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Affiliation: A.K. Choudhary is with the Department of Physiology, School of Medicine, Faculty of Health Sciences, University of Pretoria,
Pretoria, South Africa. E. Pretorius is with the Department of Physiological Sciences, Faculty of Health Sciences, Stellenbosch University,
Stellenbosch, South Africa.
Correspondence: E. Pretorius, Department of Physiological Sciences, Faculty of Health Sciences, Stellenbosch University, Private Bag X1
Maiteland, 7602, South Africa. Email: resiap@sun.ac.za.
Key words: aspartame, aspartic acid, methanol, phenylalanine.
C The Author(s) 2017. Published by Oxford University Press on behalf of the International Life Sciences Institute.
V
All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.
doi: 10.1093/nutrit/nux035
718 Nutrition ReviewsV Vol. 75(9):718–730
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drug reactions; or (5) interfere with drug action. Safety
issues associated with the use of aspartame include po-
tential toxicity from aspartame metabolites, including
methanol and/or its metabolite, formaldehyde.12,13
This review examines the existing literature (pub-
lished 2000–2016) describing the effects of aspartame
on cells and organ systems when used within the safe
dosage range. The interactions between aspartame and
cells and organ systems are examined, and extensive
references for current recommended safe dosages are
provided. Finally, literature that considers the effects of
aspartame on different cells in the body is reviewed.
Aspartame is an L-aspartyl-L-phenylalanine methyl es- further metabolized in the liver into L-tyrosine by the en-
ter14 (Figure 1) and is very stable under dry conditions zyme phenylalanine hydroxylase. L-Tyrosine in turn, is
when stored at temperatures ranging from 30 C to converted into L-dopa (L-3,4-dihydroxyphenylalanine) by
80 C.15 It degrades at high temperatures and in aqueous the enzyme tyrosine hydroxylase.23 L-Dopa is further
solutions. The rate of degradation in aqueous solutions, converted into the catecholamines—dopamine, norepi-
however, depends on pH as well as temperature.16 At nephrine (noradrenaline), and epinephrine (adrena-
room temperature, aspartame is most stable at pH 4.3, line)—by the amino acid decarboxylase enzyme.
with a half-life of 300 days. Degradation is minimal Phenylalanine can cross the blood–brain barrier.24,25
when pH ranges between 4.0 and 5.0 and reaches a maxi- Furthermore, elevations in plasma concentrations of phe-
mum when heated under conditions of high humidity at nylalanine and aspartic acid result in increased transport
a pH greater than 6.0.16 Under strongly acidic of these amino acids into the brain, modifying the brain’s
(pH < 4.0) or alkaline conditions (pH > 6.0), aspartame neurochemical composition.26 Neuroendocrine changes,
may generate methanol by hydrolysis. Under more se- particularly increased concentrations of catecholamine
vere conditions, such as elevated temperature or high resulting from phenylalanine and its hydroxylation prod-
pH, the peptide bonds are also hydrolyzed.17 This results uct, tyrosine, have been observed in the brain.26
in the release of free amino acids (particularly phenylala- Phenylalanine is a large neutral amino acid that
nine and aspartic acid). It should also be noted that the
competes with other important large neutral amino
pH of diet sodas—a major vehicle for aspartame
acids for binding on the large neutral amino acid trans-
consumption—tends to be somewhere between 3.0 and
porter.26 However, excess phenylalanine concentrations
4.0. Interestingly, following breakdown in the gut or ex-
are associated with decreased concentrations of cate-
posure to temperature changes, aspartame and its metab-
cholamine, serotonin, and dopamine.26 Aspartic acid is
olites lose their sweetness.18,19 Conditions during storage
metabolized in the liver into L-lysine and L-methionine
vs during ingestion are thus different and may determine
by the enzyme aspartate kinase. At high concentrations,
the formation of aspartame metabolites.20
aspartic acid may cross the blood–brain barrier and
Aspartame during storage. Aspartame can be stored in a bind to the N-methyl-D-aspartate receptor (also known
dry or an aqueous form. In the dry form, stability as the NMDA receptor or NMDAR)27 or to other gluta-
depends mainly on temperature, and stability decreases mate binding sites, causing an influx of calcium ions
at temperatures < 30oC or > 80oC. In the aqueous into cells (Figure 2). Increased firing of action potentials
form, stability is greatest at a pH of 4.3; beyond this pH, and higher rates of neuron depolarization can potentiate
aspartame degrades into its 3 known metabolites (phe- neurodegeneration.26 The enzyme responsible for me-
nylalanine, aspartic acid, and methanol) and loses some tabolism of methanol (Ch3OH) is species dependant.28
