Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Effects on fertility

Description of key information

Reproductive toxicity study

According to the data available for the test chemicals, No Observed Adverse Effect Level (NOAEL) was considered to be in range between 800-1000mg/kg bw as No effects on reproductive parameters were observed .When male and female rats or mice were treated with the test chemical.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data of read across substances
Justification for type of information:
Data for the target chemical is summarized based on the structurally similar read across chemicals
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
equivalent or similar to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on two reproductive toxicity studies via oral and dermal route on rats and mice
1. Reproductive and developmental toxicity study of test chemical was performed on Sprague-Dawley CD rats.
2. Reproductive toxicity study of test chemical was performed on male and female B6C3F mice.
3. Combined Repeated Dose and Reproductive/Developmental Toxicity study of test chemical by Oral Administration in Rats
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available
Species:
other: 1. rat 2. mouse 3. rat
Strain:
other: 1. Sprague-Dawley 2.B6C3F1 3.Crj: CD(SD)
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals or test system and environmental conditions:
Study 2.
Details on test animals and env. conditions
TEST ANIMALS
- Source: Taconic Farms (Germantown, NY)
- Age at study initiation: 7 weeks
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g
- Fasting period before study:
- Housing: housed individually, Polycarbonate (Lab Products, Inc., Maywood, NJ), changed weekly
and rotated every 2 weeks
bedding :Sani-Chip® heat-treated hardwood chips (P.J. Murphy Forest Products Corp., Montville, NJ), changed weekly
Rack: Stainless steel drawer-type (Lab Products, Inc., Maywood, NJ),changed and rotated every 2 weeks
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum):feed ad libitum
- Water (e.g. ad libitum):water ad libitum
- Acclimation period:16 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.1-23.9°C
- Humidity (%):42%-57%
- Air changes (per hr): 10/hour
- Photoperiod (hrs dark / hrs light): 12 hours/day

Study 4: Details on test animals and env. conditions
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: 8 weeks old
- Weight at study initiation: male 312.1 to 363.7 g, female 205.3 -230.8 g).
- Fasting period before study: No Data Available
- Housing: individually housed in a wire mesh floor cage (Nihon Cage ), raised, and allowed to take solid feed (CE - 2, Japan Clea )
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): No Data Available
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period:7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 1 ° C.
- Humidity (%):50 to 65%,
- Air changes (per hr): about 15 times / hour
- Photoperiod (hrs dark / hrs light): a lighting condition set at 12 hours of lighting (7 am to 7 pm)

Route of administration:
other: 1. oral: gavage 2.dermal 3. oral: gavage
Vehicle:
other: 1.arachis oil, DAB 9 2.ethanol 3. corn oil
Details on exposure:
Study1.
Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in arachis oil, DAB 9
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): arachis oil, DAB 9
- Concentration in vehicle: 0, 100, 300 and 1000 mg/kg/day
- Amount of vehicle (if gavage): 5ml/kg
- Lot/batch no. (if required): No data available
- Purity: No data available

Study 2
Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in ethanol
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): Test material dissolved in ethanol
- Concentration in vehicle: 0, 50, 100, 200, 400, or 800 mg/kg bw
- Amount of vehicle (if gavage):
- Lot/batch no. (if required): No data available
- Purity: No data available

Study 3: Details on exposure
PREPARATION OF DOSING SOLUTIONS:
The test chemicalwas prepared by suspending it in corn oil and used as a test sample
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): corn oil
- Concentration in vehicle: 0 ,100, 300 and 1,000 mg/kg bw/day
- Amount of vehicle (if gavage): 5ml/kg bw
- Lot/batch no. (if required): (Lot No. V 6 H 2050, Nacalai Tesque ),
- Purity: No data available
Details on mating procedure:
Study 1. Females were mated at the supplier and received at the testing facility on day 0 of gestation.
Study 4: - M/F ratio per cage:1/1
- Length of cohabitation: at the most 14 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy : until proof of pregnancy (formation of vaginal closing or sperm detection in vagina)
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
No Data Available
Duration of treatment / exposure:
Study 2: 10 days ( Days 6 - 15 of gestation)
Study 3: 14 weeks
Study 4: Males: 42 days
Females: from 14 days prior to mating to day 3 of lactation
Frequency of treatment:
Daily
Details on study schedule:
No data available
Remarks:
Study 2: 0, 100, 300 and 1000 mg/kg/day
Study 3: 0, 50, 100, 200, 400,or 800 mg/kg bw
Study 4: Doses / Concentrations:
0 (vehicle), 100, 300 or 1000 mg/kg/day.
No. of animals per sex per dose:
Study 2: No data available
Study 3: Total :100
0 mg/kg bw : 10 male and 10 female
50 mg/kg bw : 10 male and 10 female
100 mg/kg bw : 10 male and 10 female
200 mg/kg bw : 10 male and 10 female
400 mg/kg bw : 10 male and 10 female
Study 4: Total :104 animals
0 (Vehicle): 13 male and 13 female
100 mg/kg/day: 13 male and 13 female
300 mg/kg/day: 13 male and 13 female
1000 mg/kg/day: 13 male and 13 female
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
Study 2.
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes
DETAILED CLINICAL OBSERVATIONS: Yes
Time schedule: Animals were observed at least twice daily for signs of reaction to treatment and/or symptoms of illness.
BODY WEIGHT: Yes
Time schedule for examinations: Body weights were recorded on day 0, 6, 16 and 20 of gestation.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

