Secondary Direct Food Additives Permitted in Food for Human Consumption; Periodic Acid and Polyethylenimine

DEPARTMENT OF HEALTH AND HUMAN SERVICES

Food and Drug Administration

21 CFR Part 173

[Docket No. 92F-0447]

Secondary Direct Food Additives Permitted in Food for Human

Consumption; Periodic Acid and Polyethylenimine

AGENCY: Food and Drug Administration, HHS.

ACTION: Final rule.

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SUMMARY: The Food and Drug Administration (FDA) is amending the food

additive regulations to provide for the safe use of periodic acid (PA)

and polyethylenimine (PEI) as fixing agents for the immobilization of

glucoamylase enzyme preparations from Aspergillus niger for use in the

manufacture of beer. This action is in response to a petition filed by

Enzyme Bio-Systems, Ltd.

DATES: Effective February 9, 1996; written objections and requests for

a hearing by March 11, 1996.

ADDRESSES: Submit written objections to the Dockets Management Branch

(HFA-305), Food and Drug Administration, rm. 1-23, 12420 Parklawn Dr.,

Rockville, MD 20857.-

FOR FURTHER INFORMATION CONTACT: Andrew D. Laumbach,- Center for Food

Safety and Applied Nutrition (HFS-217), Food and Drug Administration,

200 C St. SW., Washington, DC 20204, 202-418-3071.

SUPPLEMENTARY INFORMATION: In a notice published in the Federal

Register of December 1, 1993 (58 FR 63381), FDA announced that a food

additive petition (FAP 1A4288) had been filed by Enzyme Bio-Systems,

Ltd., 2600 Kennedy Dr., Beloit, WI 53511, proposing that the food

additive regulations be amended to provide for the safe use of periodic

acid and polyethyleneimine as fixing agents for immobilizing those

enzymes that are generally recognized as safe (GRAS) or approved as

food additives.

In a letter of February 2, 1994 (Ref. 1), the petition was amended

by the petitioner to provide for the use of PA and PEI as fixing agents

for the immobilization of glucoamylase enzyme preparations from A.

niger for use in the manufacture of light beer. The Bureau of Alcohol,

Tobacco, and Firearms, the Federal agency responsible for the

regulation of alcoholic beverages such as beer, has informed FDA that

the term ``light,'' with respect to the description of beer, is not

defined by regulation or any other regulatory standards (Ref. 2).

Accordingly, FDA has omitted the term ``light'' in the regulation

responding to this petition because there are no applicable Federal

standards defining ``light beer.''

Although the filing notice refers to polyethyleneimine as one of

the two petitioned additives under agency evaluation, it became

apparent during the review of the petition that the name of the

additive should be changed to be consistent with the name of the

substance that is currently listed in Sec. 173.357(a)(2) (21 CFR

173.357(a)(2)), ``polyethylenimine reaction product with 1,2-

dichloroethane.'' While the name of the additives differ, the additives

share the same Chemical Abstract Service (CAS) Registry Number (CAS

Reg. No. 68130-97-2) and are thus considered chemically identical by

the agency. The petitioner has agreed to the name change. Therefore,

the petitioned additive is identified as a PEI reaction product with

1,2-dichloroethane (DCE) in the regulation set forth below. However,

for purposes of discussion, this preamble will use the term

``polyethylenimine'' to refer to the additive, PEI reaction product

with 1,2-dichloroethane.

Glucoamylase enzyme preparation from A. niger is the substance

that is to be immobilized with the fixing agents set forth in the

regulation below; the regulatory status of that enzyme preparation is

not addressed by this action. The agency is, however, concurrently

evaluating this particular enzyme preparation, along with a variety of

other enzymes from other sources, in its review of petition GRASP

3G0016 (Docket No. 84G-0257) for the affirmation of the GRAS status of

certain enzymes. (Eight enzyme preparations included in GRASP 3G0016

were recently affirmed as GRAS (60 FR 32904, June 26, 1995).) The

petition GRASP 3G0016 contains published data and information to

support the view that the enzyme preparation glucoamylase from A. niger

has had a long history of use prior to 1958 in the preparation of food

as well as fermentable materials that are used in the production of

alcoholic beverages (Refs. 3 and 4). Further, FDA is not aware of any

data or information showing that glucoamylase from A. niger poses a

safety concern. Finally

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FDA acknowledges that under the Federal Food, Drug, and Cosmetic Act

(the act), a food manufacturer may market a substance for use in food

on the basis of the manufacturer's independent determination that the

substance is GRAS and thus exempt from the definition of food additive

in section 201(s) of the act (21 U.S.C. 321(s)).

