4-Ethoxy-4'-(trans-4-ethylcyclohexyl)-2,3-difluor-1,1'-biphenyl

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

For the test item 1000 mg/kg bw/d was considered the parental NOAEL and 1000 mg/kg bw/d was considered the NOEL for reproductive / developmental toxicity.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 06, 2015 - Oct 29, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

1995-07-27

Deviations:

no

Principles of method if other than guideline:

none

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Hrl: Sprague Dawley (SD)

Details on species / strain selection:

The Hrl: Sprague Dawley (SD) rat is a rodent of proven experimental validity for the purpose of the study.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories, Zona Industriale Azzida, 57, 33049 San Pietro al Natisone (UD), Italy on July 14th, 2015 (shipping slip No15003754).
- Age at study initiation: 9 weeks
- Weight at study initiation: (P) Males: 248.2 - 270.1 g; Females 174.0 - 193.4 g
- Fasting period before study: -
- Housing: single
- Use of restrainers for preventing ingestion (if dermal): no
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 15
- Air changes (per hr): 20 air changes per hour filtered on HEPA
- Photoperiod (hrs dark / hrs light): 12 / 12 hours

Route of administration:

oral: gavage

Vehicle:

other: Methocel KM4 Premium 0.25 % in Milli-Q

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0.25 % in pure water (Milli-Q)
- Amount of vehicle (if gavage): 5 mL/kg
- Lot/batch no. (if required): 2K27012N01

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 7 evenings/week with a maximum of 14 evenings
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentration of the formulated test item was checked twice during the dosing period (i.e., on formulations prepared on one day of the pre mating and post mating periods).The HPLC analytical method was validated. A stability and homogeneity study in 0.25 % Methocel® K4M Premium in water was performed.

Duration of treatment / exposure:

Oral, by gavage
F0 Males: The test item or the vehicle was administered daily for 2 weeks before the mating period and throughout the same. Males were further dosed after the mating period to achieve a minimum total dosing period of 28 days.
F0 Females: The test item or the vehicle was administered daily for 2 weeks before the start of the mating period and throughout the same. Treatment continued during pregnancy and until Day 3 of lactation.

Frequency of treatment:

daily

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

80 (40m / 40f)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on 28 day study with similar chemical
- Rationale for animal assignment (if not random): random

Parental animals: Observations and examinations:

Mating
The rats were mated, one male to one female. Each male was placed and left with the same female for 7 days/week, for a maximum of 14 days, up to detection of pregnancy. Every morning, a vaginal smear from each female was examined at the microscope. The day on which the presence of spermatozoa was found (positive smear) was considered Day 0 of
pregnancy for that female.
The slides of vaginal smears were disposed of after evaluation.
Females that never showed a positive smear were separated from the male at the end of the mating period.

Clinical Signs
Physical appearance, behavior and clinical signs were observed twice a day. Any deviation from the norm was recorded.
The duration of gestation was recorded and calculated from Day 0 of pregnancy (day of positive smear).
The day of parturition was defined as Day 0 of lactation.
Observations for birth were scheduled three times a day during working hours. If births occurred after working hours, the following day was defined as Day 0 of lactation.
Each litter was examined as soon as possible after delivery to establish the number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than corresponding control pups) and the presence of gross abnormalities.

Body Weight and Food and Water Consumption
Males and females were weighed on the day before the start of dosing (Day -1 of the study), on the first day of dosing (Day 1 of the study) and then weekly thereafter up to the end of treatment.
During pregnancy, females were weighed on days 0, 7, 14 and 20 and within 24 hours of parturition (Day 0 of lactation) and on Day 4 of lactation.
Weights were reported individually for each adult animal.
Body weight of Female No. 32 of Group 2 was recorded on Days 0, 1, 2, 3 and 4 of lactation, after dystocic parturition, to closely monitor the health of the animal.
During the pre-mating period and pregnancy, food consumption was measured weekly and during lactation on Day 4.
Water consumption was measured twice a week on the following days:
- Pre-mating: Days -1, 4, 7, 11, 14 of the study;
- Pregnancy: Days 0, 4, 7, 11, 14, 17 and 20;
- Lactation: Days 0 and 4.

Sperm parameters (parental animals):

A detailed qualitative examination of one testis and the equilateral epididymis was done in 10 male rats of the control (vehicle) group, as well as in 10 males of the high dose group, taking into account the tubular stages of the spermatogenic cycle. The examination was conducted in order to identify treatment-related effects such as missing germ cell layers or types, retained spermatids, multinucleate or apoptotic germ cells and disorganization of the germinal epithelium.