Study 3.
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes
DETAILED CLINICAL OBSERVATIONS:
Time schedule: Observed twice dailyBODY WEIGHT: Yes
Time schedule for examinations: the animals
were weighed initially, weekly, and at the end of the
studies.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.
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

Study 4:
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes
DETAILED CLINICAL OBSERVATIONS:
Time schedule: Daily
BODY WEIGHT: Yes
Time schedule for examinations: For all males, measurements were made on 1, 8, and 15 days of administration 1 (administration start date), 8, 15, 22, 29, 36, 42 days and postmenopausal days and females for all cases,
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes For all males, measurements were made on 1, 8, and 15 days of administration (administration start date), 8, 15,22, 29, 36, 42 days and postmenopausal days and females for all cases,
Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
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
Time schedule for examinations
Oestrous cyclicity (parental animals):
Study 2: For 12 consecutive days prior to scheduled terminal sacrifice, the vaginal vaults of the females were moistened with saline, if necessary, and samples of vaginal fluid and cells were stained. Relative numbers of leukocytes, nucleated epithelial cells, and large squamous epithelial cells were determined and used to ascertain estrous cycle stage (i.e., diestrus, proestrus, estrus, and metestrus).
Study 4: Estrous cycle length and pattern not performed.
Sperm parameters (parental animals):
Study 3: Male animals were evaluated for sperm count and motility. The left testis and left epididymis were isolated and weighed. The tail of the epididymis (cauda epididymis) was then removed from the epididymal body (corpus epididymis) and weighed. modified Tyrode’s buffer (mice) was applied to slides and a small incision was made at the distal border of the cauda epididymis. The sperm effluxing from the incision were dispersed in the buffer on the slides, and the numbers of motile and nonmotile spermatozoa were counted for five fields per slide by two observers. Following completion of sperm motility estimates, each left cauda epididymis was placed in buffered saline solution. Caudae were finely minced, and the tissue was incubated in the saline solution and then heat fixed at 65°C. Sperm density was then determined microscopically with the aid of a hemacytometer. To quantify spermatogenesis, the testicular spermatid head count was determined by removing the tunica albuginea and homogenizing the left testis in phosphate-buffered saline containing 10% dimethyl sulfoxide. Homogenization-resistant spermatid nuclei were counted with a hemacytometer.

Study 4: Sperm examination were not performed.
Litter observations:
2. No data available
3. Live fetuses were weighed individually including placenta and examined for external abnormalities
4. Litter size at birth, neonatal death, sex ratio of pups were observed
Postmortem examinations (parental animals):
Study 2.
Postmortem examinations (Parent Animal)
SACRIFICE : Females were sacrificed by on overdose of
ether on day 20 of gestation
GROSS NECROPSY: The uterus was weighed and the fetuses were removed by caesarean section. Corpora
lutea were counted and the number and distribution of intrauterine implantations were classified as live or dead fetuses, late intrauterine deaths (resorptions), or early intrauterine deaths (resorption sites). Intrauterine deaths were classified on the basis of the presence (late) or absence (early) of fetal or decidual tissue in addition to placental tissue.
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.:
macroscopic examination was performed

Study 3.
SACRIFICE : Carbon dioxide asphyxiation
GROSS NECROPSY: yes
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.:
macroscopic examination was performed

