INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY WORLD HEALTH ORGANIZATION Toxicological evaluation of some food colours, thickening agents, and certain other substancse WHO FOOD ADDITIVES SERIES NO. 8 The evaluations contained in this publication were prepared by the Joint FAO/WHO Expert Committee on Food Additives which met in Geneva, 14-23 April 19751 World Health Organization, Geneva 1975 1 Nineteenth Report of the Joint FAO/WHO Expert Committee on Food Additives, Wld Hlth Org. techn. Rep. Ser., 1975, No. 576; FAO Nutrition Meetings Report Series, 1975, No. 55. The monographs contained in the present volume are also issued by the Food and Agriculture Organization of the United Nations, Rome, as FAO Nutrition Meetings Report Series, No. 55A ISBN 92 4 166008 2 (C) FAO and WHO 1975 AMARANTH Explanation This substance was evaluated for acceptable daily intake for man by the Joint FAO/WHO Expert Committee (see Annex 1, Refs No. 10 and No. 30) in 1964 and 1972. Since the previous evaluation, additional data have become available and are summarized and discussed in the following monograph. The previously published monographs have been expanded and are reproduced in their entirety below. BIOLOGICAL DATA BIOCHEMICAL ASPECTS Adult rats were given intravenous injections of the colour. The bile was collected for six hours and an average of 53% (43-79%) of the quantity of the colour administered was found (Ryan & Wright, 1961). Six adult rats were given a single oral dose of 100 mg as aqueous solution per animal. Only 0.45% of the dose administered was found in the faeces collected over a period of 48 hours. A single oral dose of 50 mg per animal was administered to four rats. Only 2.8% was absorbed from the gastro-intestinal tract; the metabolites in the urine and bile were predominantly products resulting from the reductive fission of the azo-linkage, such as 1-amino-4-naphthalene sulfonic acid and 1-amino-2-hydroxy-3,6-naphthalene disulfonic acid. The former compound was found also in the faeces. The liver enzyme that reduces azo-linkages plays little part in the metabolism, as was shown in experiments in which the colour was given by intrasplenic infusion. Reduction of the compound is therefore most probably affected by the intestinal bacteria (Radomski & Mellinger, 1962). After a prolonged administration of a daily dose of 50 mg/rat the vitamin A content of the liver showed a threefold to fourfold decrease. The glutathione content of the liver and spleen was increased (Galea, 1962). Rats fed amaranth showed no change in liver phosphatase, glucose-6-phosphate dehydrogenase or gluconate-6-phosphate dehydrogenase (Manchon & Albrecht, 1972) but an increase in catalase, succinic dehydrogenase (Rubenchik, 1962) and azoreductase (Albrecht et al., 1973). The absorption and elimination of 1-amino-4-naphthalene-sulfonic acid, one of the azo-reduction products in rats were examined after dosing by gavage, in drinking-water and mixed into the diet. Little is metabolised by mammalian liver azo-reductase systems, almost all reduction occurring through gut microflora. 1-amino-2-hydroxy-3, 6-naphthalene sulfonic acid was not studied as it is not well absorbed. After intravenous administration there is rapid clearance from the serum within two hours, 42% being excreted in the urine within 24 hours, only 11% in the faeces (accounting for 53%) orally administered compound was rapidly absorbed but cleared within 24 hours. Fifteen per cent. appeared in the urine, 69% in the faeces (accounting for 84%). Of FD & C Red No.2 administered orally 10% appear in the urine and 43% in the faeces (accounting for 53%). Part of the absorbed reduction product is further metabolised to unknown products (Pritchard et al., 1974). TOXICOLOGICAL STUDIES Special studies on reproduction and teratogenicity Mouse Groups of eight to 10 pregnant mice were given by gavage once daily either from day 0-7 or day 6-18 of pregnancy doses of amaranth corresponding to 0, 7.5, 30 or 100 mg/kg body weight. Mothers were sacrificed on day 18 and litter parameters, resorptions and malformations examined. No significant effects were noted on resorption incidence or foetal deaths and foetal growth. No specific teratogenic effects related to