Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2009-12-17 to 2010-07-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed according to the OECD guideline No 421, and in compliance with the Good Laboratory Practice There were no circumstances that may have affected the quality or integrity of the data.
Reason / purpose:
reference to same study
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
In compliance with the Swiss Ordinance relating to Good Laboratory Practice adopted May 18th, 2005 [SR 813.112.1]. This Ordinance is based on the OECD Principles of GLP, as revised in 1997 and adopted on Nov. 26th, 1997 [OECD Council C(97)186/Final].
Limit test:
no
Test material information:
Composition 1
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
Animals: Rat, HanRcc: WIST(SPF)
Rationale: Recognized by international guidelines as a recommended test system.
Breeder: Harlan Laboratories, B.V., Kreuzelweg 53, 5961 NM Horst / Netherlands
Number of Animals: 40 males (10 per group) and 40 females (10 per group)
Age (at Start of Treatment): 11 weeks
Body Weight Range (at Start of Treatment): Males (301 to 330 g) and females (174 to 206 g)
Identification: Cage card and individual animal number (ear tattoo).
Randomization: Computer-generated random algorithm. In addition body weights (recorded on the day of allocation) were taken into consideration in order to ensure similar mean body weights in all groups.
Acclimatization: Under test conditions after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
Conditions: Standard laboratory conditions. Air-conditioned with 10 - 15 air changes per hour, continuously monitored environmental conditions (temp. range: 22 ± 3 °C; relative humidity range: 18.8 – 71.8%). There was 12 hour fluorescent light / 12-hour dark cycle with music during the light period.
Accommodation: Individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding (‘Lignocel’ J.Rettenmaier & Söhne GmbH & CoKG, 73494 Rosenberg / Germany, imported by Provimi Kliba SA, 4303 Kaiseraugst / Switzerland). During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular estrus cycles.
Diet: Pelleted standard Kliba Nafag 3433 rodent maintenance diet (Provimi Kliba SA, 4303 Kaiseraugst / Switzerland) was available ad libitum (batch no. 60/09).
Water: Community tap-water from Füllinsdorf was available ad libitum in water bottles.

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
DOSE FORMULATIONS

The dose formulations were prepared weekly using the test item as supplied by the Sponsor and using a correction factor of 2.42 so the dose actually ingested corresponds to the dose adjusted for the main constituent.

TRIQUAT MONOMER was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration.

Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.

STORAGE OF DOSE FORMULATIONS

Dose formulations were stored in refrigerator (2 - 8 °C) in glass beakers.

Based upon the results of stability analyses performed within the Harlan Laboratories study C46062 (non-GLP Study), dose formulations were stable for at least one week.

TREATMENT

Method: Oral, by gavage
Rationale for Method: Administration by gavage is a common and accepted route of exposure for this type of studies.
Frequency of Administration: Once daily
Target Dose Levels (for the main constituent): Group 1: 0 mg/kg/day (control group); Group 2: 100 mg/kg/day; Group 3: 300 mg/kg/day; Group 4: 1000 mg/kg/day
Rationale for Dose Level Selection: The dose levels were selected based on a previous 28-Day Oral (Gavage) toxicity study in Han Wistar, Harlan Laboratories Study C46073, using dose levels of 100, 300, and 1000 mg/kg/day, resulting in a NOAEL of 1000 mg/kg/day.
Dose Volume: 10 mL/kg body weight
Duration of Acclimatization Period: 7 days
Duration of Treatment Period: Males (minimum 4 weeks); females (approximately 7 weeks)
Details on mating procedure:
MATING, GESTATION AND LACTATION

During the pairing period, females were housed with sexually mature males (1:1) until evidence of copulation was observed. The females were removed and housed individually if:

- the daily vaginal smear was sperm positive, or
- a copulation plug was observed.

The day of mating was designated day 0 post coitum.

If a female did not mate during the 14-day pairing period, this female was paired with a male of the same group which had already mated successfully. If mating was not recorded during this additional pairing period of a maximum of 14 days, the female was sacrificed and, if indicated, the reproductive organs examined histopathologically in order to ascertain the reason for the infertility.

