Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, NOAEL for test material was considered to be of 616.0mg/kg bw/day for F0, F1 generation. When male and female rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Link to relevant study records

Referenceopen allclose all

Endpoint:
one-generation reproductive toxicity
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 from peer reviewed journal
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Reproductive toxicity study of test material was performed on CFE-S rats
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available
Species:
rat
Strain:
other: CFE-S
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 15 week old
Route of administration:
oral: feed
Vehicle:
not specified
Details on exposure:
Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material mixed with feed .
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0,97.5,616 mg/kg bw/day (0,1950,7800 ppm)
- Amount of vehicle (if gavage): No data available

- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
- M/F ratio per cage:1:1
- Length of cohabitation: No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The presence of sperm during daily vaginal inspections (day 0 of pregnancy). .
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]No data available
- After successful mating each pregnant female was caged (how): individual
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
9 days ( from day 6 through day 15 of gestation )
Frequency of treatment:
Daily
Details on study schedule:
No data available
Remarks:
0, 97.5,616 mg/kg bw/day (0,1950,7800ppm)
No. of animals per sex per dose:
Total:120
0 mg/kg bw/day:20male and 20 female
97.5mg/kg bw/day:20male and 20 female
616mg/kg bw/day:20male and 20 female
Control animals:
yes
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule:


BODY WEIGHT: Yes
Time schedule for examinations: biweekly
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):

Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
Each fetus was weighed, measured, and examined for gross abnormalities
Postmortem examinations (parental animals):
Postmortem examinations (Parent Animal)
SACRIFICE : All females were killed by chloroform inhalation on the 19th day of pregnancy and the fetuses delivered by Caesarian section.

Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]: No data
Maternal animals: yes

GROSS NECROPSY: The number and distribution of fetuses, and the number of corpora lutea, live and
stillboin fetuses, and early and late resorptions were recorded
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.:
macroscopic examination was performed
Postmortem examinations (offspring):
Each fetus was weighed, measured, and examined for gross abnormalities. One-third of each litter was examined for visceral abnormalities by employing the slicing method of Wilson (1965). The remaining two-thirds were cleared and the bone structure stained with alizarin red S in order to define any skeletal abnormalities
Statistics:
Statistical analysis using the 95% confidence level. The methods used included chi square test, analysis of variance and t test, and the Fisher exact probability test (Snedecor, 1962)
Reproductive indices:
No data available
Offspring viability indices:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
The 616 mg/kg /day dose group excreted blue-brown colored urine
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not specified
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
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No dose related significant deference were observed in the average number of implantation sites, live pups, or early and late resorptions per litter, or in the number of females with one or more resorption sites.
Dose descriptor:
NOAEL
Effect level:
616 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
reproductive performance
Remarks on result:
other: overall no toxic effects on reproductive parameters
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not specified
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
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No grossly abnormal pups were noted in the 97.5 mg/kg /day dose group (244 pups); there was one in control group (244 pups). One grossly abnormal pup was noted out of the 262 examined in the 616 mg/kg /day dose group.
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
616 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
gross pathology
other: overall no developmental toxic effects observed
Remarks on result:
other: overall no developmental toxic effects observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
No Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 616 mg/kg/day (7800ppm. When male and female CFE-S rats were treated with test material orally.
Executive summary:

Reproductive and developmental toxicity study of test material was performed on male and female CFE-Srats .20 male and 20 female rats /dose group were used.One male was mated with one female until copulation was confirmed by the presence of sperm during daily vaginal inspections (day 0 of pregnancy).The females then were weighed, transferred into individual cages.The test materialmixed with feed were administers in dose concentration 0, 97.5,616 mg/kg bw/day (0, 1950,7800 ppm)from day from day 6 through day 15. Pregnancy was further confirmed by biweekly weighing of the females. All females were killed by chloroform inhalation on the 19th day of pregnancy and the fetuses delivered by Caesarian section. The number and distribution of fetuses, and the number of corpora lutea, live and stillboin fetuses, and early and late resorptions were recorded. Each fetus was weighed, measured, and examined for gross abnormalities. One-third of each litter was examined for visceral abnormalities by employing the slicing method of Wilson (1965). The remaining two-thirds were cleared and the bone structure stained with alizarin red S in order to define any skeletal abnormalities (Murphy, 1965)

 

No dose-related significant differences were observed in the parameters examined. The 616mg/kg /day (7800 ppm) dose group excreted blue-brown colour urine.No grossly abnormal pups were noted in the 97.5 mg/kg /day dose group (244 pups); there was one in control group (244 pups). One grossly abnormal pup was noted out of the 262 examined in the 616 mg/kg /day dose group.however, the average numbers of implantation sites, live pups, and early or late resorptions were not significantly different among the groups. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 616 mg/kg/day (7800ppm.When male and femaleCFE-Srats were treated with test material orally.

