2-[(2-nitrophenyl)amino]ethanol

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1974

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

This no GLP compliant study used the test item in formulation with other hair dyes instead of pure recommended by REACh regulation. Raw data including food consumption was not provided.

Data source

Reference

Reference Type:

publication

Title:

Chronic Toxicity, Teratologic and Reproduction Studies with Hair Dyes

Author:

T. Wernick, B.M. Lanman, and J.L. Fraux

Year:

1975

Bibliographic source:

Toxicol. Appl. Pharmacol., 32: 450-460

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

A fertility and reproductive performance study was performed on Sprague-Dawley CD rats. Sixty males and 120 females were divided into six groups. Levels of 0, 1950 and 7800 ppm of a hair dye composite material containing 15 hair dye substances which were incorporated in the diet and provided ad libitum. The concentration of HC Yellow n° 2 in the composite material was 0.28%. The average doses were calculated as 0, 86 and 351 mg/kg bw/d which corresponds to doses of 0, 0.24 and 0.98 mg/kg bw/d HC Yellow n° 2. In part I of the study the females received the basal diet from week 8 prior to mating through the weaning of their litters. Males were treated 8 weeks before mating period and during this period. In part II of the study males received the basal diet from week 8 prior to mating and during mating while the females received the test diets from week 8 prior to mating and during gestation and lactation. Uterus was examined for number and distribution of embryos, presence of empty implantation sites, number of embryos undergoing resorption. Necropsy was performed on female which had not deliver litter. Duration of pregnancy was recorded, litters were examined for numbers of of live and stillborn pups and gross abnormalities.

GLP compliance:

no

Limit test:

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: Dye Base composite was derived from Clairol, Inc. No batch was given

FORM AS APPLIED IN THE TEST (if different from that of starting material)
The test item was used at 0.28% in formulation with : Acid Orange 3 (0.24%), H.C. Blue No. 2 (1.63%), Celliton Fast Navy Blue BRA (0.64%), 2-Nitro-p-phenylenediamine (0.24%), 4-Nitro-a-phenylenediamine (0.16%), 2-amino-4-nitrophenol (0.05%), HC Yellow No.5 (0.05%), HC Yellow No. 4 (0.31%), Disperse Violet 11 (0.40%), Disperse Blue 1 (0.61%), Disperse Black 9 (0.13%), HC Blue 1 (1.54%), HC Red No. 3 (0.02%), HC Yellow No. 3 (0.65%), Lauramide DEA (20.22%), Imino-bis propylamine (9.64%), Cellulose ether (17.94%), Citric acid (16.02%), BHT (1.61%), Sodium m-nitrobenzene sulfonate (2.25%), TEA-Dodecylbenzene sulfonate (1.61%), Monoethanolamine (22.42%), Perfume (0.67%), Perfume (0.67%)

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: from Charles River Breeding Laboratories, Inc
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: not specified
- Weight at study initiation: (P) Males: 240-280 g; Females: 180-220 g
- Fasting period before study: no
- Housing: not specified
- Diet (e.g. ad libitum): Basal diet Purina laboratory chow ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: not specified

Administration / exposure

Route of administration:

oral: feed

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency):Prepared twice weekly
- Mixing appropriate amounts with (Type of food): basal diet of Purina laboratory chow
- Storage temperature of food: not specified

Details on mating procedure:

- M/F ratio per cage: 1/2
- Length of cohabitation: 10 days
- Proof of pregnancy: sperm in vaginal smear referred to as[day 0 of pregnancy
- After 10 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): individually in cage

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

Part 1 : 8 weeks before mating and males were treated beofre and during the mating
Part 2 : Females were treated 8 weeks prior mating and during gestation , male were treated with basal chow diet only

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 ppm (nominal)

Doses / concentrations 2

Dose / conc.:

1 950 ppm (nominal)

Doses / concentrations 3

Dose / conc.:

7 800 ppm (nominal)

No. of animals per sex per dose:

10 males and 20 females per conditions for each part of the study.

Control animals:

yes

yes, plain diet

Examinations

Parental animals: Observations and examinations:

Uterus was examined for number and distribution of embryos, presence of empty implantation sites, number of embryos undergoing resorption. Pregnant females, unproductive males, duration of pregnancy were recorded

Postmortem examinations (parental animals):

Necropsy was performed on female which had not deliver litter.

Postmortem examinations (offspring):

litters were examined for numbers of of live and stillborn pups and gross abnormalities.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

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:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Details on results (P0)

There were no dose related significant differences in any of the parameters examined which included male and female fertility, length of gestation, numbers of females with resorption sites, live pups per litter, pup bodyweight, and pup survival. The female fertility index in the high dose group in the Part 1 and the average pup weight in the high dosage group in Part 2 were lower than the control values, but the differences were not statistically significant at the 95% confidence level. At the dietetary concentrations fed, there were no effects on food consumption and body weight gains of either males or females. No abnormal pups were seen upon dissection of embryos after 13 days of gestation or upon gross examination at weaning after 21 days.