Study 4.
Postmortem examinations (Parent Animal)
SACRIFICE : Males were killing under pentobarbital sodium deep anesthesia the next day (day corresponding to 43 days of administration). Females delivered were sacrificed on the fourth day of nursing, females that did not deliver to the 25th day of pregnancy, females who did not copulate were exsanguinated and lethal under pentobarbital sodium anesthesia at the end of the mating period,

GROSS NECROPSY: Yes

HISTOPATHOLOGY / ORGAN WEIGHTS: Yes
Weights of the heart, liver, kidney, thymus, testis and epididymis were measured for all cases ,the testes and epididymis were fixed and stored in Bouin's solution, and other organs and brain, spleen, adrenal gland and bladder were fixed and fixed in 10% formalin. For these high-dose groups and control groups, paraffin sections were prepared according to a conventional method, and hematoxylineosin staining was performed to perform histopathological examination.
Postmortem examinations (offspring):
Study1.
Postmortem examinations (offspring)
SACRIFICE
One half of the fetuses for each litter were fixed in Bouin’s solution to examine the viscera and brain by Wilson’s slicing technique. After examination these tissues were discarded. The remaining fetuses were processed (alizarin red staining), examined for skeletal abnormalities and retained.
Study 2.No data available
Statistics:
Study1.
If normal distribution, Dunnett-Test comparing treated groups to control. The Steel-Test was applied when the data could not be assumed to follow normal distribution. Fisher’s Exact Test for 2 x 2 tables was applied if the variables could be dichotomized without loss of information (Bonferroni-Holm-corrected).

Study 2.Statistical analyses for possible dose-related effects on survival used Cox’s (1972) method for testing two groups for equality and Tarone’s (1975) life table test to identify dose-related trends. All reported P values for the survival analyses are two sided.
Reproductive indices:
No data available
Offspring viability indices:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
2. Compound-related symptoms were observed in all treatment groups as salivation (severe in the 1000 mg/kg/day group) and propulsion of the head
3. no effects observed
4. In males, no abnormalities in general conditions were observed. In females, there was only one change from day 42 of administration (day of delivery) to the one in 100 mg / kg administration group (administration 42 days: piloerection, blood-like fluid discharge from the vaginal opening; administration 43 to 44 days: coat contamination) But it recovered on day 45 of administration. In other females, no abnormalities in the general condition were observed
Dermal irritation (if dermal study):
effects observed, treatment-related
Description (incidence and severity):
3.The primary clinical finding was irritation of the skin at the site of application, which was noted in all males and females administered 400 or 800 mg/kg.
Mortality:
no mortality observed
Description (incidence):
2.No deaths occurred in any dams in the control or treated groups.
3.All mice survived until the end of the study
4.No deaths in male and female was observed.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2.Body weight, body weight gains and corrected body weight gains were comparable across all groups
3.Final mean body weights and body weight gains of dosed males and females were generally similar to those of the vehicle control groups
4.In male, the body weight gain increased from 8 to 15 days in the 100 mg / kg administration group significantly (p <0.01) compared with the control group, However, in the group administered with 300 mg / kg or more, there was no significant difference between the control group and treated group. In females, there was no significant difference in body weight and body weight gain between control group and treated group before breeding, pregnancy period and nursing period.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
4. There was no significant difference between the control group and treated group for males and female at any time of feeding intake.
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:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
4. Atrophy of seminiferous tubules was found in 2 animals in the 1000 mg / kg administration group, one on both sides and one on one side, but both were localized and extremely mildly changed. There were no other abnormalities. Sperm granuloma was observed in one of the control group, and there was no abnormality. There were no abnormalities in the ovaries of the unexpanded and infertile cases.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
3.There were no significant differences between dosed and vehicle control groups in estrous cycle characterization.
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
3.There were no significant differences between dosed and vehicle control groups in male reproductive tissue evaluations.
Reproductive performance:
no effects observed
Description (incidence and severity):
3.Post-implantation loss and total embryonic deaths were statistically significantly increased in all treated groups compared to the control group. These findings were considered incidental because in each group there was one single female with a high incidence of embryonic deaths and the incidence of post weight loss was not dose dependent.
Dose descriptor:
NOAEL
Effect level:
> 800 - <= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
dermal irritation
body weight and weight gain
reproductive function (oestrous cycle)
reproductive function (sperm measures)
Remarks on result:
other: overall no effects on reproductive function
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Clinical signs:
no effects observed
Description (incidence and severity):
4. no effects observed
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
4. No mortality observed. There was no significant difference between the control group and treated group for the number of births, delivery rate, birth rate, fertility rate, and survival rate on 4th day of newborn.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
3.There were no significant differences in the body weights of live fetuses (on a litter or individual basis) between the treated and control groups.