In this scientific and legal context, FDA believes that it is

appropriate to proceed with a final rule approving the use of PEI and

PA as fixing agents for immobilizing glucoamylase from A. niger for

use in the manufacture of beer even though the agency has not completed

the GRAS affirmation process for all of the enzymes that are the

subject of GRASP 3G0016, including glucoamylase enzyme preparation.

In its evaluation of PA and PEI for the proposed use, FDA reviewed

the safety of the additives and the chemical impurities that may be

present in the additives resulting from the manufacturing processes.

Although neither PA nor PEI has been shown to cause cancer, PEI may

contain minute amounts of unreacted ethylenimine (EI) and 1,2-

dichloroethane because these reactants are used in the manufacture of

the additive. EI and 1,2-dichloroethane have been shown to be

carcinogens in bioassays with mice and rats (Refs. 5, 6, and 7). The

presence of such impurities is not unique to this additive. Residual

amounts of reactants and manufacturing aids are commonly found as

contaminants in chemical products, including food additives.

I. Determination of Safety

Under section 409(c)(3)(A) of the act (21 U.S.C. 348(c)(3)(A)), the

so-called ``general safety clause,'' a food additive cannot be approved

for a particular use unless a fair evaluation of the data and

information available to FDA establishes that the additive is safe for

that use. FDA's food additive regulations (21 CFR 170.3(i)) define safe

as ``a reasonable certainty in the minds of competent scientists that

the substance is not harmful under the intended conditions of use.''

The anticancer or Delaney clause (section 409(c)(3)(A)) of the act)

further provides that no food additive shall be deemed to be safe if it

is found to induce cancer when ingested by man or animal. Importantly,

however, the Delaney clause applies to the additive itself and not to

the constituents of the additive. That is, where an additive has not

been shown to cause cancer, but contains a carcinogenic impurity, the

additive is properly evaluated under the general safety clause using

risk assessment procedures to determine whether there is a reasonable

certainty that no harm will result from the proposed use of the

additive. (See Scott v. FDA, 728 F.2d 322 (6th Cir. 1984).)

II. Evaluation of Safety of Petitioned Use of the Additives

FDA estimates that the petitioned use of the additives PA and PEI

to fix glucoamylase enzyme preparations would result in mean exposures

to these additives of 0.7 micrograms per person per day (g/

person/day) for iodate, which is formed from the decomposition of PA

(Ref. 8), and no greater than 330 g/person/day for PEI (Ref.

9).

FDA does not ordinarily consider chronic toxicological testing to

be necessary to determine the safety of additives whose use will result

in such low exposure levels (Ref. 10), and the agency has not required

such testing here. The agency has reviewed the available toxicological

data from acute toxicity studies on the additives. No adverse effects

were reported in these studies (Ref. 11).

FDA has evaluated the safety of PEI under the general safety

clause, considering all available data and using risk assessment

procedures to estimate the upper-bound limit of risk presented by EI

and DCE that may be present as impurities in the additive. This risk

evaluation of EI and DCE has two aspects: (1) Assessment of the

exposure to the impurities from the proposed use of the additive, and

(2) extrapolation of the risk observed in animal bioassays to the

conditions of probable exposure to humans.

A. Ethylenimine (EI)

Using estimates of the average intake of beer, FDA estimates the

potential exposure to EI from the petitioned use of PEI as an

immobilizing agent for glucoamylase enzyme preparations from A. niger

used in the production of beer to be 0.33 nanograms (ng)/person/day

(Ref. 9). To estimate the risk from EI (Ref. 5), the agency used data

from a carcinogen bioassay with the B6C3F1 strain of mice using the

oral route of exposure. EI treatment caused an increased incidence of

both lung and liver tumors that were neoplastic (Ref. 5).

