INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY
WORLD HEALTH ORGANIZATION
SUMMARY OF TOXICOLOGICAL DATA OF CERTAIN FOOD ADDITIVES
WHO FOOD ADDITIVES SERIES NO. 12
The data contained in this document were examined by the
Joint FAO/WHO Expert Committee on Food Additives*
Geneva, 18-27 April 1977
Food and Agriculture Organization of the United Nations
World Health Organization
* Twenty-first Report of the Joint FAO/WHO Expert Committee on Food
Additives, Geneva, 1977, WHO Technical Report Series No. 617
ORANGE I
EVALUATION FOR ACCEPTABLE DAILY INTAKE
BIOLOGICAL DATA
BIOCHEMICAL ASPECTS
In dogs it was found that 6 to 41% of the administered colour was
excreted in the faeces. No colour was found in the urine. Incubating
the colour with a dog's fresh intestinal content destroyed 80% of the
colour in one hour (DFG Dye Commission, 1957; Vos et al., 1953).
The urine of a dog given 5 g Orange I contained a small quantity
of the unchanged colour together with sulphanilic acid and 4-amino-
1-naphthol after 48 hours (Sisley and Porcher, 1911). (The unchanged
colour in the urine could be due to contamination.)
Oral administration of 100 mg of Orange I to a dog with a bile
fistule resulted in the recovery of only 0.4% in the bile, compared to
a 47% recovery after s.c. administration of the dye (Radomski and
Deichmann, 1956). Low excretion of Orange I in animals not showing
diarrhoea is due to its degradation by intestinal flora, as
demonstrable by in vitro tests, rather than to its absorption
through the intestinal wall (Radomski and Deichmann, 1956).
Acute toxicity
Animal Route LD50 mg/kg Reference
mg/kg bw
Rat i.p. 1 000 mg Truhaut, 1955
Concentration of 1% of the dye in the diet caused some deaths,
spleen enlargement, leucocytosis, anaemia, diarrhoea and growth
depression in rats; 0.5% of the diet increased the incidence of
nephritis and enlargement of the spleen with hyperplasia and increased
pigmentation (Bourke et al., 1956).
Cathartic effect
Orange I induces catharsis in the rat at a concentration of
20 mg/kg body weight. The cathartic action of 200 mg of Orange I was
reported when administered orally in dogs (Radomski and Deichmann,
1956).
Mutagenicity
Mutagenicity testing of Orange I gave questionable or variable
responses following reduction and microsomal activation (Brown et al.,
1977).
Short-term studies
Rat
Ten rats were fed this colour at a level of 4% in the diet. The
animals lived only a few months. Gross staining of the glandular
stomach and the small intestine, and granular deposits in the stomach
and small intestine were observed (Willheim and Ivy, 1953).
Weanling male rats were fed diets containing 1, 2 or 5% of the
colour for 16 weeks. The animals with the 2 and 5% died all within the
first two weeks of the feeding period. A mortality of 80% occurred in
the rats fed 1% and the growth rate of the surviving animals in this
group was markedly retarded. The inclusion of the colour in the diet
caused severe diarrhoea, enlargement of the spleen and an anaemia
(Hallesy and Doull, 1956).
Dog
This colour had a significant cathartic effect when 100 to 200 mg
were given to groups of five dogs (Vos et al., 1953).
Long-term studies
Mouse
No liver tumours were observed in a group of 20 mice (sex and
strain unspecified) which received weekly doses of 15-20 mg Orange I
for life, given over five days a week in the form of a solution in tap
water added to 20 g brown bread. The last surviving mouse died
409 days after the start of treatment; the median lifespan was not
reported. Autopsies were carried out on 14 mice and sections examined
for seven mice (Cook et al., 1940). [The short duration of the
experiment was noted by the working group.]
Rat
This colour was fed to 85 rats at a level of 0.1% in the diet.
The daily intake varied from 10 to 15 mg for a period of 400 days. No
tumours were observed (DFG, 1957).