Endpoint:
two-generation reproductive toxicity
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 from peer reviewed journal
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Reproductive and developmental toxicity study of test material was performed on Sprague Dawley rats.
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available
Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals and environmental conditions:
No data available
Route of administration:
dermal
Vehicle:
not specified
Details on exposure:
Details on exposure
The hair on the back of the rats (of both the treated and control groups) was clipped short to ensure maximum skin contact with the applied materials. The test materials, prepared fresh for each application by mixing equal volumes of the dye solution and 6% hydrogen peroxide, were topically applied to the shaved areas, (approximately 1 in. diameter) at a twice-weekly dose of 0.5 ml application. Successive applications were made to adjacent areas in order to minimize skin irritation. During the mating and lactation phase in the reproduction study, the dyes were allowed to dry on the skin before the rats were returned to their cages with their mates or offspring.
Details on mating procedure:
- M/F ratio per cage:
- Length of cohabitation: 15 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The day on which sperm or the copulatory plug was found was termed day 0 of gestation

- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- After successful mating each pregnant female was caged (how):
- Any other deviations from standard protocol:
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
100 days
Frequency of treatment:
daily
Details on study schedule:
No data available
Remarks:
0 and 0.5 ml (0 and 0.001418 mg/kg )
No. of animals per sex per dose:
Total: 480
F0 geneartion:
Control- 1: 40 male, 40 female
Control- 2: 40 male, 40 female
Control- 3: 40 male, 40 female
0.001418 mg/kg : 40 male, 40 female

F1 geneartion
Control- 1: 20 male, 20 female
Control- 2: 20 male, 20 female
Control- 3: 20 male, 20 female
0.001418 mg/kg : 20 male, 20 female
Control animals:
yes
Details on study design:
After the first (F1a) litters had been weaned the number of F0 parents was reduced from 40 to 20 in each group. After a 10-day rest period the remaining animals were re-mated to produce the F1b litters, following the same procedure as described for the reproduction of the F1a litters. After weaning, 60 male and 60 female F1a pups were selected from each group.The remaining F1a pups were killed. The F0 parents were killed once the F1b litters had been weaned. Twenty male and 20 female rats per group were selected from the F1b litters to become the F0 parents Of the next generation After 100 days these rats were mated, avoiding brother/sister pairing, to produce the F2a and F2b litters. After the F2b litters had been weaned, five male and five female F1 parents were necropsied .
Positive control:
No data available
Parental animals: Observations and examinations:
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule:


BODY WEIGHT: Yes
Time schedule for examinations:
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):Yes

Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
On day 21 of lactation the pups were counted, sexed, and weighed individually and observed for signs of pharmacological effects.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]

GROSS NECROPSY: yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGHTS: yes
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY: yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGTHS: yes
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Statistics:
In the reproduction study fertility, and foetal and postnatal survival were compared using the chi-square criterion with the Yates' corrections for 2 x 2 contingency tables and/or Fisher's exact probability test as described by Siegel (1956) to judge the significance of the differences. Foetal and postnatal survival were also compared using the Mann Whitney U test as described by Siegel (1956) and Weil (1970) to judge the significance of the differences. The mean number of live pups per litter and the mean litter weights at days 0, 4 and 14 in the reproduction study, body weights (at wk 13, 27, 40, 53, 66, 79, 92 and 105), and haematological and biochemical parameters (at months 3, 12, 18 and 24)in the toxicity-carcinogenicity study were compared by one-way analysis of variance, Bartlett's test for homogeneity of variances and the appropriate t test (for equal or unequal variances) as described by Steel & Torrie (1960) using Dunnett's multiple comparison tables (Dunnett, 1964) to judge the significance of the differences. The mean body weights of the pups at day 21 (of the reproduction study) were compared by analysis of variance (hierarchical classification) and a t test as described by Steel & Torrie (1960) using Dunnett's multiple comparison tables to judge the significance of the differences.
Reproductive indices:
No data available
Offspring viability indices:
offspring viability indices were observed
Clinical signs:
no effects observed
Description (incidence and severity):
The topical application of hair-dyeing formulations had no effect on the general health of any of the animals in any of the generations tested.
Dermal irritation (if dermal study):
effects observed, non-treatment-related
Description (incidence and severity):
the skin irritation seen in all of the treated groups which consisted of mild dermatitis seen intermittently throughout the treatment period in each generation No pharmacotoxicological signs were observed to be related to the application of the dyes.
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body-weight gains was comparable among the test and control groups in each generation
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
food consumption was comparable among the test and control groups in each generation
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Clinical biochemistry findings:
not specified
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
Fertility, gestation, survival and live birth indices and the mean numbers weaned and mean weaning weights for each litter in each generation were comparable among the test and control groups.
Dose descriptor:
NOAEL
Effect level:
0.001 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
gross pathology
reproductive performance
Remarks on result:
other: No effects on reproductive performance was observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Clinical signs:
no effects observed
Description (incidence and severity):
The topical application of hair-dyeing formulations had no effect on the general health of any of the animals in any of the generations tested.
Dermal irritation (if dermal study):
effects observed, non-treatment-related
Description (incidence and severity):
the skin irritation seen in all of the treated groups which consisted of mild dermatitis seen intermittently throughout the treatment period in each generation No pharmacotoxicological signs were observed to be related to the application of the dyes.
Mortality / viability:
no mortality observed
Description (incidence and severity):
survival was comparable among the test and control groups in each generation
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body-weight gains was comparable among the test and control groups in each generation
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
food consumption was comparable among the test and control groups in each generation
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Clinical biochemistry findings:
not specified
Urinalysis findings:
no effects observed
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
0.001 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
Remarks on result:
other: No toxic effects were observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Clinical signs:
no effects observed
Description (incidence and severity):
The topical application of hair-dyeing formulations had no effect on the general health of any of the animals in any of the generations tested
Dermal irritation (if dermal study):
no effects observed
Description (incidence and severity):
the skin irritation seen in all of the treated groups which consisted of mild dermatitis seen intermittently throughout the treatment period in each generation No pharmacotoxicological signs were observed to be related to the application of the dyes.
Mortality / viability:
no mortality observed
Description (incidence and severity):
survival was comparable among the test and control groups in each generation
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body-weight gains was comparable among the test and control groups in each generation
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
food consumption were comparable among the test and control groups in each generation
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
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
0.001 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
Remarks on result:
other: No toxic effects were observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Mean litter and reproduction data for the F1a litters of rats topically treated with hair-dyeing formulations

