Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference 1

Reference Type:

publication

Title:

A chronic toxicity/carcinogenicity study of FD&C Yellow No. 5 (Tartazine) in mice.

Author:

Borzelleca J. and Hallagan J.

Year:

1988

Bibliographic source:

Food and Chemical Toxicology.Vol. 26, No. 3, pp. 189-194, 1988

Reference 2

Reference Type:

other: Authoritative data base

Title:

GCID: 133984

Author:

ACToR database

Year:

2011

Bibliographic source:

ACToR (Aggregated Computational Toxicology Resource):- Borzelleca J. and Hallagan J. (1988b) A chronic toxicity/carcinogenicity study of FD & C Yellow No. 5 (Tartazine) in mice. Fd Chem Toxic 26, 189-194.

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

The chronic study was conducted to evaluate the toxic effects of repeated administration of trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate [FD & C Yellow No. 5 (tartrazine)] to Charles River CD-1 mice by the oral route.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

EC Number:

217-699-5

EC Name:

Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

Cas Number:

1934-21-0

Molecular formula:

C16-H12-N4-O9-S2.3Na

IUPAC Name:

trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

Details on test material:

- Name of test material (as cited in study report):trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate [FD & C Yellow No. 5 (tartrazine)]
- Molecular formula :C16H12N4O9S2.3Na
- Substance type:organic
- Physical state:Soild

Specific details on test material used for the study:

- Name of test material : trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate [FD & C Yellow No. 5 (tartrazine)]
- Molecular formula: C16H12N4O9S2.3Na
- Moleculat weight: 534.3681 g/mol
- Substance type: organic
- Physical state: Solid
- Purity: 90%

Test animals

Species:

mouse

Strain:

CD-1

Details on species / strain selection:

No data

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington, MA
- Age at study initiation: 42 days
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: The mice were housed individually in stainless-steel cages kept in a separate room phase
- Diet (e.g. ad libitum): Purina Rodent Chow , ad libitum
- Water (e.g. ad libitum): N/A
- Acclimation period: N/A

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 2 0-21°C
- Humidity (%): 40--60%
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle.

Administration / exposure

Route of administration:

oral: feed

Details on route of administration:

No data

Vehicle:

other: diet as Purina Rodent Chow

Details on oral exposure:

Fresh diets were prepared and presented weekly at dose levels of 0, 0, 0.5, 1.5 or 5.0% (Actual Dose levels: Males: 714, 2173 or 8103 mg/kg/d; Females: 870, 2662 or 9735 mg/kg/day). The diets were blended in a twin-shell blender, and assays were performed to determine the homogeneity and stability of FD & C Yellow No. 5 in the prepared diets prior to the start of the study. Dietary concentrations of the compound were determined weekly during the first 13 wk of the study, and then monthly thereafter. Analyses of the basic feed for heavy metals, chlorinated hydrocarbons, and aflatoxin were conducted on all lots of feed used during the study.

DIET PREPARATION
- Rate of preparation of diet (frequency): Weekly
- Mixing appropriate amounts with (Type of food): The basal diet (Purina Rodent Chow)
- Storage temperature of food: The basal diet (Purina Rodent Chow) and the experimental diets were stored in an environmentally-controlled room with limited access. The test compound was stored in sealed containers in a locked closet.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Purina Rodent Chow
- Concentration in vehicle: 0, 0, 0.5, 1.5 or 5.0% (Actual Dose levels:Males: 714, 2173 or 8103 mg/kg/d; Females: 870, 2662 or 9735 mg/kg/day)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analyses of the basic feed for heavy metals, chlorinated hydrocarbons and aflatoxin were conducted on all lots of feed used during the study. These analyses demonstrated that the basic feed contained acceptably low levels of contaminants, that the diets were prepared properly, and that the dietary content of the test material was stable.

Duration of treatment / exposure:

104 weeks

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 0, 0.5, 1.5 or 5.0% (Actual Dose levels:Males: 714, 2173 or 8103 mg/kg/d; Females: 870, 2662 or 9735 mg/kg/day)
Basis:
no data

No. of animals per sex per dose:

Total: 600
0 % (control) - 60 male and 60 female mice
0 % (control) - 60 male and 60 female mice
0.5 % - 60 male and 60 female mice
1.5 % - 60 male and 60 female mice
5.0 % - 60 male and 60 female mice

Control animals:

yes, concurrent vehicle

Details on study design:

No data

Positive control:

No data

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily with at least 5 hr between observations.
- Cage side observations checked in table [No.?] were included. Death and mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Twicw weekly for gross signs of toxicity and Detailed physical examinations for signs of toxicity and palpation for masses were conducted weekly.

BODY WEIGHT: Yes
- Time schedule for examinations: weekly for the first 14 wk, bi-weekly for wk 16-26, and monthly thereafter

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, weekly for the first 14 wk, bi-weekly for wk 16-26, and monthly thereafter
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at months 3, 6, 12, 18 and 24
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 10 animals/sex from each group
- Parameters checked in table [No.?] were examined. haemoglobin, haematocrit, erythrocyte and total and differential leucocyte counts, and erythrocyte morphology.