Litter observations:

On Day 0 of lactation, the pups of each litter will be identified with a mark made by appropriately coloring different sides of the body; this mark will be renewed when necessary.
Live pups will be counted and sexed and litters weighed within 24 hours of parturition (Day 0 of lactation) and on Day 4 post partum.
In addition to the observations on parent animals, any abnormal behavior of the offspring will be recorded.
At birth and during the lactation period, all the young will be individually observed for:
- Live and stillbirths;
- Mortality ensuing after live birth, ascertained daily. Whenever possible, any pup found dead will be examined externally and internally in an attempt to determine the cause of death;
- Sex (on Day 0 of lactation by an external examination and measurement of the ano-genital distance and by internal examination, at sacrifice, on Day 4 of lactation);
- External abnormalities at birth;
- Pup weight (on Days 0 and 4 of lactation).

Postmortem examinations (parental animals):

Necropsy and gross pathological examination were performed on all rats.
Males and females were sacrificed by cervical dislocation after preliminary CO2 anesthesia.

Histopathology
Reproductive organs (ovary, epididymis, prostate, seminal vesicle, testis) and all tissues showing abnormality were fixed.
Detailed histological examination was performed on the ovaries, testes and epididymides (with special emphasis on stages of spermatogenesis and histopathology of interstitial testicular cell structure) of the animals of the highest dose group and the control group.

Postmortem examinations (offspring):

Necropsy and gross pathological examination were performed on all rats.
Pups were sacrificed by a pain-free method (i.e.. overdose of sodium pentothal i.p.).
Dead pups and pups killed on Day 4 of lactation were examined externally for gross abnormalities.

Statistics:

Body weight, food and water consumption, mortality, clinical signs, dosing, reproduction data,and fetal weight data were recorded and stored in the PROVANTIS™ system.

Reproductive indices:

-Mating index: percent ratio between the animals with positive smear + females found to bepregnant but without positive smear and the animals mated.
- Fertility index: percent ratio of females having evident signs of pregnancy with respect to the females that had positive vaginal smear and females found to be pregnant but without positive smear.
- Pregnancy index: the percent ratio of females with live births with respect to the pregnant females.
- Pregnancy period: the duration of pregnancy was determined for all those dams that reached pregnancy term as being the time that elapsed between the positive vaginal smear and the start of parturition.
- Mean mating time: mean mating time will be calculated on the dams proved pregnant and will be expressed for each group as the mean time lapse (in days) between the beginning of the mating period and the ascertainment that copulation occurred. The females found to be pregnant but without positive smear will be excluded from this calculation.

Offspring viability indices:

- The mean F1 body weight was obtained by averaging the mean weight of each litter at the various times.
- The F1 probability of survival was analyzed per group. Animals that died owing to accidental causes or were sacrificed at the end of lactation were statistically censored.
- The mean value per litter of live pups (number of males and females and total) was calculated at different times (Days 0, 4 of lactation).
- The following indices of each litter were calculated:
Birth index: (No. of live newborns at birth/No. of implantations) x 100
Viability index on Day 4: (No. live pups on Day 4 after birth/No. of live births) x 100
Group mean values were calculated from individual data in two ways:
Mean A: calculated on all the surviving females having evident signs of pregnancy, including those that presented 100 % post-implantation losses.
Mean B: calculated only on those females with viable fetuses at term.
The external, visceral and skeletal malformations and variations found were described for each litter.
All data were recorded and stored in PROVANTIS TM System.

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

One Female of the 1000 mg/kg/day treated group showed piloerection and hunched posture on
Day 10-12 of the study. On Day 11 of the study, due to the severe clinical signs, this female did
not receive treatment. This female was found dead on day 13 of the study; at histopathology
evaluation this animal showed severe inflammatory reactions around the esophagus and lungs
(thoracic cavity), typical of a gavage error.

One Female of the 100 mg/kg bw/day had a dystocic parturition after showing pale ear and tail
with piloerection and slight blood discharge from the vagina on Day 21-22 of pregnancy up to
Day 2 of lactation.

