[Federal Register Volume 79, Number 98 (Wednesday, May 21, 2014)]
[Rules and Regulations]
[Pages 29078-29085]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 2014-11584]


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DEPARTMENT OF HEALTH AND HUMAN SERVICES

Food and Drug Administration

21 CFR Part 172

[Docket No. FDA-2009-F-0303]


Food Additives Permitted for Direct Addition to Food for Human 
Consumption; Advantame

AGENCY: Food and Drug Administration, HHS.

ACTION: Final rule.

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SUMMARY: The Food and Drug Administration (FDA or we) is amending the 
food additive regulations to provide for the safe use of advantame as a 
non-nutritive sweetener and flavor enhancer in foods generally, except 
meat and poultry. This action is in response to a petition filed by 
Ajinomoto Co., Inc.

DATES: This rule is effective May 21, 2014. See section IX for further 
information on the filing of objections. Submit either electronic or 
written objections and requests for a hearing by June 20, 2014. The 
Director of the Office of the Federal Register approves the 
incorporation by reference of certain publications listed in the rule 
as of May 21, 2014.

ADDRESSES: You may submit either electronic or written objections and 
requests for a hearing identified by Docket No. FDA-2009-F-0303, by any 
of the following methods:

Electronic Submissions

    Submit electronic objections in the following way:
     Federal eRulemaking Portal: http://www.regulations.gov. 
Follow the instructions for submitting comments.

Written Submissions

    Submit written objections in the following way:

[[Page 29079]]

     Mail/Hand delivery/Courier (for paper submissions): 
Division of Dockets Management (HFA-305), Food and Drug Administration, 
5630 Fishers Lane, Rm. 1061, Rockville, MD 20852.
    Instructions: All submissions received must include the Agency name 
and Docket No. FDA-2009-F-0303 for this rulemaking. All objections 
received will be posted without change to http://www.regulations.gov, 
including any personal information provided. For detailed instructions 
on submitting objections, see the ``Objections'' heading of the 
SUPPLEMENTARY INFORMATION section.
    Docket: For access to the docket to read background documents or 
objections received, go to http://www.regulations.gov and insert the 
docket number, found in brackets in the heading of this document, into 
the ``Search'' box and follow the prompts and/or go to the Division of 
Dockets Management, 5630 Fishers Lane, Rm. 1061, Rockville, MD 20852.

FOR FURTHER INFORMATION CONTACT: Felicia M. Ellison, Center for Food 
Safety and Applied Nutrition (HFS-265), Food and Drug Administration, 
5100 Paint Branch Pkwy., College Park, MD 20740-3835, 240-402-1264.

SUPPLEMENTARY INFORMATION: 

Table of Contents

I. Background
II. Evaluation of Safety of Advantame
    A. Chemistry and Intake Considerations of Advantame
    B. Overview of Advantame Safety Studies
    C. Toxicology/Safety Assessment of Advantame
    D. Estimating an Acceptable Daily Intake of Advantame
III. Comments
IV. Conclusion
V. Public Disclosure
VI. Environmental Impact
VII. Paperwork Reduction Act of 1995
VIII. Section 301(ll) of the FD&C Act
IX. Objections
X. References

I. Background

    In a notice published in the Federal Register of July 21, 2009 (74 
FR 35871), we announced that Ajinomoto Co., Inc., c/o Ajinomoto 
Corporate Services LLC, 1120 Connecticut Ave. NW., Suite 1010, 
Washington DC, 20036, had filed a food additive petition (FAP 9A4778). 
The petition proposed to amend the food additive regulations in part 
172 Food Additives Permitted for Direct Addition to Food for Human 
Consumption (21 CFR part 172), to provide for the safe use of advantame 
as a non-nutritive sweetener in tabletop applications and powdered 
beverage mixes.
    In a letter dated August 24, 2012, the petitioner informed us that 
the care of FAP 9A4778 had been transferred from Ajinomoto Corporate 
Services LLC to Ajinomoto North America, Inc., One Parker Plaza, 400 
Kelby St., Fort Lee, NJ 07024.
    In an amended notice published in the Federal Register of October 
26, 2012 (77 FR 65340), we announced that Ajinomoto Co., Inc., c/o 
Ajinomoto North America, Inc., One Parker Plaza, 400 Kelby St., Fort 
Lee, NJ 07024, had amended its food additive petition to also provide 
for the safe use of advantame as a non-nutritive sweetener and flavor 
enhancer in foods generally, except in meat and poultry.

II. Evaluation of Safety of Advantame

    Under section 409(c)(3)(A) of the Federal Food, Drug, and Cosmetic 
Act (the FD&C Act) (21 U.S.C. 348(c)(3)(A)), a food additive cannot be 
approved for a particular use unless a fair evaluation of the data 
available to FDA establishes that the additive is safe for that use. 
``Safe'' or ``safety'' in the context of food additives means that 
there is ``a reasonable certainty in the minds of competent scientists 
that the substance is not harmful under the intended conditions of 
use'' (21 CFR 170.3(i)). To establish with reasonable certainty that a 
food additive is not harmful under its intended conditions of use, we 
consider the projected human dietary exposure to the additive, the 
additive's toxicological data, and other relevant information (such as 
published literature) available to us. We compare an individual's 
estimated daily intake (EDI) of the additive from all food sources to 
an acceptable intake level established by toxicological data. The EDI 
is determined by projections based on the amount of the additive 
proposed for use in particular foods and on data regarding the amount 
consumed from all food sources of the additive. We commonly use the EDI 
for the 90th percentile consumer of a food additive as a measure of 
high chronic dietary intake.