4. There was no significant difference between the control group and the treated group on body weight at 0 and 4 days of nursing.
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
Anogenital distance (AGD):
no effects observed
Description (incidence and severity):
4. no effects observed
Nipple retention in male pups:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
2.There were no external macroscopic findings noted in any fetus that were considered to be an effect of the treatment with the test article. Visceral examinations of the preserved fetuses did not reveal any treatment-related abnormalities. Statistically significant retardation in ossification was observed in the 300 and 1000 mg/kg/day groups compared to the controls. The incidence of two sternebrae unossified was significantly increased in the 300 and 1000 mg/kg/day groups compared to the control group. The incidence of incomplete ossification of the skull bones was also significantly increased in the 1000 mg/kg/day group compared to the control group but was essentially due to two dams, which had a total of 10 incomplete ossified skull bones of the 17 observed for this group. The skeletal retardation effects were considered to be incidental because the values were within the normal range of variation for this strain.
4. No abnormalities were found in all pups examined in either the external observation at day 0 or the autopsy performed at day 4 after birth.
Histopathological findings:
not specified
Other effects:
no effects observed
Description (incidence and severity):
2.The sex ratio of the fetuses was not affected by the treatment with the test substance.
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
viability
mortality
body weight and weight gain
gross pathology
other: not specified
Remarks on result:
other: overall no developmental toxic effects observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified

 Summary of Reproductive Tissue Evaluations for Male Mice in the 14-Week Dermal Study

 

 

Vehicle Control

200 mg/kg

400 mg/kg

800 mg/kg

n

10

10

10

10

Weights (g)

 

 

 

 

Necropsy body wt

36.4±0.9

33.4±0.6**

34.7±0.4

34.4±0.4

L. cauda epididymis

0.0132±0.0006

0.0147±0.0007

0.0149±0.0006

0.0152±0.0006

L. epididymis

0.0427 ±0.0009

0.0401±0.0021

0.0425±0.0016

0.0376 ±0.0025

L. testis

0.1213±0.0018

0.1156±0.0015

0.1189±0.0017

0.1229±0.0022

Spermatid measurements

 

 

 

 

Spermatid heads (107/g testis)

20.17±0.71

21.06±0.53

20.90±0.64

19.53±0.26

Spermatid heads (107/testis)

2.45 ±0.09

2.43±0.06

2.48±0.07

2.40±0.04

Spermatid count

(mean/10-4 mL suspension)

76.43 ±2.83

75.98±1.94

77.58±2.34

74.95± 1.36

Epididymal spermatozoal measurements

 

 

 

 

Motility (%)

69.04 ±4.11

68.68± 1.63

68.51±2.79

63.62±3.45

Concentration

 

 

 

 

(106/g cauda epididymal tissue)

1683± 119

1786 ±91

1390±151

1523± 123

** Significantly different (P#0.01) from the vehicle control group by Dunnett’s test

a Data are presented as mean ± standard error. Differences from the vehicle control group are not significant by Dunnett’s test (tissue weights) or Dunn’s test (spermatid and epididymal spermatozoal measurements).

 

Estrous Cycle Characterization for Female Mice in the 14-Week Dermal Study

 

Vehicle Control

200 mg/kg

400 mg/kg

800 mg/kg

n

10

10

10

10

Necropsy body wt (g)

31.2 ± 0.8

30.7 ± 0.8

30.3 ± 0.4

30.1± 0.5

Estrous cycle length (days)

4.10 ±0.10

4.00 ±0.00

4.30 ±0.13

4.06 ±0.06b

Estrous stages (% of cycle)

 

 

 

 

Diestrus

26.7

30.8

29.2

30.0

Proestrus

22.5

22.5

18.3

19.2

Estrus

26.7

25.0

29.2

28.2

Metestrus

24.2

21.7

23.3

22.5

 

a Necropsy body weight and estrous cycle length data are presented as mean ± standard error. Differences from the vehicle control group are not significant by Dunnett’s test (body weights) or Dunn’s test (estrous cycle length). By multivariate analysis of variance, dosed females did not differ significantly from the vehicle control females in the relative length of time spent in the estrous stages.

b Estrous cycle was longer than 12 days or unclear in 1 of 10 animals.