All dams were allowed to give birth and rear their litters (F1 pups) up to day 4 post partum. Day 0 was designated as the day on which a female had delivered all her pups.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
ANALYSIS OF DOSE FORMULATIONS

On the first treatment day samples from the control group as well as three samples (top, middle and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of concentration and homogeneity. Samples of about 2 g of each concentration were taken from the middle only to confirm stability (3 hrs and 7 days). Before the end of treatment, samples were taken from the middle to confirm concentration. The aliquots for analysis of dose formulations were frozen (-20 ± 5 °C) and delivered on dry ice to Analytic Department (Harlan Laboratories Ltd., Itingen / Switzerland) and stored there at -20 ± 5 °C until analysis.

The samples were analyzed by HPLC coupled to an UV detector following an analytical procedure provided by the Sponsor and adapted at Harlan Laboratories. The test item was used as the analytical standard. Analyzed samples were not discarded without written consent from the study director.

Samples were considered accurately prepared and sufficiently stable if the following acceptance criteria were met: ±20% of nominal for sample content, ±15% deviation from mean calculated from top, middle and bottom samples for homogeneity and ±10% from time-zero reference (content or mean of homogeneity samples).

Application formulations were found to be homogeneously prepared and sufficient formulation stability under storage conditions was approved.

The application formulations investigated during the study were found to comprise TRIQUAT MONOMER in the range of 83.2% to 100.6% and, thus, the required content limit of ±20% with reference to the nominal concentration was met. The homogeneous distribution of TRIQUAT MONOMER in the preparations was approved because single results found did not deviate more than 7.2% (<15%) from the corresponding mean.

In addition, the test item was found to be stable in application formulations when kept 7 days in the refrigerator (2 - 8 °C) due to recoveries which met the variation limit of 10% from the time-zero (homogeneity) mean.
Duration of treatment / exposure:
Males: Minimum 4 weeks
Females: Approximately 7 weeks
Frequency of treatment:
Once daily
Details on study schedule:
Acclimatization: 7 days (males and females)
First Test Item Administration : Day 1 of pre-pairing (males and females)
Pre-Pairing: 14 days (males and females)
Pairing: 15 days maximum (females); 15 days (males)
Gestation: approximately 21 days (females)
Treatment Ends: On day 3 post partum (females), on day before sacrifice (males)
Necropsy: On day 4 post partum (females); after a minimum of 28 days treatment (males)
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Parental animals: Observations and examinations:
Viability / Mortality: Twice daily
Clinical Signs: Daily cage-side clinical observations (once daily, during acclimatization and up to day of necropsy). Additionally females were observed for signs of difficult or prolonged parturition, and behavioral abnormalities in nesting and nursing.
Food Consumption: Males (weekly during pre-pairing and after pairing periods; females (pre-pairing period days 1 - 8 and 8 - 14, gestation period days 0 - 7, 7 - 14 and 14 - 21 post coitum and lactation period days 1 - 4 post partum). No food consumption was recorded during the pairing period.
Body Weights: Recorded daily from treatment start to day of necropsy.
Estrous cyclicity (parental animals):
Not examined [see Study C46073 - 28-day repeated dose toxicity study]
Litter observations:
The litters were examined for litter size, live births, still births and any gross anomalies. The sex ratio of the pups was recorded. Pups were weighed individually (without identification) on days 0 (if possible), 1 and 4 post partum.
Postmortem examinations (parental animals):
TERMINATION OF THE STUDY

Males were sacrificed after they had been treated for at least 28 days. Dams were sacrificed on day 4 post partum.

If birth did not occur on the expected date (day 21 post coitum), the dam was sacrificed on day 25 post coitum.

NECROPSY

All parent animals sacrificed or found dead were subjected to a detailed macroscopic examination to establish, if possible, the cause of death. Specimens of abnormal tissue were fixed in neutral phosphate buffered 4% formaldehyde solution.

At the scheduled sacrifice, all parent animals were killed by an injection of sodium pentobarbital. All P generation animals were exsanguinated.

All parent animals were examined macroscopically for any structural changes, either at the scheduled necropsy or during the study if death occurred.