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, non-treatment-related

Description (incidence):

One Female of the 1000 mg/kg bw/day treated group showed piloerection and hunched posture on Day 10-12 of the study. On Day 11 of the study, due to the severe clinical signs, this female did
not receive treatment. This female was found dead on day 13 of the study; at histopathology
evaluation this animal showed severe inflammatory reactions around the esophagus and lungs
(thoracic cavity), typical of a gavage error.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Males did not show any compound-related effects on body weight at any time point during the
study at 100, 300 and 1000 mg/kg bw/d in comparison to controls.
During the pre-mating period, pregnancy and lactation no compound-related effects were
observed on body weight and body weight gain in females at 100, 300 and 1000 mg/kg bw/d in
comparison to controls.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No changes were observed in the mean food consumption of the 100, 300 and 1000 mg/kg bw/day treated males, in comparison to controls, except for a statistically significantly lower mean food consumption observed at 300 and 1000 mg/kg bw/d during the Day 1-7of the pre-mating period.
No effects were seen on food consumption of females at 100, 300 and 1000 mg/kg bw/d in comparison to the control group during the pre-mating period, gestation and lactation.

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

effects observed, treatment-related

Description (incidence and severity):

Higher mean water consumption was observed in males treated at 100, 300 and 1000 mg/kg bw/day during the study, reaching statistically significantly higher mean values in the Day 7-11 and 11- 14 periods.
During the 2-week pre-mating period higher mean water consumption was observed at 100, 300
and 1000 mg/kg bw/d in the Day 7-11 period for females. This observation was not confirmed thereafter since during pregnancy and lactation no differences were observed in the mean water consumption of females treated at 100, 300 and 1000 mg/kg bw/day in comparison to control females.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Endocrine findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

The histopathological findings noted occurred either incidentally as single cases or were equally
distributed over the control and high dose group and are considered spontaneous in nature. An
organ-specific effect on the testis structure caused by treatment can be ruled out.
Mononuclear (inflammatory) cell infiltrates are commonly observed in the epididymidal
interstitium of male rats. The etiology is unknown (Foley, 2001).
All the described lesions occur in very low numbers of both control and treated animals and may
be considered as part of spontaneous background pathology.
The high dose female no. 75 that was found dead on day 13 of the study showed severe
inflammatory reactions around esophagus and lungs (thoracic cavity), typical for a gavage error.

Histopathological findings: neoplastic:

not examined

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Description (incidence and severity):

No differences were seen in the mating index (ratio between mated and paired animals) or fertility index (ratio between pregnant and mated animals) among the various experimental groups. The mean pre-coital time was similar in all groups. Pregnancy length and pregnancy index (ratio between females with live births and pregnant females) among the different experimental groups showed no differences. The slightly lower pregnancy index observed in the 100 mg/kg bw/day treated group was due to female No. 32, the one with a dystocic parturition, which delivered one stillborn pup. No compound-related changes were observed in the mean number of corpora lutea in Groups 2, 3 and 4 in comparison to the control group. Indeed, in comparison to controls, higher mean values per litter were noted in the low and mid dose group while lower values were noted in the highest dose group.
A slightly lower mean value of implantations was observed in the treated groups, in comparison to controls, though without any dose-relationship or statistical difference. Due to the above results higher mean pre-implantation losses were observed in all the treated groups in comparison to controls, without any dose-relationship. Slightly lower mean number of live pups was observed at 100, 300 and 1000 mg/kg bw/d in comparison to the control group, even if with no dose-relationship. No stillborn pups were found in the control group and at 300 mg/kg bw/day. Two stillborn pups were observed at 100 mg/kg bw/d, one in female No. 31 and one in female No. 32, and two at 1000 mg/kg bw/d, one in female No. 77 and one in female No.80. The post implantation losses observed at 100, 300 and 1000 mg/kg bw/d showed slightly higher mean values per litter in comparison to controls, due to the slightly lower mean number of implantations. The birth index values showed no relevant differences in the treated group in comparison to controls. Indeed the slightly lower birth index observed in the 100 mg/kg bw/day treated group was due, as for the pregnancy index, to female No. 32, which delivered one stillborn pup. No relevant changes in the mean values per litter of pups found dead was observed in any experimental group nor in the mean viability index on day 4.
No runts (pups significantly smaller than others) were observed in any group.

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: higher mean water consumption was observed in males of the treated groups in the Day 7-11 and 7-14 periods in comparison to controls

Key result

Critical effects observed:

no

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not examined

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

The mean weight of male and female pups at birth and on Day 4 was similar in treated and control groups.

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

no effects observed

Description (incidence and severity):

No differences were seen in the sex ratio among the different experimental groups.

