2-[(2-nitrophenyl)amino]ethanol

Carcinogenicity

Administrative data

Endpoint:

carcinogenicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1984

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

This no GLP compliant study was performed with a formulation of different hair dyes including the registered substance. REACh regulation required the use of pure test item. Only one formulation was tested instead of at least 3 different concentrations of the test item as required in the OECD guideline. No certificate of analysis was provided, the batch of the test item, purity, solubility were not reported.

Data source

Reference

Reference Type:

publication

Title:

Evaluation of the Toxicity and Carcinogenicity of Hair Dyes in Swiss Mice

Author:

M.M. Jacobs, C.M. Burnett, A.J. Penicnak, J.A. Herrera, W.E. Morris, P. Shubik, M.
Apaja, and G. Granroth

Year:

1984

Bibliographic source:

19 Drug and Chemical Toxicology, 7: 573-586, 1984

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Swiss mice (8 weeks old), groups of 60 males and 60 females, were painted three times weekly with a hair dye formulation for 20 months. Aliquots of 0.05 ml were delivered to an area of skin (1 cm2) in the interscapular region. The mice were shaved 24 hours before treatment as needed. Two control groups of were shaved only and received no treatments. The oxidative dye solutions were mixed with an equal volume of 6% H2O2 just prior to application. One of the non-oxidative hair dye formulations contained 0.5% HC Yellow n° 2. A gross necropsy was performed on all mice.

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

2-[(2-nitrophenyl)amino]ethanol

EC Number:

225-555-8

EC Name:

2-[(2-nitrophenyl)amino]ethanol

Cas Number:

4926-55-0

Molecular formula:

C8H10N2O3

IUPAC Name:

2-[(2-nitrophenyl)amino]ethanol

Test material form:

not specified

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: supplied from Clairol, no more information
- Expiration date of the lot/batch: not specified
- Purity test date: not specified

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
No information

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
No information

FORM AS APPLIED IN THE TEST (if different from that of starting material) :
Test item HC Yellow No. 2 at 0.5% in formulation with : Lauris Diethanolamine (1.5%), Carbitol (5.0%), Oleic Acid (1.0%), Diethanolamine (2.0%), Hydroxyethyl Cellulose (2.4%), Iriethanolamine Dodecylbenzene Sulfonate (0.5%), PEG-50 Tallow amide (1.9%), Disperse Black 9 (0.5%), H.C. Red 3 (0.3%), Disperse Blue 1 (0.3%), HC Yellow 3 (0.3%), 1-N-(Trishydroxymethyl)-Methylamino-2-Methoxy-4-Hydroxy-Anthraquinone (0.3%), water (q.s. 100)

Test animals

Species:

mouse

Strain:

Swiss

Remarks:

Swiss Webster mice from the Eppley Colony

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Roscoe B. Jackson Memorial Laborator, bred from Chicago Medical school and then at Eppley Institute
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation:six weeks old
- Weight at study initiation: not specified
- Fasting period before study: not specified
- Housing: individually housed in plastic cages with granular cellulose bedding
- Diet (e.g. ad libitum): Wayne Lab-blox pellets ad libitum
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: two weeks

DETAILS OF FOOD AND WATER QUALITY:No details

ENVIRONMENTAL CONDITIONS
No information

IN-LIFE DATES: No information

Administration / exposure

Route of administration:

dermal

Vehicle:

water

Details on exposure:

TEST SITE
- Area of exposure: intercapsular region
- % coverage: not specified
- Type of wrap if used: not specified
- Time intervals for shavings or clipplings: shaved 24 hours before treatment

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no information

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.05 ml
- Concentration (if solution): 0.5% in formulation
- Constant volume or concentration used: yes
- For solids, paste formed: no

VEHICLE
Formulation was used.

USE OF RESTRAINERS FOR PREVENTING INGESTION: Not specified

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

20 months

Frequency of treatment:

three weekly

Post exposure period:

Not specified

Doses / concentrations

Dose / conc.:

0.5 other: % in formulation with other hair dyes

No. of animals per sex per dose:

60 mice were used per sex

Control animals:

yes

yes, concurrent no treatment

Details on study design:

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

Examinations

Observations and examinations performed and frequency:

Mortality, behavior, and physical appearance, especially dermal changes were observed daily. On development, skin lesions were charted weekly and a continuous record of gross appearance was maintened. Diagnosis of benign or malignant tumor was made on histopathologic examination. Mice were weighed weekly.

After nine months of treatment , 10 males and 10 females were randomly selected from each group for clinical tests, hematology and necropsy. Urine samples were analyzed for colorn pH, occult blood, albumin and glucose. The blood samples obtained by cardiac puncture were analized for complete blood counts. Differential white cell counts were also determined. Following the 20 months of exposure, blood samples were taken at the terminal sacrifice from 5 mice per sex for complete blood count and differential count.