For the parent animals, special attention was directed at the organs of the reproductive system.
The number of implantation sites and corpora lutea was recorded for all dams with litters. The uteri of non-pregnant females were placed in a solution of ammonium sulfide to visualize possible hemorrhagic areas of implantation sites.

ORGAN WEIGHTS

At the scheduled sacrifice, the testes and epididymides of all parental males were weighed as pairs.

TISSUE PRESERVATION

The ovaries from all parental females were preserved in neutral phosphate buffered 4% formaldehyde solution.

The testes and epididymides from all parental males were preserved in Bouin’s fixative. The prostate and seminal vesicles from all males were fixed in neutral phosphate buffered 4% formaldehyde solution.

In addition, all organs showing macroscopic lesions of all adult animals were preserved.

HISTOTECHNIQUE

All organ and tissue samples to be examined by the study pathologist were processed, embedded and cut at an approximate thickness of 2 - 4 micrometers and stained with hematoxylin and eosin. Additionally, the testis was stained by PAS-hematoxylin. Special stains were used at the discretion of the study pathologist.

HISTOPATHOLOGY

Slides of all organs and tissues collected at terminal sacrifice from the animals of the control and high-dose groups were examined by the study pathologist.

Special emphasis was made on the stages of spermatogenesis and histopathology of interstitial cell structure.

Histological examination of ovaries was carried out on one female (no. 51) that did not give birth. In addition, microscopic examination of the reproductive organs was made in one infertile male (No. 11).

A peer review was carried by a pathologist (Harlan Laboratories Ltd, Switzerland).
Postmortem examinations (offspring):
TERMINATION OF THE STUDY
Pups were sacrificed on day 4 post partum.

NECROPSY
All pups sacrificed or found dead were subjected to a detailed macroscopic examination to establish, if possible, the cause of death. Specimens of abnormal tissue were fixed in neutral phosphate buffered 4% formaldehyde solution.

At the scheduled sacrifice, all pups were killed by an injection of sodium pentobarbital.

Dead pups, except those excessively cannibalized, were examined macroscopically.

All pups were examined macroscopically for any structural changes, either at the scheduled necropsy or during the study if death occurred.
Statistics:
The following statistical methods were used to analyze food consumption, body weights and reproduction data:
- Means and standard deviations of various data were calculated.
- The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test was applied if the variables could be dichotomized without loss of information.
Reproductive indices:
From the on-line recorded reproduction data, the following parameters were calculated: fertility indices, mean precoital time, post-implantation losses, mean litter size.
Offspring viability indices:
From the on-line recorded reproduction data, the following parameters were calculated: dead/live pups at first litter check, pup sex ratios and postnatal loss (up to day 4 post partum).
Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Description (incidence and severity):
In females at 300 mg/kg, dark-red nodules on left uterine horn and hemorrhagic vagina were noted in one female. This was not considered to be test item-related.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
All findings recorded were within the range of normal background which may be recorded in animals of this strain and age.
Other effects:
not examined
Reproductive function: estrous cycle:
not examined
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
[sperm staging]
Reproductive performance:
no effects observed
1 IN-LIVE DATA PARENTAL ANIMALS

1.1 MORTALITY
All animals survived until the scheduled necropsy.

1.2 CLINICAL SIGNS OR OBSERVATIONS
No clinical signs or symptoms were observed during the study.

1.3 FOOD CONSUMPTION OF MALES
Pre-pairing Period

No statistically significant alterations were observed in mean food consumption.

In order of ascending dose levels, the overall differences in food consumption were: 2.1%, -3.9% and -5.0% during the pre-pairing period (percentages refer to the respective values of the control group). These differences were not meaningful as the food consumption was stable in all groups over the pre-pairing period.

1.4 FOOD CONSUMPTION OF FEMALES
Pre-pairing, Gestation and Lactation Periods

No statistically significant alterations of mean food consumption were observed at any dose level when compared to the respective values in the control group during the pre-pairing, gestation and lactation periods.

In order of ascending dose levels, the overall differences in food consumption were: 3.2%,
-2.2% and +1.1% during the pre-pairing period, +2.6%, +0.9% and +3.1% during the gestation period and ±0.0%, -5.7% and -12.2% during the lactation period (percentages refer to the respective values of the control group). Although the mean food consumption was lower during the lactation period, body weights were not affected. Thus no adverse effect occurred.