Anogenital distance (AGD):

no effects observed

Description (incidence and severity):

No effects were observed on the mean ano-genital distance of males and females pups observed at 100, 300 and 1000 mg/kg bw/d in comparison to controls. The correspondence between internal and external sex was 100 % in all experimental groups.

Nipple retention in male pups:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Description (incidence and severity):

No abnormalities at birth were seen in any pup.

Histopathological findings:

no effects observed

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

Offspring development was not affected by the test-item.

Key result

Dose descriptor:

NOEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Conclusions:

On the basis of the above findings 1000 mg/kg bw/d was considered the parental NOAEL and 1000 mg/kg bw/d was considered the NOEL for reproductive / developmental toxicity.

Executive summary:

In this study the assessment and evaluation of the toxic characteristics of a chemical test item was performed in Crl:CD (SD) rats according to OECD 421. The test material was administered to the animals daily by oral route at 100, 300 and 1000 mg/kg bw/day as follows:

F0 Males were treated for 2 weeks before the start of the mating period and throughout the same. Males were further dosed after the mating period until the minimum total dosing period of 28 days was completed.

F0 Females were treated for 2 weeks before the start of the mating period and throughout the same. Treatment continued throughout pregnancy and until Day 3 of lactation.

A concurrent control group received 0.25 % Methocel® K4M Premium aqueous solution with the same dosing regimens.

All groups consisted of 10 males and 10 females. All animals were observed for survival and clinical signs; in addition to the observations on parents, any abnormal behavior of the offspring was recorded.
Body weight and food and water consumption were recorded at scheduled times during the premating, pregnancy and lactation periods.

On Day 0 of lactation pups were counted, sexed (by an external examination and measurement of the ano-genital distance, confirmed later by an internal examination at sacrifice) and weighed. All pups were individually observed for live and stillbirths; mortality ensuing after live birth was ascertained daily. Any pup found dead was examined externally and internally in an attempt to determine the cause of death.

Males were sacrificed at the end of the treatment period (Day 29 of the study), females were sacrificed on Day 4 of lactation with their litters. Females showing no evidence of copulation were killed 24-26 days after the last day of the mating period.
The uteri of apparently non-pregnant females were stained using the Salewski method and examined for the presence of implantation sites.
At the time of sacrifice all animals were examined macroscopically for any abnormalities or pathological changes. Special attention was paid to the organs of the reproductive system of adult animals. The number of implantation sites and corpora lutea was recorded in females. The testes and epididymides of all male adult animals were weighed. Detailed histological examination was performed on the ovaries, testes and epididymes (with special emphasis on stages of spermatogenesis and histopathology of interstitial testicular cell structure) of the animals of the highest dose group and the control group.  

No treatment-related deaths occurred.

One female only in group 2 had a dystocic parturition, showing pale ear and tail with piloerection and slight blood discharge from the vagina from Day 21-22 of pregnancy up to Day 2 of lactation. No clinical signs were observed during the study in any other animal.

No relevant changes were seen in body weight, and in food or water consumption of females at any time point of the study.

Statistically significantly higher mean water consumption was observed in males of the treated groups in the Day 7-11 and 7-14 periods in comparison to controls.

At necropsy no relevant macroscopic changes were seen in any animal and no differences were noted in the mean weight of testes and epididymides of treated males in comparison to controls.

No treatment-related changes were recorded during histological examination performed on the testes, epididymides, prostate, seminal vesicles, and ovaries.

No effects were observed on the reproductive performance of animals, i.e. no differences were noted comparing the mating and fertility indices (ratio between mated and paired animals and ratio between pregnant and mated animals, respectively) of the treated groups with the control group. In addition the mean pre-coital time was similar in all experimental groups.

No differences were seen in the pregnancy length and in the pregnancy index (ratio between females with live births and pregnant females) between treated and control groups.

Offspring development was not affected by the test-item.

No changes in the anogenital distance of male and female pups on Day 0 was observed, and the examination on Day 4 confirmed a 100 % correspondence between internal and external sex for all the animals.

Necropsy of pups on Day 4 did not evidence any compound-related changes.

In conclusion, the test material given orally to male rats for twenty-eight consecutive days and to females during the pre-mating, mating and gestation periods and up to day 3 of lactation at the doses of 100, 300 and 1000 mg/kg bw/day induced only a statistically significantly higher mean water consumption in treated males in comparison to controls.