Conclusions:
No Observed Adverse Effect Level (NOAEL) was considered to be 800-1000 mg/kg bw for reproductive toxicity, when male and female rats and mice were treated with the test chemical by oral and dermal application.
Executive summary:

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of the test chemical. The studies are as mentioned below:

Study 2:

The reproductive and prenatal development toxicity study of test material was performed on female Sprague Dawley CD(SD)rats. The test material dissolved in arachis oil, DAB 9were administered in dose concentration 0, 100, 300 and 1000 mg/kg/day from day 6 through day 15 of gestation by oral gavage route. Dose volume was 5 ml/kg body weight, adjusted for body weighed on day 6 of gestation. Females were mated at the supplier and received at the testing facility on day 0 of gestation. Animals were observed at least twice daily for signs of reaction to treatment and/or symptoms of illness. Body weights were recorded on day 0, 6, 16 and 20 of gestation. Females were sacrificed by on overdose of ether on day 20 of gestation. The uterus was weighed and the fetuses were removed by caesarean section. Corpora lutea were counted and the number and distribution of intrauterine implantations were classified as live or dead foetuses. Live fetuses were weighed individually including placenta and examined for external abnormalities. One half of the fetuses for each litter were fixed in Bouin’s solution to examine the viscera and brain by Wilson’s slicing technique. After examination these tissues were discarded. The remaining fetuses were processed (alizarin red staining), examined for skeletal abnormalities and retained. No deaths occurred in any dams in the control or treated groups. Compound-related symptoms were observed in all treatment groups as salivation (severe in the 1000 mg/kg/day group) and propulsion of the head. Body weight, body weight gains and corrected body weight gains were comparable across all groups. There were no significant macroscopic findings in any of the control or treated animals. Post-implantation loss and total embryonic deaths were statistically significantly increased in all treated groups compared to the control group. These findings were considered incidental because in each group there was one single female with a high incidence of embryonic deaths and the incidence of post weight loss was not dose dependent. The sex ratio of the fetuses was not affected by the treatment with the test substance. There were no significant differences in the body weights of live fetuses (on a litter or individual basis) between the treated and control groups. There were no external macroscopic findings noted in any fetus that were considered to be an effect of the treatment with the test article. Visceral examinations of the preserved fetuses did not reveal any treatment-related abnormalities. Statistically significant retardation in ossification was observed in the 300 and 1000 mg/kg/day groups compared to the controls. The incidence of two sternebrae unossified was significantly increased in the 300 and 1000 mg/kg/day groups compared to the control group. The incidence of incomplete ossification of the skull bones was also significantly increased in the 1000 mg/kg/day group compared to the control group but was essentially due to two dams, which had a total of 10 incomplete ossified skull bones of the 17 observed for this group. The skeletal retardation effects were considered to be incidental because the values were within the normal range of variation for this strain. Hence No Observed Adverse Effect Level (NOAEL) was considered to be 1000mg/kg bw for reproductive and embryo-foetal toxicity. When female Sprague Dawley rats were treated with test material orally.

Study 2:

In toxicology and Carcinogenesis study reproductive organ were evaluated after dermal application of test material for 14 weeks. The test material dissolved in ethanol and applied to shaved skin in dose concentration 0, 50, 100, 200, 400, or 800 mg/kg. Groups of 10 male and 10 female mice received dermal applications daily for 14 weeks. Feed and water were available ad libitum. Rats and mice were housed individually. Clinical findings were recorded daily, and the animals were weighed initially, weekly, and at the end of the studies. At the end of the studies, samples were collected for sperm motility or vaginal cytology from mice in the 0, 200, 400, and 800 mg/kg groups for evaluations. The following sperm motility parameters were evaluated: spermatid heads per gram of testis, spermatid heads per testis, spermatid count, motility, and concentration in cauda epididymis. The left cauda, epididymis, and testis were weighed. Vaginal samples were collected for 12 consecutive days prior to the end of the studies for vaginal cytology evaluations. The length of the estrous cycle and the length of time spent in each stage of the cycle were evaluated. All mice survived until the end of the study. Final mean body weights and body weight gains of dosed males and females were generally similar to those of the vehicle control groups. The primary clinical finding was irritation of the skin at the site of application, which was noted in all males and females administered 400 or 800 mg/kg.. There were no significant differences between dosed and vehicle control groups in male reproductive tissue evaluations or in estrous cycle characterization. Hence No Observed Adverse Effect Level (NOAEL) was considered to be 800mg/kg bw for reproductive toxicity. When male and female mice were treated with test material by dermal application for 14 weeks.

