Reaction products of diazotised sodium 2-amino-5-{[2-(sulfonatooxy)ethyl]sulfonyl}benzenesulfonate coupled with sodium 2-(4-aminobenzene-1-sulfonyl)ethyl sulfate, subsequently coupled with diazotised sodium 2-(4-aminobenzene-1-sulfonyl)ethyl sulfate

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from publication

Data source

Reference

Reference Type:

publication

Title:

CHRONIC TOXICITY STUDIES ON FOOD COLOURS, PART IV. OBSERVATIONS ON THE TOXICITY OF TARTRAZINE, AMARANTH AND SUNSET YELLOW IN RATS.

Author:

BY W. A. MANNELLH, . C. GRICE,F . C. Lu AND M. G. ALLMARK

Year:

1982

Bibliographic source:

J. Pharm. Pharmacol. 10, 625

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

Chronic toxicity study of food colours TARTRAZINE, AMARANTH AND SUNSET YELLOW were observed in rats.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 6-hydroxy-5-[(4-sulphonatophenyl)azo]naphthalene-2-sulphonate

EC Number:

220-491-7

EC Name:

Disodium 6-hydroxy-5-[(4-sulphonatophenyl)azo]naphthalene-2-sulphonate

Cas Number:

2783-94-0

Molecular formula:

C16H12N2O7S2.2Na

IUPAC Name:

disodium 6-hydroxy-5-[(4-sulfonatophenyl)diazenyl]naphthalene-2-sulfonate

Test material form:

other: solid

Details on test material:

- Name of test material (as cited in study report):SUNSET YELLOW
- Molecular formula (if other than submission substance): C16H12N2O7S2.2Na
- Molecular weight (if other than submission substance): 452.4 g/mole
- Substance type: organic
- Physical state: solid
- Impurities (identity and concentrations):Not available

Test animals

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Not available
- Age at study initiation: 6 and 7 weeks
- Weight at study initiation: 129 g-males and 105 g-females(appx)
- Fasting period before study: Not available
- Housing: Individual cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Not available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not available
- Humidity (%):Not available
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): Not available

Administration / exposure

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: Alphacel

Remarks:

a non-nutritive cellulose

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:Alphacel (a non-nutritive ce!lulose) was added so that the level of dye plus alphacel was 1.5 per cent. The control diet contained 1.5 per cent alphacel.

DIET PREPARATION
- Rate of preparation of diet (frequency):Not available
- Mixing appropriate amounts with (Type of food):Alphacel (a non-nutritive ce!lulose) diet
- Storage temperature of food: Not available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Alphacel(a non nutritive cellulose)-No justification available.
- Concentration in vehicle: 0, appx 15,150 and 750 mg/kg
- Amount of vehicle (if gavage): Not available
- Lot/batch no. (if required): Not available
- Purity: Not available

Details on mating procedure:

Mating was not performed.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

64 weeks

Frequency of treatment:

Daily

Details on study schedule:

not specified

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

15 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

150 mg/kg bw/day (nominal)

Doses / concentrations 4

Dose / conc.:

750 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Total: 120
Control - 15 males and 15 females
15 ma/kg bw - 15 males and 15 females
150 mg/kg bw - 15 males and 15 females
750 mg/kg bw- 15 males and 15 females

Control animals:

other: control diet contained 1.5 per cent alphacel.

Details on study design:

After 26 weeks on test, five rats of each group receiving 1.5 per cent colourwere killed for histological examination. Haemoglobin estimations were done using a slight modification of the
pyridine-haemochromogen method of Rimington. At the end of the experiment, electrocardiograms and electroencephalograms were recorded from six rats of each sex on the control diet and three rats of each sex on the 1.5 per cent level of each colour,

Positive control:

not specified

Examinations

Parental animals: Observations and examinations:

Mortality, Body weight, food consumption and food efficiency were examined.

Oestrous cyclicity (parental animals):

No data

Sperm parameters (parental animals):

No data

Litter observations:

No data

Postmortem examinations (parental animals):

Necroscopies were performed on rats that died during the test. After 26 weeks on test, five rats of each group receiving 1.5 per cent colour were killed for histological examination. Haemoglobin estimations were done(Hb,RBC,WBC,Differential cell counts)
Gross examination-After 64 weeks all surviving rats were anaesthesized with ether on all organs and tissues gross examined .
Organ weights recorded.
Histopathological examination-Tissues with gross pathology change was studied for histopathology.
Tissues examined -Lung, liver, heart, spleen, thyroid, pancreas, stomach, small intestine, kidney, urinary bladder, adrenal, testis, prostate, coagulating gland, ovary, uterus, and thymus.