amaranth administration were seen (Larsson, 1974). The administration of 27, 90, 300 or 1000 mg/kg body weight of amaranth to pregnant CD-1 outbred mice for 10 consecutive days from day six of gestation by gavage had no apparent effects on midation, maternal or foetal survival. No difference in abnormalities were seen in skeletal or soft tissues compared with controls (Anonymous, 1972a). Rat Groups of one male and four female rats were fed diets supplying 1.5 and 15 mg/kg body weight/day of amaranth. There were two control groups of a similar size. Each group was mated three times, at four to five months, seven to eight months and 10-12 months after the start of the study. The following were studied: female fertility, gestation period, numbers of live and dead pups born and numbers surviving for four days and one month. Abnormalities in the pups were noted. Similar observations were carried out on first (F1) and second (F2) generations of rats fed on the same diets as their parents. The authors considered that their results showed amaranth decreased fertility, increased the number of stillbirths, produced deformities in the young and reduced survival of the young (Shtenberg & Gavrilenko, 1970). Groups of 13-15 pregnant rats were given 0, 7.5, 15, 30, 100 or 200 mg/kg/day amaranth by stomach tube from day 0-19 and sacrificed on day 20. No adverse effects were noted on implantation. The percentage of dead foetuses increased in a dose-related manner. At 200 mg/kg/day there was a foetotoxic effect. Resorptions increased from 15 mg/kg upwards with total resorption of occasional litters at 100 and 200 mg/kg/day. No teratogenic abnormality related to amaranth was seen. No obvious effect was noted on foetal sex (Collins et al., 1972). To elucidate the effect of gavage versus diet incorporation on teratological findings a number of comparative studies using the same protocol and material were carried out on Osborne-Mendel and Charles River CD-1 rats in three different laboratories using a uniform protocol. Six control and four experimental groups of 20 pregnant rats were used. All experimental females were treated by gavage with 200 mg/kg amaranth/day either from days 0-19 or days 6-15 or days 7-9 and the fourth group were given 0.2% of the dye in drinking-water from day 0-20 corresponding to an intake of 200 mg/kg. The controls received by gavage a solution from days 0-19, days 6-15, days 7-9, by intubation on days 0-19, distilled water gavage days 0-19 and a group entirely untreated and unhandled. No significant differences in embryotoxicity were noted between experimental rats and controls for the Osborn-Mendal strain or the CD-1 strain. The observed significant findings, reimplants, viable foetuses, percentage resorption/litter and weights of male and female foetuses were overall negative. The original gavage finding was not reproduced and the earlier study had an unusually low incidence of resorptions compared with the normal incidence (Anonymous, 1974a). Eight groups of 16-22 pregnant rats were given 0, 15, 50 or 150 mg/kg/day of amaranth by stomach tube on days 6-15 of pregnancy and Caesarean section was performed on day 20. No compound-related adverse effects were noted in terms of implantation, foetal mortality, foetal weight or reproductive performance in any of the experimental groups as compared to the control animals. No evidence of compound- induced teratogenic effects was noted (Keplinger et al., 1974). In another study five groups of four pregnant Charles River rats received from gestation, days 6-15 inclusive, 0, 15, 150, 450 and 1500 mg/kg body weight amaranth per day by gavage. Females were killed on day 20. No abnormal effects were seen as regards maternal weight gain, litter size, average foetal weight and number of resorptions. No gross abnormalities related to the compound were seen (Burnett et al., 1974). The administration of 27, 90, 300 or 1000 mg/kg body weight of amaranth to pregnant rats for 10 consecutive days (6-15 of gestation) had no clearly discernible effect on nidation or on maternal or foetal survival. The incidence of abnormalities in soft and skeletal tissues of the test groups did not differ significantly from those of control animals (Anonymous, 