A. Chemistry and Intake Considerations of Advantame

    Advantame is the common or usual name for the chemical N-[N-[3-(3-
hydroxy-4- methoxyphenyl)propyl]-[alpha]-aspartyl]-L-phenylalanine 1-
methyl ester, monohydrate (CAS Reg. No. 714229-20-6). The additive is a 
white to yellowish crystalline powder that is an N-substituted 
derivative of the sweetener aspartame (21 CFR 172.804), with the amino 
nitrogen of aspartame alkylated with a 3-hydroxy-4-methoxy phenyl 
moiety. Advantame also is similar to the sweetener neotame, another N-
substituted derivative of aspartame that is approved as a sweetener in 
foods generally, except meat and poultry, in accordance with current 
good manufacturing practice, in an amount not to exceed that reasonably 
required to accomplish the intended technical effect, in foods for 
which standards of identity established under section 401 of the FD&C 
Act (21 U.S.C. 341) do not preclude such use (21 CFR 172.829). Data in 
the petition show that advantame has a sweetening potency that is 
approximately 20,000 times that of sucrose, depending on its food 
application (Ref. 1). The petitioner has proposed the use of advantame 
in food at levels not in excess of that reasonably required to produce 
its intended technical effect. We have reviewed results from taste 
panel studies that investigated the sweetness profile of advantame as a 
function of concentration in a variety of foods, and these data 
demonstrate that advantame can be used at self-limiting levels in food 
(Refs. 1 and 2).
    Based upon data from stability studies on advantame, we concluded 
that advantame is stable under normal storage and use conditions. The 
stability studies show that degradation of advantame is pH-, time-, and 
temperature-dependent and is more likely to occur from its use in low 
pH foods (i.e., acidic foods) during extended storage conditions. Under 
such extreme conditions, the principal degradation product is N-[N-[3-
(3-hydroxy-4-methoxyphenyl)propyl]-[alpha]-aspartyl]-L-phenylalanine 
(ANS-acid), which is the de-esterified form of advantame. As is the 
case with neotame, the N-alkyl substituent effectively prevents the 
common dipeptide cyclization reaction that results in the formation of 
a diketopiperazine derivative (Refs. 1 to 3).
    Further, there is no concern from exposure to these degradation 
products under either normal or extended storage and use conditions 
(Refs. 2 and 4).
    The petitioner determined the eaters-only EDI of advantame (i.e., 
the EDI for the population of study subjects that consumed one or more 
of the foods containing the additive) from its proposed use as a 
general-purpose sweetener and flavor enhancer at the 90th percentile of 
consumption to be 10 milligrams per person per day (mg/p/d) for the 
total U.S. population (all ages) and 8.1 mg/p/d for children (3 to 11 
years old). The corresponding mean estimated intakes are 4.9 mg/p/d and 
4.6 mg/p/d, respectively. We concur with the petitioner's exposure 
estimate for advantame (Ref. 2).
    We also estimated the eaters-only EDI of the principal degradation 
product

[[Page 29080]]

(ANS-acid), related impurities that may be formed during the 
manufacture of advantame, and related degradation products that may be 
formed under certain conditions in food. The eaters-only EDI of the 
principal degradation product, related impurities, and related 
degradation products at the 90th percentile of consumption is 0.10 mg/
p/d, 0.15 mg/p/d, and 0.20 mg/p/d, respectively, for the total U.S. 
population (all ages); and 0.08 mg/p/d, 0.12 mg/p/d, and 0.16 mg/p/d, 
respectively for children (3 to 11 years old) (Ref. 2).
    We also estimated the eaters-only dietary exposure to both 
advantame and its degradation products for other subpopulations, 
including various age groups of children, and have concluded that the 
exposure estimated for the U.S. population (all ages) represents the 
upper-bound cumulative dietary exposure to advantame and its 
degradation products from food (Ref. 2).

B. Overview of Advantame Safety Studies

    In support of the safety of advantame, the petitioner submitted 37 
preclinical (animal), clinical (human subjects), and specialty 
toxicology studies, along with several additional exploratory or 
screening studies. All pivotal preclinical studies were conducted in 
accordance with our Good Laboratory Practice (GLP) regulations 
appearing in 21 CFR part 58, or in accordance with other 
internationally accepted GLP standards.
    The preclinical studies included in vivo short-term, sub-chronic, 
and chronic studies in the rat, mouse, rabbit, and dog, including 
reproductive and developmental studies in the rat and rabbit. The 
safety data also included neurotoxicity and immunotoxicity studies in 
the rat; pharmacokinetic studies in the mouse, rat, and dog; 
carcinogenicity studies in the mouse and rat; and a series of in vitro 
mutagenicity and genotoxicity studies. The petitioner also submitted 
studies assessing tolerance in the rabbit and dog, and palatability in 
the mouse.
    In addition, the petitioner submitted four clinical studies that 
examined tolerance, absorption, distribution, metabolism, and excretion 
(ADME) of advantame in human subjects. Subjects in the ADME studies 
included healthy adult males and females, as well as adult males and 
females with type 2 diabetes.

