CARNAUBA WAX
First draft prepared by
Dr Karen B. Ekelman and Dr Hui-Chen Chang
US Federal Administration
Washington DC, USA
1. EXPLANATION
Carnauba wax has not been evaluated previously by the Joint
FAO/WHO Expert Committee on Food Additives. Carnauba wax is obtained
from the leaves and buds of the Brazilian wax palm, Copernica
cerifera. The wax is hard, brittle, sparingly soluble in cold
organic solvents and insoluble in water.
2. BIOLOGICAL DATA
2.1 Biochemical aspects
No information available.
2.2 Toxicological studies
2.2.1 Acute toxicity studies
No information available.
2.2.2 Short-term studies
2.2.2.1 Rats
Three experiments have been conducted. In the first experiment,
groups of 15 male and 15 female Wistar rats were fed diets
containing 0 (control group), 1, 5, or 10% carnauba wax or 10%
cellulose powder for 13 weeks. The rats fed diets containing
cellulose powder acted as a control group for possible effects due
to the replacement of a significant proportion of the diet by a
non-nutrient test material. In the second and third experiments,
groups of five rats of each sex were fed diets containing 1%
carnauba wax then 10% carnauba wax or 10% cellulose powder for 2 and
6 weeks, respectively. Food consumption and body weights were
recorded weekly only in the 13-week study. Haematological, serum,
and urinalysis and gross and microscopic examinations of tissues
were used to evaluate the toxicity of carnauba wax.
Results from the 13-week study indicated that rats given 10%
carnauba wax consumed more food than the controls, but showed no
differences in body weight. Similar results were also seen in the
rats given 10% cellulose powder in the diet. The calculated mean
intakes of carnauba wax were 0, 800, 4200 and 8800 mg/kg bw/day for
males and 0, 900, 4600 and 10 200 mg/kg bw/day for females.
Blood samples were collected at weeks 2, 6 and 13 for
haematological examination and serum analyses; results of these
analyses were similar for treated and control animals. Urinalysis at
weeks 2, 6 and 13 of the study did not show any treatment-related
changes. Organ weights (brain, pituitary, thyroid, heart, liver,
spleen, kidneys, adrenal glands, gonads, stomach, small intestine
and caecum) showed no compound-related changes, nor were there any
dose-related histological changes (Rowland, 1982).
2.2.2.2 Dogs
Four groups of 6 male and 6 female Beagle dogs were exposed to
0 (control), 0.1, 0.3, or 1% carnauba wax in their diets for 28
weeks. Food consumption, body weights and behavioural effects were
recorded weekly. Blood and urinary samples were collected at weeks
11 and 26. Organs were weighed (brain, pituitary gland, thyroid
gland, heart, liver, spleen, kidneys, and adrenal glands) and gross
and microscopic examinations of tissues were performed at the end of
study.
No significant differences in body weights nor food consumption
were noted between treatment and control groups of animals. Data
from analyses of blood, urine and organ weights did not show any
treatment-related effects in dogs consuming carnauba wax. The only
significant (p <0.05) clinical observation at 26 weeks was higher
free fatty acid levels in male dogs at all dietary levels of
carnauba wax as compared to controls. However, free fatty acid
levels in all male dogs fed carnauba wax were well within the normal
range for Beagle dogs (200-800 µM/l) whereas values for control dogs
were unusually low (138 ± 38 µM/l). Ophthalmic, gross and
histopathologic examinations revealed no significant
treatment-related effects for up to 1% carnauba wax in the diet
(Parent, 1983a).
2.2.3 Long-term/carcinogenicity studies
No information available.