Based on a potential exposure of 0.33 ng/person/day, FDA estimates

that the upper-bound limit of individual lifetime risk from the

potential exposure to EI from the use of PEI is 1.2 x 10-7 , or

less than 1.2 in 10 million (Refs. 12 and 13). Because of the numerous

conservative assumptions used in calculating the exposure estimate,

actual lifetime averaged individual exposure to EI is expected to be

substantially less than the worst-case exposure, and therefore, the

calculated upper-bound limit of risk would be less. Thus, the agency

concludes that there is a reasonable certainty that no harm will result

from the proposed use of the additive as a result of exposure to EI.

B. 1,2-Dichloroethane (DCE)

Again, using estimates of average intake of beer, FDA estimates the

potential exposure to DCE to be 0.33 ng/person/day (Ref. 9). The agency

used data from two bioassays sponsored by the National Cancer Institute

to estimate risk; the bioassays showed that DCE is carcinogenic to mice

and rats at multiple tissue sites (Ref. 6). Based on the potential

exposure of 0.33 ng/person/day, FDA estimates that the upper-bound

limit of individual lifetime risk from the potential exposure to DCE

from the use of PEI is 6.4 x 10-11, or less than 6.4 in 100

billion (Refs. 12 and 13). Because of the numerous conservative

assumptions used in calculating this exposure estimate, actual lifetime

averaged individual exposure to DCE is expected to be substantially

less than the worst-case exposure, and therefore, the calculated upper-

bound limit of risk would be less. Thus, the agency concludes that

there is a reasonable certainty that no harm will result from the

proposed use of PEI as a result of exposure to DCE.

C. Need for Specifications

The agency has also considered whether specifications are necessary

to control the amount of EI and DCE impurities in PEI. The agency finds

that specifications for PEI are necessary, and that the specifications

in Sec. 173.357(a)(2) should be retained. The PA does not require

specifications for its use (Ref. 14) because it does not contain

impurities that need to be controlled.

III. Conclusions

FDA has evaluated data in the petition and other relevant material

and concludes that the proposed use of PA and PEI as fixing agents for

the immobilization of glucoamylase enzyme preparations from A. niger

used in the manufacture of beer is safe (Ref. 15). Based on this

information, the agency has also concluded that the additives will

function as intended. Therefore, Sec. 173.357(a)(2) should be amended

as set forth below.

FDA is also amending Sec. 173.357(a)(2) to revise the division name

and address listed in the regulation as a source of methods

incorporated by reference. The change results from a reorganization of

[[Page 4873]]

the Center for Food Safety and Applied Nutrition announced in a final

rule published in the Federal Register of April 1, 1993 (58 FR 17091).

In accordance with Sec. 171.1(h) (21 CFR 171.1(h)), the petition

and the documents that FDA considered and relied upon in reaching its

decision to approve the petition are available for inspection at the

Center for Food Safety and Applied Nutrition by appointment with the

information contact person listed above. As provided in Sec. 171.1(h),

the agency will delete from the documents any materials that are not

available for public disclosure before making the documents available

for inspection.

IV. Environmental Impact

In the notice of filing for this petition that published in the

Federal Register of December 1, 1993, FDA gave interested parties an

opportunity to submit comments on the petitioner's environmental

assessment by January 3, 1994, to the Dockets Management Branch

(address above). FDA received no comments in response to that notice.

The agency has carefully considered the potential environmental

effects of this action. FDA has concluded that the action will not have

a significant impact on the human environment, and that an

environmental impact statement is not required. The agency's finding of

no significant impact and the evidence supporting that finding,

contained in an environmental assessment, may be seen in the Dockets

Management Branch (address above) between 9 a.m. and 4 p.m., Monday

through Friday.