C. Toxicology/Safety Assessment of Advantame

1. Pharmacokinetics and Metabolism of Advantame
    The petitioner conducted pharmacokinetic and metabolism studies in 
the rat, dog, and humans to support the safety of advantame for human 
use. The studies were designed to address the metabolic fate (i.e., 
absorption, distribution, metabolism, and excretion) of advantame.
    a. Absorption of advantame. Pharmacokinetic parameters estimated 
from advantame study data show that absorption of advantame and its 
metabolites occurs almost entirely in the small intestine, and that the 
amount absorbed can approach 15 percent in humans. Advantame absorption 
rates varied 2- to 4-fold between individuals. The rat and dog appeared 
to absorb less advantame than humans (8 to 15 percent as compared to 
humans). Absorption of advantame was limited by rapid intestinal 
hydrolysis of the methyl ester in all species.
    b. Distribution of advantame. The petitioner conducted studies with 
radiolabelled advantame to identify which organs might accumulate 
advantame or its metabolites if absorbed. In the rat, the radiolabelled 
advantame was found primarily in the organs of absorption 
(gastrointestinal (GI) tract), metabolism (liver), and excretion (GI 
tract, kidneys, and urinary bladder). Low levels of radioactivity were 
observed in all other tissues. Distribution of radiolabelled advantame 
in the dog was studied after oral dosing and was dominated by high 
concentrations of radioactivity in the organs of absorption, followed 
by excretory organs, such as the liver and kidneys. There was very 
little radioactivity detected in other tissues. In a study using 
radiolabelled advantame in pregnant rats, low levels of radioactivity 
were observed in the placenta, with no radioactivity observed in the 
fetuses. Based on these findings, we conclude there is no concern for 
possible accumulation of advantame or its metabolites at expected human 
intake levels.
    c. Metabolism of advantame. Data from metabolism studies using 
radiolabelled advantame in the rat, dog, and human volunteers showed 
five metabolites: (1) The methyl ester hydrolysis product (ANS9801-
acid); (2) a sulfate conjugate of ANS9801-acid (ANS-a-SO4), N-[N-[3-(3-
sulfoxy-4-methoxyphenyl)propyl]-L-[alpha]-aspartyl]-L-phenylalanine; 
(3) de-methyoxylated metabolite of ANS9801-acid (RF-1), N-[N-[3-(3,4-
dihydroxyphenyl)propyl]-L-[alpha]-aspartyl]-L-phenylalanine; (4) the 
phenylalanine cleavage product of ANS9801-acid (HF-1), N-[3-(3-hydroxy-
4-methoxyphenyl)propyl]-L-[alpha]-aspartic acid; and (5) 3-(3-hydroxy-
4-methoxyphenyl) propylamine (HU-1).
    ANS9801-acid represented 40 percent or more of the excreted 
metabolic products in all species tested. HF-1 and HU-1 were other 
minor metabolites. These metabolites likely are derived from ANS9801-
acid in the intestines. In humans, HF-1 and ANS9801-acid were the only 
metabolites identified in feces, at 30  12 percent and 52 
 13 percent of the dose, respectively. Other 
(uncharacterized) metabolites accounted for 0 to 3 percent of the dose 
in feces. ANS9801-acid represented 43 percent of urinary radioactivity, 
with HU-1 and HF-1 representing 35 percent and 19 percent of the 
urinary radioactivity, respectively. The remaining 2 to 3 percent of 
urinary radioactivity consisted of uncharacterized metabolites. 
Overall, 82 to 100 percent of the radioactivity was accounted for in 
these studies, which is within the acceptable range of recoveries for 
pharmacokinetic studies.
    Methanol and phenylalanine both are released during the metabolism 
of advantame. The metabolism studies provided by the petitioner 
indicated that most advantame residues excreted in the feces and urine 
are in the form of the metabolite ANS9801-acid. At the EDI for 
advantame, it is unlikely that even 100 percent conversion of advantame 
to methanol or phenylalanine would affect physiological levels of 
methanol or phenylalanine. Therefore, we conclude that the amounts of 
methanol and phenylalanine released from metabolism of advantame do not 
represent a safety concern (Ref. 5).
    d. Excretion of advantame. Advantame and its metabolites were 
rapidly eliminated from the rat and human. The findings were similar in 
dogs, with the exception of the excretion of the metabolite ANS-a-SO4, 
which was eliminated more slowly. Advantame has an approximate half-
life (the amount of time required for a quantity of a substance to fall 
to half its initial value) of less than 60 minutes after absorption in 
humans. The metabolite ANS9801-acid has a half-life of 3 to 5 hours in 
humans. Ultimately, 90 to 95 percent of absorbed advantame is excreted 
in the feces and urine within 24 hours of absorption. Based on the 
review findings from the metabolism and pharmacokinetic studies on 
advantame, there is no indication that advantame or its metabolites 
will accumulate in humans. In addition,

[[Page 29081]]