2.2.4 Reproduction studies
Four groups of 25 female (F0) and 25 male Wistar rats were
administered 0, 0.1, 0.3, or 1% carnauba wax in the diet for 4 weeks
prior to mating and throughout the remainder of the study, including
gestation and lactation. Each (F1) pup was supplied with the same
diet as its dam through weaning and for an additional 13 weeks. Food
consumption and body weights were recorded weekly for the F0
generation (except during mating) and the F1 generation. F0
females were weighed on days 0, 6, 11, 15 and 20 of gestation. Total
litter weights were measured at birth and at 4, 14 and 21 days after
birth. The number of pregnant females, number of pups born alive or
dead, and survival of the progeny were recorded. Ophthalmic
examinations were performed on the F1 animals at the time of
weaning and at the end of the 13-week study. Haematological and
clinical chemical measurements were performed on F1-generation
animals at weaning and at 6 and 13 weeks thereafter. Complete gross
necropsies were performed an all F0 animals, F1 animals that
were killed after weaning, and F1 animals used for the
post-weaning 13-week feeding study. Organs (adrenal glands,
epididymides, heart, liver, kidneys, ovaries, spleen, testes,
thyroid with parathyroid glands, and uterus) from animals used in
the post-weaning 13-week feeding study were weighed, and
histopathological examination of the weighed organs and other organs
(brain, eyes, large and small intestine, lungs, lymph nodes,
pancreas, pituitary gland, sciatic nerve, seminal vesicles, skeletal
muscle, skin, spinal cord, sternum and marrow, stomach, urinary
bladder, and grossly abnormal lesions) for animals in control and
high-dose groups was performed. For animals in the low and
middle-dose groups, only grossly abnormal tissues were examined
microscopically.
No effects on reproduction parameters (fertility indices,
gestation indices, viability indices, lactation indices and pup
weights) were observed after feeding carnauba wax to pregnant
F0-generation female rats at levels up to 1% of the diet. Although
scattered significant differences (p <0.05) between groups in body
weights and food consumption of male and female rats in both the
F0 and F1 generations were noted (data not shown in report), the
magnitude of the changes was small and did not show a clear dose
relationship indicative of a compound-related effect. This study
reported that mean intake of carnauba wax calculated on the basis of
food consumption during the 13 weeks of the study was 0.08, 0.25,
and 0.81 g/kg bw/day for males and 0.09, 0.27, and 0.67 g/kg bw/day
for females treated at levels of 0.1, 0.3, and 1.0% carnauba wax,
respectively.
A statistically significant increase (p <0.05) in haematocrit
of female rats fed diets containing 0.1% and 1% carnauba wax was
noted at end of study, but was not observed in animals fed 0.3%
carnauba wax. Blood urea nitrogen levels were significantly elevated
(p <0.05) in males fed diets containing 0.1 and 1% carnauba wax for
6 weeks and in males fed diets containing 1% carnauba wax for 13
weeks. Although serum glutamate-pyruvate transaminase decreased and
chloride increased in male rats fed 0.3 and 1% carnauba for 13
weeks, the values were reported to be within the normal
physiological ranges for Wistar rats. A decrease in serum free fatty
acids was seen in male and female rats fed carnauba wax at dietary
levels of 0.3% and 1% at week 13. Those haematology and clinical
chemistry changes did not appear to be related to consumption of
carnauba wax.
There were no treatment-related differences in histological,
ophthalmic and gross pathology findings in either F0 or F1
animals at any dietary level of carnauba wax used in this study
(Parent et al., 1983b).
2.2.5 Special studies on mutagenicity
Results of mutagenicity studies on carnauba wax are summarized
in Table 1.
Table 1. Results of Mutagenic Studies on Carnauba Wax
Test System Test Object Concentration of Results References
Carnauba Wax
Ames test1 S. typhimurium 3.3-1000 µg in Negative Mortelmans and Griffin,
TA1537, TA1538, plate tests 1981
TA98
Ames test1 S. typhimurium 0.01-0.5% in Negative Mortelmans, and Griffin
TA1537, TA1538, suspension tests 1981
TA98
Ames test1 S. typhimurium 0.1-2.5% in Negative Mortelmans, and Griffin
TA1537, TA1538, suspension tests 1981
TA98
Gene mutation2 S. typhimurium 0.01% in plate Negative Litton Bionetics,
TA1535, TA1537, tests Inc., 1975
TA1538
Gene mutation2 S. typhimurium 0.005 and 0.01% in Inconsistent Litton Bionetics,
TA1535, TA1537, suspension tests changes3 Inc., 1975
TA1538
Gene mutation2 S. cerevisiae 0.3 and 1.75% in Negative Litton Bionetics,
D4 suspension tests Inc., 1975
1 The Ames/ Salmonella assays in the presence and absence of an Aroclor 1254-stimulated,
rat-liver homogenate metabolic activation system, were used in this study.
2 A series of in vitro microbial assays with and without metabolic activation were used.
In the activation assays, the tissue homogenate of liver, lung and testes were
prepared from either mouse, rat or monkey.
