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
COCHINEAL, CARMINE AND CARMINIC ACID
Cochineal is a red colouring matter consisting of the dried
bodies of the female insect Coccus cacti (Dactylopius coccus Costa,
Fam. Coccidae), containing eggs and larvae. The insect grows on
various species of the cactus, Nopalea (Fam. Cactacea), in the
Canary Islands and Central and South America. The colouring matter is
carminic acid. Carmine is the aluminium/calcium lake of carminic acid
(Meloan et al., 1971).
EVALUATION FOR ACCEPTABLE DAILY INTAKE
No information available.
Special studies on skin sensitization
Three subjects with lip lesions gave positive patch tests when
tested with red lip salve containing carmine (the Al/Ca lake of
carminic acid), but negative reactions to colourless lip salve. Since
the coloured product had previously been used by each individual, the
symptoms of allergic sensitivity were ascribed to carmine (Sarkany et
Special studies on mutagenicity
Carminic acid was negative in the Bacillus subtilis rec-assay
for DNA-damaging ability (Kada et al., 1972). Carminic acid was not
mutagenic for several strains of Salmonella typhimuriam in the
presence of liver microsomal preparations or enzymatic extracts of rat
cecal micro-flora (Brown and Brown, 1976; Brown et al., 1977).
The embryotoxicity and teratogenicity of carmine has been studied
in mice. Mice were killed on day 19 of gestation, after i.p.
injections of lithium carmine or sodium carmine on day 8. Treated
animals of both groups showed resorption rates (20%) higher than those
of control groups (2%). The malformation rate was about 16% in the
lithium carmine group and 2.5% after injection of sodium carmine. Only
animals given sodium carmine showed an increase in the number of
retarded foetuses (Schluter, 1970).
Groups of mice were injected once with 2.5% lithium carmine at a
dose of 150 mg earmine/kg on day 6, 8, 10, 12 or 14 of pregnancy. A
teratogenic effect was observed on the first three treatment days,
with the maximal response on day 8 (Schluter, 1971a and b).
No information available.
Mice (number not stated) were given intraperitoneal injections of
a 1 to 2% aqueous solution of the lithium salt of carminic acid for a
period of 60 days. The only abnormality observed was proliferation of
spleen tissue (Harada, 1931).
Groups of 40 rats, equally divided by sex, received ammoniacal
cochineal carmine in 0.4% aqueous gum tragacanth by intubation at
dosage levels of 0, 2.5, 5.0 and 10.0 mg/kg five days per week for
13 weeks. Body weight was recorded bi-weekly. Blood counts were made
three times. Gross and microscopic findings were not remarkable aside
from a dose-related accumulation of colour in the tissues of the rats
receiving the two higher dosage levels. No haematological effects were
noted. At the two highest levels some decreased growth was apparent.
Urine and faeces of the treated rats were coloured during the period
of administration (Battelle, 1962).
Groups of 50 weanling rats equally divided by sex were fed
carmine in the diet at levels of 0, 50, 250 and 500 mg/kg bw/day for
90 days. Blood counts, blood glucose, blood urea nitrogen and
urinalyses were performed three times. No effects due to the carmine
were reported in terms of growth, haematology and other clinical
findings. Gross and microscopic pathology were not remarkable (FDRL,
Five rabbits were given intravenous injections every five to
seven days, of 3 to 10 ml of a 2 to 4% aqueous solution of the lithium
salt of carminic acid. The treatment was continued for periods
varying from 130 to 529 days. No tumours were observed, but great
proliferation of the tissue of the spleen was noted (Harada, 1931).
Four groups of 30 mated female rats received daily 0, 200, 500 or
1000 mg/kg bw of carmine by gastric intubation as aqueous solution
during pregnancy days 0 to 20. A group of 17 similar animals received
a solution of chlorides to provide an intake of sodium, potassium and
ammonium equal to that resulting from the highest dose level of
carmine. No adverse effects were noted on body weight, pregnancy rate
pre-implantation losses, the average number of live young litter
weight or foetal weight. The group given the highest dose of carmine
and the cations control had an increased number of implantation sites
and of post-implantation losses. The latter was considered to be due
to an inability to maintain the increased numbers of implantations
rather than to an embryotoxic effect. No teratogenic effects were
noted in the foetuses and the degree of ossification of those from the
carmin treated rats tended to be more advanced than those from the
control (Gaunt et al., 1976).
Battelle Memorial Institute (1962) Unpublished report submitted to WHO
Brown, J.P. and Brown, R. J. (1976) Mutation Res., 40, 203
Brown, J.P., Roehm, G. W. & Brown, R. J. (1977) Environ. Mutagen
Sec. 8th Ann. Meet., Abst., p. 33
Food and Drug Research Laboratories (1962) Unpublished report
submitted to WHO
Gaunt, I. F., Clode, S. A. and Lloyd, A. G. (1976) Unpublished report
from B.I.B.R.A., submitted to WHO - Studies of the teratogenicity and
embryotoxicity of carmine in the rat. Report 162/1/76, July 1976
Harada, M. (1931) cited by Hartwell, J. L.: Survey of compounds which
have been tested for carcinogenic activity, second edition, 1951, p.
Meloan, S. N., Valentine, L. S. and Puchtler, H. (1971) Histochemie,
Kada, T., Tutikawa, K. and Sadaie, Y. (1972) Mutation Res., 16,
Sarkany, R. H., Meara, R. H. and Everall, J. (1961) Trans. St.
John's Hosp. Derm. Soc., 48, 39
Schluter, G. (1970) Z. Anat. Entwickl.-Gesch., 131, 228
Schluter, G. (1971a) Naunyn-Schmiedebergs Arch. Pharmak., 270, 56
Schluter, G. (1971b) ibid, 270, 316