Effect levels (P0)

Dose descriptor:

NOEL

Effect level:

7 800 ppm (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

body weight and weight gain

reproductive performance

Any other information on results incl. tables

Table 1 :Summary of results

Concentration in diet(%)		0	0.195	0.78	0	0.195	0.78
		Part 1			Part 2
	Summary of mating performance
mg/kg/day(average)		0	86	351	0	124	554
No. Males		10	10	10	10	10	10
No.Females		20	20	20	20	20	20
No.Pregnancies		17	19	13	19	18	19
Femalefertilityindex		85	95	65	95	90	95
No.Unproductivemales		1	0	2	0	0	0
Malefertilityindex		90	100	80
	Summary of treated male induced pregnancies terminated at the 13th day of gestation
No.Pregantssacrified		8	9	8	9	8	9
Average No. Ofembryos
Lefthorn		5.3	6	5.7	6.2	6.3	7.3
righthorn		6.6	6.1	6.3	6	5.4	5.8
total		11.9	12.1	12	12.2	11.7	13.1
No.Emptyimplant sites		0	0	0	1	0	0
No.Undergoingresorption		7	5	6	3	2	6
No.Abnormalities		0	0	0	0	0	0
No.Litterswithresorption		5	5	4	3	2	4
	Summary of treated male induced pregnancies allowed to litter normally
No.Females		9	10	5	10	10	10
Average length of gestation (days)		21.6	21.7	21.4	21.7	21.4	21.1
Gestation index		100	100	100	100	100	100
Average No. Pups per litter		10.5	11.1	10.8	10.5	11.6	11.6
Percent alive atBirth		94	98	98	95	91	96
Percent alive at 4days		92	98	94	88	78	88
Precentalive at 21Days		89	98	93	87	77	86
Viability index		98	99	96	92	86	92
lactation index		98	100	98	99	98	98
average bodyweight of pups at birth		6.5	6	6.4	6.5	6.3	6.1
average bodyweight of pups at 4 days		10.6	9.3	9.7	10.1	9.6	9.6
average bodyweight of pups at 21 days		46.8	48.1	48.2	47.1	49.3	44.3

 

Applicant's summary and conclusion

Conclusions:

Under the experimental conditions of the study, the test item at 0.28% with other hair dyes did not led to alteration of female or male fertility and reproductive parameters. However, this study cannot be used for classification. This no GLP compliant study used the test item in formulation with other hair dyes instead of pure recommended by REACh regulation. Raw datas including food consumption were not provided.

Executive summary:

The purpose of this no GLP-compliant study was to assess the potential reprotoxic effect of the test item in formulation with other hair dyes in Sprague Dawley rats exposed by food.

A fertility and reproductive performance study was performed on Sprague-Dawley CD rats. Sixty males and 120 females were divided into six groups. Levels of 0, 1950 and 7800 ppm of a hair dye composite material containing 15 hair dye substances which were incorporated in the diet and provided ad libitum. The concentration of HC Yellow n° 2 in the composite material was 0.28%. The average doses were calculated as 0, 86 and 351 mg/kg bw/d which corresponds to doses of 0, 0.24 and 0.98 mg/kg bw/d HC Yellow n° 2. In part I of the study the females received the basal diet from week 8 prior to mating through the weaning of their litters. The males were fed with the test diets from week 8 prior to mating and during the mating period. In part II of the study males received the basal diet from week 8 prior to mating and during mating while the females received the test diets from week 8 prior to mating and during gestation and lactation. Uterus was examined for number and distribution of embryos, presence of empty implantation sites, number of embryos undergoing resorption. Necropsy was performed on female which had not deliver litter. Duration of pregnancy was recorded, litters were examined for numbers of of live and stillborn pups and gross abnormalities.

There were no dose related significant differences in any of the parameters examined which included male and female fertility, length of gestation, numbers of females with resorption sites, live pups per litter, pup bodyweight, and pup survival. The female fertility index in the high dose group in the Part 1 and the average pup weight in the high dosage group in Part 2 were lower than the control values, but the differences were not statistically significant at the 95% confidence level. At the dietetary concentrations fed, there were no effects on food consumption and body weight gains of either males or females. No abnormal pups were seen upon dissection of embryos after 13 days of gestation or upon gross examination at weaning after 21 days.

Under the experimental conditions of the study, the test item at 0.28% with other hair dyes did not led to alteration of female or male fertility and reproductive parameters. However, this study cannot be used for classification. This no GLP compliant study used the test item in formulation with other hair dyes instead of pure recommended by REACh regulation. Raw datas including food consumption were not provided.