Sacrifice and pathology:

A necropsy was performed on all mice found dead or sacrificed in moribund condition and those sacrificed at the termination of the study by carbon dioxide inhalation. Livers and kidneys were weighed (from mice which were sacrificed at the termination fo the exposure period), mean and absolute weights were calculated. Gross description of all pathology were recorded. Tissues were preserved in 10% buffered formalin, embedded in paraffin, and stained with Eosine & Hematoxilin. All tumors and pathological lesions were observed histopathologically.

Tissues selected for microscopic evaluation : adrenals, bone marrow, brain, bronchi, colon, esophagus, eyes, gallbladder, heart; kidneys, liver, lungs, lymphnodes, mammary gland, ovaries, pancreas, parathyroids, pituitary, prostate, salivary gland, skin, small intestine, spinal cord, spleen, stomach, testes, thymus, thyroids, trachea, urinary bladder and uterus.

Statistics:

Chi-square and Fisher Exact Test.

Results and discussion

Results of examinations

Clinical signs:

not specified

Dermal irritation (if dermal study):

no effects observed

Mortality:

mortality observed, non-treatment-related

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:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, non-treatment-related

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Histopathological findings: neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

The predominant tumors diagnosed were liver hemangioma, lung adenoma and malignant lymphoma. However it was considered as non toxicologically relevant. The predominant tumours seen were those that occur commonly in the Eppley Swiss mouse. The incidence was within the range of control values.

Other effects:

not examined

Relevance of carcinogenic effects / potential:

Under the experimental condition of the study, no carcinogenic effects of the hair dyes was clearly indicated.

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

0.5 other: % in formulation with hair dyes

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical biochemistry

clinical signs

haematology

histopathology: neoplastic

histopathology: non-neoplastic

mortality

urinalysis

Any other information on results incl. tables

Table 1 :Summary of the results

	Control 1		Control 2		Test group
	M	F	M	F	M	F
Number of animals	60	60	60	60	60	60
Lung adenoma	13	19	9	19	13	10
Liver Hemangiomas	1	8	1	7	3	7
Malignant Lymphomas	13	6	7	6	16	6

 

Applicant's summary and conclusion

Conclusions:

Under the experimental conditions of the study, the test item HC Yellow No. 2 in formulation with other hair dyes (including Disperse Blue 1 considered as EU Carcinogen Category 2) did not induce unusual tumors or increasing incidence of tumor development. Although the formulation contained Disperse Blue 1, formulation did not led to tumours induction. This non GLP compliant study was performed with a formulation of different hair dyes including the registered substance. REACh regulation required the use of pure test item. Only one formulation was tested instead of at least 3 different concentrations of the test item as required in the OECD guideline. No certificate of analysis was provided, the batch of the test item, purity, solubility were not reported.

Executive summary:

The purpose of this No GLP-compliant study was to assess the potential carcinogenicity effect of hair dyes including the test item by dermal application on Eppley Swiss mice (60 males and 60 females) during 20 months.

Swiss mice (8 weeks old), groups of 60 males and 60 females, were painted three times weekly with a hair dye formulation for 20 months. Aliquots of 0.05 ml were delivered to an area of skin (1 cm2) in the interscapular region. The mice were shaved 24 hours before treatment as needed. Two control groups of were shaved only and received no treatments. The oxidative dye solutions were mixed with an equal volume of 6% H2O2 just prior to application. One of the non-oxidative hair dye formulations contained 0.5% HC Yellow n° 2. A gross necropsy was performed on all mice.

The application of hair dyes did not have an adverse effect on average body weight gains or survival of any group. Body weights were not depressed more than 10% in any group compared to the controls. The predominant tumours seen were those that occur commonly in the Eppley Swiss mouse, namely lung adenomas, liver haemangiomas, and malignant lymphomas. No unusual tumours developed in any of the groups.

Under the experimental conditions of the study, the test item HC Yellow No. 2 in formulation with other hair dyes (including Disperse Blue 1 considered as EU Carcinogen Catgery 2) did not induce unusual tumors or increasing incidence of tumor development. Although the formulation contained Disperse Blue 1, this method did not led to increase of tumours induction incidence compared to control groups. This non GLP compliant study was performed with a formulation of different hair dyes including the registered substance. REACh regulation required the use of pure test item. Only one formulation was tested instead of at least 3 different concentrations of the test item as required in the OECD guideline. No certificate of analysis was provided, the batch of the test item, purity, solubility were not reported.