V. Objections

Any person who will be adversely affected by this regulation may at

any time on or before March 11, 1996, file with the Dockets Management

Branch (address above) written objections thereto. Each objection shall

be separately numbered, and each numbered objection shall specify with

particularity the provisions of the regulation to which objection is

made and the grounds for the objection. Each numbered objection on

which a hearing is requested shall specifically so state. Failure to

request a hearing for any particular objection shall constitute a

waiver of the right to a hearing on that objection. Each numbered

objection for which a hearing is requested shall include a detailed

description and analysis of the specific factual information intended

to be presented in support of the objection in the event that a hearing

is held. Failure to include such a description and analysis for any

particular objection shall constitute a waiver of the right to a

hearing on the objection. Three copies of all documents shall be

submitted and shall be identified with the docket number found in

brackets in the heading of this document. Any objections received in

response to the regulation may be seen in the Dockets Management Branch

between 9 a.m. and 4 p.m., Monday through Friday.

VI. References

The following references have been placed on display in the Dockets

Management Branch (address above) and may be seen by interested persons

between 9 a.m. and 4 p.m., Monday through Friday.

1. Letter from McKenna & Cuneo to the Direct Additives Branch

(HFS-217), amending the filing of Food Additive Petition No. 1A4288,

February 2, 1994.

2. Memorandum from Wine and Beer Branch, Bureau of Alcohol,

Tobacco, and Firearms to the Direct Additives Branch (HFS-217),

regarding regulations for ``light'' beer, October 6, 1994.

3. Underkofler, L. A., R. R. Barton, and S. S. Rennert,

``Microbiological Process Report--Production of Microbial Enzymes

and their Applications,'' Applied Microbiology, 6:212-221, 1958.

4. Beckhorn, E. J., M. D. Labbee, and L. A. Underkofler,

``Production and Use of Microbial Enzymes for Food Processing,''

Journal of Agricultural Food Chemistry, 13:30-34, 1965.

5. Memorandum from the Quantitative Risk Assessment Committee to

the Office of Toxicological Sciences (HFS-100), concerning risk

estimate for ethyleneimine, August 22, 1985.

6. Memorandum from the Cancer Assessment Committee, Color and

Cosmetics Evaluation Branch (HFF-158) to Division of Food and Color

Additives (HFF-330), Preliminary Risk Assessment on 1,2-

Dichloroethane (DCE) Migrating from Food and Beverage Contact Paper,

June 23, 1982.

7. Memorandum from Quantitative Risk Assessment Committee to the

Office of Toxicological Sciences (HFF-100), Epichlorohydrin, 1,2-

Dichloroethane, and 2,4-Toluenediamine in Reverse Osmosis Membranes

(FAP 6B3955), February 2, 1988.

8. Memorandum from the Chemistry Review Branch (HFS-247) to the

Direct Additives Branch (HFS-217), concerning letter dated October

20, 1993, and submission dated October 27, 1993, from McKenna &

Cuneo, January 11, 1994.

9. Memorandum from the Chemistry Review Branch (HFS-247) to the

Direct Additives Branch (HFS-217), Enzyme Bio-Systems Ltd.,

Submission of September 12, 1991; February 17, 1993.

10. Kokoski, C. J., ``Regulatory Food Additive Toxicology'', in

Chemical Safety Regulation and Compliance, edited by Homburger, F.

and J. K. Marquis, S. Karger, New York, NY, pp. 24-33, 1985.

11. Memorandum from the Additive Evaluation Branch (HFF-158) to

the Direct Additives Branch (HFF-217), concerning evaluation of Food

Additive Petition No. 1A4288, February 25, 1992.

12. Memorandum from the Chemistry Review Branch (HFS-247) to the

Direct Additives Branch (HFS-217), Exposure estimate for QRAC

evaluation, February 8, 1994.

13. Memorandum from the Direct Additives Branch (HFS-217) to the

Quantitative Risk Assessment Committee (HFS-308), Estimation of the

upper-bound lifetime risk for ethyleneimine (EI) and 1,2-

dichlorethane (DCE) for uses requested in FAP 1A4288 (Enzyme Bio-

Systems Ltd.), April 15, 1994.