of its sweetness. The aqueous form also becomes In primates, methanol is metabolized into formaldehyde
sweeter with increased temperature.21 (HCHO) in the liver by alcohol dehydrogenase.29 In
rodents, on the other hand, methanol is mainly metabo-
Aspartame during ingestion. Upon ingestion, aspartame lized by alcohol catalase and differences in the embry-
is metabolized by gut enzymes (esterase and peptidase) onic metabolism of CH3OH may determine species
into 3 amino acid isolates, phenylalanine (50%), aspartic sensitivity, in which mouse embryos were more sensitive
acid (40%), and methanol (10%).22 Phenylalanine is than the rat.30 Formaldehyde is oxidized into formic
Effect of aspartame on blood cells and fibrin Effect of aspartame on the brain
packaging
Once the cellular antioxidant capacity is overcome by
Injury to cell membranes by free radicals can lead to a the generation of ROS and reactive nitrogen species
change in cell membrane fluidity, impairing the vital (RNS), cellular damage may occur.92 Neuronal cells are
functions of blood cells (erythrocytes, neutrophils, and especially vulnerable to oxidative stress because of high
lymphocytes).92 Most importantly, it can affect immu- concentrations of polyunsaturated fatty acids, which
nity by altering neutrophil and lymphocyte function.93 render them much more susceptible to lipid peroxida-
Oral administration of aspartame at both a higher dos- tion compared with other tissues.113 The excess free
age (> 40 mg/kg/d)55 and a safe dosage ( 40 mg/kg/ radicals can attach to fatty acids in the neuronal cell
d)55,70,71 can increase the production of free radicals membrane, thereby interfering with neuronal cell func-
and induce oxidative stress in blood cells (erythrocytes, tion.97 The production of excess free radicals may also
neutrophils, and lymphocytes) by altering the oxidant/ increase permeability of the blood–brain barrier in a
antioxidant balance. Oxidative stress in erythrocytes time- and concentration-dependent manner.98
can lead to damage of the erythrocyte membrane108; The consumption of higher dosages of aspartame
impair the flow of erythrocytes through the microcircu- (> 40 mg/kg/d) on the brain was also previously stud-
lation and the delivery of oxygen to the tissues91; induce ied.56–60,62–64 Results suggest that higher aspartame dos-
erythrocyte aging91; and induce inflammation.109 ages may result in changed enzyme activities.56–60,63–64
Reactions between excessive amounts of reactive It was also found, in an in vivo voltammetry study that
oxygen species (ROS) and superoxide radicals from ac- aspartame decreases evoked extracellular dopamine lev-
tivated neutrophils can exert cytotoxic effects and may els in the rat brain.62 Brain areas affected by aspartame
induce overactivation of the nuclear repair enzyme may include the cerebral cortex, hypothalamus and hip-
poly-(adenosine diphosphate [ADP]-ribose) polymer- pocampus, as shown in a paper by Iyyaswamy and
ase, which may cause adenosine triphosphate depletion Rathinasamy in 2012.60 Areas like the hippocampus and
and cellular injury.96 The activation of poly-(ADP-ri- medial prefrontal cortex play important roles in mem-
bose) polymerase is also known to upregulate multiple ory and decision making.114,115 Even the US Food and
pathways of proinflammatory signaling.96 Hence, oxida- Drug Administration–approved ADI of aspartame
tive stress in blood cells after aspartame consumption ( 40 mg/kg) has been shown to be toxic to the
can modify the expression or activation of inflamma- brain.13,74,116 Inside neuronal cells, intense or prolonged
tory mediators. The T lymphocytes (T cells) are ren- oxidative stress causes overexpresssion of proinflamma-
dered hyporesponsive to activating stimuli, but both tory cytokines (interleukin 6 [IL-6], interleukin 1b
exposure to ROS produced by activated neutrophils110 [IL-1b], interleukin 8 [IL-8]), considered a hallmark of
and prolonged exposure to high ROS concentrations94 neuroinflammation.100,101 The proinflammatory cytokines
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of exposure
Arbind et al. (2014)71 Wistar albino male rats 40 mg/kg Oral for 15, 30, and 90 d Induced oxidative stress in blood
cells (RBCs, neutrophils, and
lymphocytes). Altered neutro-
phil function, irrespective of
duration of exposure
723
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Table 3 Continued
724
Reference Cells/tissue In vitro or animal model Aspartame dosage Route and duration Results
of administration
Christian et al. (2004)78 Male Sprague–Dawley rats Rat dosage (250 mg/kg) Oral, via the drinking Altered T-maze performance and
water, for 3–4 mo increased muscarinic choliner-
gic receptor densities or
enzymes in certain brain
regions
Abd El-Samad (2010)79 Wistar albino male rats Rat dosage (250 mg/kg) Oral for 8 wk Harmful effects (condensed nu-
clei and loss of characteristic
pyriform shape of Purkinje