Postmortem examinations (offspring):

No data

Statistics:

Statistics –significant differences between mean values were determined by Student’s“t”test (forgrowth, food consumption and food efficiency).

Reproductive indices:

No data

Offspring viability indices:

No data

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Description (incidence):

No effect on survival of treated male and female rats were observed at 15, 150 and 750 mg/kg bw as compared to control.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Decrease in mean body weight in females at 4, 32 and 16 weeks.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

Decreased in food consumption of females were observed with no correlation with the concentration given in diet.

Food efficiency:

no effects observed

Description (incidence and severity):

No significant difference in male or female food efficiency was observed as compared to control.

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

The only significant difference observed was lowered white cell count for females at 750 mg/kg bw as compared to controls. No adverse effect on blood cells was observed.

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:

effects observed, non-treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Middle ear and respiratory infections were observed, considered to be in older rats of the colony. Chronic otitis media was observed in nearly 50 per cent of the animals. The disease was evenly distributed in the various groups.
Pathological changes in the adrenal cortex were observed in 22 animals. This particular pathology was acute in nature and was not considered to be an effect of the food colours since it was observed in two of the control animals and since there was no correlation between the incidence and the concentration of colour fed.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

18 per cent tumour incidence was observed in treated rats. The differences in tumour incidence are not significant according to chi-square tests.

Other effects:

no effects observed

Description (incidence and severity):

The tracings of ECG were essentially normal. No significant deviation of electrical axis observed in treated rats.

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

750 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

mortality

body weight and weight gain

food consumption and compound intake

food efficiency

haematology

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

histopathology: neoplastic

other: No significant effect on reproductive organ weight and histopathology of sexual organs testis,ovary and uterus.

Remarks on result:

other: No toxic effect were observed.

Target system / organ toxicity (P0)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Effect levels (F1)

Dose descriptor:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

other: not specified

Target system / organ toxicity (F1)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Overall reproductive toxicity

Reproductive effects observed:

not specified

Treatment related:

not specified

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 750 mg/kg bw when male and female rats were treated with test chemical orally in feed Alphacel (a non-nutritive cellulose) for 64 weeks.

Executive summary:

In a chronic toxicity study, male and female rats were treated with test chemical in the concentration of 0, 15, 150 and 750 mg/kg bw orally in feed Alphacel (a non-nutritive cellulose) for 64 weeks. No effect on survival of treated male and female rats were observed at 15, 150 and 750 mg/kg bw as compared to control. Decrease in mean body weight in females at 4, 32 and 16 weeks. Decreased in food consumption of females were observed with no correlation with the concentration given in diet. No significant difference in male or female food efficiency was observed as compared to control. The only significant difference observed was lowered white cell count for females at 750 mg/kg bw as compared to controls. No adverse effect on blood cells was observed. Similarly, Decrease in liver weight at 150 mg/kg bw and spleen weight at 750 mg/kg bw in female rat. These changes were not correlated with the level of food colour in the diet and are difficult to interpret. No effects on reproductive organs were observed in treated male and female rats as compared to control. Respiratory tract infections accounted for 28 deaths. Two animals died of starvation, one of meningitis, one of a ruptured right auricle and three as the result of neoplasms. In addition, Middle ear and respiratory infections were observed, considered to be in older rats of the colony. Chronic otitis media was observed in nearly 50 per cent of the animals. The disease was evenly distributed in the various groups. Pathological changes in the adrenal cortex were observed in 22 animals. This particular pathology was acute in nature and was not considered to be an effect of the food colours since it was observed in two of the control animals and since there was no correlation between the incidence and the concentration of colour fed. 18 per cent tumour incidence was observed in treated rats. The differences in tumour incidence are not significant according to chi-square tests. The tracings of ECG were essentially normal. No significant deviation of electrical axis observed in treated rats. Therefore, NOAEL was considered to be 750 mg/kg bw when male and female rats were treated with test chemical orally in feed Alphacel (a non-nutritive cellulose) for 64 weeks.