CLINICAL CHEMISTRY: No data
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

IMMUNOLOGY: No data
- Time schedule for examinations: No data
- How many animals: No data
- Dose groups that were examined: No data
- Parameters checked in table [No.?] were examined. No data

OTHER: No data

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, Necropsies were conducted on all animals dying spontaneously, killed in a moribund condition, or killed on schedule. Both absolute and relative organ weights were recorded for brain, gonads, kidneys, liver, spleen and thyroid.

HISTOPATHOLOGY: Yes, complete histological study was conducted on all animals from the two control groups and from the high-dose (5.0%) group.The following tissues were examined histologically for all control and high-dose mice: adrenals (two), aorta (abdominal), bone and marrow (femur), blood smear, brain (three sections: through the frontal cortex and basal ganglia, parietal cortex and thalamus, and cerebellum and pons), oesophagus, eye (two, with optic nerve), gall bladder, heart (with coronary vessels), liver, lung and mainstem bronchi (lungs inflated with formalin), lymph nodes (mesenteric and mediastinal), mammary gland (inguinal), nerve (sciatic), ovaries, pancreas, pituitary, prostate, salivary gland (mandibular), seminal vesicles (two), skeletal muscle (biceps femoris), skin, spinal cord (cervical), spleen, stomach, testes (with epididymides), thymus, thyroid with parathyroid, trachea, urinary bladder (inflated with formalin), uterus, any tissue with gross changes of an uncertain nature (with a section of an area of normal appearance from the same tissue), and any
tissue or masses or suspect tumours with regional lymph nodes.

Other examinations:

No data

Statistics:

The variances of the two control groups were statistically tested for equality by using the the F test. If the variances were equal a standard independent two-sample test was used to determine equality of means. If the variances differed, Welch's t test was used to determine the equality of means,using the Smith-Satterthwaite correction for unequal variances.

Statistical evaluation of the equality of means of control and treated groups was made by the one- way analysis of variance (ANOVA) technique.

Bartlett's test was performed to determine whether groups had equal variance and parametric procedures were used if they did not. The one-way ANOVA was applied using the F distribution to assess significance.

Dunnett's test was used to determine which means were significantly different from the controls.If a non-parametric procedure for testing equality of means was indicated, the Kruskal-Wallis test was used. If differences from the controls were indicated,a summed rank test was used to determine which groups differed significantly.A statistical test for trend among the dose levels was also performed. In the parametric case standard regression techniques were used with a test for trend and lack of fit.In the non-parametric case,Jonckheer's test for monotonic trend was used.

The statistical analysis of tumour incidence data was performed using contingency table techniques.For each comparison, a 2 × 2 table was constructed from the numbers of animals with and without the event of interest in both the control and treated groups.The table was evaluated by the Fisher exact test.
The data on time to neoplastic lesion, and survival data were analyzed for each sex separately by the series of programs included in the National Cancer Institute package for histopathologically proven tumours,time to tumour, and time to death.Total benign neoplastic lesions, total malignant neoplastic lesions and animals bearing benign and/or malignant neoplastic lesions were also analyzed.

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Physical observations noted throughout the study included hair loss (due to friction against the cage), lacrimation, nasal discharge, staining of hair in the anogenital region and soft stools. These observations occurred randomly and in low incidence throughout all groups and were not related to compound administration. Amber or yellow-brown coloured urine was noted at all treatment levels within 1 wk of the start of the study. The faeces of mice fed at dietary levels of 1.5 and 5.0% were purple and yellow-brown, respectively. Yellow hair and skin were noted in all treatment groups.

Mortality:

no mortality observed

Description (incidence):

The survival of the mice was found to be similar for the treated and control groups throughout the study.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Mean body weights of male and female mice in the 5.0% treatment group, and male mice in the 1.5% treatment group were lower than the controls at a number of sampling intervals. These differences, while usually slight, were statistically significant at several intervals. This decrease is not considered toxicologically significant and may be due to the non-nutritive character of FD & C Yellow No. 5.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Mean food consumption was increased in male mice that consumed 5.0% FD & C Yellow No. 5. The differences from control values were slight but statistically significant at several sampling intervals. The mean food consumptions among other treatment groups and controls were similar

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Description (incidence and severity):

No consistent statistically significant differences between control and treated animals were observed for the haematological parameters evaluated

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

A variety of common neoplastic, inflammatory and degenerative lesions were observed macroscopically and histologically. These lesions occurred in similar
incidence among control and treated mice and were not considered related to treatment with the compound. There was no significant difference among groups in the number of tumour-bearing animals

Other effects:

not specified

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

8 103 other: mg/kg/day

Based on:

test mat.

Sex:

male

Basis for effect level:

other: No consistent, significant compound-related adverse effects were noted on clinical signs and mortality,body weight and weight gain,food consumption and compound intake,haematology, urinalysis, gross pathology and histopathology: neoplastic.