Group

No. (%) ofpregnancies*

Mean littersize (range)at birtht

Gestation index‡

Weaningindex§

No. ofpupsborn dead

Average no.weanedll

Mean (range) weights (g)at weaning

 

 

 

 

 

 

 

Male

Female

Control-1

28 (70)

10.8 (5-16)

98

100

6

9.1

49.9 (41-76)

46.2 (35-72)

Control-2

27 (89)

11.1 (1-15)

98

99

7

8.9

46.5 (27-61)

44.5 (28-60)

Control-3

31 (78)

11.5 (6-17)

98

99

8

9.3

48.1 (40-2)

46.7 (37-61)

7405

34 (85)

12.2 (3-18)

 97

99

15

9.2

47.1 (37-80)

47.8 (37-79)

 

*The no. (%) of pregnant rats from 40 matings (or from 39 for control group 2).

ɫ Litter size was reduced to ten rats at day 4 if necessary.

‡Gestation index, no. of newborn pups alive/total no. born (%),

§Weaning index, no. of pups surviving to day 21/no. of pups retained at day 4 (%).

ll The average no. weaned is the total no. weaned at day 21/no. of litters born, including those in which all were born dead.

Mean litter and reproduction data for the F1b litters of rats topically treated with hair-dyeing formulations

Group

No. (%) ofpregnancies*

Mean littersize (range)at birtht

Gestation index‡

Weaningindex§

No. ofpupsborn dead

Average no.weanedll

Mean (range) weights (g)at weaning

 

 

 

 

 

 

 

Male

Female

Control-1

19 (95)

11.1 (3-15)

99

100

3

8.9

54.6

53.3

Control-2

13 (65)

12.6 (7-19)

99

100

2

9.3

55.6

55.9

Control-3

14 (70)

13.5 (8-16)

99

98

2

9.6

54.0

51.0

7405

14 (70)

12.3 (5-18)

93

100

13¶

8.9

52.0

50.7

*The no. (%) of pregnant rats from 20 matings.

 ɫ Litter size was reduced to ten rats at day 4 if necessary.

‡ Gestation index, no. of newborn pups alive/total no. born (%).

§Weaning index, no. of pups surviving to day 21/no. of pups retained at day 4 (%).

ll The average no. weaned is the total no. weaned at day 21/no. of litters born, including those in which all were born dead.

¶One litter born with ten dead pups.

Mean litter and reproduction data for the F2, litters of rats topically treated with hair-dyeing formulations

Group

No. (%) ofpregnancies*

Mean littersize (range)at birtht

Gestation index‡

Weaningindex§

No. ofpupsborn dead

Average no.weanedll

Mean (range) weights (g)at weaning

 

 

 

 

 

 

 

Male

Female

Control-1

17 (85)

11.6 (1-15)

99

98

2

8.9

48.7

46.6

Control-2

18 (80)

11.8 (2-18)

98

99

4

9.2

48.8

45.8

Control-3

16 (84)

12.8 (4-17)

100

100

0

9.4

49.4

48.8

7405

16 (80)

12.1 (9-16)

96

100

8

9.3

49.5

47.6

*No (%) of pregnant rats from 20 matings (or from 19 for control group 3).

ɫLitter size was reduced to ten rats at day 4 if necessary.

‡Gestation index, no. of newborn pups alive/total no. born (%).

§Weaning index, no. of pups surviving to day 21/no. of pups retained at day 4 (%).

ll the average no. weaned is the total no. weaned at day 21/no. of litters born, including those in which all were born dead.

 

Mean litter and reproduction data for the F2b litters of rats topically treated with hair-dyeing formulations

Group

No. (%) ofpregnancies*

Mean littersize (range)at birtht

Gestation index‡

Weaningindex§

No. ofpupsborn dead

Average no.weanedll

Mean (range) weights (g)at weaning

 

 

 

 

 

 

 

Male

Female

Control-1

16 (80)

12.6 (5-18)

99

99

3

9.2

54.5

52.7

Control-2

14 (74)¶

1 12.7 (2-18)

99

100

1

9.3

54.7

54.4

Control-3

14 (74)¶

12.6 (5-20)

95

100

10

9.4

54.6

55.4

7405

15 (79)¶

10.8 (1-15)

98

100

4

8.6

56.7

55.8

*No. (%) of pregnant rats from 20 matings (19¶) matings.

ɫ Litter size was reduced to ten rats at day 4 if necessary.

‡Gestation index, no. of newborn pups alive/total no. born (%).

§Weaning index, no. of pups surviving to day 21/no. of pups retained at day 4 (%).

ll the average no. weaned is the total no. weaned at day 21/no. of litters born, including those in which all were born dead.