Study 4:

Combined Repeat Dose and Reproductive / Developmental Toxicity Screening Test of test chemical were performed on male and female Sprague Dawley rats. The test chemical was suspended in corn oil in dose concentration 0 ,100, 300 and 1,000 mg/kg bw/day and administered via oral gavage route in dose volume 5 ml/kg bw. Dose selected on the bases of preliminary test by the previously repeated 14-day oral administration.13 male and 13 female each were placed in each group. Administration period for each males, 2 weeks before mating, and 42 days from the end of the mating period to the day before the necropsy for males, and for female 2 weeks before mating and a maximum of 2 weeks mating period (Until mating) and once daily for the entire gestation period and 3 days after nursing (delivery day = nursing 0 day) in mating females. Matings were made with the same sex of the same group living together within the same group for up to two weeks from the evening on 15th day of administration. Mating was confirmed every morning by examining the presence of sperm in the vaginal plug and vaginal smear and females confirmed to be mated were separated from male starting from that day as the 0th day of pregnancy and raised individually. All the animals were observed forClinical signs, body weight and food consumption.No deaths in male and female were observed. In males, no abnormalities in general conditions were observed. In females, there was only one change from day 42 of administration (day of delivery) to the one in 100 mg / kg administration group (administration 42 days: piloerection, blood-like fluid discharge from the vaginal opening; administration 43 to 44 days: coat contamination) But it recovered on day 45 of administration. In other females, no abnormalities in the general condition were observed. In male, the body weight gain increased from 8 to 15 days in the 100 mg / kg administration group significantly (p <0.01) compared with the control group. However, in the group administered with 300 mg / kg or more, there was no significant difference between the control group and treated group. In females, there was no significant difference in body weight and body weight gain between control group and treated group before breeding, pregnancy period and nursing period. There was no significant difference between the control group and treated group for males and female at any time of feeding intake. There was no significant difference between the control group and treated group in the mating rate, conception rate, the number of days taken from the start of cohabitation to mating and the number of estrus periods recurred during that period. An abnormality in labor condition (not treated with placenta) and abnormality in nursing condition (not collecting children, poor protrusion of nipple, poor protrusion of infant, child's fetus) was administered to one of 100 mg / kg administration group, The decrease in body surface temperature) was observed, and all children died on nursing day 2, but no abnormality was observed in other mother animals. For male In the 100 mg / kg administration group, the actual weight and the specific body weight value of the liver increased significantly (p <0.05) compared to the control group, but not the dose-dependent change. Atrophy of seminiferous tubules was found in 2 animals in the 1000 mg / kg administration group, one on both sides and one on one side, but both were localized and extremely mildly changed. There were no other abnormalities. Sperm granuloma was observed in one of the control group, and there was no abnormality. There were no abnormalities in the ovaries of the unexpanded and infertile cases. For female In the 100 mg / kg administration group, the actual weight of the kidney decreased significantly (p <0.05) compared to the control group, but not the dose-dependent change. For other organs in male and female, no significant difference was observed between the control group and treated group. Birth rate was 100% in all administration groups, and no significant difference was observed between gestation period between control group and treated group. No significant difference was observed between the control group and treated group for the number of luteinism, the number of implantation and the implantation rate of pregnant animals.There was no significant difference between the control group and treated group for the number of births, delivery rate, birth rate, fertility rate, and survival rate on 4th day of newborn. In addition, no significant difference was observed between the control group and treated group for sex ratio.There was no significant difference between the control group and thetreated group on body weight at 0 and 4 days of nursing. Births that showed morphological abnormalities were not observed in the external table observation on the day of birth, necropsy of the dead child, and necropsy on the 4th day of nursing. Hence, No Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 1000mg/kg bw ,when male and female Sprague Dawley rats were treated with test chemical orally.

 

Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to bein range between 800-1000mg/kg bw as No effects on reproductive parameters were observed .When male and female rats or mice were treated with the test chemical orally and by dermal application .Thus, comparing this value with the criteria of CLP regulation the test chemical is not likely to classify as reproductive toxicant.