given the rapid rates of excretion, there is no indication that 
advantame or its metabolites will accumulate in the body from the 
proposed uses of advantame (Ref. 3). The potential intake of the 
primary metabolite, the ANS9801-acid, as well as other minor 
metabolites is of no toxicological consequence. Therefore, we conclude 
that the metabolism and pharmacokinetic studies of advantame do not 
raise any safety concerns (Ref. 5).
2. Neurotoxicity and Immunotoxicity Assessment of Advantame
    The petitioner investigated the potential neurotoxicity of 
advantame in rats. Within each of the standard toxicology studies 
submitted, the petitioner also reported physical, behavioral, and 
clinical observations for each animal, followed by extensive 
histological evaluations of brain, spinal cord, and peripheral nerves. 
Data on critical prenatal neurological development were examined in the 
in utero phase of the carcinogenicity/chronic toxicity studies in rats. 
No treatment-related neurotoxicological effects or abnormal behaviors 
were seen in animals that were exposed to advantame in these studies.
    In addition to examining various general endpoints related to 
neurological systems within standard toxicology studies, the petitioner 
conducted a neurobehavioral study in which rats were fed diets 
containing 10, 100, or 1,000 milligrams per kilogram body weight (mg/kg 
bw) of advantame. One group of rats was fed a diet containing 3 mg/kg 
bw of amphetamine sulfate as a positive control. Locomotor activity of 
the rats was measured for 10 minutes at each dose interval beginning 
with the pre-dose period followed by measurements performed at 30, 60, 
180, and 300 minutes post-dose. The study authors concluded that there 
were no significant effects of advantame on spontaneous locomotor 
activity at any dose level under the conditions of the study.
    Based on the lack of effect on rat locomotor activity of advantame 
given at the highest dose, we concluded that the No Observed Effect 
Level (NOEL) under the conditions of this study was 1,000 mg/kg bw 
(Ref. 6). Given the lack of signs of neurotoxicity, as well as an 
absence of histopathological change in the central nervous system 
(brain and spinal cord) and peripheral nerves in any of the treated 
animals, we conclude that the neurotoxicity studies of advantame do not 
raise safety concerns (Ref. 4).
    The petitioner presented data for two general, repeat-dose 
toxicology studies, a 4-week and a 13-week rat study, that evaluated 
the immunotoxicity potential of advantame. Findings related to various 
immune responses in these rat studies initially appeared to represent 
potential immunotoxicity responses (Ref. 7). After further evaluation, 
we determined that the lymphocyte reduction observed in the studies was 
due to individual animal variations and not to treatment with 
advantame. We also evaluated the reported low thymic weights in the 
high-dose groups of both sexes for the 4-week study and concluded that 
this change was consistent with a non-specific high-dose stress 
response because it was limited to the high-dose groups and affected 
only a few animals. We reviewed the seemingly dose-related degenerative 
changes in the thymuses of the 13-week female groups and determined 
that this change likely was incidental because it was not reported in 
either the 4-week or 2-year rat studies. Overall, we concluded that the 
immunological findings observed in the two rat studies did not have any 
toxicological significance as there was no evidence of a treatment-
related immunotoxic response (Ref. 8). Based on these evaluations, we 
concluded that advantame did not cause immunotoxicological effects 
within the context of these rat studies (Ref. 4).
    The petitioner conducted an additional immunotoxicity study in the 
same rat strain used in the 4-week and 13-week rat studies. In this 
study, rats were fed diets containing 0 mg/kg bw (control); 1,500 mg/kw 
bw; 5,000 mg/kg bw; and 15,000 mg/kg bw of advantame for 4 weeks. 
Groups of 10 rats of each sex were examined at the end of treatment, as 
well as after a 30-day recovery period. No treatment-related effects 
were detected in the various immunological parameters examined, 
including lymphocyte counts, thymus weights, immunophenotyping of 
lymphocytes, and lymphocyte proliferation assay, in the study. Based on 
these data, we concluded that advantame did not produce any immunotoxic 
effects under the conditions of this study (Ref. 9).
3. Human Clinical Studies
    The petitioner submitted four human clinical studies as part of the 
safety data for advantame to demonstrate tolerance of the sweetener in 
humans. The first clinical study was conducted to investigate the 
tolerability of advantame when administered orally to healthy adult 
males at dose levels of 0.1 mg/kg bw, 0.25 mg/kg bw, and 0.35 mg/kg bw. 
The study also investigated the pharmacokinetic profile of advantame in 
the same volunteers. We concluded that the oral administration of 
advantame was tolerable in healthy adult male subjects when 
administered as a single dose at each dose level without the occurrence 
of any treatment-related adverse events during a subsequent 7-day 
observation period (Ref. 10). Based on this study, we concluded that 
advantame is well tolerated in healthy human males.
    The second clinical study was conducted to characterize the 
metabolic profile of advantame in urine and feces in human subjects. 
This study investigated the absorption, metabolism, and excretion of 
radiolabelled advantame after a single oral dose at 0.25 mg/kg bw in 
six healthy adult male volunteers. In this study, systemic absorption 
of advantame was reported to be in the range of 9 to 30 percent (Ref. 
10). We concluded that data on the pharmacokinetic profile of advantame 
from this study, although limited, was useful in our evaluation of the 
safety of advantame. Based on this study, we have no safety concerns 
with the absorption, metabolism, or excretion of advantame as it was 
well tolerated in human subjects.
    The third clinical study was conducted to investigate the 
tolerability of repeated daily consumption of a 30 mg dose of advantame 
(equivalent to 0.375 mg/kg bw/day to 0.5 mg/kg bw/day) over a period of 
4 weeks using six healthy subjects of each sex. The study also included 
a placebo control group consisting of six healthy subjects of each sex 
that received diets without advantame. Based on results of the study, 
we concluded that, although there were apparent small differences in 
blood plasma values of the main metabolite of advantame, ANS9801-acid, 
the differences were not due to randomization procedures of the study 
and, instead, were reflective of within-subject variability inherent in 
the subjects of the study (Ref. 10). We concluded advantame was well 
tolerated in these subjects and that there were no safety concerns.
    The fourth clinical study was conducted as a double blind, placebo-
controlled study in diabetic subjects designed to investigate the 
tolerability of repeated daily consumption of a 30 mg dose (equivalent 
to 0.375 mg/kg bw/day to 0.5 mg/kg bw/day) of advantame fed daily for 
12 weeks, using 18 diabetic subjects of each sex per group. Diabetic 
subjects in the placebo-controlled group received diets without 
advantame. Based on the results of this study, we noted that there were 
no clinically significant changes identified. We concluded that 
advantame was tolerated

