INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY
WORLD HEALTH ORGANIZATION
TOXICOLOGICAL EVALUATION OF SOME
FOOD COLOURS, EMULSIFIERS, STABILIZERS,
ANTI-CAKING AGENTS AND CERTAIN
FAO Nutrition Meetings Report Series
No. 46A WHO/FOOD ADD/70.36
The content of this document is the result of the deliberations of the
Joint FAO/WHO Expert Committee on Food Additives which met in Rome,
27 May - 4 June 19691
Food and Agriculture Organization of the United Nations
World Health Organization
1 Thirteenth report of the Joint FAO/WHO Expert Committee on Food
Additives, FAO Nutrition Meetings Report Series, in press;
Wld Hlth Org. techn. Rep. Ser., in press.
CHLOROPHYLL COPPER COMPLEX AND CHLOROPHYLLIN COPPER COMPLEX,
SODIUM AND POTASSIUM SALTS
Oil soluble chlorophyll copper complex has part of the magnesium
replaced by copper. Water soluble chlorophyllin copper complex has
part of the magnesium replaced by copper and its methyl and phytyl
ester groups replaced by sodium and potassium. Any toxic effects are
therefore, in part, due to free ionisable copper present in the
complex. Potassium sodium chlorophyllin copper complex (4 per cent.
total Cu, 0.25 per cent. ionic Cu), in concentrations above 0.1 per
cent. of the diet, appears as chlorophyllin and Cu ions in the plasm.
Phylloerythrine is probably not formed in the rat; ingested
chlorophyll was excreted in the faeces as calcium complex. No copper
storage occurred in liver, kidney or spleen of rats at dietary levels
of 0.1 per cent, or 1 per cent. of sodium and potassium chlorophyllin
copper complex. There was no effect on iron storage at these levels.
Guinea-pigs fed 0.5 per cent. or rats fed 3 per cent. of the complex
in their diet showed no evidence of scurvy (Harrison et al., 1954).
Compound Animal Route LD50 Reference
chlorophyllin mouse oral 7000 Harrison et al., 1954
Chlorophyllin i.p. 190 Harrison et al., 1954
copper complex mouse i.v. > 400 Worden et al., 1955
i.m. > 500 Worden et al., 1955
i.p. > 1000 Worden et al., 1955
rat i.v. > 250 Worden et al., 1955
i.m. > 250 Worden et al., 1955
i.p. >1000 Worden et al., 1955
rabbit i.v. > 200 Worden et al., 1955
i.m. > 60 Worden et al., 1955
i.p. > 500 Worden et al., 1955
cat i.p. >60 Worden et al., 1955
dog i.v. > 200 Worden et al., 1955
i.m. > 50 Worden et al., 1955
i.p. > 200 Worden et al., 1955
pig i.v. > 10 Worden et al., 1955
i.m. > 20 Worden et al., 1955
i.p. > 50 Worden et al., 1955
Six mice were given 2500 mg/kg body-weight sodium chlorophyllin copper
complex orally for 7 days without any ill effects (Worden et al.,
1955). Five male and 4 female rats were fed a diet containing 15 per
cent. Na K C.C. for 10 days without any adverse effects except weight
loss related to food refusal (Harrison et al., 1954). Two guinea-pigs,
2 rabbits, 2 cats and 1 dog were given sodium chlorophyllin copper
complex 1000 mg/kg body-weight, orally daily for 7 days without any
adverse effects (Worden et al., 1955).
Mouse. Sodium chlorophyllin copper complex in 0.05, 0.1, 1.0 and 5
per cent. aqueous solutions was injected daily for 10 days
subcutaneously at points along the spine followed by exposure of the
mice to sunlight, UV lamp, or darkness. Many mice died and liver and
kidney changes, more extensive in irradiated animals, were seen
Rat. Thirty rats received oral doses of 2000 mg/kg body-weight
sodium chlorophyllin copper complex for 18 weeks without any adverse
effects (Worden et al., 1955). Offsprings of 6 female rats fed 1 per
cent. of potassium disodium chlorophyllin copper complex for 19 weeks
exhibited locomotory difficulties and skeletal muscle defects (Reber &
Guinea-pig. Five female guinea-pigs received 0.5 per cent.
potassium sodium chlorophyllin copper complex in their drinking water
for 11 weeks without ill effects or pathological change. There was no
evidence of scurvy (Harrison et al., 1954).
Fowl. Sixty-day-old chickens received orally 70 mg/kg body-weight
sodium chlorophyllin copper complex for 6 weeks and 8-year-old fowls
received 500 mg/kg body-weight for 3 weeks without gross adverse
effects. The yolk of all eggs laid was coloured an intense green
(Worden of al., 1955).
Rat. Groups of 40 rats were fed diets containing 0, 0.1, 1.0 and 3
per cent. of potassium sodium chlorophyllin copper complex (4 -5 per
cent. total Cu, 0.25 per cent. ionic Cu) over their life span. Growth
rate, feed efficiency, haematology and urinalysis were comparable to
the controls. Reproduction showed no impairment of conception. No
gross or histopathological changes attributable to the potassium
sodium chlorophyllin copper complex were seen. There was no evidence
of Cu toxicity or deposition in liver, kidney or spleen (Harrison et
The copper in these complexes is firmly bound. Though increased plasma
levels of copper have been reported there is no significant tissue
storage nor is there any evidence of destruction of ascorbic acid.
Chlorophyll copper complex has higher toxicity when given parenterally
but this has no toxicological significance if this colour is used by
the oral route, No significant chronic effects were seen in the
long-term tests in rats. Changes in the copper levels in the plasma
may be of importance for people suffering from Wilson's disease. At
present the limit for free copper in the sodium and potassium salts of
chlorophyllen copper complex is much higher than that in the complex.
Level causing no toxicological effect in the rat
3 per cent. (= 30 000 ppm) in the diet equivalent to 1500 mg/kg
body-weight per day.
Estimate of acceptable daily intake of chlorophyll copper complex for
0 - 15
Estimate of acceptable daily intake of chlorophyllin copper complex,
sodium and potassium salts, for man
0 - 15
Further work required by June 1972
See tentative specifications, Ref. 19, Annex I.
Harrison, J. W. E., Leoni, S. E. & Trabin, B. (1954) J. Arner.
Pharm. Assoc., Sc. Ed., 43, 722
Reber, E. F. & Willigan, D. A. (1954) Amer. J. vet. Res, 15, 643
Tomino, U. (1958) Kobe Ika Daigaku Kiyo, 14, 98
Worden, A. N., Bunyan, J. & Kleissner, M. (1955) Brit. Vet. J.,