Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalenesulphonate

Carcinogenicity

Administrative data

Endpoint:

carcinogenicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Long-Term Toxicity Studies of Carmoisine in Mice

Author:

P. L. Mason, I. F. Gaunt, K. R. Butterworth, Joan Hardy, Ida S. Kiss and P. Grasso

Year:

1974

Bibliographic source:

Fd Cosmet. Toxicol. Vol. 12 pp. 601-607, 1974

Materials and methods

Principles of method if other than guideline:

Carmoisine was tested for its carcinogenic activity using mouse as the test animal

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalenesulphonate

EC Number:

222-657-4

EC Name:

Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalenesulphonate

Cas Number:

3567-69-9

Molecular formula:

C20H12N2Na2O7S2

IUPAC Name:

disodium 4-hydroxy-3-[(4-sulfonato-1-naphthyl)diazenyl]naphthalene-1-sulfonate

Details on test material:

- Name of test material (as cited in study report): Carmoisine
- Molecular formula (if other than submission substance): C20H14N2O7S2.2Na
- Molecular weight (if other than submission substance): 502.4338 g/mol
- Substance type: organic
- Physical state: No data available
Purity No data available
- Impurities (identity and concentrations): Dye content, min 85%; matter volatile at 135°C, max 10%; matter insoluble in water, max 0.1%; matter soluble in diisopropyl ether, max 2.0%;
subsidiary dyes, max 2"0~o; chlorides and sulphates (as sodium salts), max 5%; copper,
max 10 ppm; arsenic, max 1 ppm; lead, max 10 ppm; heavy metals (as sulphides) not producing more intense colour than the reference standard.

Test animals

Species:

mouse

Strain:

other: ASH/CS1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: No data available
- Age at study initiation: No data available
- Weight at study initiation: No data available
- Fasting period before study: No data available
- Housing: The mice were housed in cages of five
- Diet (e.g. ad libitum): Oxoid pasteurized breeding diet supplemented with 80 ppm vitamin K3 ad libitum
- Water (e.g. ad libitum): Water ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 1°C
- Humidity (%): 50-60%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

IN-LIFE DATES: From: To: No data available

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: Feed

Details on exposure:

No data available

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data available

Duration of treatment / exposure:

Duration of exposure: 80 weeks

Frequency of treatment:

Daily

Post exposure period:

No data available

Doses / concentrations

Remarks:

Doses / Concentrations:
0.01, 0.05, 0.25 or 1.25 % (14.3, 71.4, 357.1 or 1785.7 mg/kg bw)
Basis:

No. of animals per sex per dose:

Total: 300
0 mg/kg bw: 60/sex/dose
14.3 mg/kg bw: 30/sex/dose
71.4 mg/kg bw: 30/sex/dose
357.1 mg/kg bw: 30/sex/dose
1785.7 mg/kg bw: 30/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Examinations

Observations and examinations performed and frequency:

Observation and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: under continual surveillance
- Cage side observations checked in table [No.?] were included. any abnormalities in condition
or behaviour

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data

DERMAL IRRITATION (if dermal study): No data - Time schedule for examinations: No data

BODY WEIGHT: Yes
- Time schedule for examinations: weighed every 2 wk up to wk 57 and at the end of the study

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

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:

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: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: ten animals from the control group and those given the two highest dietary levels (0.25 and 1.25%) at wk 12, 24 and 52
- Parameters checked in table [No.?] were examined. The blood was examined for haemoglobin concentration and packed cell volume, and the numbers of erythrocytes and leucocytes were counted. In addition, differential leucocyte and reticulocyte counts were made on the blood samples from the mice fed the control diet and those fed the highest level of carmoisine. At the end of the study the blood samples from all surviving animals were examined for haemoglobin and differential leucocytes only.

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

OTHER: No data

Sacrifice and pathology:

Scarifies And Pathology:
GROSS PATHOLOGY:
Yes, An autopsy was conducted at which any macroscopic abnormalities were noted
HISTOPATHOLOGY:
Yes, the brain, heart, liver, spleen and kidneys were weighed. Samples of these tissues, together with salivary gland, thyroid, adrenal glands, lymph nodes, pancreas, pituitary, ovaries, uterus, urinary bladder, trachea, lungs, oesophagus, stomach, small intestine, colon, rectum, spinal cord and skeletal muscle and any other tissue that appeared to be abnormal, were preserved in 10% buffered formalin. Paraffin-wax sections of these tissues were stained with
haematoxylin and eosin. All tissues from the mice fed control diet and those fed 1.25%
carmoisine were examined microscopically, while at the lower dietary levels examination was confined to the heart, liver and kidney together with any tissue that appeared to be abnormal at autopsy.