Conclusions:
NOAEL was considered to be 0.001418 mg/kg as the frequent topical application of test material does not appear to have an adverse effect on reproductive performance or on the health and survival of the developing foetus and postnatal animal. When male and female rats were treated with test material for two generation .
Executive summary:

The two-generation reproductive and developmental toxicity study of test material was performed on male and female Sprague Dawley rats . The test material in dose concentration  0.001418 mg/kg was applied The hair on the back of the rats (of both the treated and control groups) was clipped short to ensure maximum skin contact with the applied materials. The test materials, prepared fresh for each application by mixing equal volumes of the dye solution and 6% hydrogen peroxide, were topically applied to the shaved areas, (approximately 1 in. diameter) at a twice-weekly dose of 0.05 ml application. Successive applications were made to adjacent areas in order to minimize skin irritation. During the mating and lactation phase in the reproduction study, the dyes were allowed to dry on the skin before the rats were returned to their cages with their mates or offspring. After the first (F1a) litters had been weaned the number of F0 parents was reduced from 40 to 20 in each group. After a 10-day rest period the remaining animals were re-mated to produce the F1b litters, following the same procedure as described for the reproduction of the F1a litters. After weaning, 60 male and 60 female F1a pups were selected from each group.The remaining F1a pups were killed. The F0 parents were killed once the F1b litters had been weaned. Twenty male and 20 female rats per group were selected from the F1b litters to become the F0 parents Of the next generationAfter 100 days these rats were mated, avoiding brother/sister pairing, to produce the F2a and F2b litters. After the F2b litters had been weaned, five male and five female F1 parents were necropsied .The topical application of hair-dyeing formulations had no effect on the general health of anyof the animals in any of the generations tested.Body-weight gains, food consumption and survivalwere comparable among the test and control groups in each generationthe skin irritation seen in all of thetreated groups which consisted of mild dermatitisseen intermittently throughout the treatment period in each generation No pharmacotoxicological signswere observed to be related to the application ofthe dyes. Fertility, gestation, survival and live birthindices and the mean numbers weaned and meanweaning weights for each litter in each generationwere comparable among the test and control groups. Hence NOAEL was considered to be  0.001418 mg/kg as the frequent topical application of test material does not appear to have an adverse effect on reproductive performance or on the health and survival of the developing foetus and postnatal animal. When male and female rats were treated with test material for two generation .

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
616 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from authoritative database
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
0.001 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from authoritative database
Additional information

Reproductive toxicity study

Data available from different studies were reviewed to determine the reproductive toxicity of test chemical.The studies are as mentioned below:

Study 1

Reproductive and developmental toxicity study of test material was performed on male and female CFE-Srats .20 male and 20 female rats /dose group were used.One male was mated with one female until copulation was confirmed by the presence of sperm during daily vaginal inspections (day 0 of pregnancy).The females then were weighed, transferred into individual cages.The test materialmixed with feed were administers in dose concentration 0, 97.5,616 mg/kg bw/day (0, 1950,7800 ppm)from day from day 6 through day 15. Pregnancy was further confirmed by biweekly weighing of the females. All females were killed by chloroform inhalation on the 19th day of pregnancy and the fetuses delivered by Caesarian section. The number and distribution of fetuses, and the number of corpora lutea, live and stillboin fetuses, and early and late resorptions were recorded. Each fetus was weighed, measured, and examined for gross abnormalities. One-third of each litter was examined for visceral abnormalities by employing the slicing method of Wilson (1965). The remaining two-thirds were cleared and the bone structure stained with alizarin red S in order to define any skeletal abnormalities (Murphy, 1965)

No dose-related significant differences were observed in the parameters examined. The 616mg/kg /day (7800 ppm) dose group excreted blue-brown colour urine.No grossly abnormal pups were noted in the 97.5 mg/kg /day dose group (244 pups); there was one in control group (244 pups). One grossly abnormal pup was noted out of the 262 examined in the 616 mg/kg /day dose group.however, the average numbers of implantation sites, live pups, and early or late resorptions were not significantly different among the groups. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 616 mg/kg/day (7800ppm.When male and femaleCFE-Srats were treated with test material orally.

Study 2

In a Teratology study,New Zealand White female rabbits were treated with test material in the concentration of0, 19.5 and 97.5 mg/kg/day orally by gavage fromday 6 to day 18 of gestation. Bluebrown colored urine were observed in all treated rabbits. No adverse effect onbody weight gain, Number of pregnancies, numbers of corpora lutea, implantations, resorptions, and live and stillborn fetuses were observed in treated female rabbits. No effect on fetusesweight were observed in treated female rabbits. In addition, No gross abnormalities, visceral and skeletal abnormalities were observed in fetusesof treated female rabbits. Therefore, NOAEL was considered to be 97.5 mg/kg/day for F0 and F1 geneartion when New Zealand White female rabbits were treated with test material orally by gavage for 13 days.

Study 3

The two-generation reproductive and developmental toxicity study of test material was performed on male and female Sprague Dawley rats . The test material in dose concentration  0.001418 mg/kg was applied The hair on the back of the rats (of both the treated and control groups) was clipped short to ensure maximum skin contact with the applied materials. The test materials, prepared fresh for each application by mixing equal volumes of the dye solution and 6% hydrogen peroxide, were topically applied to the shaved areas, (approximately 1 in. diameter) at a twice-weekly dose of 0.05 ml application. Successive applications were made to adjacent areas in order to minimize skin irritation. During the mating and lactation phase in the reproduction study, the dyes were allowed to dry on the skin before the rats were returned to their cages with their mates or offspring. After the first (F1a) litters had been weaned the number of F0 parents was reduced from 40 to 20 in each group. After a 10-day rest period the remaining animals were re-mated to produce the F1b litters, following the same procedure as described for the reproduction of the F1a litters. After weaning, 60 male and 60 female F1a pups were selected from each group.The remaining F1a pups were killed. The F0 parents were killed once the F1b litters had been weaned. Twenty male and 20 female rats per group were selected from the F1b litters to become the F0 parents Of the next generationAfter 100 days these rats were mated, avoiding brother/sister pairing, to produce the F2a and F2b litters. After the F2b litters had been weaned, five male and five female F1 parents were necropsied .The topical application of hair-dyeing formulations had no effect on the general health of anyof the animals in any of the generations tested.Body-weight gains, food consumption and survivalwere comparable among the test and control groups in each generationthe skin irritation seen in all of thetreated groups which consisted of mild dermatitisseen intermittently throughout the treatment period in each generation No pharmacotoxicological signswere observed to be related to the application ofthe dyes. Fertility, gestation, survival and live birthindices and the mean numbers weaned and meanweaning weights for each litter in each generationwere comparable among the test and control groups. Hence NOAEL was considered to be  0.001418 mg/kg as the frequent topical application of test material does not appear to have an adverse effect on reproductive performance or on the health and survival of the developing foetus and postnatal animal. When male and female rats were treated with test material for two generation .