cells) on cells in the cerebellar
cortex
Ashok & Sheeladevi Liver Wistar albino male rats 40 mg/kg Oral for 90 d Altered antioxidant status, ex-
(2015)80 pression of stress protein, and
induced apoptotic changes in
the liver
Choudhary & Devi Wistar albino male rats 40 mg/kg Oral for 15 d and 30 d Induced oxidative stress in liver,
(2014),70 Kumar regardless of duration of ex-
Choudhary et al. posure. Levels of serum pro-
(2014)81 tein and bilirubin that reflect
liver function were altered af-
ter 30 d of aspartame
administration
Iman (2011)82 Wistar albino male rats 40 mg/kg Oral for 2, 4, and 6 wk Induced oxidative stress in liver
after 4 wk and 6 wk of
treatment
Kim et al. (2011)77 Zebrafish (10 wk old) 3mM Oral for 12 d Inflammatory cells in the liver
were infiltrated
Choudhary & Devi Kidney Wistar albino male rats 40 mg/kg Oral for 15 d and 30 d Induced oxidative stress in kid-
(2014),70 Kumar ney and serum values that re-
Choudhary et al. flect kidney function (such as
(2014)81 creatinine, urea, and uric acid)
after 30 d of aspartame
administration
Iman (2011)82 Wistar albino male rats 40 mg/kg Oral for 2, 4, and 6 wk Induced oxidative stress in kid-
ney after 6 wk
Martins & Azoubel Wistar female rats (14 mg/kg) heated Intragastric on days 9, 10, Morphometric alterations in all
(2007)83 to 40 C and 11 of pregnancy renal structures (glomerulus,
proximal and distal convo-
luted tubules, and collecting
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ducts) of the rat fetal kidney
during organogenesis
(continued)
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(2016),75 Choudhary & tion, and reduced heart rate
Sundareswaran variability, as evidenced by
(2016)84 sympathetic dominance and
loss of vagal tone, but could
not induce structural change
Gudadhe et al. (2013)85 Neonatal mice 100 mg/g Intraperitoneal for 2 wk Compensatory hypertrophy of
725
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Table 4 Effects of aspartame (higher and safe dosages) on different cells and organ systems
Cells/tissue At higher dosages At safe dosages Effects
(> 40 mg/kg) ( 40 mg/kg)
Blood Affected Affected Impaired delivery of oxygen to the tissues by red
blood cells91,92; red blood cell aging91; altered
neutrophil function93; decreased T-cell prolifera-
tion94; platelet hyperactivity and hyperaggreg-
ability95; upregulation of proinflammatory
signaling96
Brain Affected Affected Dysfunction of neuronal cells97; disruption of
blood–brain barrier98; impaired neurobehavio-
ral parameters (learning and memory)99; upre-
gulation of neuroinflammation, which may
initiate neurotropic effects100,101
are important regulators of matrix metalloproteinases,98 regenerative processes,121 resulting in hepatic dam-
which are zinc-containing enzymes produced by acti- age.102 The cytotoxic effects of ROS and RNS in the
vated microglia. They are responsible for breaking down liver may also lead to inactivation of the heme group
the extracellular matrix in cerebral blood vessels, which and nitrosylation of iron–sulfur group.103,104 In addi-
leads to damage of neurons117 and a disruption in the tion, the inflammation caused by the presence of ROS
blood–brain barrier.118 results in the modulation of hepatocyte metabolism.105
Neurobehavioral parameters (like learning and Furthermore, Kupffer cells activated by free radicals are
memory) were impaired not only with higher dosages responsible for the release of proinflammatory cyto-
of aspartame65,67 but also with safe dosages.75–78 At kines.122 The toxicity of methanol released during the
high concentrations, ROS and RNS may lead to de- metabolism of aspartame was also shown to lead to apo-
creased synaptic plasticity by attenuating long-term po- ptotic changes in the liver of Wistar albino rats.80
tentiation and synaptic neurotransmission.99
Effect of aspartame on the kidney
Effect of aspartame on the liver
The kidneys are vital organs, necessary for maintaining
The liver is the principal detoxifying organ and main- the composition and volume of body fluids, the acid–
tains metabolic homeostasis.119 Free radical reactions base balance, and the redox status.106 Oxidative stress
are widely known to be involved in liver injury.120 Both may lead to kidney injury.107 Both higher dosages and
higher dosages of aspartame68,69 and safe dos- safe dosages of aspartame may induce oxidative stress
ages70,77,81,82 have been shown to impair the antioxidant in the kidneys of rats.68,70,81–83 It was previously sug-
status of the liver, which may lead to hepatocellular in- gested that in the kidney, oxidative stress may contrib-
jury. During oxidative stress, the balance between ROS ute to the progression of kidney fibrosis,123 and chronic
and antioxidants shifts toward increased production kidney disease may be associated with inflammation
of the former, which may affect the energetic and and upregulated inflammatory cytokines.124