3 The results from non-activation suspension tests were negative. The results from
activation suspension tests showed scattered increased mutation responses in the
presence of rat-liver or testes homogenate with strain TA1537, and in the
presence of monkey-lung homogenate with TA1538.
2.2.6 Special studies on teratogenicity
Four groups of Wistar rats, each consisting of 25 females, were
mated to test for teratogenic effects; they were fed 0 (control),
0.1, 0.3 or 1% carnauba wax in the diet for two weeks before mating
and throughout gestation. Body weights of pregnant dams were
recorded on days 0, 6, 11, 15, and 20 of gestation. On day 20 of
gestation, pregnant females were killed, caesarian sections were
performed, and gross pathological changes were noted. The uterine
contents were examined and the following litter reproduction data
were recorded: number of corpora lutea, implantation sites,
resorption sites, live and dead fetuses; gross malformations; and
body weights of live fetuses. One-half of fetuses were examined for
signs of visceral pathological changes and the other half were
examined for signs of skeletal abnormalities.
Results from this study indicated that there were no
significant changes in body weights of pregnant dams during
gestation; no significant differences in reproduction data among
test groups; and no dose-related effects of carnauba wax on skeletal
or soft tissue development in fetuses (Food and Drug Research
Laboratories, Inc., 1977).
2.3 Observations in man
No information available.
3. COMMENTS
Short-term feeding studies of carnauba wax at 10% in the diet
of rats showed no significant compound-related toxic effects. In a
28-week study with beagle dogs in which carnauba wax was fed at
levels of 0/1, 0/3, and 1% in the diet, no compound-related toxic
effects were identified.
No adverse effects were observed in fetuses in a teratogenicity
study in rats following dietary exposure 0/1, 0/3, and 1% carnauba
wax during gestation.
A reproduction study combined with a 13-week oral toxicity
study with F1 animals was conducted in Wistar rats. During both
phases of the study, animals (parents and offspring) were fed diets
containing 0/1, 0/3, or 1% carnauba wax. No adverse effects
associated with the consumption of carnauba wax at levels up to 1%
of the diet (equal to 700 mg/kg bw/day for female rats) were
observed in this experiment.
Although the results of mutagenicity studies were largely
negative, scattered positive responses with S. typhimurium strains
TA1537 and TA1538 were observed in the presence of metabolic
activation.
4. EVALUATION
The Committee allocated an ADI of 0-7 mg/kg bw for carnauba
wax.
5. REFERENCES
FASEB (1975) Evaluation of the health aspects of carnauba wax as a
food ingredient. SCOGS-47.
FOOD AND DRUG RESEARCH LABORATORIES, INC. (1977) Evaluation of the
effects of carnauba wax in FDRL/Wistar derived rats after dietary
exposure through one full generation. Unpublished report of July 8,
1977 submitted to Brazilian Embassy, Washington, D.C.
LITTON BIONETICS INC. (1975) Mutagenic evaluation of compound
MX8015869, carnauba wax (73-48). Unpublished report of April 15,
1975, submitted to Food and Drug Administration, Rockville, MD, 33
pp.
MORTELMANS, K.E. & GRIFFIN, ANN. F. (1981) Microbial mutagenesis
testing of substances compound F73-048: carnauba wax, yellow.
Unpublished SRI Report LSU-6909 from Frank B. Ross & Co. Submitted
to Food and Drug Administration by Frank B. Ross & Co. Report
(LSU-6909), 19 pp.
PARENT, R.A., COX, G.E., BABISH, J.G., GALLO, M.A., HESS, F.G. &
BECCI, P.J. (1983a) Subchronic feeding study of carnauba wax in
beagle dogs. Fd. Chem. Toxicol., 21(1): 85-87.
PARENT, R.A., RE, T.A., BABISH, J.G., COX, G.E., VOSS, K.A. & BECCI,
P.J. (1983b) Reproduction and subchronic feeding study of carnauba
wax in rats. Fd. Chem. Toxicol., 21(1): 89-93.
ROWLAND, I.R., BUTTERWORTH, K.R., GAUNT, I.F., GRASSO, P. &
GANGOLLI, S.D. (1982) Short-term toxicity study of carnauba wax in
rats. Fd. Chem. Toxicol., 20: 467-471.
See Also:
Toxicological Abbreviations
CARNAUBA WAX (JECFA Evaluation)