Study 4

In a Teratology study, pregnent female  CD rats were treated wtih test material in the concentration of 0 and 0.001418 mg/kg dermally in6% hydrogen peroxide on days 1, 4, 7, 10, 13, 16, and 19 of gestation. Acetylsalicylic acid were used as positive control. Changes in the color of the skin and hair at the site of dye application were observed in treated rats. No irritation or other changes were observed in treated rats.No effect on body weight and food consumption of treated rats was observed as compared to control. Similarly,No significant effects on number of corpora lutea, implantation sites, live fetuses, sex ratio, resorption sites or mean resorptions per pregnancy of treated female rats wereobserved as compared to control. In addition, No effect on body weight offetuseswas observedas compared to control. No significant soft-tissue anomalies were observed infetuses of treated female rats as compared to control.Normally occurring accessory ribs variations wereobserved infetuses of treated female rats as compared to control. Therefore, NOAEL was considered to be 0.001416 mg/kg for F0 and F1 geneartion when pregnent female  CD rats were treated wtih test material dermallyon days 1, 4, 7, 10, 13, 16, and 19 of gestation.

 Based on the data available from different studies ,test material did not showedreproductive toxicityat dose concentration 616.0 mg/kg body weight/day,when male and female rats were treated with test material orally,Thus, comparing this value with the criteria ofCLP regulationtest materialis not likely to classify as reproductive toxicant.

 

 

 

 

eproductive toxicity study

Data available from different studies were reviewed to determine the reproductive toxicity of testchemical.The studies are as mentioned below:

Study 1

Reproductive and developmental toxicity study of test material was performed on male and female CFE-Srats .20 male and 20 female rats /dose group were used.One male was mated with one female until copulation was confirmed by the presence of sperm during daily vaginal inspections (day 0 of pregnancy).The females then were weighed, transferred into individual cages.The test materialmixed with feed were administers in dose concentration 0, 97.5,616 mg/kg bw/day (0, 1950,7800 ppm)from day from day 6 through day 15. Pregnancy was further confirmed by biweekly weighing of the females. All females were killed by chloroform inhalation on the 19th day of pregnancy and the fetuses delivered by Caesarian section. The number and distribution of fetuses, and the number of corpora lutea, live and stillboin fetuses, and early and late resorptions were recorded. Each fetus was weighed, measured, and examined for gross abnormalities. One-third of each litter was examined for visceral abnormalities by employing the slicing method of Wilson (1965). The remaining two-thirds were cleared and the bone structure stained with alizarin red S in order to define any skeletal abnormalities (Murphy, 1965)

No dose-related significant differences were observed in the parameters examined. The 616mg/kg /day (7800 ppm) dose group excreted blue-brown colour urine.No grossly abnormal pups were noted in the 97.5 mg/kg /day dose group (244 pups); there was one in control group (244 pups). One grossly abnormal pup was noted out of the 262 examined in the 616 mg/kg /day dose group.however, the average numbers of implantation sites, live pups, and early or late resorptions were not significantly different among the groups. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 616 mg/kg/day (7800ppm.When male and femaleCFE-Srats were treated with test material orally.

Study 2

In a Teratology study,New Zealand White female rabbits were treated with test material in the concentration of0, 19.5 and 97.5 mg/kg/day orally by gavage fromday 6 to day 18 of gestation. Bluebrown colored urine were observed in all treated rabbits. No adverse effect onbody weight gain, Number of pregnancies, numbers of corpora lutea, implantations, resorptions, and live and stillborn fetuses were observed in treated female rabbits. No effect on fetusesweight were observed in treated female rabbits. In addition, No gross abnormalities, visceral and skeletal abnormalities were observed in fetusesof treated female rabbits. Therefore, NOAEL was considered to be 97.5 mg/kg/day for F0 and F1 geneartion when New Zealand White female rabbits were treated with test material orally by gavage for 13 days.

Study 3

The two-generation reproductive and developmental toxicity study of test material was performed on male and female Sprague Dawley rats . The test material in dose concentration  0.001418 mg/kg was applied The hair on the back of the rats (of both the treated and control groups) was clipped short to ensure maximum skin contact with the applied materials. The test materials, prepared fresh for each application by mixing equal volumes of the dye solution and 6% hydrogen peroxide, were topically applied to the shaved areas, (approximately 1 in. diameter) at a twice-weekly dose of 0.05 ml application. Successive applications were made to adjacent areas in order to minimize skin irritation. During the mating and lactation phase in the reproduction study, the dyes were allowed to dry on the skin before the rats were returned to their cages with their mates or offspring. After the first (F1a) litters had been weaned the number of F0 parents was reduced from 40 to 20 in each group. After a 10-day rest period the remaining animals were re-mated to produce the F1b litters, following the same procedure as described for the reproduction of the F1a litters. After weaning, 60 male and 60 female F1a pups were selected from each group.The remaining F1a pups were killed. The F0 parents were killed once the F1b litters had been weaned. Twenty male and 20 female rats per group were selected from the F1b litters to become the F0 parents Of the next generationAfter 100 days these rats were mated, avoiding brother/sister pairing, to produce the F2a and F2b litters. After the F2b litters had been weaned, five male and five female F1 parents were necropsied .The topical application of hair-dyeing formulations had no effect on the general health of anyof the animals in any of the generations tested.Body-weight gains, food consumption and survivalwere comparable among the test and control groups in each generationthe skin irritation seen in all of thetreated groups which consisted of mild dermatitisseen intermittently throughout the treatment period in each generation No pharmacotoxicological signswere observed to be related to the application ofthe dyes. Fertility, gestation, survival and live birthindices and the mean numbers weaned and meanweaning weights for each litter in each generationwere comparable among the test and control groups. Hence NOAEL was considered to be  0.001418 mg/kg as the frequent topical application of test material does not appear to have an adverse effect on reproductive performance or on the health and survival of the developing foetus and postnatal animal. When male and female rats were treated with test material for two generation .