14. Memorandum from the Chemistry Review Branch (HFS-247) to the

Direct Additives Branch (HFS-217) concerning Food Additive Petition

No. 1A4288, April 5, 1994.-

15. Memorandum from the Additives Evaluation Branch No. 1 (HFS-

226) to the Direct Additives Branch (HFS-217), final evaluation

memorandum, May 31, 1994.

List of Subjects in 21 CFR Part 173

Food additives.

Therefore, under the Federal Food, Drug, and Cosmetic Act and under

authority delegated to the Commissioner of Food and Drugs, 21 CFR part

173 is amended as follows:

PART 173--SECONDARY DIRECT FOOD ADDITIVES PERMITTED IN FOOD FOR

HUMAN CONSUMPTION

1. The authority citation for 21 CFR part 173 continues to read as

follows:

-Authority: Secs. 201, 402, 409 of the Federal Food, Drug, and

Cosmetic Act (21 U.S.C. 321, 342, 348).

2. Section 173.357 is amended in the table in paragraph (a)(2)

under the headings ``Substances'' and ``Limitations'' by alphabetically

adding a new entry for ``periodic acid'' and by revising the entry for

``polyethylenimine reaction product with 1,2-dichloroethane'' to read

as follows:

Sec. 173.357 Materials used as fixing agents in the immobilization of

enzyme preparations.

* * * * *

(a) * * *

(2) * * *

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Substances Limitations

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Periodic acid (CAS Reg. No. 10450-60-9).

Polyethylenimine reaction product with 1,2-dichloroethane May be used as a fixing material in the

(CAS Reg.No. 68130-97-2) is the reaction product of immobilization of glucoamylase enzyme preparations

homopolymerization of ethylenimine in aqueous from Aspergillus niger for use in the manufacture

hydrochloric acid at 100 deg.C and of cross-linking with of beer.

1,2-dichloroethane. The finished polymer has an average May be used as a fixing material in the

molecular weight of 50,000 to 70,000 as determined by gel immobilization of:

permeation chromatography. The analytical method is 1. Glucose isomerase enzyme preparations for use in

entitled ``Methodology for Molecular Weight Detection of the manufacture of high fructose corn syrup, in

Polyethylenimine,'' which is incorporated by reference in accordance with Sec. 184.1372 of this chapter.

accordance with 5 U.S.C. 552(a) and 1 CFR part 51. Copies 2. Glucoamylase enzyme preparations from

may be obtained from the Division of Petition Control, Aspergillus niger for use in the manufacture of

Center for Food Safety and Applied Nutrition (HFS-215), beer. Residual ethylenimine in the finished

200 C St. SW., Washington, DC 20204, and may be examined polyethylenimine polymer will be less than 1 part

at the Center for Food Safety and Applied Nutrition's per million as determined by gas chromatography-

Library, 200 C St. SW., rm. 3321, Washington, DC, or at mass spectrometry. The residual ethylenimine is

the Office of the Federal Register, 800 North Capitol St. determined by an analytical method entitled

NW., suite 700, Washington, DC. ``Methodology for Ethylenimine Detection in

Polyethylenimine,'' which is incorporated by

reference in accordance with 5 U.S.C. 552(a) and 1

CFR part 51. Residual 1,2-dichloroethane in the

finished polyethylenimine polymer will be less than

1 part per million as determined by gas

chromatography. The residual 1,2-dichloroethane is

determined by an analytical method entitled,

``Methodology for Ethylenedichloride Detection in

Polyethylenimine,'' which is incorporated by

reference in accordance with 5 U.S.C. 552(a) and 1

CFR part 51. Copies may be obtained from the

Division of Petition Control, Center for Food

Safety and Applied Nutrition (HFS-215), 200 C St.

SW., Washington, DC 20204, or may be examined at

the Center for Food Safety and Applied Nutrition's

Library, 200 C St. SW., rm. 3321, Washington, DC,

or the Office of the Federal Register, 800 North

Capitol St. NW., suite 700, Washington, DC.

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Dated: January 17, 1996.

William K. Hubbard,

Associate Commissioner for Policy Coordination.

[FR Doc. 96-2747 Filed 2-8-96; 8:45 am]

BILLING CODE 4160-01-F