2

Dose descriptor:

NOAEL

Effect level:

9 735 other: mg/kg/day

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No consistent, significant compound-related adverse effects were noted on clinical signs and mortality,body weight and weight gain,food consumption and compound intake,haematology, urinalysis, gross pathology and histopathology: neoplastic.

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table 1. Survival and group mean body weights and percentage difference from control values at study termination for mice fed FD & C Yellow No. 5 in the diet

Dose level (%)	Survival	Group body weight (g)	Difference from control
Males
0	30/60	39	+1.3
0	28/60	38	-1.3
0.5	31/60	39	0
1.5	21/60	38	-2.6
5.0	29/60	37	-5.1
Females
0	20/60	33	0
0	34/60	33	0
0.5	18/60	32	-3.0
1.5	24/60	32	-3.0
5.0	33/60	30	-9.1

 

Table 2. Mean food and compound consumption for mice fed FD &C Yellow No. 5 in the diet.

Dose level (%)	Food consumption (mg/mouse/day)		Compound comsumption (mg/kg/day)
	Male	Female	Male	Female
0	5100	5000	-	-
0	5000	4900	-	-
0.5	5000	4800	714	870
1.5	5000	5000	2173	2662
5.0	5500	5200	8103	9735

 

Table 3. Summary of neoplasms in male and female rats fed FD & C Yellow No. 5 in the diet

Observation	Dose level (%)	No. (%) of rats affected
		0	0	0.5	1.5	5.0
Males
Total no. of rats examined		60	60	27*	33*	60
No neoplasms		30 (50)	32 (53)	-	-	34 (56)
Benign neoplasms+		20 (33)	23 (38)	21 (77)	13 (39)	22 (36)
Malignant neoplasms+		11 (18)	16 (26)	6 (22)	22 (66)	20 (33)
Females
Total no. of rats examined		60	60	28*	30*	60
No neoplasms		39 (65)	35 (58)	-	-	28 (46)
Benign neoplasms+		38 (46)	24 (40)	14 (50)	10 (33)	15 (25)
Malignant neoplasms+		16 (26)	18 (30)	15 (53)	20 (66)	15 (25)

*Only grossly detected masses and lesion were examined from animals in the 0.5 and 1.5% groups.

+ Some rats had benign and malignant neoplasms.

Applicant's summary and conclusion

Conclusions:

The no-observed-adverse-effect level established was 5.0% providing an average intake of 8103 and 9735 mg/kg/day for males and females mice, respectively. Lifetime exposure of mice to FD & C Yellow No. 5 (tartrazine) as a dietary admixture at levels up to 5.0% did not demonstrate carcinogenic or toxic effects.

Executive summary:

Chronic toxicity study was conducted to evaluate the toxic effects of repeated administration of trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophe nylazo)pyrazole-3-carboxylate [FD & C Yellow No. 5 (tartrazine)] to Charles River CD-1 mice by the oral route.Charles River CD-1 mice were fed FD & C Yellow No. 5 in the diet at levels of 0.0, 0.0, 0.5,1.5 or 5.0% (Actual Dose levels:Males: 714, 2173 or 8103 mg/kg/d; Females: 870, 2662 or 9735 mg/kg/day) in a long-term toxicity study for a maximum of 104 weeks. Each group consisted of 60 males and 60 females. The animals were observed for death, mortality, clinical signs of toxicity, hemotology, body weight and food consumption and were subjected to groos and histopathology. No consistent, significant compound-related adverse effects were noted. Physical observations noted throughout the study included hair loss (due to friction against the cage), lacrimation, nasal discharge, staining of hair in the anogenital region and soft stools. These observations occurred randomly and in low incidence throughout all groups and were not related to compound administration. Amber or yellow-brown coloured urine was noted at all treatment levels within 1 wk of the start of the study. The faeces of mice fed at dietary levels of 1.5 and 5.0% were purple and yellow-brown, respectively. Yellow hair and skin were noted in all treatment groups. The survival of the mice was found to be similar for the treated and control groups throughout the study. Mean body weights of male and female mice in the 5.0% treatment group, and male mice in the 1.5% treatment group were lower than the controls at a number of sampling intervals. These differences, while usually slight, were statistically significant at several intervals. This decrease is not considered toxicologically significant and may be due to the non-nutritive character of FD & C Yellow No. 5. Mean food consumption was increased in male mice that consumed 5.0% FD & C Yellow No. 5. The differences from control values were slight but statistically significant at several sampling intervals. The mean food consumptions among other treatment groups and controls were similar. No consistent statistically significant differences between control and treated animals were observed for the haematological parameters evaluated. A variety of common neoplastic, inflammatory and degenerative lesions were observed macroscopically and histologically. These lesions occurred in similar incidence among control and treated mice and were not considered related to treatment with the compound. There was no significant difference among groups in the number of tumour-bearing animals. Based on these considerations, the no- observed - adverse effect level (NOAEL) established in this study was 5.0% (8103 mg/kg/day and 9735 mg/kg/day for male and female mice, respectively). Lifetime exposure of mice to FD & C Yellow No.5 as a dietary admixture at levels up to 5.0% did not demonstrate carcinogenic or toxic effects.