[[Page 29082]]

at daily doses up to 0.5 mg/kg bw/day in people with type 2 diabetes.
    We raised concerns about the experimental design (e.g., sample size 
and the randomization procedures) in some of the clinical studies (Ref. 
10). However, overall, we ultimately concluded that advantame was well-
tolerated in healthy males when fed a single dose of advantame at dose 
levels of 0.1 mg/kg bw/day to 0.5 mg/kg bw/day. The third and fourth 
clinical studies showed that advantame was tolerated in healthy males 
and females and type 2 diabetic males and females when repeatedly fed a 
dose of 0.375mg/kg bw/day to 0.5 mg/kg bw/day for 4 weeks. The doses 
administered in the third and fourth studies were approximately 3-fold 
higher than the EDI for consumers of all ages at the 90th percentile of 
consumption (Ref. 10).
    Pharmacokinetic evaluations of advantame were conducted on blood 
plasma samples from the human subjects that received single and repeat 
dose administrations of advantame. Data from these analyses showed that 
advantame was undetectable in plasma samples 4 hours after its 
administration. The repeat dosing studies showed variation in the 
plasma levels of ANS9801-acid for some subjects. The significance of 
this variability could not be determined because of the small number of 
subjects examined. However, the variable ANS9801-acid levels were not 
associated with any clinically significant, treatment-related toxicity 
in these subjects.
    Clinically significant treatment-related toxicities or adverse 
events were not noted in the advantame-treated groups in any of these 
clinical studies. Overall, the clinical studies showed that oral 
administration of advantame was tolerated in humans fed up to 30 mg per 
day (Ref. 10).
4. Critical Toxicology Studies
    We reviewed all studies and supplemental information submitted by 
the petitioner. During our review, we determined that certain studies 
were more pivotal in supporting a regulatory decision on the petitioned 
uses of advantame. We based our determination on the experimental 
design of the studies as well as the types of the studies' endpoints. 
We gave greater weight to the studies that examined the reproductive 
and developmental effects, long-term exposure, chronic toxicity, 
carcinogenicity potential, and investigations of specific toxicological 
issues presented by these studies. The critical studies were: (1) A 
two-generation reproduction study in rats; (2) a chronic (52-week) dog 
study; (3) a 104-week mouse carcinogenicity study, and (4) a combined 
104-week rat carcinogenicity feeding study with in utero and chronic 
(52-week) phases.
    a. Two-generation reproduction study in the rat. Reproductive 
performance and fertility were assessed over two generations in rats 
fed diets containing advantame at levels of 2,000 ppm, 10,000 ppm, or 
50,000 ppm. The parental rats received the advantame diet for 10 weeks 
before pairing and during mating. Parental and first generation female 
rats continued to receive the advantame treatment throughout gestation, 
lactation, and until death. A control group of rats received the 
untreated basal diet for the same period of time. The first generation 
contained 25 male and 25 females from each of the parent groups and 
received advantame at the same dietary concentrations as their parents 
throughout the study until termination. Direct treatment of the first 
generation rats began at 4 weeks of age for 10 weeks before pairing and 
mating for the second generation litters. The first generation 
continued treatment until termination after the second generation 
litters were weaned.
    Under the conditions of this study, advantame administration to 
rats did not produce any effects on mortality, body weight, estrous 
cycle, sperm motility, mating, fertility, duration of gestation, 
outcome of parturition, litter size, sex ratio, pup birth weights, 
survivability of pups, motor activity of pups, organ weights, or 
histopathology in either generation. However, at the 50,000 ppm dose 
level, statistically significant increased feed consumption in the 
advantame treated rats compared to the control rats during the 
maturation phases (before pairing) of parental males and first 
generation males and females was reported. This increased feed 
consumption, in the absence of any effect on feed conversion efficiency 
and body weight gain, was not considered toxicologically significant 
(Refs. 4 and 11). Based upon the findings, we established a No Observed 
Adverse Effect Level (NOAEL) at the 50,000 ppm dose for advantame-
treated rats in this study.
    b. Chronic (52-week) study in dogs. Chronic toxicity of advantame 
was evaluated in beagle dogs that were fed diets containing advantame 
at levels of 0 ppm, 2,000 ppm, 10,000 ppm, and 50,000 ppm over a 52-
week period using four dogs/per sex/per group. Two additional dogs per 
sex were assigned to each dose group as part of a 6-week recovery phase 
without advantame. This study was performed to evaluate systemic 
toxicity of advantame in non-rodent species. The only clinical sign 
related to advantame treatment was the observation of pale feces in all 
high-dose and some mid-dose dogs of both sexes. We established a NOAEL 
for this study at the 50,000 ppm dose of advantame, the highest dose 
tested, equivalent to 2,058 mg/kg bw/day in male dogs and 2,139 mg/kg 
bw/day in female dogs (Refs. 4 and 12). We also concluded that systemic 
toxicity in the test animals associated with advantame administration 
was not apparent.
    c. The 104-week mouse carcinogenicity study. The carcinogenicity 
potential of advantame was evaluated in mice (64/sex/group). The mice 
were fed diets containing advantame at levels of 0 ppm, 2,000 ppm, 
10,000 ppm, or 50,000 ppm for 104 weeks beginning when they were 
approximately 6 weeks old. One hundred seventy-three male and 177 
female mice died or were euthanized at the point of near death over the 
study period. A statistically significant effect of treatment on the 
distribution of deaths in the various dosing groups compared to the 
controls was not reported. The study's authors noted that the high 
death rate was not altered by the administration of advantame and that 
no specific factors that contributed to this rate were greater in 
number in the experimental groups compared to the control groups.
    We noted a low survival rate of the test animals, a common finding 
in 2-year bioassays using the CD-1 mouse, and a number of various 
clinical signs in both the control and treated mice (Ref. 13). Our 
evaluation of the mouse survival data revealed no evidence of premature 
deaths that were due to treatment and none of the findings indicated a 
proliferative response as the cause of early death in these mice. We 
considered the data available up to the 92-week observation period and 
determined that 25 or more surviving animals per group was adequate to 
evaluate the carcinogenic potential for advantame. We concluded that 
none of the clinical signs observed correlated consistently with a 
histomorphological diagnosis or were an indication of treatment-related 
toxicity (Ref. 14).
    The Center for Food Safety and Applied Nutrition's Cancer 
Assessment Committee (CAC) evaluated data from the 104-week mouse study 
for the carcinogenic potential of advantame. The CAC concluded that 
oral administration of advantame at doses up to 50,000 ppm for 104 
weeks did not produce any treatment-related tumors or any evidence of 
increased incidences of tumors in mice (Ref. 15). We established a NOEL 
for female mice of 10,000 ppm

