[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