Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

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 publication

Data source

Reference

Reference Type:

publication

Title:

Repeated dose toxicity test of test chemical

Author:

J. F. BORZELLECA et. al.

Year:

1991

Bibliographic source:

Fd Chem. Toxic., 1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

Toxicity of test chemical was assessed in HaM/ICR (CD-1) mice in life time study.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

EC Number:

247-368-0

EC Name:

Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

Cas Number:

25956-17-6

Molecular formula:

C18H16N2O8S2.2Na

IUPAC Name:

disodium 6-hydroxy-5-[(2-methoxy-3-methyl-4-sulfonatophenyl)diazenyl]naphthalene-2-sulfonate

Test material form:

solid

Details on test material:

IUPAC name: Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate
Mol. formula: C18H14N2Na2O8S2
Molecular Weight: 496.4266 gm/mol
Smiles: c12c(cc(cc2)S(=O)(=O)[O-])ccc(c1/N=N/c1c(cc(c(c1)C)S(=O)(=O)[O-])OC)O.[Na+].[Na+]
InChI: 1S/C18H16N2O8S2.2Na/c1-10-7-14(16(28-2)9-17(10)30(25,26)27)19-20-18-13-5-4-12(29(22,23)24)8-11(13)3-6-15(18)21;;/h3-9,21H,1-2H3,(H,22,23,24)(H,25,26,27);;/q;2*+1/p-2/b20-19+;;

Test animals

Species:

mouse

Strain:

other: HaM/ICR (CD-1)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: 50 days old
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: Five mice were housed in each polycarbonate box cage. Heat-treated wood chips served as the bedding material.
- Diet (e.g. ad libitum): Purina Laboratory Chow ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: No data

DETAILS OF FOOD AND WATER QUALITY:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-21°C
- Humidity (%): 40-60%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:

oral: feed

Details on route of administration:

Purina Laboratory Chow

Vehicle:

other: Purina Laboratory Chow

Details on oral exposure:

Fresh diets were prepared and presented weekly. The test material was incorporated into the basal diet using a Patterson- Kelley twin shell blender, and assays were performed to determine the homogeneity and stability of test chemical in the prepared diets.

Duration of treatment / exposure:

104 weeks

Frequency of treatment:

1 wk prior to breeding, throughout a 2- or 3-wk breeding period, and during the gestation and lactation periods. When the last litter reached 21 days of age, weanling mice were randomly selected to populate the F1 phase.

Doses / concentrations

Doses / concentrations 1

Remarks:

Males: 0.37, 1.39 or 5.19% (0, 507, 1877, 7422 mg/kg bw/day)

Doses / concentrations 2

Remarks:

Females: 0.37, 1.39 or 5.19% (0, 577, 2043, 8304 mg/kg bw/day)

No. of animals per sex per dose:

50 mice/sex/group

Control animals:

yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly (wk 0-10) and every 4 wk thereafter

BODY WEIGHT: Yes
- Time schedule for examinations: weekly (wk 0-10) biweekly (wk 11-26) and every 4 wk thereafter

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):Yes
- Time schedule for examinations: weekly (wk 0-10) and every 4 wk thereafter

FOOD EFFICIENCY: No data

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

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood collected from the tail vein during the study, and from the
abdominal aorta at termination at wk 13, 26, 52 and termination
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 5 mice/sex/group
- Parameters checked: haematocrit, haemoglobin, erythrocyte count, and total and differential leucocyte counts.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood collected from the tail vein during the study, and from the
abdominal aorta at termination at wk 52 and at termination
- Animals fasted: No data
- How many animals: 5 mice/sex/group
- Parameters checked: aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, blood urea nitrogen, fasting glucose, total protein, serum calcium and total bilirubin.

URINALYSIS: Yes
- Time schedule for collection of urine: Urine samples were obtained by housing mice overnight in metabolism cages (5/cage) at wk 52 and at termination.
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No data
- Parameters checked: specific gravity, pH, glucose, ketones, total protein, bilirubin and sediment.