[[Page 29083]]

advantame in the diet (based on decreased weight gain at 50,000 ppm) 
and a NOEL of 50,000 ppm advantame in the diet for male mice, 
equivalent to 5,693 mg/kg bw/day (Ref. 16).
    d. Combined 104-week rat carcinogenicity study with in utero phase 
and toxicity phase. This study included three phases: (1) An in utero 
reproduction phase; (2) a 52-week chronic toxicity phase; and (3) a 
104-week oral carcinogenicity phase. In each of the study phases, rats 
were fed diets containing advantame at levels of 2,000 ppm, 10,000 ppm, 
or 50,000 ppm. The control groups of rats received a similar diet 
without advantame for the same period of time. The in utero 
reproduction phase of this study was designed to generate and assess 
populations of rats that had been exposed to advantame prior to mating, 
during mating, and throughout gestation and lactation up to weaning and 
the start of the main chronic and carcinogenicity studies. Four-week-
old offspring produced during the parent mating were used to populate 
the first generation that was subsequently used in the 104-week 
carcinogenicity study and in the 52-week chronic toxicity rat study. 
Offspring that did not meet the survival criteria or had abnormal 
bodyweights were not used, and where possible, the numbers of surviving 
offspring per litter were reduced by random selection to four males and 
four females per litter. Adult parent males were killed after mating; 
adult parent females were killed after litters were weaned. Body 
weights, feed consumption, and survival rates were evaluated in the 
parent rats. The abilities to mate and give birth also were evaluated 
in the parent rats. The numbers of offspring, sex ratios, and litter 
weights were recorded for the first generation offspring.
    Results from the in utero phase of the rat study showed that: (1) 
Fertility, growth, and survival in the parent rats was unaffected by 
advantame treatment; (2) body weights and feed consumption in the 
treated parent groups were similar to that seen in the control rats; 
and (3) initial body weights of the first generation rats that were 
selected for either the carcinogenicity study or the 52-week toxicity 
study were not affected by exposure to advantame during preconception, 
in utero, or during weaning.
    The chronic toxicity phase of this study consisted of three 
advantame treatment groups of first generation rats selected from the 
in utero study, with 20 of each sex per group. The rats were fed diets 
containing advantame at levels of 2,000 ppm, 10,000 ppm, and 50,000 
ppm. A group of untreated first generation rats not exposed to 
advantame was selected to serve as controls for this 52-week phase of 
the study. An additional 10 rats of each sex were added to the control 
group and the 10,000 ppm and 50,000 ppm treatment groups to provide 
animals for a 6-week recovery phase without advantame following their 
initial advantame exposure period (week 0 to week 52).
    The study's authors reported no effect of the administration of 
advantame on mortality, maternal body weight gain and feed consumption, 
fertility, or on the growth and survival of offspring during the in 
utero phase. (``In utero,'' in this context, refers to the exposure of 
the developing embryo-fetus within the womb (uterus) of the mother 
(Parental F0 females).) Two animals died during the course of the 
treatment phase. These deaths, however, were not dose related. One male 
in the high-dose group died during the recovery phase.
    The CAC evaluated data from the 104-week rat carcinogenicity study 
for the carcinogenic potential of advantame. The CAC concluded that 
oral administration of advantame at doses up to 50,000 ppm for 104 
weeks did not produce any treatment-related tumors or any evidence of 
increased incidences of tumors in rats (Ref. 15). We established a 
NOAEL for this study of 50,000 ppm advantame in the diet, equivalent to 
an achieved dose of 3,279 and 4,025 mg/kg bw/day in males and females, 
respectively (Ref. 16). We also concluded that advantame treatment did 
not result in an increased incidence of tumors in rats.
    Based on our review of the previously mentioned critical studies, 
we concluded that there is no cause for concern regarding the 
carcinogenicity potential of advantame as proposed for its use as a 
non-nutritive sweetener and flavor enhancer in foods.

D. Estimating an Acceptable Daily Intake of Advantame

    In determining an acceptable daily intake (ADI) for a new 
ingredient, we rely on a comprehensive evaluation of all relevant 
studies and information submitted by the petitioner. Four studies had 
the greatest impact in our reaching a safety decision. These studies 
are highlighted in table 1.

     Table 1--Summary of Study Data Pertinent to Establishing an Acceptable Daily Intake Value for Advantame
----------------------------------------------------------------------------------------------------------------
                                                                                           NOEL \2\    NOAEL \3\
                 Study                       Dose range (ppm)       Pivotal \1\ Endpoint     (ppm)       (ppm)
----------------------------------------------------------------------------------------------------------------
Rat two-generation reproductive study.  0, 2,000, 10,000, 50,000.  ND                         10,000      50,000
Dog 52-week study.....................  0, 2,000, 10,000, 50,000.  ND                         10,000      50,000
Mouse 2-year bioassay.................  0, 2,000, 10,000, 50,000.  ND                         10,000      50,000
Rat 2-year bioassay with in utero and   0, 2,000, 10,000, 50,000.  ND                         10,000      50,000
 1-year chronic phase.
----------------------------------------------------------------------------------------------------------------
\1\ ND = None Detected.
\2\ NOEL = No Observed Effect Level.
\3\ NOAEL = No Observed Adverse Effect Level.