1.5 BODY WEIGHTS OF MALES
Pre-pairing and Pairing Periods

No statistically significant alterations were noted in mean body weight and mean body weight gain of males throughout the entire duration of the study.

In the order of ascending dose levels, the overall mean body weight gains were: +15%, +14%, +13% and +14% during the pre-pairing period, +9%, +10%, +8% and +8% during the pairing period (percentages refer to the respective time intervals).

1.6 BODY WEIGHTS OF FEMALES
Pre-pairing, Gestation and Lactation Periods

No statistically significant alterations of mean body weights and mean body weight gain were observed at any dose level when compared to the respective values in the control group.

In the order of ascending dose levels, the overall mean body weight gain was +9%, +7%, +8% and +7% during the pre-pairing period, +60%, +61%, +59% and +56% during the gestation period and +4%, +4%, +3% (0 to 21 days) and +4% during the laction period (percentages refer to the respective time intervals).


2 REPRODUCTION AND BREEDING DATA

2.1 MATING PERFORMANCE AND FERTILITY
One female (No. 75) in group 4 mated during the second pairing period. No statistically significant alterations were observed in median and mean precoital times and all values were within the range of the historical control data. For females which mated during the first pairing period, mean precoital times were 2.1, 2.7, 2.3 and 4.1 days and median precoital time was 2, 3, 3 and 4 days in order of ascending dose level.

Female no. 44 in group 1, female no. 51 in group 2 and female no. 75 in group 4 were not pregnant. Thus the fertility indices were 90.0%, 90.0%, 100.0% and 90.0% in groups 1, 2, 3 and 4.

2.2 DURATION OF GESTATION
See attached tables on pp. 43-46

The mean duration of gestation was unaffected by treatment with the test item and within the range of the historical control data. Mean duration of gestation was 21.4, 21.7, 21.2 and 21.3 days, in order of ascending dose level.

2.3 CORPORA LUTEA COUNT

The mean number of corpora lutea per dam (determined at necropsy) was not affected by the treatment with the test item and no statistical significances were observed. Mean corpora lutea count was 13.3, 12.8, 13.7 and 14.1 in order of ascending dose level.

2.4 IMPLANTATION RATE AND POST-IMPLANTATION LOSS
See attached tables on pp. 43-46

No statitistically significant alteration was noted in the mean number of implantations per dam and in post-implantation losses.

The mean numbers of implantations per litter were 13.2, 12.2, 13.6 and 11.2 in order of ascending dose level. In group 4, mean number of implantations was outside the lower limit of the historical control data. This was not considered to be a test item-related effect since there was only a marginal difference when compared to the control group. Mean incidence of post-implantation loss as a percentage of total implantations was 12.6, 9.1, 17.6 and 8.9% in order of ascending dose level. All these values were within the range of the historical control data.

2.5 LITTER SIZE AT FIRST LITTER CHECK
See attached tables on pp. 43-46

The number of live pups at first litter check was unaffected by treatment with the test item. The mean number of live pups per litter was 11.6, 11.1, 11.2 and 10.2 in order of ascending dose level. In group 4, litter size was outside the range of the historical control data. Since this was secondary to the lower number of implantations, it was not considered to a test item-related effect.

2.6 POSTNATAL LOSS DAYS 0 - 4 POST PARTUM
See attached tables on pp. 43-46

Postnatal loss was not affected by the treatment with the test item. In group 1, female No. 50 lost the whole litter (two pups) on day 2 post partum, female No. 45 lost two pups on day 1 post partum at first litter check and female No. 49 lost one pup on day 3. In group 2, female No. 55 lost one pup on day 2 post partum. No postnatal loss occurred in groups 3 and 4.

The resulting viability indices were 97.1%, 99.0%, 100.0% and 100.0% in order of ascending dose levels.


3 TERMINAL FINDINGS - PARENTAL ANIMALS

3.1 ORGAN WEIGHTS
In males, weights (absolute and relative to body weight) of testes and epididymides in all test item-treated groups were comparable to those of the control groups and thus were not affected by the treatment with the test item.