1972). Five groups of 10 male and 20 female rats (Fo) were fed 0, 1.5, 15, 45 or 150 mg colour in their diet for two weeks and then mated twice to produce the F1 generation. This was mated three times, the F2b litters being used for producing the F2 generation. The F2 parents were mated once to produce an F3a generation. No significant findings were seen as regards growth of the parent generations or in the litter, weaning and teratological observations on all three generations (Haley et al., 1972; Smith et al., 1974a and 1974b). In another three-generation reproduction and teratology study groups of Osborne-Mendel rats were fed 0, 30, 300, 3000 or 30 000 ppm colour from weaning to three months and then mated to produce an F1, F2, and F3 generation. No significant effects were seen on parent or offspring parameters, e.g. survival, weight gain, fertility, litter size, viability of pups, weaning performance and offspring survival. No dye effect could be linked distinctly to any adverse effects seen except weanling weights were significantly decreased at the highest dose level in females of the F1d generation and in males and females of the F2a x F2c generation. No cumulative effects were seen (Collins et al., 1975a). Progeny from the F1a and F3b generation of Osborne-Mendel rats treated with amaranth in the Fo generation at levels of 0, 30, 300, 3000 or 30 000 ppm were used for teratology studies. In the F1a generation the number of corpora lutea was lower at 30 000 ppm but preimplantation loss per litter did not differ from controls. Resorption and decreased mean foetal weight of progeny in F1a were not dose-related. Implantation and survival parameters in the F3b generation were similar to controls. No specific skeletal or soft tissue abnormality were seen which could be correlated to dose levels of the colour (Collins et al., 1975b). Samples of amaranth obtained from three manufacturers were administered to rats by gavage, or mixed in a semi-synthetic diet to provide in each case dosage levels of 0, 15, 30, 100 or 200 mg/kg body weight/day, from days 1-19 of pregnancy. Prenatal values for corpora lutea, live foetuses, deciduomas, dead foetuses and foetal weight gave no evidence of any amaranth-related deleterious effect on implantation or embryonic survival (Khera et al., 1974). In a study on two metabolites (sodium naphthionate and the R-amino salt) and one intermediate of amaranth, dose levels of 15, 30, 100 and 200 mg/kg/day were administered by stomach tube to pregnant rats during days 0-19 of pregnancy. No adverse effects were noted on implantation, but at the two highest dose levels of naphthionate and R-salt the multiple resorption rate was significantly higher than that among control litters. Sodium naphthionate at the 100 mg/kg dose level induced a significant increase in percentage of foetuses with sternebral abnormalities, but this was not observed with the R-salt. The latter increased the number of skeletal abnormalities at the 30 mg/kg level (considered an aberrant finding) and adversely influenced skeletal development at the higher dose levels (Collins et al., 1973). Rabbit Eight groups of 10-14 pregnant rabbits were given 0, 1.5, 5.0 and 15.0 mg/kg/day of amaranth by capsule on days 6-16 of pregnancy and sacrificed on day 29 by Caesarean section. No evidence of compound- induced teratogenic effects were noted, nor were significant effects seen in terms of implantation, pup weight, young born alive and total numbers of resorptions. Average numbers of early resorptions per litter revealed an increase which approaches statistical significance (P = 0.05) for the 1.5 and 15.0 mg/kg groups while for the 5.0 mg/kg group the increase is significant (Keplinger et al., 1974). The administration of 27, 90, 300 or 1000 mg/kg (body weight) of amaranth to pregnant rabbits from days 6-18 of gestation had no clearly discernible effect on nidation or on maternal or foetal survival. The incidence of abnormalities in soft or skeletal tissues of the test groups did not differ significantly from those of control groups (Anonymous, 1972b). Hamster The administration of up to 1000 mg/kg (body weight) of amaranth to pregnant hamsters from days 6-10 of gestation had no discernible effect on nidation