁽²⁾ is used in the transesterification. The immobilization process is carried out in accordance with good manufacturing practice using materials approved for food use. Following reaction and separation from the catalyst, the resultant crude mixture of transesterified oil may undergo winterization (optional) to remove saturated fatty acids. This crude mixture then undergoes steam deodorization or molecular distillation,⁽³⁾ followed by addition of antioxidants (tocopherols). The enzymatic method results in the regular-grade product with approximately 25% phytosterol esters and 10 - 20% diglycerides. Enzymotec states that epichlorohydrin (used in the production of the immobilized enzyme) and 1,3-dichloro-2-propanol (formed by reaction of water with epichlorohydrin), if present, would be removed under the conditions (elevated temperature, vacuum) of the distillation process.

In the chemical method, sodium methoxide is used as the catalyst in the transesterification. Following chemical transesterification, the mixture is neutralized by addition of aqueous citric acid solution. The transesterified oil then undergoes processing similar to that used for traditional vegetable oils, including bleaching, winterization (optional), and steam deodorization or molecular distillation.⁽³⁾ Antioxidants (tocopherols) are added to the distilled ingredient. The chemical method results in the regular-grade product with approximately 25% or the high-grade product having approximately 60% phytosterol esters and 5 - 18% diglycerides. Enzymotec states that methanol (resulting from use of sodium methoxylate), if present, would be removed under the conditions (elevated temperature, vacuum) of the distillation process.

Enzymotec provides specifications for regular-grade and high-grade PSEDGV, including those for identity, purity, and microbial specifications. Specifications for regular-grade PSEDGV include triglyceride (35 - 65%), diglyceride (10 - 20%), monoglyceride (< 5%), phytosterol esters (> 20%), free fatty acids (< 10%), and lead (< 0.1 mg/kg). Specifications for high-grade PSEDGV include triglyceride (7 - 40%), diglyceride (5 - 18%), monoglyceride (< 5%), phytosterol esters (> 50%), free fatty acids (< 0.5%), and lead (< 0.1 mg/kg). Enzymotec provides results of batch analyses that support compliance with specifications.

Enzymotec discusses the intended use of, and provides estimated intakes for, PSEDGV. Enzymotec intends to use the regular-grade PSEDGV at a level of 2.6 g per RACC and the high-grade PSEDGV at a level of 1.1 g per RACC. The intended use of either grade of PSEDGV results in a level of 0.65 g phytosterol ester per RACC. Enzymotec provides estimates of intake of PSEDGV using the United States Department of Agriculture 1994-1996 Continuing Survey of Food Intakes by Individuals and the 1998 Supplemental Children's Survey. Enzymotec estimates that the eaters-only intakes of phytosterol esters (from regular- and high-grade PSEDGV) are 5.1 g/person/day (g/p/d) at the mean and 8.4 g/p/d at the 90th percentile. Enzymotec further estimates that intake of PSEDGV, in addition to current uses of phytosterols and phytosterol esters, would result in total intakes, expressed on a phytosterol ester basis, of 7.3 g/p/d at the mean and 12.9 g/p/d at the 90th percentile. The notice also includes corresponding estimates of intake of diglycerides, which Enzymotec notes are normal components of the diet through consumption of edible oils. Enzymotec considers these estimates (phytosterol esters, diglycerides) to be conservative, in part because of the assumption that all foods within a product category contain PSEDGV and in part because of the use of 2-day survey data, which are known to overestimate consumption of infrequently consumed foods.

Enzymotec considers the safey of PSEDGV to be supported by safety of its components: phytosterol esters and diglycerides.⁽⁴⁾ The notifier discusses published studies for phytosterols and diglycerides. Published studies for phytosterols include: absorption, distribution, metabolism, and elimination (ADME); subchronic and chronic animal; developmental and reproductive toxicity; genotoxicity; estrogenic potential; and clinical. Published studies for diglycerides include: ADME, subchronic and chronic animal, genotoxicity, and clinical. In addition, Enzymotec describes two unpublished studies with PSEDGV, a 10-week gavage feeding study in mice (regular-grade, canola oil-based PSEDGV) and a 4-week clinical study (regular-grade, olive oil-based PSEDGV). Although these two studies were not formally designed to investigate the safety of PSEDGV, Enzymotec concludes that these studies showed no adverse effect from the consumption of PSEDGV.

Standards of Identity

In the notice, Enzymotec states its intention to use PSEDGV in several food categories for which standards of identity exist 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.

Allergen Labeling

The Food Allergen Labeling and Consumer Protection Act of 2004 (FALCPA) amends the Federal Food, Drug, and Cosmetic 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. Issues associated with labeling food are the responsibility of the Office of Nutritional Products, Labeling, and Dietary Supplements in the Center for Food Safety and Applied Nutrition.

Conclusions

Based on the information provided by Enzymotec, and other information available to FDA, the agency has no questions at this time regarding Enzymotec's conclusion that PSEDGV 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 PSEDGV. As always, it is the continuing responsibility of Enzymotec 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 000206, as well as a copy of the information in this notice that conforms to the information in the proposed GRAS exemption claim (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,

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

---

⁽¹⁾The acronym "PSEDGV" denotes phytosterol esters and diglycerides produced using vegetable oils/fats as sources of fatty acids. There is a distinction between this ingredient and another ingredient produced using fish oils as sources of fatty acids. While Enzymotec's GRAS Panel reviewed both ingredients, only PSEDGV is the subject of GRN 000206.

⁽²⁾The lipase enzyme preparation from C. rugosa is the subject of GRN 000081.

⁽³⁾The notifier collectively terms the processes steam deodorization or molecular distillation as "distillation."

⁽⁴⁾Enzymotec notes FDA's previous responses to GRAS notices for phytosterols/phytosterol esters and diglycerides.