Effect on fertility: via oral route
Endpoint conclusion:
no study available
Dose descriptor:
NOAEL
800 mg/kg bw/day
Study duration:
subchronic
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 study available
Additional information

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of the test chemical. The studies are as mentioned below:

Study 2:

The reproductive and prenatal development toxicity study of test material was performed on female Sprague Dawley CD(SD)rats. The test material dissolved in arachis oil, DAB 9were administered in dose concentration 0, 100, 300 and 1000 mg/kg/day from day 6 through day 15 of gestation by oral gavage route. Dose volume was 5 ml/kg body weight, adjusted for body weighed on day 6 of gestation. Females were mated at the supplier and received at the testing facility on day 0 of gestation. Animals were observed at least twice daily for signs of reaction to treatment and/or symptoms of illness. Body weights were recorded on day 0, 6, 16 and 20 of gestation. Females were sacrificed by on overdose of ether on day 20 of gestation. The uterus was weighed and the fetuses were removed by caesarean section. Corpora lutea were counted and the number and distribution of intrauterine implantations were classified as live or dead foetuses. Live fetuses were weighed individually including placenta and examined for external abnormalities. One half of the fetuses for each litter were fixed in Bouin’s solution to examine the viscera and brain by Wilson’s slicing technique. After examination these tissues were discarded. The remaining fetuses were processed (alizarin red staining), examined for skeletal abnormalities and retained.

 No deaths occurred in any dams in the control or treated groups. Compound-related symptoms were observed in all treatment groups as salivation (severe in the 1000 mg/kg/day group) and propulsion of the head. Body weight, body weight gains and corrected body weight gains were comparable across all groups. There were no significant macroscopic findings in any of the control or treated animals. Post-implantation loss and total embryonic deaths were statistically significantly increased in all treated groups compared to the control group. These findings were considered incidental because in each group there was one single female with a high incidence of embryonic deaths and the incidence of post weight loss was not dose dependent. The sex ratio of the fetuses was not affected by the treatment with the test substance. There were no significant differences in the body weights of live fetuses (on a litter or individual basis) between the treated and control groups. There were no external macroscopic findings noted in any fetus that were considered to be an effect of the treatment with the test article. Visceral examinations of the preserved fetuses did not reveal any treatment-related abnormalities. Statistically significant retardation in ossification was observed in the 300 and 1000 mg/kg/day groups compared to the controls. The incidence of two sternebrae unossified was significantly increased in the 300 and 1000 mg/kg/day groups compared to the control group. The incidence of incomplete ossification of the skull bones was also significantly increased in the 1000 mg/kg/day group compared to the control group but was essentially due to two dams, which had a total of 10 incomplete ossified skull bones of the 17 observed for this group. The skeletal retardation effects were considered to be incidental because the values were within the normal range of variation for this strain. Hence No Observed Adverse Effect Level (NOAEL) was considered to be 1000mg/kg bw for reproductive and embryo-foetal toxicity. When female Sprague Dawley rats were treated with test material orally.

Study 3:

In toxicology and Carcinogenesis study reproductive organ were evaluated after dermal application of test material for 14 weeks. The test material dissolved in ethanol and applied to shaved skin in dose concentration 0, 50, 100, 200, 400, or 800 mg/kg. Groups of 10 male and 10 female mice received dermal applications daily for 14 weeks. Feed and water were available ad libitum. Rats and mice were housed individually. Clinical findings were recorded daily, and the animals were weighed initially, weekly, and at the end of the studies. At the end of the studies, samples were collected for sperm motility or vaginal cytology from mice in the 0, 200, 400, and 800 mg/kg groups for evaluations. The following sperm motility parameters were evaluated: spermatid heads per gram of testis, spermatid heads per testis, spermatid count, motility, and concentration in cauda epididymis. The left cauda, epididymis, and testis were weighed. Vaginal samples were collected for 12 consecutive days prior to the end of the studies for vaginal cytology evaluations. The length of the estrous cycle and the length of time spent in each stage of the cycle were evaluated.

All mice survived until the end of the study. Final mean body weights and body weight gains of dosed males and females were generally similar to those of the vehicle control groups. The primary clinical finding was irritation of the skin at the site of application, which was noted in all males and females administered 400 or 800 mg/kg.. There were no significant differences between dosed and vehicle control groups in male reproductive tissue evaluations or in estrous cycle characterization. Hence No Observed Adverse Effect Level (NOAEL) was considered to be 800mg/kg bw for reproductive toxicity. When male and female mice were treated with test material by dermal application for 14 weeks.