or on maternal or foetal survival. The incidence of abnormalities seen in soft or skeletal tissues of the test groups did not differ significantly from those of control groups (Anonymous, 1972c). Cat A teratogenic and reproduction study was performed in four groups of 12 adult female shorthair cats given 0, 300, 900 and 3000 ppm amaranth in their diet prior to and during breeding and gestation. All females were bred with males fed a diet of 3000 ppm amaranth. Six females were delivered by Caesarian section at approximately 60 days gestation, the other six were allowed to queen litters normally. Difficulties were experienced with standardizing the pregestation feeding period because of uncertainty about natural oestrus onset. For similar reasons the c.s. delivered kittens were at various stages of gestation. Implantation and resorption sites, corpora lutea, stillborn and viable progeny were examined for 24 hours after c.s. Normally queened kittens were weaned after eight weeks and examined. The number of resorption sites was increased at 3000 ppm and 24-hour viability after c.s. was reduced in the 300 ppm group. The mean body weight at queening was lower in the 900 ppm group only but no effects were seen at eight weeks postnatally. None of the parameters examined could be interpreted with certainty as evidence of adverse effects (Korinke et al., 1974). Another study is under way in four groups of 20 female cats using 0, 80, 160 and 300 mg/kg per day of amaranth in gelatin capsules for 15-45 pregestation days. Oestrus was synchronized and untreated males were used for mating (Khera, 1975). Dog A teratogenic and reproduction study was performed in four groups of 12 adult female beagles given 0, 300, 900 and 3000 ppm amaranth in their diet and mated with males treated with 3000 ppm. Pregestation treatment varied from 45 to 382 days for the first litters and from 132 to 572 days for the second litters. Caesarian section was performed on six females in each group after 60 days gestation. The remaining females were allowed to deliver spontaneously for two litters. Pups were weaned until eight weeks of age. Two consecutive litters were studied. No significant effects were noted on breeding, body weight, food consumption of dams or on litter size, viability, pathology and skeletal development of pups (Mastalki et al., 1975). Special studies on mutagenicity The colour was tested for mutagenic action in a concentration of 0.5 g/100 ml in cultures of Escherichia coli. No mutagenic effect was found (Lück & Rickerl, 1960). FD & C Red No.2 was shown to have a mutagenic response in the host-mediated assay for the two strains of Salmonella typhimurium G-46 and TA-1530, particularly when the compound was administered by multiple injection over five days. Apparently this mutagenic response is induced by some metabolite rather than by the compound itself, since in vitro exposure of the compound to the bacteria did not induce mutations. Similar results were obtained in recombination tests with yeast (Legator, 1972; Newell & Maxwell, 1972a. No consistent responses occurred to suggest that FD & C Red No. 2 is mutagenic to the rat as a result of a dominant lethal test. The positive reference control compound, TEM, produced mutagenic responses from the second through the fifth weeks of the experiment, as would be expected from this known mutagen (Newell & Maxwell, 1972b). Other special studies Chick Chick embryo studies were performed with the dye and sodium naphthionate being administered to the yolk and air cell of varying doses. No definite dose/response relationship was seen the compound being less toxic at intermediate levels than at low and high dose levels (Winbush, 1972). Guinea-pig In experiments with guinea-pigs, it was found that this colour had no sensitization activity (Bar & Griepentrog, 1960). Cat The test for Heinz bodies was negative after injecting four cats with a 5% aqueous solution of this colour at a level of 1 g on the first day and 0.1 g on the ninth and on the eighteenth day (Anonymous, 1957). Rabbit Six groups of nine rabbits were used in dermal irritation and percutaneous toxicity tests employing 0.1% × 1% dye in ointment or water vehicle. No significant dermal or systemic toxicity