Neither reproductive performance nor offspring development was affected by the test item. No treatment-related changes were recorded during histological examination performed on the ovaries, testes and epididymis.

On the basis of the above findings 1000 mg/kg bw/d was considered the parental NOAEL and 1000 mg/kg bw/d was considered the NOEL for reproductive / developmental toxicity.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

This study was performed according to GLP and the methods applied are fully compliant with OECD TG 421.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

In this study the assessment and evaluation of the toxic characteristics of a chemical test item was performed in Crl:CD (SD) rats according to OECD 421. The test material was administered to the animals daily by oral route at 100, 300 and 1000 mg/kg bw/day as follows:

F0 Males were treated for 2 weeks before the start of the mating period and throughout the same. Males were further dosed after the mating period until the minimum total dosing period of 28 days was completed.

F0 Females were treated for 2 weeks before the start of the mating period and throughout the same. Treatment continued throughout pregnancy and until Day 3 of lactation.

A concurrent control group received 0.25 % Methocel® K4M Premium aqueous solution with the same dosing regimens.

All groups consisted of 10 males and 10 females. All animals were observed for survival and clinical signs; in addition to the observations on parents, any abnormal behavior of the offspring was recorded.
Body weight and food and water consumption were recorded at scheduled times during the premating, pregnancy and lactation periods.

On Day 0 of lactation pups were counted, sexed (by an external examination and measurement of the ano-genital distance, confirmed later by an internal examination at sacrifice) and weighed. All pups were individually observed for live and stillbirths; mortality ensuing after live birth was ascertained daily. Any pup found dead was examined externally and internally in an attempt to determine the cause of death.

Males were sacrificed at the end of the treatment period (Day 29 of the study), females were sacrificed on Day 4 of lactation with their litters. Females showing no evidence of copulation were killed 24-26 days after the last day of the mating period.
The uteri of apparently non-pregnant females were stained using the Salewski method and examined for the presence of implantation sites.
At the time of sacrifice all animals were examined macroscopically for any abnormalities or pathological changes. Special attention was paid to the organs of the reproductive system of adult animals. The number of implantation sites and corpora lutea was recorded in females. The testes and epididymides of all male adult animals were weighed. Detailed histological examination was performed on the ovaries, testes and epididymes (with special emphasis on stages of spermatogenesis and histopathology of interstitial testicular cell structure) of the animals of the highest dose group and the control group.  

No treatment-related deaths occurred.

One female only in group 2 had a dystocic parturition, showing pale ear and tail with piloerection and slight blood discharge from the vagina from Day 21-22 of pregnancy up to Day 2 of lactation. No clinical signs were observed during the study in any other animal.

No relevant changes were seen in body weight, and in food or water consumption of females at any time point of the study.

Statistically significantly higher mean water consumption was observed in males of the treated groups in the Day 7-11 and 7-14 periods in comparison to controls.

At necropsy no relevant macroscopic changes were seen in any animal and no differences were noted in the mean weight of testes and epididymides of treated males in comparison to controls.

No treatment-related changes were recorded during histological examination performed on the testes, epididymides, prostate, seminal vesicles, and ovaries.

No effects were observed on the reproductive performance of animals, i.e. no differences were noted comparing the mating and fertility indices (ratio between mated and paired animals and ratio between pregnant and mated animals, respectively) of the treated groups with the control group. In addition the mean pre-coital time was similar in all experimental groups.

No differences were seen in the pregnancy length and in the pregnancy index (ratio between females with live births and pregnant females) between treated and control groups.

Offspring development was not affected by the test-item.

No changes in the anogenital distance of male and female pups on Day 0 was observed, and the examination on Day 4 confirmed a 100 % correspondence between internal and external sex for all the animals.

Necropsy of pups on Day 4 did not evidence any compound-related changes.

In conclusion, the test material given orally to male rats for twenty-eight consecutive days and to females during the pre-mating, mating and gestation periods and up to day 3 of lactation at the doses of 100, 300 and 1000 mg/kg bw/day induced only a statistically significantly higher mean water consumption in treated males in comparison to controls.

Neither reproductive performance nor offspring development was affected by the test item. No treatment-related changes were recorded during histological examination performed on the ovaries, testes and epididymis.

On the basis of the above findings 1000 mg/kg bw/d was considered the parental NOAEL and 1000 mg/kg bw/d was considered the NOEL for reproductive / developmental toxicity.

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

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on toxicity to reproduction, the test item does not require classification according to Regulation (EC) No 1272/2008 (CLP).