INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY
WORLD HEALTH ORGANIZATION
TOXICOLOGICAL EVALUATION OF SOME
FOOD COLOURS, ENZYMES, FLAVOUR
ENHANCERS, THICKENING AGENTS, AND
CERTAIN FOOD ADDITIVES
WHO FOOD ADDITIVES SERIES 6
The evaluations contained in this publication were prepared by the
Joint FAO/WHO Expert Committee on Food Additives which met in Rome,
4-13 June 19741
World Health Organization Geneva 1975
1 Eighteenth Report of the Joint FAO/WHO Expert Committee on
Food Additives, Wld Hlth Org. techn. Rep. Ser., 1974, No. 557.
FAO Nutrition Meetings Report Series, 1974, No. 54.
CHLOROPHYLLIN COPPER COMPLEX, POTASSIUM AND SODIUM SALTS
Explanation
These compounds have been evaluated for acceptable daily intake
by the Joint FAO/WHO Expert Committee on Food Additives (see Annex 1,
Ref. No. 20) in 1969.
Since the previous evaluation additional data have become
available and are summarized and discussed in the following monograph.
The previously published monograph has been expanded and is reproduced
in its entirety below.
BIOLOGICAL DATA
BIOCHEMICAL ASPECTS
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% total Cu,
0.25% ionic Cu), in concentrations above 0.1% of the diet, appears as
chlorophyllin and Cu ions in the plasma. 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% or 1% of sodium and potassium
chlorophyllin copper complex. There was no effect on iron storage at
these levels. Guinea-pig fed 0.5% or rats fed 3% of the complex in
their diet showed no evidence of scurvy (Harrison et al., 1954).
TOXICOLOGICAL STUDIES
Acute toxicity
Six mice were given 2500 mg/kg bw sodium chlorophyllin copper
complex orally for seven days without any ill effects (Worden et al.,
1955). Five male and four female rats were fed a diet containing 15%
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, two
rabbits, two cats and one dog were given sodium chlorophyllin copper
complex 1 000 mg/kg bw orally daily for seven days without any adverse
effects (Worden et al., 1955).
Acute toxicity
LD50
Compound Animal Route (mg/kg bw) Reference
Potassium sodium Mouse Oral 7 000 Harrison et al., 1954
chlorophyllin
copper complex
Chlorophyllin i.p. 190 Harrison et al., 1954
copper complex
Sodium Mouse i.v. > 400 Worden et al., 1955
chlorophyllin
copper i.m. > 500 Worden et al., 1955
complex
i.p. > 1 000 Worden et al., 1955
Rat i.v. > 250 Worden et al., 1955
i.m. > 250 Worden et al., 1955
i.p. > 1 000 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
Short-term studies
Mouse
Sodium chlorophyllin copper complex in 0.05, 0.1, 1.0 and 5%
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 (Tomino, 1958).
Rat
Thirty rats received oral doses of 2000 mg/kg bw sodium
chlorophyllin copper complex for 18 weeks without any adverse effects
(Worden et al., 1955). Offspring of six female rats fed 1% of
potassium disodium chlorophyllin copper complex for 19 weeks exhibited
locomotory difficulties and skeletal muscle defects (Reber & Willigan,
1954).
Guinea-pig
Five female guinea-pigs received 0.5% 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 bw sodium
chlorophyllin copper complex for six weeks and eight-year-old fowls
received 500 mg/kg bw for three weeks without gross adverse effects.
The yolk of all eggs laid was coloured an intense green (Worden et
al., 1955).
Long-term studies
Rat
Groups of 40 rats were fed diets containing 0, 0.1, 1.0 and 3% of
potassium sodium chlorophyllin copper complex (4-5% total Cu, 0.25%
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 al.,
1954).
Comments:
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.
EVALUATION
Level causing no toxicological effect
Rat: 3% (= 30 000 ppm) in the diet equivalent to 1500 mg/kg bw.
Estimate of acceptable daily intake for man
0-15 mg/kg bw*,**
FURTHER WORK OR INFORMATION
Required by June 1978
Revision of tentative specification.
REFERENCES
Harrison, J. W. E., Iconi, 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., 111,
385
* Temporary.
** As Na + K chlorophyllin copper complex.