related to treatment was seen (Carson, 1962). Acute toxicity Animal Route LD50 References per kg body weight Rat I.p. >1 g Anonymous, 1957 Rat I.v. >1 g Anonymous, 1957 Mouse Oral >10 g Anonymous, 1959 C.p. >0.5 g Anonymous, 1952 Short-term studies Rat A group of five young rats were given the colour subcutaneously twice daily for three days. The rats were killed on the fourth day. The colour was administered in aqueous solution at a level of 250 mg per kg body weight each injection. No oestrogenic activity (normal uterine weight) was detected in comparison with a control group. No other abnormalities were found (Graham & Allmark, 1959). Six groups of six male and six female rats were fed either basal stock diet, a highly purified basal diet, stock diet containing 2.5% or 5% amaranth and purified basal diet containing 2.5% or 5% amaranth for 21 days. As the 5% level amaranth had practically no adverse effect on weight gain or appearance but in the purified diet growth ceased and animals died within two weeks. Vitamin supplements had no protective effect but 10% cellulose, 10% alfalfa meal or watercress powder protected against this dietary effect (Ershoff & Thurston, 1974). Long-term studies Mouse Twenty mice were fed 15-20 mg of this colour five days a week for periods up to 477 days. Autopsies were conducted on 18 of the mice. No lesions were observed in the one liver examined (Cook et al., 1940). Feeding studies were conducted with C3Hf and C57B1 mice; 100 of each strain were fed at 1.0 and 2.0% and 200 of each strain served as controls. No tumours were observed in the mice of either strain (Anonymous, 1964a). Two groups of 100 mice received either 0 or 0.01 g amaranth paste (= 0.004 g amaranth) by gavage daily. One drop of either 9, 10-dimethyl-2-benzanthracene or 3, 4-benzopyrene was applied once per week to interscapular skin. Papillomata appeared 3.5 weeks earlier in test animals and in a greater number of animals. A larger percentage became malignant in test animals (Baigusheva, 1968). Fifty male and 50 female Swiss-Webster albino mice were treated weekly for 18 months with 0.1 ml 1% colour suspension in water. One hundred males x 100 females were controls. No carcinogenic effects on skin were noted (Carson, 1963, 1966). Rat Three groups of 15 male and 15 female rats were given diets containing 0.03%, 0.3% and 1.5% of the colour for 64 weeks. The mortality of the rats was the same as that in a similar control group. At a level of 1.5% a significant decrease in growth rate was found in female rats but not in male rats. This was attributed to an effect on food utilization rather than on food consumption. Female rats fed the colour at 0.3% and 1.5% showed an increase in liver weight. At the higher concentration there was also an increase in kidney weight. No influence on food intake, histopathology and blood-picture and no significant difference in tumour incidence was found (Mannel et al., 1958). Ten rats were fed the colour at a level of 0.2% in the diet. Each animal received an average of 0.1 g/kg body weight per day for 417 days. The total intake of the colour was 11 g/animal. The observation period was 830 days. One intestinal carcinoma was observed (Anonymous, 1957). No tumours were noted in 11 rats subjected to the subcutaneous injection of 0.5 ml of 1% solution twice weekly for 365 days. The observation period was 879 days. The total dose administered was 0.5 g/animal (Anonymous, 1957). At a level of 4% in the diet the colour was fed to five male and five female rats for periods up to 18 months. Gross staining of the flandular stomach and small intestine was observed. Granular deposits were noted in the stomach, small intestine and in some cases in the colon. A lymphosarcoma was observed. No tumours occurred in 50 control animals surviving 20 months or more (Willheim & Ivy, 1953). This colour was injected subcutaneously into 18 rats of both sexes for 94 to 99 weeks. In general 1 ml of a 2-3% solution was injected weekly for 693 days. No tumours were observed (Nelson & Hagan, 1953). Rats fed the dye at 20 mg/day for 78 weeks showed 68% mortality compared with 13% for controls, a lowering of the vitamin A content of liver and vacuolar dystrophy with eventual fatty degeneration