    Based on our review of the studies summarized in table 1, we 
determined the most appropriate study for establishing an ADI for 
advantame was the combined 104-week rat carcinogenicity study with in 
utero and chronic (52-week) phases. This study was of sufficient length 
and overall complexity to produce information on chronic exposure, 
potential toxicity, and potential carcinogenicity of advantame. 
Therefore, the data from the 1-year chronic phase of this study was 
chosen to determine the ADI. The primary reasons for selecting it were 
its length (52-weeks) and the inclusion of a 6-week recovery phase 
(control, 10,000 ppm, and 50,000 ppm dose groups), the total number of 
animals in each dose group (20 animals of each sex per group for the 
chronic phase with 10 additional animals of each sex for groups in the 
recovery phase), and the high overall animal survival rate. In 
addition, the results from the 2-year phase showed no indication that 
advantame is carcinogenic.

[[Page 29084]]

    Based on the NOAEL for the 1-year chronic toxicity study, we 
concluded that the appropriate ADI for advantame is 1,970 mg/p/d (Ref. 
4). This level is significantly higher than the EDI for advantame of 10 
mg/p/d for humans of all ages at the 90th percentile.

III. Comments

    We received two comments in response to the advantame food additive 
petition. One comment merely expressed support for the petitioned use 
of advantame, providing that safety is shown and the substance is 
properly declared when used as an ingredient in food. The other comment 
stated that they did not object to the petition, but rather to the use 
of advantame as a flavoring substance in food prior to a premarket 
approval for use as a sweetener and flavor enhancer without declaring 
advantame as an ingredient on the food label. Because this comment is 
not relevant to the safety of advantame, it has no bearing on our 
evaluation of the advantame petition.

IV. Conclusion

    We have evaluated the data and other information submitted by the 
petitioner in support of the safe use of advantame as a general-purpose 
sweetener and flavor enhancer in food and conclude that there is a 
reasonable certainty that the substance is not harmful under the 
petitioned conditions of use. Therefore, we conclude that the food 
additive regulations should be amended as set forth in this document.

V. Public Disclosure

    In accordance with Sec.  171.1(h) (21 CFR 171.1(h)), the petition 
and the documents that we considered and relied upon in reaching our 
decision to approve the petition will be made available for public 
disclosure (see FOR FURTHER INFORMATION CONTACT). As provided in Sec.  
171.1(h), we will delete from the documents any materials that are not 
available for public disclosure.

VI. Environmental Impact

    We have carefully considered the potential environmental effects of 
this action and have concluded that it will not have a significant 
impact on the human environment, and that an environmental impact 
statement is not required. FDA's finding of no significant impact and 
the evidence supporting that finding, contained in an environmental 
assessment, may be seen in the Division of Dockets Management (see 
ADDRESSES) between 9 a.m. and 4 p.m., Monday through Friday.

VII. Paperwork Reduction Act of 1995

    This final rule contains no collection of information. Therefore, 
clearance by the Office of Management and Budget under the Paperwork 
Reduction Act of 1995 is not required.

VIII. Section 301(ll) of the FD&C Act

    Our review of this petition was limited to section 409 of the FD&C 
Act. This final rule is not a statement regarding compliance with other 
sections of the FD&C Act. For example, the Food and Drug Administration 
Amendments Act of 2007, which was signed into law on September 27, 
2007, amended the FD&C Act to, among other things, add section 301(ll) 
of the FD&C Act (21 U.S.C. 331(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 (21 U.S.C. 355), a biological product licensed under 
section 351 of the Public Health Service Act (42 U.S.C. 262), or a drug 
or biological product for which substantial clinical investigations 
have been instituted and their existence has been made public, unless 
one of the exemptions in section 301(ll)(1) to (ll)(4) of the FD&C Act 
applies. In our review of this petition, we did not consider whether 
section 301(ll) of the FD&C Act or any of its exemptions apply to food 
products containing this food additive. Accordingly, this final rule 
should not be construed to be a statement that a product containing 
this food additive, if introduced or delivered for introduction into 
interstate commerce, would not violate section 301(ll) of the FD&C Act. 
Furthermore, this language is included in all food additive final rules 
that pertain to food and therefore should not be construed to be a 
statement of the likelihood that section 301(ll) of the FD&C Act 
applies.

IX. Objections

    If you will be adversely affected by one or more provisions of this 
regulation, you may file with the Division of Dockets Management (see 
ADDRESSES) either electronic or written objections. You must separately 
number each objection, and within each numbered objection you must 
specify with particularity the provision(s) to which you object, and 
the grounds for your objection. Within each numbered objection, you 
must specifically state whether you are requesting a hearing on the 
particular provision that you specify in that numbered objection. If 
you do not request a hearing for any particular objection, you waive 
the right to a hearing on that objection. If you request a hearing, 
your objection must include a detailed description and analysis of the 
specific factual information you intend to present in support of the 
objection in the event that a hearing is held. If you do not include 
such a description and analysis for any particular objection, you waive 
the right to a hearing on the objection.
    It is only necessary to send one set of documents. Identify 
documents 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 Division of Dockets Management between 9 a.m. and 4 
p.m., Monday through Friday, and will be posted to the docket at http://www.regulations.gov.

X. References

    The following references have been placed on display in the 
Division of Dockets Management (see ADDRESSES) and may be seen by 
interested persons between 9 a.m. and 4 p.m., Monday through Friday, 
and are available electronically at http://www.regulations.gov.

