[Federal Register Volume 61, Number 28 (Friday, February 9, 1996)]
[Rules and Regulations]
[Pages 4871-4874]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 96-2747]



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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 

[[Page 4872]]
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) *  *  *

[[Page 4874]]


                                                                                                                
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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