of liver cells (Galea et al., 1972). Groups of 24 weanling rats were fed the colour of 0.5%, 1.0%, 2.0% and 5.0% in the diet. A similar group served as controls. The animals on 5.0% showed slight growth inhibition. Gross and microscopic examinations revealed a questionable increase in the number of mammary tumours; two tumours were observed in the control group and three, three, six and four tumours respectively in the groups receiving the colour. Additional feeding studies for two years were carried out to repeat this result with Osborne-Mendel and Sprague-Dawley rats at 0.0, 1.0, and 2.0% with 50 male and 50 female animals of each strain. There was no statistically significant influence on the formation of tumours. One hundred males and 100 females were used as controls (Anonymous, 1964b). One group of 50 outbred rats was fed a diet containing amaranth paste (40% amaranth) for 25 months. A control group of 35 rats was used. Dietary levels ranged from 0.8% to 1.6% amaranth. Tumours of the peritoneum and intestine began to appear in 18 survivors at 19 months. By 25 months a total of 11 tumours was found. No tumours were seen in controls. All tumours were histologically malignant (Baigusheva, 1968). Two groups of 50 outbred male rats were fed on a diet containing either 0% or 2% amaranth for 33 months, that is until all animals died. A small depression of body weight, compared with controls was observed but this was not statistically significant. The 15 tumours found (in 13 rats out of 48 survivors) included three lymphosarcomas, four sarcomas, one adenofibroma, three intestinal carcinomas, one hepatoma and three skin carcinomas. The first tumour appeared after six months, most of the others at 21-23 months. The author reported that not one of the 50 control rats developed any kind of tumour during the whole course of the experiment (Andrianova, 1970). When amaranth was fed to rats for up to 18 months at a dietary level of 0.12% growth was markedly depressed, mortality was increased and liver damage was evident. The level of vitamin A was decreased by about 50% within a few days and continued to decrease progressively throughout the study. A rise in serum albumin and gamma-globulin was noted, but there were no marked increases in serum or liver glutamic-oxaloatic or glutamic pyruvic transaminases (Galea et al., 1962). However, feeding of 1200, 3000, 10 000 and 20 000 ppm of amaranth in the diet did not confirm the reduction of vitamin A reserves in the rat liver in another experiment (Truhaut & Ferrando, 1975). In another study unmated rats of both sexes were given by gavage either 1.5 or 15 mg/kg amaranth for 12 months. Females showed inhibition of oestrus cycle, increased foetal deaths, impaired lactation. Males had decreased length of sperm life and mortality and reduced resistance (Shtenberg & Gavrilenko, 1972). Groups of animals from the F2a generation of a multi-generation study using dose levels of 0, 30, 300, 3000 or 30 000 ppm in the diet are being used for a two-year long-term study. No results are yet available (Anonymous, 1975). Dog A seven-year toxicity study was carried out on female beagles. Five dogs were fed 2% of the colour in the diet, three dogs were used as controls. No histopathological or other abnormalities were found (Anonymous, 1974b). OBSERVATIONS IN MAN Of seven patients with recurrent urticaria or angio-oedema suspected to be sensitive to azo-dyes such as amaranth, one reacted with urticaria to amaranth provocation. Objective signs of a reaction were observed in one further case and in three other patients subjective symptoms were noted (Michaelson & Juhlin, 1973). Comments: Many long-term studies have been carried out in rats, mice and dogs. Only two of the long-term studies indicated a carcinogenic potential not seen in any of the other studies. These studies were also evaluated in a report of the International Agency for Research on Cancer (Lyon). This Committee, as well as the IARC, concluded that because of the uncertainty about the impurity content of the amaranth employed in these two studies, the carcinogenicity of this compound could not be evaluated. Several new studies on reproduction and teratology were available for evaluation. These gave