1. Memorandum from H. Lee, Division of Petition Review, CFSAN, FDA 
to F. Ellison, Division of Petition Review, CFSAN, FDA, July 28, 
2009.
2. Memorandum from H. Lee, Division of Petition Review, CFSAN, FDA 
to F. Ellison, Division of Petition Review, CFSAN, FDA, December 26, 
2012.
3. Memorandum from H. Lee, Division of Petition Review, CFSAN, FDA 
to F. Ellison, Division of Petition Review, CFSAN, FDA, November 24, 
2009.
4. Memorandum from T. S. Thurmond, Division of Petition Review, 
CFSAN, FDA to F. Ellison, Division of Petition Review, CFSAN, FDA, 
May 14, 2013.
5. Memorandum from W. Roth, Division of Food Contact Notification, 
CFSAN, FDA to F. Ellison, Division of Petition Review, CFSAN, FDA, 
January 22, 2013.
6. Memorandum from T. Walker, Division of Petition Review, CFSAN, 
FDA to F. Ellison, Division of Petition Review, CFSAN, FDA, December 
8, 2011.
7. Memorandum from S.K. Park, Division of Petition Review, CFSAN, 
FDA to F. Ellison, Division of Petition Review, CFSAN, FDA, April 
18, 2011.
8. Memorandum from S. Francke-Carroll and S. Mog, Senior Science and 
Policy Staff, CFSAN, FDA to T.S. Thurmond, S.K. Park, and C. 
Whiteside, Division of Petition Review, CFSAN, FDA, March 17, 2011.
9. Memorandum from S. Francke-Carroll and S. Mog, Senior Science and 
Policy Staff, CFSAN, FDA to C. Whiteside, T.S. Thurmond, and S.K. 
Park, Division of Petition Review, CFSAN, FDA, March 1, 2013.
10. Memorandum from C. Whiteside, Division of Petition Review, 
CFSAN,

[[Page 29085]]

FDA to F. Ellison, Division of Petition Review, CFSAN, FDA, February 
7, 2013.
11. Memorandum from A. Khan, Division of Petition Review, CFSAN, FDA 
to F. Ellison, Division of Petition Review, CFSAN, FDA, June 22, 
2010.
12. Memorandum from T. Walker, Division of Petition Review, CFSAN, 
FDA to F. Ellison, Division of Petition Review, CFSAN, FDA, December 
27, 2011.
13. Memorandum from I. Chen, Division of Petition Review, CFSAN, FDA 
to F. Ellison, Division of Petition Review, CFSAN, FDA, May 24, 
2010.
14. Memorandum from S. Francke-Carroll and S. Mog, Senior Science 
and Policy Staff, CFSAN, FDA to C. Whiteside and A. Khan, Division 
of Petition Review, CFSAN, FDA, March 17, 2011.
15. Memorandum from A. Khan, Division of Petition Review, CFSAN, FDA 
to F. Ellison, Division of Petition Review, CFSAN, FDA, March 3, 
2012.
16. CFSAN Cancer Assessment Committee Full Committee Review, 
Carcinogenicity Evaluation of Advantame, April 27, 2012.

List of Subjects in 21 CFR Part 172

    Food additives, Incorporation by reference, Reporting and 
recordkeeping requirements.

    Therefore, under the Federal Food, Drug, and Cosmetic Act and under 
authority delegated to the Commissioner of Food and Drugs, 21 CFR part 
172 is amended as follows:

PART 172--FOOD ADDITIVES PERMITTED FOR DIRECT ADDITION TO FOOD FOR 
HUMAN CONSUMPTION

0
1. The authority citation for 21 CFR part 172 continues to read as 
follows:


    Authority:  21 U.S.C. 321, 341, 342, 348, 371, 379e.


0
2. Add Sec.  172.803 to subpart I to read as follows:


Sec.  172.803  Advantame.

    (a) Advantame is the chemical N-[N-[3-(3-hydroxy-4-
methoxyphenyl)propyl]-[alpha]-aspartyl]-L-phenylalanine 1-methyl ester, 
monohydrate (CAS Reg. No. 714229-20-6).
    (b) Advantame meets the following specifications when it is tested 
according to the methods described or referenced in the document 
entitled ``Specifications and Analytical Methods for Advantame'' dated 
April 1, 2009, by the Ajinomoto Co. Inc., Sweetener Department 15-1, 
Kyobashi 1-chome, Chuo-ku, Tokyo 104-8315, Japan. The Director of the 
Office of the Federal Register approves this incorporation by reference 
in accordance with 5 U.S.C. 552(a) and 1 CFR part 51. Copies are 
available from the Office of Food Additive Safety (HFS-200), Center for 
Food Safety and Applied Nutrition, 5100 Paint Branch Pkwy., College 
Park, MD 20740. Copies may be examined at the Food and Drug 
Administration's Main Library, 10903 New Hampshire Ave., Bldg. 2, Third 
Floor, Silver Spring, MD 20993, 301-796-2039, or at the National 
Archives and Records Administration (NARA). For information on the 
availability of this material at NARA, call 202-741-6030 or go to: 
http://www.archives.gov/federal-register/cfr/ibr-locations.html.
    (1) Assay for advantame, not less than 97.0 percent and not more 
than 102.0 percent on a dry basis.
    (2) Free N-[N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-[alpha]-
aspartyl]-L-phenylalanine, not more than 1.0 percent.
    (3) Total other related substances, not more than 1.5 percent.
    (4) Lead, not more than 1.0 milligram per kilogram.
    (5) Water, not more than 5.0 percent.
    (6) Residue on ignition, not more than 0.2 percent.
    (7) Specific rotation, determined at 20 [deg]C 
[[alpha]]D: -45.0 to -38.0[deg] calculated on a dry basis.
    (c) The food additive advantame may be safely used as a sweetening 
agent and flavor enhancer in foods generally, except in meat and 
poultry, in accordance with current good manufacturing practice, in an 
amount not to exceed that reasonably required to achieve the intended 
technical effect, in foods for which standards of identity established 
under section 401 of the Federal Food, Drug, and Cosmetic Act do not 
preclude such use.
    (d) If the food containing the additive purports to be or is 
represented to be for special dietary use, it must be labeled in 
compliance with part 105 of this chapter.

    Dated: May 15, 2014.
Leslie Kux,
Assistant Commissioner for Policy.
[FR Doc. 2014-11584 Filed 5-19-14; 11:15 am]
BILLING CODE 4160-01-P