Sodium 4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]toluene-3-sulphonate

Repeated dose toxicity: dermal

Administrative data

Endpoint:

sub-chronic toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

May 1965

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

The test substance was used in formulation at low concentration (0.119 and 1.190%) instead of pure recommended by REAch regulation. No information about chemical substance analysis was provided (as batch, purity, stability). Only 10 animals per dose group were used instead of 20 required in Standard method (OECD Guideline 411 method)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1965

Report date:

1965

Materials and methods

Test guideline

Qualifier:

no guideline available

Version / remarks:

The test substance was used in formulation at low concentration (0.119 and 1.190%) instead of pure recommended by REAch regulation. No information about chemical substance analysis was provided (as batch, purity, stability). Only 10 animals per dose group were used instead of 20 required in Standard method (OECD Guideline 411 method)

Principles of method if other than guideline:

USP Hydropholic ointment, alone or containing 0.1 or 1% of External D&C Violet No. 2, was applied to shaved intact skin of male albino rabbit (5 animal per dose test group and 10 in control group). Doses were applied daily, 5 days per week, for a total of 65 applications over 91 days interval. Each dose of 0.5g of the appropriate mixture. All animals were observed for signs of skin irritation or systemic toxicity prior to each daily application of the ointment.

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]toluene-3-sulphonate

EC Number:

224-618-7

EC Name:

Sodium 4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]toluene-3-sulphonate

Cas Number:

4430-18-6

Molecular formula:

C21H15NO6S.Na

IUPAC Name:

sodium 2-[(4-hydroxy-9,10-dioxo-9,10-dihydroanthracen-1-yl)amino]-5-methylbenzenesulfonate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Supplied by John H. Breck, Inc, no more details
- Expiration date of the lot/batch: no information
- Purity test date: Not less than 80%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In tubes, no more details

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test item was used in two formulations containing 0.119% and 1.190% of test item.

Test animals

Species:

rabbit

Strain:

other: Albino

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: local supplier, no more details
- Females (if applicable) nulliparous and non-pregnant: not applicable
- Age at study initiation: not specified
- Weight at study initiation: 2.8 to 3.61 kg
- Fasting period before study: no information
- Housing:Animals were housed individually, no more details
- Diet (e.g. ad libitum): Standard laboratory diet (Hales and Hunter PIONEER rabbit pellets Medicated) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: period of 1-2 weeks

DETAILS OF FOOD AND WATER QUALITY:no information

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 22.2 deg Celsius
- Humidity (%): 40-60%
- Air changes (per hr): air conditionned room, no more details
- Photoperiod (hrs dark / hrs light): 9 hours of light and 15 hours of dark on weekdays, 3 hours of light and 21 hours of dark on weekends

IN-LIFE DATES: From: To: No information

Administration / exposure

Type of coverage:

open

Vehicle:

other: Test item in formulation containing : purified water (35%), white petrolatum (25%), stearyl alcohol (25%), Propylene glycol (12%), sodium lauryl sulfate (1%), propylparaben (0.015%), methylparaben (0.025%)

Details on exposure:

TEST SITE
- Area of exposure: no information
- % coverage: no information
- Type of wrap if used: no wrap used
- Time intervals for shavings or clipplings: the fur was clipped as necessary from time to time during the test period to insure intimate contact of the ointment with the bare skin.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no information
- Time after start of exposure: no information

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5g per animal per day
- Concentration (if solution): 0.119% or 1.190% in formulation
- Constant volume or concentration used: yes constant volume used 0.5g
- For solids, paste formed: yes

VEHICLE
- Justification for use and choice of vehicle (if other than water): in formulation USP Hydrophilic Ointment
- Amount(s) applied (volume or weight with unit): 0.5g of appropriate ointment
- Concentration (if solution): not applicable
- Lot/batch no. (if required): No information
- Purity: no information

USE OF RESTRAINERS FOR PREVENTING INGESTION: No

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

91 Days

Frequency of treatment:

Daily, 5 days per weeks

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0.119 other: % in formulation

Doses / concentrations 2

Dose / conc.:

1.19 other: % in formulation

No. of animals per sex per dose:

5 males per dose group and 10 animals in control group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: no information
- Rationale for animal assignment (if not random): not specified
- Rationale for selecting satellite groups: no satellite group

Examinations

Observations and examinations performed and frequency:

Animal were weighed initially, at weekly intervals thereafter, and on the day prior to sacrifice. The general physical condition of each animal was noted and recorded daily at the same time that the skin was observed for irritation

Sacrifice and pathology:

Liver and kidney weights were recorded at autopsy for all animals receiving applications of the dye and for 5 control rabbits. No other organs were weighed.
Complete autopsies were performed in treated animals and 5 control animals. Portions of liver, kidney and skin from the treated area were fixed in buffered neutral formalin, sectioned, stained with hematoxylin and eosin and examined microscopally. The tissues were examined from all animals receiving doses of the ointment containing 1% of the dye and from 5 control animals.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

Animals showed signs of nasal and ocular discharges.

Dermal irritation:

not examined

Description (incidence and severity):

Skin of control animal appeared slighty pink while skin of treated animals assumed violet coloration. No skin or edema were evident.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

The mean liver weight was significantly lower than control but the absolute organ weights between test and control showed no difference.

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

not examined

Effect levels

Dose descriptor:

NOAEL

Effect level:

0.119 other: % in formulation

Based on:

test mat.

Sex:

male

Basis for effect level:

body weight and weight gain

clinical signs

dermal irritation

gross pathology

organ weights and organ / body weight ratios

Applicant's summary and conclusion

Conclusions:

Under experimental conditions of this study, the registered item External D&C Violet No. 2 showed no adverse effect at 0.119% and 1.190% in formulation. Hence the No Observe Effect Level could be defined as 0.119%.However, this study cannot be used for classification because The test substance was used in formulation at low concentration (0.119 and 1.190%) instead of pure recommended by REACh regulation. No information about chemical substance analysis was provided (as batch, purity, stability). Only 5 animals per dose group were used instead of 20 required in Standard method (OECD Guideline 411 method). The dosing procedure was performed daily only 5 days per week instead of 7 days per week.

Executive summary:

This no-GLP compliant study was performed to assess the potential toxic effect of the test item External D&C Violet no. 2 on albino rabbits dermally exposed during thirteen weeks.

USP Hydropholic ointment, alone or containing 0.1 or 1% of External D&C Violet No. 2, was applied to shaved intact skin of male albino rabbit (5 animal per dose test group and 10 in control group). Doses were applied daily, 5 days per week, for a total of 65 applications over 91 days interval. Each dose of 0.5g of the appropriate mixture. All animals were observed for signs of skin irritation or systemic toxicity prior to each daily application of the ointment.

Under experimental conditions of this study, the registered item External D&C Violet No. 2 showed no adverse effect at 0.119% and 1.190% in formulation. Hence the No Observe Effect Level could be defined as 0.119%.However, this study cannot be used for classification because The test substance was used in formulation at low concentration (0.119 and 1.190%) instead of pure recommended by REACh regulation. No information about chemical substance analysis was provided (as batch, purity, stability). Only 5 animals per dose group were used instead of 20 required in Standard method (OECD Guideline 411 method). The dosing procedure was performed daily only 5 days per week instead of 7 days per week.