Study 4

In a Teratology study, pregnent female  CD rats were treated wtih test material in the concentration of 0 and 0.001418 mg/kg dermally in6% hydrogen peroxide on days 1, 4, 7, 10, 13, 16, and 19 of gestation. Acetylsalicylic acid were used as positive control. Changes in the color of the skin and hair at the site of dye application were observed in treated rats. No irritation or other changes were observed in treated rats.No effect on body weight and food consumption of treated rats was observed as compared to control. Similarly,No significant effects on number of corpora lutea, implantation sites, live fetuses, sex ratio, resorption sites or mean resorptions per pregnancy of treated female rats wereobserved as compared to control. In addition, No effect on body weight offetuseswas observedas compared to control. No significant soft-tissue anomalies were observed infetuses of treated female rats as compared to control.Normally occurring accessory ribs variations wereobserved infetuses of treated female rats as compared to control. Therefore, NOAEL was considered to be 0.001416 mg/kg for F0 and F1 geneartion when pregnent female  CD rats were treated wtih test material dermallyon days 1, 4, 7, 10, 13, 16, and 19 of gestation.

Study 5

In a Reprodevelopmental toxicity study, CD male rats were treatre wtih test material l in the concentration of 0 and 20mg/kg intraperitoneally in sterile water3 times a week for 8 weeks. Male rats were treated and effect on female rats were observed.No effect on body weight gain of female rats was observed as compared to control.No effect on Implantation and resorption sites, dead and live fetuses and percentage of resorptions per litter of femalerats were observed as compared to control. Therefore, NOAEL was considered to be 20 mg/kg for F0 and F1 geneartion when male  CD rats were treatre wtih test material for 8 weeks.

 Based on the data available from different studies ,test material did not showedreproductive toxicityat dose concentration 616.0 mg/kg body weight/day,when male and female rats were treated with test material orally,Thus, comparing this value with the criteria ofCLP regulationtest materialis not likely to classify as reproductive toxicant.

 

 

 

 

Effects on developmental toxicity

Description of key information

Developmental toxicity study

Based on the various studies available for the test chemical were reviewed to determine the developmental toxicity, NOAELfor test chemical was considered to be 616.0 mg /kg bw/day .When rats were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
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 from peer reviewed journal
Qualifier:
according to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Developmental toxicity study of test material was performed on rabbits.
GLP compliance:
not specified
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
No data available
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 0.5 % aqueous methylcellulose
Details on exposure:
Details on exposure

PREPARATION OF DOSING SOLUTIONS:test material dissolved in 0.5 % aqueous methylcellulose

DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food):No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): 0.5 % aqueous methylcellulose
- Concentration in vehicle:0, 19.5 and 97.5mg/kg/day)
- Amount of vehicle (if gavage):1 ml/kg.
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Female were artificially inseminated and day of insemination was designated day 0 of pregnancy.
Duration of treatment / exposure:
13 days (on days 6 to 18 of gestation)
Frequency of treatment:
Daily
Duration of test:
30 day
Remarks:
0, 19.5 and 97.5mg/kg bw/day
No. of animals per sex per dose:
Total : 48
0 mg/kg bw/ day: 12 female
19.5 mg/kg bw/ day: 12 female
97.5 mg/kg bw/ day: 12 female

With omposite:
97.5 mg/kg bw/ day: 12 female
Control animals:
not specified
Details on study design:
No data available
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations:

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes / No / No data
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data
- Time schedule for examinations:

POST-MORTEM EXAMINATIONS: Yes / No / No data
- Sacrifice on gestation day #
- Organs examined:

OTHER:
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes
- Skeletal examinations: Yes
- Head examinations: No data
Statistics:
Statistical analysis were performed by using the 95 % confidence level. The methods used included square test, analysis of variance and
t test, and the Fisher exact probability test (Snedecor, 1962).
Indices:
No data available
Historical control data:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Bluebrown colored urine were observed in all treated rabbits.
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No adverse effect on body weight gain of treated rabbits were observed as compared to control.
Food consumption and compound intake (if feeding study):
not specified
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
Number of abortions:
not specified
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
No adverse effect onNumber of pregnancies,
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
97.5 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
clinical signs
body weight and weight gain
pre and post implantation loss
total litter losses by resorption
early or late resorptions
dead fetuses
changes in number of pregnant
Remarks on result:
other: No effects on reproductive performance was observed
Abnormalities:
not specified
Localisation:
not specified
Fetal body weight changes:
no effects observed
Description (incidence and severity):
No effect on fetusesweight were observed in treated female rabbits.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
97.5 mg/kg bw/day
Based on:
test mat.
Sex:
not specified
Basis for effect level:
external malformations
skeletal malformations
visceral malformations
Remarks on result:
other: no developmental toxicity was observed
Abnormalities:
not specified
Localisation:
other: not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
NOAEL was considered to be 97.5 mg/kg/day for F0 and F1 geneartion when New Zealand Whitefemale rabbits were treated with test material orally.
Executive summary:

In a Teratology study,New Zealand White female rabbits were treated with test material in the concentration of 0, 19.5 and 97.5 mg/kg/day orally by gavage fromday 6 to day 18 of gestation. Bluebrown colored urine were observed in all treated rabbits.