some conflicting results with regard to foetotoxicity, although none of them produced any evidence of teratogenic effects related to amaranth administration. Further studies to elucidate the observations generating concern over reproductive effects have shown that, in retrospect, a study showed apparent adverse effects because of the unexpectedly low foetal resorption in control animals compared with non-contemporary controls. An extensive comparative study has failed to reproduce these effects in the same strain. The single positive teratogenic study also suffered from an inadequate specification for the dye employed. The Committee took account of the fact that the new data in addition to the new information on the deficiency of the specification of amaranth in the positive studies previously reviewed, allowed for more precise assessment of the ADI. EVALUATION Level causing no toxicological effect Rat: 0.3% in the diet equivalent to 150 mg/kg body weight. Estimate of acceptable daily intake for man 0-0.75 mg/kg body weight.* FURTHER WORK OR INFORMATION Required by 1978. The result of long-term feeding study on the progeny of rats that were fed amaranth during the gestation and lactation period. REFERENCES Albrecht, R., Manchon, Ph., Keko-Pinto, C. & Lowy, R. (1973) Evolution, au Cours du Temps, de Quelques Effets du Bordeaux S et du Jaune de Beurre sur l'Organisme du Rat, Fd. Cosmet. Toxicol., 11, 175-184 Andrianova, M. M. (1970) Carcinogenous properties of red food pigments - amaranth, SX purple and 4R purple, Vop. Pitan., 29, 61-65 Anonymous (1952) Unpublished report submitted to the World Health Organization by the Food and Drug Administration, United States of America Anonymous (1957) DFG (Deutsche Forschungsgemeinschaft) Farbstoff Kommission, Toxikologische Daten von Farbstoffen und ihre Zulassung fur Lebensmittel in verschiedenen Lärdern, Mitt. 6(2), Wiesbaden, Steiner Verlag, p. 40 Anonymous (1959) Unpublished report submitted to the World Health Organization by the Food and Drug Administration, United States of America Anonymous (1964a) Unpublished report submitted to the World Health Organization by the Food and Drug Administration, United States of America Anonymous (1964b) Unpublished report submitted to the World Health Organization by the Food and Drug Administration, United States of America Anonymous (1972a) Unpublished report from the Food and Drug Research Labs submitted to the World Health Organization by the Food and Drug Administration, United States of America * Temporary. Anonymous (1972b) Unpublished report from the Food and Drug Research Labs submitted to the World Health Organization by the Food and Drug Administration, United States of America Anonymous (1972c) Unpublished report from the Food and Drug Research Labs submitted to the World Health Organization by the Food and Drug Administration, United States of America Anonymous (1974a) Memorandum of Meeting on FD & C Red No.2 held on 6 June 1974 at the Food and Drug Administration, Washington, D.C. Unpublished report submitted to the World Health Organization by the Food and Drug Administration, United States of America Anonymous (1974b) Unpublished report submitted to the World Health Organization by the Food and Drug Administration, United States of America Anonymous (1975) Unpublished report submitted to the World Health Organization by the Food and Drug Administration, United States of America Baigusheva, M. M. (1968) Carcinogenic properties of the amaranth paste, Vop. Pitan., 27, 46-50 Bär, F. & Griepentrog, F. (1960) Die Allergenwirkung von Fremden Stoffen in den Lebensmitteln, Med. U. Ernaehr., 1, 99 Burnett, C. M., Agersborg, H. P. K., jr, Borzelleca, J. F., Eagle, E., Ebert, A. G., Pierce, E. C., Kirschman, J. C. & Scala, R. A. (1974) Teratogenic studies with certified colors in rats, Toxicol. appl. Pharmacol., 29, (1), 121, Abstract 118 Carson, S. (1962) The safety evaluation of FD & C Red 2 by dermal application to rabbits. Unpublished report from the Food and Drug Research Labs, Inc. to the Toilet Goods Association, Inc. submitted to the World Health Organization by the United States Food and Drug Administration Carson, S. (1963) Skin painting studies in mice. Unpublished report from the Food