INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY
WORLD HEALTH ORGANIZATION
TOXICOLOGICAL EVALUATION OF SOME
FOOD COLOURS, EMULSIFIERS, STABILIZERS,
ANTI-CAKING AGENTS AND CERTAIN
OTHER SUBSTANCES
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.
AMMONIUM SALTS OF PHOSPHATIDIC ACIDS
Biological Data
Biochemical aspects
This compound was readily absorbed from the rat gastro-intestinal
tract in the presence of fats without changes in fat absorption or
gastro-intestinal function (Frazer, 1954). No in vitro haemolytic
effect was seen with this product nor was it any stronger than natural
lecithin (Frazer, 1954). Incubation of the ammonium salt of
32P-phosphatidic acid with simulated gastric juice, homogenate of
intestinal mucosa, trypsin or chymotrypsin in vitro resulted in the
liberation of inorganic phosphate. The ammonium salt of
32Pphosphatidic acid was given to eight male and seven female rats at
a level of 60-30 mg/kg body weight. Urine and faeces were collected at
intervals of 1, 2, 4, 6, 7 and 24 h and at 10 and 16 days after
administration rats were killed for examination. Similarly, inorganic
phosphate labelled with 32P was given to three male and three female
rats by intubation and rats were killed 1, 2 and 4 h after
administration. Many organs were examined for radioactivity. In
addition, incorporation of 32P into phospholipids of tissues at
various intervals after dosing was determined. Percentage recoveries
after 24 h were 91.4 per cent. males, 91.9 per cent. females, after 16
days 93.4 per cent. males, 98.1 per cent. females. Thus 79 per cent.
of the labelled material was excreted in the faeces within 24 h while
some four per cent, was eliminated in the urine. The remaining 17 per
cent. were stored in bone and muscle as inorganic phosphate and
phosphate ester while the rest of the tissue contained oily traces
mostly in the phospholipid fraction. Comparison with uptake of
labelled inorganic phosphate showed similar distribution of the
phosphate to bone, muscle, liver, gut content and urine. The
radioactive phospholipid fraction present in the rat 4 h after
administration was found to be nearly identical whether 32P was
administered as phosphatidic salt or as inorganic orthophosphate. The
radioactive phospholipid fraction in the liver of test rats was
chromatographically identical with the liver phospholipid of rats fed
32P orthophosphate. The observed tissue radioactivity is due to
breakdown to inorganic phosphate which enters the phosphate and
phospholipid pools. There was no evidence of storage in tissues of any
P containing moiety of the phosphatidic salt. The triglyceride moiety
was probably hydrolyzed and absorbed by normal physiological routes
(Feuer, 1967).
Acute toxicity
Animal Route LD50 Reference
mg/kg body weight
Rat Oral 5 000 Frazer, 1954
Rat I.m. 2 000 Frazer, 1954
Guinea-pig I.m. 2 000 Frazer, 1954
Rabbit Oral 5 000 Frazer, 1954
Dog Oral 2 000 Frazer, 1954
No abnormal pathological findings or behaviour patterns were seen
(Frazer, 1954).
Short-term studies
Rat. After twice-weekly intraperitoneal injections in rats of 2 g/kg
for five weeks, there was no deleterious effect on growth, relative
spleen weight, haematology or corpuscular fragility (Gaunt et al.,
1967).
Groups of 15 male and 15 female rats each received diets containing 0
per cent., 0.75 per cent., 1.5 per cent., 3.0 per cent. and 6.0 per
cent. of the compound for 90 days. No adverse effects were seen on
appearance, growth, food consumption, haematological indices, liver
and kidney function, relative organ weights and gross and histological
appearance of the organs. Similar results were obtained in rats given
a dietary level of six per cent. soya lecithins for 90 days except
that a slight transient anaemia was seen in females (Gaunt et al.,
1967).
Groups of 50 rats each received 0 per cent., 1 per cent. or 2.5 per
cent. compound in their diet for 45 weeks. No adverse effects were
seen with regard to mortality, weight gain, liver function, kidney
function and histopathology in test groups compared with controls
(Frazer, 1954).
Long-term studies
None available.
Comments
The biochemical studies show that the ammonium salts of phosphatidic
acids break down into normal food constituents. The available rat
studies show this material to be non-toxic at the level of six per
cent. in the diet, the highest concentration tested. Because of the
possible substantial intake of this material by children it is
necessary to have a long-term study on one species and desirable to
determine the metabolic fate in man. A long-term study in the rat is
now in progress.
EVALUATION
Level causing no significant toxicological effect in the rat
Six per cent. (= 60 000 ppm) in the diet equivalent to 3000 mg/kg body
weight/day.
Estimate of acceptable daily intake for man
mg/kg body weight
Temporary acceptance 0-15
Further work required by June 1974
Submission of the results of the long-term study in the rat and
metabolic studies in several species.
For further work on specifications, see ref. 19 Annex 1.
REFERENCES
Feuer, G. (1967) Fd Cosmet, Toxicol., 5, 631
Frazer, A. C. (1954) Unpublished report dated July 1954
Gaunt, I. F., Grasso, P. & Gangolli, S. D, (1967) Fd Cosmet.
Toxicol., 5, 623
See Also:
Toxicological Abbreviations
Ammonium salts of phosphatidic acids (WHO Food Additives Series 5)