NEUROBEHAVIOURAL EXAMINATION: No data

OTHER:

Sacrifice and pathology:

Sacrifice and pathology
GROSS PATHOLOGY: Yes
Gross autopsies were conducted on all mice that died spontaneously, were killed in a moribund condition, or were killed at termination. Mice were killed by exsanguination under sodium pentobarbital. Absolute and relative organ weights were determined for the heart, liver, spleen, kidneys, testes with epididymides, thyroid and adrenal glands.

HISTOPATHOLOGY: Yes
Mice killed at 42 wk for histopathological evaluation and complete histology was conducted on all mice from all groups

Statistics:

Gain in group mean body weight and total food consumption, group mean body weight at termination, and absolute and relative organ weights were analysed by the methods of Bartlett (1937), Scheffe (1953) and Snedecor and Cochran (1967a). Survival data were analysed by the methods of Sachs (1959) and Snedecor and Cochran (1967b).

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

There were no compound-related effects on survival.
Localized alopecia, laboured respiration, coloured hair coat, lacrimation and thinness were noted in similar incidences among control and treated mice.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no consistent statistically significant compound-related effects on mean body weights or survival.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption decreased among the mid- and high-dose males for wk 50-104. There were no consistent statistically significant compound-related effects on mean food consumption.

Food efficiency:

not specified

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Description (incidence and severity):

Few of the haematological parameters differed significantly between control and treated mice, and none of the differences was compound related.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

Few of the Clinical chemistry parameters differed significantly between control and treated mice, and none of the differences was compound related.

Urinalysis findings:

no effects observed

Description (incidence and severity):

Few of the Urinanalysis parameters differed significantly between control and treated mice, and none of the differences was compound related.

Behaviour (functional findings):

not examined

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

No significant compound related gross changes, including changes in organ weights, were noted in mice that died on test or were killed during the study or at termination.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No significant compound related gross changes, including changes in organ weights, were noted in mice that died on test or were killed during the study or at termination.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

The mean thyroid weights in the high-dose males and females were 0.0133 and 0.0099 g, respectively, compared with 0.00879 and 0.0078 g in controls (P < 0.05). The respective thyroid/body-weight ratios were 3.51 and 3.23 compared with 2.26 and 2.36 in controls (P ~< 0.05). The significance of these findings
is unclear; there was no histopathology, they were apparently not dose-dependent but were observed at the highest level only and appear to be species-specific. The appearance of lymphocytic lymphoma occurred in study A earlier among treated groups than among controls. Lymphomas were observed in one low-dose male, one mid-dose female and two males and females from the high-dose group between wk 31 and 37. Lymphomas were not observed among controls until wk 85 and 70 for males and females, respectively

Other effects:

not specified

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

7 300 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Remarks on result:

other: No adverse effect observed

2

Dose descriptor:

NOAEL

Effect level:

8 300 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Remarks on result:

other: No adverse effect observed

Target system / organ toxicity

Critical effects observed:

no

System:

other: not specified

Organ:

not specified

Applicant's summary and conclusion

Conclusions:

The NOAEL value of test chemical to Male and female HaM/ICR (CD-1) mice in life time study was observed at dose concentration of 7300 and 8300 mg/kg bw/day respectively.

Executive summary:

Toxicity of test chemical was assessed in HaM/ICR (CD-1) mice in life time study. 50 CD1 mice per sex per dose group were fed test chemical in diet at dose concentration of Males: 0.37, 1.39 or 5.19% (0, 507, 1877, 7422 mg/kg bw/day), Females: 0.37, 1.39 or 5.19% (0, 577, 2043, 8304 mg/kg bw/day), for1 wk prior to breeding, throughout a 2- or 3-wk breeding period, and during the gestation and lactation periods. When the last litter reached 21 days of age, weanling mice were randomly selected to populate the F1 phase. No significant compound related adverse effect were observed on mortality, clinical signs, body weight, food consumption, hematology, clinical chemistry,gross and histopathology. Hence no-observable-adverse effect level in these studies was 5.19%; approximately 7300 and 8300 mg/kg body weight/day for male and female mice, respectively.