No adverse effect onbody weight gain, Number of pregnancies, numbers of corpora lutea, implantations, resorptions, and live and stillborn fetuses were observed in treated female rabbits. No effect on fetusesweight were observed in treated female rabbits. In addition, No gross abnormalities, visceral and skeletal abnormalities were observed in fetusesoftreated female rabbits. Therefore, NOAEL was considered to be 97.5 mg/kg/day for F0 and F1 geneartion when New Zealand White female rabbits were treated with test material orally by gavage for 13 days.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data from peer reviewed journal
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Teratology and percutaneous toxicity of test material was performed on rats
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
CD-1
Details on test animals and environmental conditions:
No data available
Route of administration:
dermal
Vehicle:
other: 6% hydrogen peroxide
Details on exposure:
Details on exposure

PREPARATION OF DOSING SOLUTIONS: test material with ammoniacal soap base mixed with an equal volume of 6% hydrogen peroxide just prior to use.

DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food):No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water):No data available
- Concentration in vehicle:0 and 0.4 % (0 and 0.001418 mg/kg)
- Amount of vehicle (if gavage):1 ml/kg (1 ml day formulations which contine 0.4 % of test substance and 1 ml vehicle)
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Pregnant female rats were used.Presence of sperm in the vagina considered day 0 of gestation
Duration of treatment / exposure:
7 days
Frequency of treatment:
Every 3 day (On days 1, 4, 7, 10, 13, 16, and 19 of gestation)
Duration of test:
20 day
Remarks:
0 and 0.4 % (0 and 0.001418 mg/kg)
No. of animals per sex per dose:
Total: 100
Control 1: 20 female
Control 1: 20 female
Control 1: 20 female
0.001418 mg/kg : 20 female

Positive Control : 20 female
Control animals:
yes
Details on study design:
No data available
Maternal examinations:
Clinicl sign, body weight and food consumption were observed.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes / No / No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes
- Skeletal examinations: Yes
- Head examinations: No data
Statistics:
The number of females exhibiting resorption sites, number of females exhibiting two or more resorptions, number of dead or resorbed fetuses, and the number of fetuses with soft-tissue or skeletal anomalies and accessory ribs was compared using chi-square test criterion with Yates' 2x2 contingency tables or Fisher's exact probability test.

The mean number of corpora lutea, implantation sites, live fetuses, and resorption sites was compared by analysis of variance (one-way classification) as described by Steel and Torrie using Dunnett's multiple comparison tables.

The live fetal weights were compared by analysis of variance as described by Steel and Torrie using Dunnett's multiple comparison tables.

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.
Indices:
No data available
Historical control data:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Changes in the color of the skin and hair at the site of dye application were observed in treated rats.
Dermal irritation (if dermal study):
no effects observed
Description (incidence and severity):
No irritation or other changes were observed in treated rats.
Mortality:
not specified
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effect on body weight of treated rats was observed as compared to control.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No effect on food consumption of treated rats was observed as compared to control
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
Number of abortions:
not specified
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
not specified
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified
Changes in number of pregnant:
not specified
Other effects:
not specified
Details on maternal toxic effects:
No significant effect on number of corpora lutea and implantation sites oftreated rats was observed as compared to control.No significant effect on live fetuses, sex ratio, resorption sites or mean resorptions per pregnancy oftreated rats was observed as compared to control.

Dose descriptor:
NOAEL
Effect level:
0.001 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
clinical signs
body weight and weight gain
food consumption and compound intake
pre and post implantation loss
total litter losses by resorption
early or late resorptions
Remarks on result:
other: No effects on reproductive performance was observed
Abnormalities:
not specified
Localisation:
not specified
Fetal body weight changes:
no effects observed
Description (incidence and severity):
No effect on body weight of fetuses was observed in treated female rats as compared to control.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
No effect on Live fetuses was observed in treated female rats as compared to control
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Description (incidence and severity):
No significant soft-tissue anomalies were observed infetuses of treated female rats as compared to control.
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
0.001 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
reduction in number of live offspring
changes in sex ratio
fetal/pup body weight changes
external malformations
skeletal malformations
visceral malformations
Remarks on result:
other: No effects on developmental parameters was observed
Abnormalities:
not specified
Localisation:
other: not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Summary of Teratology Study in Rats Receiving 7405

Observations

Control I

Untreated

Control II

Untreated

Control III

Untreated

Acetylsalicylic

acid (250 mg/kg-day)

7405

(2 ml/kg)

Maternal parameters

 

 

 

 

 

Total no. females

20

20

20

20

20

 Mean no. corpora lutea

15.35

13.55

15.25

16.15d

15.55

Mean no. implantation sites

12.40

12.10c

13.90

13.25

14.30a,b

No. females exhibiting resorption

Sites

13

14

12

15

11

No. females exhibiting 2 or more resorption sites

9

7

6

15d

7

No. females aborting

0

0

0

0

0

Fetal parameters

 

 

 

 

 