Other examinations:

No data available

Statistics:

No data available

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

no effects observed

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:

effects observed, treatment-related

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
Clinical signs: There was external contamination of the fur by the coloured diet. The faeces in the mice given dietary levels of 357.1 or 1785.7 mg/Kg bw were normally coloured when voided, but rapidly (in 1-2 min) darkened to a purple colour on exposure to the air. There was some red colouring of the urine at most dose levels but samples taken with precautions against contamination showed no red colour, although at the 1785.7 mg/Kg bw level there was a slight brownish colour. The ingestion of carmoisine had no effect on either the condition or the behaviour of the animals.

Mortality:
Although deaths occurred in all treatment groups during the study, there was no association between the death rate or the total number of animals dying and the presence of carmoisine in the diet.

BODY WEIGHT AND WEIGHT GAIN carmoisine was not found to have any adverse effect on the rate of body-weight gain

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study) No data available

FOOD EFFICIENCY No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study) No data available

OPHTHALMOSCOPIC EXAMINATION No data available

HAEMATOLOGY In female mice given 1785.7 mg/kg bw, the haemoglobin concentration was significantly lower than the controls after treatment for 12 and 52 wk. In male mice, the same effect was seen at wk 52 with dietary levels of 357.1 or 1785.7 mg/kg bw. In both sexes given 1785.7 mg/kg bw carmoisine for 12 wk, this finding was associated with a lower
reticulocyte count and at wk 52 the packed cell volume was lower in the females. The only
significant difference in the leucocytes was a reduced total count in the females given the
highest dietary level of carmoisine for 24 wk.

CLINICAL CHEMISTRY No data available

URINALYSIS No data available

NEUROBEHAVIOUR No data available

ORGAN WEIGHTS There were no differences in absolute or relative organ weights, between the treated and control mice

GROSS PATHOLOGY At autopsy the contents of the small intestine, but not the caecum or colon, were coloured red in mice given 357.1 or 1785.7 mg/kg bw carmoisine. On exposure to air the contents of the caecum and colon became purple-coloured.

HISTOPATHOLOGY: NON-NEOPLASTIC No data available

HISTOPATHOLOGY: NEOPLASTIC The histopathological changes were those normally encountered in mice of this age and the incidence was similar in all treatment groups within each sex. Pulmonary adenomas were present in both male and female groups in similar numbers. A hepatocellular adenoma and a subcutaneous fibroma occurred in two male control mice and a single granulosa-cell tumour was found in a female control. There were three renal adenomas in male mice, one in each of the groups fed 0 (control), 14.3 and 357.1 mg/Kg bw carmoisine. Several malignant tumours occurred in both male and female mice. Two fibrosarcomas and a mammary adenocarcinoma were found in two male mice fed control diet and one female mouse fed 357.1 mg/Kg bw, respectively. Neoplasms of the reticulo-endothelial system included lymphomas in two male controls and a lymphosarcoma in a male mouse fed 357.1 mg/Kg bw carmoisine. Lymphoblastomas occurred in both male and female mice, with a similar incidence in all groups.

HISTORICAL CONTROL DATA (if applicable) No data available

OTHER FINDINGS No data available

Effect levels

Dose descriptor:

NOAEL

Effect level:

1 785.7 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No carcinogenic potential noted of the test chemical

Remarks on result:

other: Effect type: carcinogenicity (migrated information)

Any other information on results incl. tables

Table: Mortality observed in male and female mice

	14.3 mg/kg bw	71.4 mg/kg bw	357.1 mg/kg bw	1785.7 mg/kg bw
Male	6	3	3	6
Female	5	3	3	7

Table: Mean body weight gain

	14.3 mg/kg bw	71.4 mg/kg bw	357.1 mg/kg bw	1785.7 mg/kg bw
Male	19	18	17	17
Female	18	19	19	15

 

A mean gain of 18 g in controls of both sexes

Applicant's summary and conclusion

Conclusions:

Carmoisine showed no carcinogenic effect when given to mice at dietary levels of up to 1.25% (equivalent to an intake of approximately 1800-2000 mg/kg body weight/day). The No Observed Adverse Effect Level (NOAEL) for the test compound carmoisine is found to be 1.25% (1785.7 mg/Kg bw).

Executive summary:

Carmoisine was tested for its carcinogenic activity using mouse as the test animal.

 

Carmoisine was fed to mice at dietary levels of 0 (control),0.01, 0.05, 0.25 or 1.25 % (14.3, 71.4, 357.1 or 1785.7 mg/kg bw)for 80 wk. There were no adverse effects on mortality, weight gain, organ weights or the incidence ofhistopathological findings, including tumours. There was a mild anaemia in the mice given 1.25%carmoisine. It is concluded that carmoisine has no carcinogenic potential in mice at dietary levelsup to 1.25% (equivalent to an intake of approximately 1800-2000 mg/kg body weight/day).

 

The No Observed Adverse Effect Level (NOAEL) for the test compound carmoisine is found to be 1.25% (1785.7 mg/Kg bw).