Study 4:

Combined Repeat Dose and Reproductive / Developmental Toxicity Screening Test of test chemical were performed on male and female Sprague Dawley rats. The test chemical was suspended in corn oil in dose concentration 0 ,100, 300 and 1,000 mg/kg bw/day and administered via oral gavage route in dose volume 5 ml/kg bw. Dose selected on the bases of preliminary test by the previously repeated 14-day oral administration.13 male and 13 female each were placed in each group. Administration period for each males, 2 weeks before mating, and 42 days from the end of the mating period to the day before the necropsy for males, and for female 2 weeks before mating and a maximum of 2 weeks mating period (Until mating) and once daily for the entire gestation period and 3 days after nursing (delivery day = nursing 0 day) in mating females. Matings were made with the same sex of the same group living together within the same group for up to two weeks from the evening on 15th day of administration. Mating was confirmed every morning by examining the presence of sperm in the vaginal plug and vaginal smear and females confirmed to be mated were separated from male starting from that day as the 0th day of pregnancy and raised individually. All the animals were observed forClinical signs, body weight and food consumption.No deaths in male and female were observed. In males, no abnormalities in general conditions were observed. In females, there was only one change from day 42 of administration (day of delivery) to the one in 100 mg / kg administration group (administration 42 days: piloerection, blood-like fluid discharge from the vaginal opening; administration 43 to 44 days: coat contamination) But it recovered on day 45 of administration. In other females, no abnormalities in the general condition were observed. In male, the body weight gain increased from 8 to 15 days in the 100 mg / kg administration group significantly (p <0.01) compared with the control group. However, in the group administered with 300 mg / kg or more, there was no significant difference between the control group and treated group. In females, there was no significant difference in body weight and body weight gain between control group and treated group before breeding, pregnancy period and nursing period. There was no significant difference between the control group and treated group for males and female at any time of feeding intake. There was no significant difference between the control group and treated group in the mating rate, conception rate, the number of days taken from the start of cohabitation to mating and the number of estrus periods recurred during that period. An abnormality in labor condition (not treated with placenta) and abnormality in nursing condition (not collecting children, poor protrusion of nipple, poor protrusion of infant, child's fetus) was administered to one of 100 mg / kg administration group, The decrease in body surface temperature) was observed, and all children died on nursing day 2, but no abnormality was observed in other mother animals. For male In the 100 mg / kg administration group, the actual weight and the specific body weight value of the liver increased significantly (p <0.05) compared to the control group, but not the dose-dependent change. Atrophy of seminiferous tubules was found in 2 animals in the 1000 mg / kg administration group, one on both sides and one on one side, but both were localized and extremely mildly changed. There were no other abnormalities. Sperm granuloma was observed in one of the control group, and there was no abnormality. There were no abnormalities in the ovaries of the unexpanded and infertile cases. For female In the 100 mg / kg administration group, the actual weight of the kidney decreased significantly (p <0.05) compared to the control group, but not the dose-dependent change. For other organs in male and female, no significant difference was observed between the control group and treated group. Birth rate was 100% in all administration groups, and no significant difference was observed between gestation period between control group and treated group. No significant difference was observed between the control group and treated group for the number of luteinism, the number of implantation and the implantation rate of pregnant animals.There was no significant difference between the control group and treated group for the number of births, delivery rate, birth rate, fertility rate, and survival rate on 4th day of newborn. In addition, no significant difference was observed between the control group and treated group for sex ratio.There was no significant difference between the control group and thetreated group on body weight at 0 and 4 days of nursing. Births that showed morphological abnormalities were not observed in the external table observation on the day of birth, necropsy of the dead child, and necropsy on the 4th day of nursing. Hence, No Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 1000mg/kg bw ,when male and female Sprague Dawley rats were treated with test chemical orally.

 

Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to bein range between 800-1000mg/kg bw as No effects on reproductive parameters were observed .When male and female rats or mice were treated with the test chemical orally and by dermal application .Thus, comparing this value with the criteria of CLP regulation the test chemical is not likely to classify as reproductive toxicant.

Effects on developmental toxicity

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation12-hydroxy-N-(2-hydroxyethyl)octadecanamide (106-15-0) is not likely to classify as reproductive toxicant.

 

Additional information