[4-[4,4'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride

Repeated dose toxicity: dermal

Administrative data

Endpoint:

short-term repeated dose toxicity: dermal

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 peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Teratology and percutaneous toxicity studies on hair dyes

Author:

C. Burnett , E. I. Goldenthal , S. B. Harris , F. X. Wazeter , J. Strausburg , R. Kapp & R. Voelker

Year:

1976

Bibliographic source:

Journal of Toxicology and Environmental Health,1, 1027-1040, 1976

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

Repeated dose dermal toxicity study was performed to determine the dermal toxic nature of Gentian violet using Charles River CD female rats

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

[4-[4,4'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride

EC Number:

208-953-6

EC Name:

[4-[4,4'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride

Cas Number:

548-62-9

Molecular formula:

C25H30ClN3

IUPAC Name:

4-{bis[4-(dimethylamino)phenyl]methylidene}-N,N-dimethylcyclohexa-2,5-dien-1-iminium chloride

Test material form:

solid

Details on test material:

- Name of test material (as cited in study report):Gentian violet
- IUPAC name: N-(4-{bis[4-(dimethylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride
- Molecular formula : C25H30N3.Cl
Molecular weight : 407.986 g/mol
- Smiles notation: C(\c1ccc(N(C)C)cc1)(c1ccc(N(C)C)cc1)=C1\C=C\C(=[N+](/C)C)C=C1.[ClH-]
- InChl:1S/C25H30N3.ClH/c1-26(2)22-13-7-19(8-14-22)25(20-9-15-23(16-10-20)27(3)4)21-11-17-24(18-12-21)28(5)6;/h7-18H,1-6H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid

Specific details on test material used for the study:

- Name of test material : Basic violet 3
- Molecular formula : C25H30ClN3
- Molecular weight : 407.986 g/mol
- Substance type: Organic
- Physical state: No data
- Impurities (identity and concentrations): No data

Test animals

Species:

rat

Strain:

other: Charles River CD

Details on species / strain selection:

No data

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: No data
- Age at study initiation: No data
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: the mated female animals were housed individually in temperature- and
humidity-controlled rooms
- Diet (e.g. ad libitum): Ralston Purina Laboratory Chow ad libitum
- Water (e.g. ad libitum): Water ad libitum
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Controlled environment
- Humidity (%):Controlled environment
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data

IN-LIFE DATES: From: To: No data

Administration / exposure

Type of coverage:

not specified

Vehicle:

not specified

Details on exposure:

TEST SITE
- Area of exposure: The dorso-scapular area
- % coverage: No data
- Type of wrap if used: No data
- Time intervals for shavings or clipplings: The hair at the site of application on the dorso-scapular area was shaved closely the day prior to application

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

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2 mL/kg of the P-23 dye formulation
- Concentration (if solution): 0.002% (2.0 mg/Kg)
- Constant volume or concentration used: No data
- For solids, paste formed: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): No data
- Amount(s) applied (volume or weight with unit): No data
- Concentration (if solution): No data
- Lot/batch no. (if required): No data
- Purity: No data

USE OF RESTRAINERS FOR PREVENTING INGESTION: No data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

Duration of treatment: 19 days
Duration of exposure: 1, 4, 7, 10, 13, 16 and 19 day of gestation

Frequency of treatment:

Once on days 1, 4, 7, 10, 13, 16 and 19 of gestation

Doses / concentrations

Remarks:

0.002% (0.00942 mg/Kg)

No. of animals per sex per dose:

Total: 100
0 mg/Kg: 60 female rats
2.0 mg/Kg: 20 female rats
Positive control: 20 female rats

Control animals:

yes, concurrent vehicle

Details on study design:

No data

Positive control:

Acetylsalicylic acid

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data
- Time schedule: No data
- Cage side observations checked in table [No.?] were included. No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data

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

BODY WEIGHT: Yes
- Time schedule for examinations: 1, 4, 7, 10, 13, 16 and 19 day of gestation

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: 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: 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:
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data - Parameters checked in table [No.?] were examined. No data

CLINICAL CHEMISTRY:
- 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: No data sensory activity / grip strength / motor activity / other: No data

OTHER: No data

Sacrifice and pathology:

No data

Other examinations:

No data

Statistics:

All statistical analyses compared the treatment groups with the control groups. Statistically significant differences between groups were judged valid only when there were significant differences between any one of the dye treated groups and each of the three untreated control groups.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Dermal irritation:

no effects observed

Mortality:

not specified

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

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

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Details on results:

Clinical signs and mortality
Clinical signs: No signs of toxicity were noted during the study period
Mortality: No data

Dermal irritation: No irritation or other changes in appearance were noted except for changes in skin and hair color at the site of topical application of the dye formulation

Body weight and weight gain: Changes in female body weights were similar for rats in the untreated controls and all dye-treated groups at 0.00942 mg/Kg.

Food consumption and compound intake: Mean food consumption for all groups throughout gestation was similar

Food efficiency: No data

Water consumption and compound intake: No data

Opthalmoscopic examination: No data

Haematology: No data

Clinical chemistry: No data

Urinanalysis: No data

Neurobehaviour: No data

Organ weights: No data

Gross pathology: No data

Histopathology: No data

Effect levels

Dose descriptor:

NOAEL

Effect level:

0.009 other: mg/Kg

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No evidence of compound-induced toxicity was observed

Remarks on result:

other: No adverse effects were observed

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The No Observed Adverse Effect Level (NOAEL) for gentian violet in female Charles River CD rats is considered to be 0.00942 mg/Kg.

Executive summary:

Repeated dose dermal toxicity study was performed to determine the dermal toxic nature of Gentian violet. The study was performed using female Charles River CD rats. 2 mL/Kg dye formulation P-23 containing 0.002% ( 0.00942 mg/Kg) test compound was applied to the dorso-scapular area. The application was made during the gestation days 1, 4, 7, 10, 13, 16 and 19. The animals were observed for clinical signs, body weight changes, dermal irritation if any and food consumption. No dye formulation related toxicity was noted. Changes in female body weights and food consumption were similar for rats in the untreated controls and all dye-treated groups. No irritation or other changes in appearance were noted except for changes in skin and hair color at the site of topical application of the dye formulation. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for gentian violet in female Charles River CD rats is considered to be 0.00942 mg/Kg.