3.2 MACROSCOPICAL FINDINGS
No abnormal findings were noted in males. In females, the only findings observed were dark red firm nodules on left uterine horn and hemorrhagic vagina in one female (No. 68) in group 3 and cervix with gray white mucus and right horn dilated in one female in group 1 (No. 44). Type and incidence of these findings did not indicate any test item-related effect.

3.3 HISTOPATHOLOGY FINDINGS
All findings recorded were within the range of normal background, which may be recorded in animals of this strain and age. No test item-related histological findings were recorded in ovary of one female (no. 51), which did not give birth and in reproductive organ of one infertile male (no. 11).

No differences on the completeness of stages or cell populations of the testes were recorded between controls and high dose animals.
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
other: Dose adjusted for content of the main constituent.
Sex:
male/female
Basis for effect level:
other: In absence of any reproduction toxicity.
Remarks on result:
other: Generation: P (reproduction) (migrated information)
Dose descriptor:
NOEL
Effect level:
2 420 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
Dose of substance as registered (including residual water necessary for the stability).
Sex:
male/female
Basis for effect level:
other: In absence of any reproduction toxicity.
Remarks on result:
other: Generation: P (reproduction) (migrated information)
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
other: Dose adjusted for content of the main constituent.
Sex:
male/female
Basis for effect level:
other: In the absence of any signs of general toxicity.
Dose descriptor:
NOAEL
Effect level:
2 420 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
Dose of substance as registered (including residual water necessary for the stability)
Sex:
male/female
Basis for effect level:
other: In the absence of any signs of general toxicity.
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
1 LITTER DATA - F1 PUPS

1.1 EXTERNAL EXAMINATION AT FIRST LITTER CHECK AND DURING LACTATION
No test item-related findings were noted.

1.2 SEX RATIOS
See attached tables on pp. 43-46

Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test item.

In all females rearing pups, the proportion of male pups on day 4 post partum was 45%, 49%, 40% and 46% in order of ascending dose level.

1.3 PUP WEIGHTS TO DAY 4 POST PARTUM
Mean pup weights were unaffected by treatment with the test item. On day 1 post partum mean pup weights were 5.9, 6.6, 6.0 and 6.0 g for combined data of male and female pups in order of ascending dose level. In group 2, the statistically significantly higher weight of pups was considered to be incidental since it was a single occurrence and without following a dose-dependent pattern.

Mean pup weight development during lactation was unaffected by treatment with the test item. Mean pup weights on day 4 post partum were 8.5, 9.6, 8.4 and 8.7 g for combined data of male and female pups in order of ascending dose level.

1.4 MACROSCOPICAL FINDINGS
No test item-related abnormal findings were noted at macroscopic examination of the pups. The only finding noted was advanced autolysis in one pup in the control group, which was found dead.
Dose descriptor:
NOEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
other: Dose adjusted for content of the main constituent.
Sex:
male/female
Basis for effect level:
other: In absence of any developmental toxicity.
Dose descriptor:
NOEL
Generation:
F1
Effect level:
2 420 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
Dose of substance as registered (including residual water necessary for the stability)
Sex:
male/female
Basis for effect level:
other: In absence of any developmental toxicity.
Reproductive effects observed:
not specified

SUMMARY OF PERFORMANCE

P Animals Breeding for F1 Litters

Group
(mg/kg/day)

1
(0)

2
(100)

3
(300)

4
(1000)

Female numbers

41-50

51-60

61-70

71-80

Number of females paired

10

10

10

10

Number of females mated

10

10

10

10

Number of pregnant females (A)

9

9

10

9

Number of females giving birth

9

9

10

9

Numbers of females which lost the whole litter (B)

1

0

0

0

Number of females which reared their pups until day 4 post partum

8

9

10

9

 

(A)  Female No. 44 in group 1, Female No. 51 in group 2 and Female No. 75 in group 4 were not pregnant.