and Drug Research Labs, Inc. (Lab. No. 81998) to the Toilet Goods Association, Inc. submitted to the World Health Organization by the United States Food and Drug Administration Carson, S. (1966) Skin painting studies in mice. Unpublished report from the Food and Drug Research Labs, Inc. (Lab. No. 81998 Addendum) to the Toilet Goods Association, Inc. submitted to the World Health Organization by the United States Food and Drug Administration Collins, T. F. X., McLaughlin, J. & Gray, G. C. (1972) Teratology studies on food colourings. Part I. Embryotoxicity of amaranth (FD & C Red No.2) in rats, Fd. Cosmet. Toxicol., 10, 619-624 Collins, T. F. X., McLaughlin, J. & Gray, G. C. (1973) Teratology studies on food colourings. Part II. Embryotoxicity of R salt and metabolites of amaranth (FD & C Red No. 2) in rats, Fd. Cosmet Toxicol., 11, 355-365 Collins, T. F. X., Keeler, H. V., Black, T. N. & Ruggles, D. I. (1975a) Long-term effects of dietary amaranth in rats. I. Effects on reproduction, Toxicology, 3, 115-128 Collins, T. F. X., Black, T. N. & Ruggles, D. I. (1975b) Long-term effects of dietary amaranth in rats. II. Effects on fetal development, Toxicology, 3, 129-140 Cook, J. W., Hewett, C. L., Kennaway, E. L. & Kennaway, N.M. (1940) Effects produced in the livers of mice by azonaphthalenes and related compounds, Amer. J. Cancer, 40, 62-77 Ershoff, B. H. & Thurston, E. W. (1974) Effects of diet on amaranth (FD & C Red No.2). Toxicity in the rat, J. Nutr., 104, (7), 937-942 Galea, V., Ariesan & Luputiu, C. (1962) Modifiari biochimice in ficatul sobolanilor albi sub influenta colorantilor organici de sinteza oranj G.G.N. si amarant, Farmacia (Buc.), 10, 531-533 Galea, V., Preda, N., Popa, L., Sendrea, D. & Simu, G. (1972) Recherches toxicologiques sur le colorant amaranthe, J. europ. Toxicol., 5, 167-173 Graham, R. C. B. & Allmark, M. G. (1959) Screening of some food colors for estrogenic activity, Toxicol. appl. Pharmacol., 1, 144-146 Haley, S., Plank, J. B., Wright, P. L. & Keplinger, M. L. (1972) Teratogenic study with FD and C Red No.2 in albino rats. Unpublished report from Industrial Bio-Test Labs, Inc. submitted to the World Health Organization by the Inter-Industry Color Committee, United States of America Keplinger, M. L., Wright, P. L., Plank, J. B. & Calandra, J. C. (1974) Teratologic studies with FD & C Red No.2 in rats and rabbits, Toxicol. Appl. Pharmacol., 28, 209-215 Khera, K. (1975) Unpublished report submitted to the World Health Organization by the Bureau of Chemical Safety, Government of Canada Khera, K. S., Przybylski, W. & McKinley, W. P. (1974) Implantation and embryonic survival in rats treated with amaranth during gestation, Fd. Cosmet. Toxicol., 12, 507-510 Korinke, J., Taff, D., Smith, R. E. & Keplinger, M. L. (1974) Teratogenic and reproduction study with FD & C Red No.2 in cats. Unpublished report from Industrial Bio-Test Labs, Inc. submitted to the World Health Organization by Pet Food Institute Larsson, K. S. (1975) A teratologic study with the dyes amaranth and ponceau 4R in mice, Toxicology, 75-82 Legator, M. S. (1972) Mutagenicity activity of FD & C Red No.2 in the host-mediated assay. Unpublished memorandum (8 February 1972) submitted to the World Health Organization by the United States Food and Drug Administration Lück, H. & Rickerl, E. (1960) Lebensmittelzusatzstoffe und mutagene Wirkung (Food Additives and mutagenic effect). VI. Report, Z. Lebensmitt.-Untersuch., 112, 157 Manchon, Ph. & Albrecht, R. (1972) Comparaison des effets du Bordeaux S et du Jaune de Beurre chez le rat, Cah. Notr. Diet., 7, 323-325 Mannell, W. A., Grice, H. C., Lu, F. C. & Allmark, M. G. (1958) Chronic toxicity studies on food colours. IV. Observations on the toxicity of tartrazine, amaranth and sunset yellow in rats, J. Pharm. Pharmacol., 10, 625-634 Mastalski, K., Jenkins, D. H., Kinoshita, F. K. & Keplinger, M. L. (1975) Teratogenic and reproduction study with FD & C Red No.2 in beagle dogs. 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See Also: Toxicological Abbreviations Amaranth (WHO Food Additives Series 4) Amaranth (WHO Food Additives Series 13) Amaranth (WHO Food Additives Series 19) AMARANTH (JECFA Evaluation) Amaranth (IARC Summary & Evaluation, Volume 8, 1975)