Groups of 24 weanling rats, equally divided by sex, were fed the
colour at 0, 0.5, 1.0 and 2.0% in the diet. No rats fed at 2.0%
survived beyond the fifth week, observations are confined to 0, 0.5
and 1.0% levels. Body weight and food intake were recorded weekly for
two years; blood counts were taken four times. Gross findings at 1.0%
were marked increase of mortality, enlargement of spleens,
leukocytosis and anaemia, diarrhoea and growth depression. At 0.5%
kidneys of test rats showed more chronic congestion than those of
controls and there was some splenic enlargement. Microscopically,
spleens showed uniformly chronic congestion and less often slight
hyperplasia and increased pigmentation. Kidneys showed nephritis of
the type common in older rats with no difference among the groups
except in incidence (Bourke et al., 1956; US FDA, 1963).
Eighteen young rats were given subcutaneous injections, 20 mg of
the colour per week (2% aqueous solution) for two years. In six cases
fibrosarcomas were found. The controls did not get tumours (Nelson and
Davidow, 1957).
Dog
The colour was given to 14 dogs at four levels in daily
quantities by capsule ranging from about 0.02% to 1.0% in the diet.
The dogs on the lowest level survived for five years without showing
any effect. At higher levels (0.2% or more of the diet) effects were
variable; some dogs survived only for short periods, others showed
little or no effect for long periods. Pathological changes were
generally non-specific; some animals were found dead with little to
explain the death; others were emaciated and showed organ changes
chiefly of inanition (Bourke et al., 1956; US FDA, 1963).
OBSERVATIONS IN MAN
This colour induces also a cathartic action in man (80 mg). The
intact molecule appears to be the active agent (Radomski and
Deichmann, 1956).
Human volunteers who ate candy containing 0.07% of this colour
exhibited diarrhoea upon ingestion of one to eight pieces of the candy
(US FDA, 1955).
Skin tests with this colour showed no reaction in patients
sensitive to paraphenylene diamine (Baer et al., 1948).
REFERENCES
Baer, R. L., Leider, M. and Mayer, R. L. (1948) Eczematous cross-
hypersensitivity between p-phenylenediamine and azo dyes certified for
use in foods, drugs and cosmetics, Proc. Soc. exp. Biol., 67, 489
Bourke, A. R., Nelson, A. A. and Fitzhugh, O. G. (1956) Chronic
toxicity of FD and C Orange No. 1, Fed. Proc., 15, 404
Brown, J.P., Roehm, G. W. and Brown, R. J. (1977) Mutagenicity testing
of certified food colors and related azo, xanthene and triphenyl-
methane dyes with the Salmonella/microsome system, 8th Ann. Meeting,
Environmental Mutagen Society, Colorado Springs, Feb. 1977 (abstract)
Cook, J. W., Hewett, C. L., Kennaway, E. L. and Kennaway, N. B. (1940)
Effects produced in the livers of mice by azo-naphthalenes and related
compounds, Amer. J. Cancer, 40, 62-77
DFG (Deutsche Forschungsgemeinschaft) (1957) Farbstoff-Kommission,
Toxikologische Daten von Farbstoffen und ihre Zulassung fur
Lebensmittel in verschiedenen Ländern, Mitt. 6(2), Wiesbaden,
Steiner Verlag, p. 25
Hallesy, D. W. and Doull, J. (1956) J. Pharmacol. exp. Ther., 116,
26
Nelson, A. A. and Davidow, B. (1957) Injection site fibrosarcoma
production in rats by food colors., Fed. Proc., 16, 367
Radomski, J. L. and Deichmann, W. B. (1956) Cathartic action and
metabolism of certain coal-tar food dyes, J. Pharmacol. exp. Ther.,
118, 322-237
Sisley, P. and Porcher, C. (1911) Du sort des matières colorantes dans
l'organisme animal, C.R. Aced. Sci. (Paris), 152, 1062-1064
Truhaut, R. (1958) Sur l'utilisation des colorants en thérapeutique et
les dangers qui peuvent en résulter pour la santé humaine, Chimie
moderne, 3, 337-350
United States Food and Drug Administration (1955) Title 21, Food and
Drugs, Part 135, Federal Register, 20, 8492
United States Food and Drug Administration (1963) Unpublished report
Vos, B. J., Radomski, J. L. and Fuyat, H. N. (1953) Fed. Proc.,
12, 376
Willheim, R. and Ivy, A. C. (1953) Dietary carcinogenesis,
Gastroenterology, 23, 1
See Also:
Toxicological Abbreviations
ORANGE I (JECFA Evaluation)
Orange I (IARC Summary & Evaluation, Volume 8, 1975)