(B)   Female No. 50 in group 1 lost the whole litter (two pups) on day 2 post partum

 

 

Conclusions:
This study is a valid investigation of the toxicological effects resulting from repeated oral-gavage administration of the test item TRIQUAT MONOMER to rats. TRIQUAT MONOMER was administered in Milli-Q-Water as vehicle at dosages of 100, 300, and 1000 mg/kg body weight/day, and controls received the vehicle only. TRIQUAT MONOMER was administered to male rats for at least 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum.

In absence of any adverse effect, the general NOAEL (No Observed Adverse Effect Level) was established at 1000 mg/kg/day.

Under the conditions of this study, the NOEL (No Observed Effect Level) for reproduction/ developmental toxicity was considered to be 1000 mg/kg/day.
Executive summary:

The purpose of this study was to generate preliminary information concerning the effects of TRIQUAT MONOMER on male and female reproductive performance such as gonadal function, mating behavior, conception and parturition.

Four groups of 10 males and 10 females were treated by gavage with TRIQUAT MONOMER once daily. Males were treated over a 14-day pre-pairing period and during the pairing period up to one day before necropsy. Females were treated throughout the pre-pairing, pairing, gestation and lactation period up to day 4 post partum.

 

The following dose levels were used (doses expressed as doses of the main constituent):

                       Group 1:                0 mg/kg body weight/day (control group)

                       Group 2             100 mg/kg body weight/day

                       Group 3:            300 mg/kg body weight/day

                       Group 4           1000 mg/kg body weight/day

 

A standard dose volume of 10 mL/kg body weight with a daily adjustment to the actual body weight was used. Control animals were dosed with the vehicle alone (highly purified water).

 

The following results were obtained:

 

PARENTAL ANIMALS

 

General Tolerability

All animals survived until the scheduled necropsy and no clinical signs were noted during the whole study.

 

Food Consumption

Mean food consumption was not considered to be affected by the treatment with the test item in males and females. At 1000 mg/kg/day, the lower mean food consumption observed in females during lactation period (not statistically significant) was not considered to be adverse since mean body weight was not affected.

 

Body Weights

Mean body weight and body weight gain were not affected in males and females for the whole duration of the treatment.

 

Reproductive Data

Mating performance, fertility, duration of gestation, mean number of corpora lutea, of implantations and post-implantation losses were not adversely affected by the treatment with the test item.

 

Organ Weights

Mean weight of testes and epididymides of the test item treated groups were compared to that of control and thus not affected by the treatment with the test item.

 

Macroscopical Findings and Histopathological Examinations

No test item-related findings were observed during the macroscopical and histological examination.

 

Sperm Staging

No differences on the completeness of stages or cell populations of the testes were recorded between controls ad high dose animals.

 

Litter Data

The number of live pups at first litter check and the mean litter size was unaffected by treatment with the test item. Mean pup weight at first litter check and on day 4 post partum was not affected.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
2 420 mg/kg bw/day
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information
Short description of key information:
A screening assay for Reproduction/Developmental toxicity was conducted in rats (OECD421) with the test material administered by the oral route. There was no effect on the fetal development and reproductive parameters at up to 2420 mg/kg/day (dose expressed as registered substance), i.e. 1000 mg/kg/day for the main constituent. Additional information provided by the repeated oral dose toxicity study showed no effect on the reproductive organs in males and females, no effects on the female estrus cycle in the 28-day study, and no effects on the sperm staging in the OECD421 study.

Justification for selection of Effect on fertility via oral route:
No effects on fertility parameters in the available study. See justification in endpoint record

Effects on developmental toxicity

Description of key information
A screening assay for Reproduction/Developmental toxicity was conducted in rats (OECD421) with the test material administered by the oral route. There was no effect on the fetal development.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Data waiving:
exposure considerations
Justification for data waiving:
other:
Abnormalities:
not specified
Developmental effects observed:
not specified
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
2 420 mg/kg bw/day
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information
Justification for selection of Effect on developmental toxicity: via oral route:
No effects observed in the available screening study. Exposure-based waiver. See justifications in the endpoint record.

Justification for classification or non-classification

Reliable studies conducted according to current guidelines showed no effects on the reproductive / fertility parameters in males and females, and no effects on the fetal development, as well as no effects on the reproductive organs, estrus cycle and sperm staging.