Mean no. live fetuses/group

10.65

10.85

12.50

8.70c

12.95

Mean live fetal weight (g)

3.38

3.58

3.62

2.89e

3.799

No. dead or resorbed fetuses (%)

35(14.11)

25(10.33)

28 (10.07)

91 (34.34)e

27 (9.44)

Mean no. resorptions/pregnancy

1.75

1.25

1.40

4.5 5h,i

1.35

Sex ratio, M:F

106:107

102:115

119:131

100:75

117:142

No. fetuses with soft-tissue anomalies (%)

4(6.35)

4(6.06)

6(7.79)

21 (36.84)e

4 (5.00)

No. fetuses with skeletal

anomalies (%)

0(0.00)

1 (0.67)

2(1.16)

40(34.19)e

1 (0.56)

No. fetuses with accessory ribs

only (%)

75 (50.00)

56(37.09)

72 (41.62)

32 (27.35)

72 (40.22)

aSignificantly different from control II at p < 0.05.

bSignificantly different from control III at p < 0.05.

cSignificantly different from control III at p < 0.01.

dSignificantly different from controls II, III atp < 0.05.

eSignificantly different from controls I, I I , III atp < 0.01.

fSignificantly different from control I at p < 0.01.

gSignificantly different from control I atp < 0.05.

^Significantly different from control II atp <0.01.

'Significantly different from controls I, III atp < 0.05.

 

Conclusions:
NOAEL was considered to be 0.001418 mg/kg for F0 and F1 geneartion when pregnent female CD rats were treated wtih test material .
Executive summary:

In a Teratology study, pregnent female  CD rats were treated wtih test material in the concentration of 0 and 0.001418 mg/kg dermally in6% hydrogen peroxide on days 1, 4, 7, 10, 13, 16, and 19 of gestation. Acetylsalicylic acid were used as positive control. Changes in the color of the skin and hair at the site of dye application were observed in treated rats. No irritation or other changes were observed in treated rats.No effects on body weight and food consumption of treated rats were observed as compared to control. Similarly,No significant effects on number of corpora lutea, implantation sites, live fetuses, sex ratio, resorption sites or mean resorptions per pregnancy of treated female rats wereobserved as compared to control. In addition, No effect on body weight offetuseswas observedas compared to control. No significant soft-tissue anomalies were observed infetuses of treated female rats were observed as compared to control. Normally occurring accessory ribs variations wereobserved infetuses of treated female rats as compared to control. Therefore, NOAEL was considered to be 0.001416 mg/kg for F0 and F1 geneartion when pregnent female  CD rats were treated wtih test material dermally on days 1, 4, 7, 10, 13, 16, and 19 of gestation.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
616 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from authoritative database
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
0.001 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from authoritative database
Additional information

Developmental toxicity study

Data available from different studies for test chemicals were reviewed to determine the developmental toxicity of test chemical. The studies are as mentioned below:

Study 1

In a Teratology study,New Zealand White female rabbits were treated with test materialin the concentration of0, 19.5 and 97.5 mg/kg/day orally by gavage fromday 6 to day 18 of gestation. Bluebrown colored urine were observed in all treated rabbits.

No adverse effect onbody weight gain, Number of pregnancies, numbers of corpora lutea, implantations, resorptions, and live and stillborn fetuses were observed in treated female rabbits. No effect on fetusesweight were observed in treated female rabbits. In addition, No gross abnormalities, visceral and skeletal abnormalities were observed in fetusesoftreated female rabbits. Therefore, NOAEL was considered to be 97.5 mg/kg/day for F0 and F1 geneartion when New Zealand White female rabbits were treated with test materialorally by gavage for 13 days.

Study 2

In a Teratology study,CFE-S female rats were treated with test material in the concentration of 0, 195 and 616 mg/kg/day orally in diet fromday 6 to day 15 of gestation. No adverse effect onaverage number of implantation sites, live pups or in the number of females with one or more resorption sites were observed in treated female rats. No effect on fetuses weight were observed in treated female rats. In addition, No were gross abnormalities, visceral and skeletal abnormalities were observed in fetusesoftreated female rats. Therefore, NOAEL was considered to be 616mg/kg/day for F0 and F1 geneartion when CFE-S female rats were treated with test material orally in diet for 10 days.

Study 3

In a Teratology study, pregnent female  CD rats were treated wtih test materialin the concentration of 0 and 0.001418 mg/kg dermally in6% hydrogen peroxide on days 1, 4, 7, 10, 13, 16, and 19 of gestation. Acetylsalicylic acid were used as positive control. Changes in the color of the skin and hair at the site of dye application were observed in treated rats. No irritation or other changes were observed in treated rats.No effects on body weight and food consumption of treated rats were observed as compared to control. Similarly,No significant effects on number of corpora lutea, implantation sites, live fetuses, sex ratio, resorption sites or mean resorptions per pregnancy of treated female rats wereobserved as compared to control. In addition, No effect on body weight offetuseswas observedas compared to control. No significant soft-tissue anomalies were observed infetuses of treated female rats were observed as compared to control. Normally occurring accessory ribs variations wereobserved infetuses of treated female rats as compared to control. Therefore, NOAEL was considered to be 0.001416 mg/kg for F0 and F1 geneartion when pregnent female  CD rats were treated wtih test materialdermallyon days 1, 4, 7, 10, 13, 16, and 19 of gestation.

Thus, based on the data available fortest chemical,No Observed Adverse Effect Level (NOAEL) was considered to be 616mg /kg bw .When rats were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulationtest chemical is not likely to classify as reproductive and developmental toxicant.

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.