Registration Dossier

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
Combined repeated dose and reproduction /developmental toxicity screening in rats
Type of information:
other: read across from similar substance
Adequacy of study:
key study
Study period:
2016
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
OECD Guideline 422 adopted on 22 March 1996
GLP compliance:
yes
Limit test:
no
Justification for study design:
The Sprague Dawley rat was the species and strain of choice because it is accepted by many regulatory authorities and there is ample experience and background data on this species and strain.
The oral route by gavage was selected as it is a possible route of exposure of the test item in man providing best internal exposure for this kind of substance.

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
The Sprague Dawley rat was the species and strain of choice because it is accepted by many regulatory authorities and there is ample experience and background data on this species and strain.
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS srl., San Pietro al Natisone (UD), Italy
- Age at study initiation: 6 to 7 weeks
- Weight at study initiation: 187.8-206.4 g for males and 150.2-171.7 g for females
- Weight ordered: 176 to 200 g for males and 151 to 175 g for females
- Assigned to test groups randomly: yes, by computerised stratified randomisation to give approximately equal initial group mean body weights
- Fasting period before study:
- Housing: 5 of one sex to a cage - main groups from arrival to pairing; main group males after pairing; recovery groups throughout the entire study period
one male to one female- main groups during pairing
single - main group females after pairing
- Diet (ad libitum): 4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI), Italy
- Water (ad libitum): tap water
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 2
- Humidity (%): 55 ± 15
- Air changes (per hr): approximately 15 to 20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: sesame oil
Details on exposure:
The required amount of the tested substance was suspended in sesame oil. The formulations were prepared daily (concentrations of 12.5, 50 and 200 mg/mL). Concentrations were calculated and expressed in terms of test item as supplied.
The test item was administered orally by gavage at a dose volume of 5 mL/kg body weight.
Control animals received the vehicle alone at the same dose volume. The dose was administered to each animal on the basis of the most recently recorded body weight and the volume administered was recorded for each animal.
Details on mating procedure:
Mating was monogamous (one male to one female). A vaginal smear was taken from the day after the start of pairing until positive identification of copulation (sperm identification, vaginal plug in situ or copulation plugs found in the cage tray). The female was paired with the same male until positive identification occurred or 14 days had elapsed. A second mating session was allowed for the following females: nos. 11, 17, 19, 25, 27, 37 and 53 which had unsuccessful mating with the first pairing combination.
These females were re-mated with a proven male of the same treatment group. The second pairing combination was monitored for mating until positive signs of copulation. For female no. 27, the mating was inadvertently not detected in both mating sessions although the animal gave birth.
Analytical verification of doses or concentrations:
no
Remarks:
the test item is neither extractable in hydrophilic nor in lipophilic solvents, hence analytical verification of test item concentrations is not possible
Details on analytical verification of doses or concentrations:
The correct preparation of the respective dose levels was monitored in the weighing record of the test item in each formulation process
Duration of treatment / exposure:
Main groups (Groups 1 to 4)

Males - Animals were dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior
to pairing, through the mating period and thereafter at least until the minimum total dosing
period of 28 days had been completed including the day before necropsy. Dose volumes were
adjusted once per week for each animal according to the last recorded body weight.

Females - Animals were dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to
pairing and thereafter during pairing, post coitum and post partum periods until at least up
to, and including, Day 3 post partum or the day before sacrifice. Dose volumes were adjusted
once per week for each animal according to the last recorded body weight.
During the gestation period, dose volumes were calculated according to individual body
weight on Days 0, 7, 14 and 20 post coitum and then on Day 1 post partum. Thereafter
individual dose volumes remained constant.
Frequency of treatment:
Once daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Group 1
Dose / conc.:
62.5 mg/kg bw/day (nominal)
Remarks:
Group 2
Dose / conc.:
250 mg/kg bw/day (nominal)
Remarks:
Group 3
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
Group 4
No. of animals per sex per dose:
Each main group comprised 10 male and 10 female rats.
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale: the test substance did not cause any adverse effects in acute oral toxicity studies up to dose levels of 10000 mg/kg bw

Examinations

Parental animals: Observations and examinations:
Clinical signs: Once before commencement of treatment (data not presented but retained and archived together with all raw data) and at least once daily during the study, each animal was observed and any clinical signs were recorded. Observations were performed at the same time interval each day.
In addition, observation of the cage tray was performed starting from the first detected sign.

Body weight: Males were weighed weekly from allocation to termination. Females were weighed weekly
from allocation to positive identification of mating and on Days 0, 7, 14 and 20 post coitum.
Dams were also weighed on Days 1 and 4 post partum.

Food consumption:
The weight of food consumed by each cage of males and females was recorded weekly during the pre-mating period starting from allocation. Individual food consumption for the females was measured on Days 7, 14 and 20 post coitum starting from Day 0 post coitum and on Day 4 post partum starting from Day 1 post partum.
Oestrous cyclicity (parental animals):
Vaginal smears were taken daily in the morning starting from two weeks before pairing until a positive identification of copulation was made. The vaginal smear data were examined to determine the following:
1. anomalies of the oestrous cycle
2. the pre-coital interval (i.e., the number of nights paired prior to the detection of mating)
Sperm parameters (parental animals):
Organ weights: Epididymides, prostate gland, seminal vesicles with coagulating glands, testes
Histopathology: Epididymides, prostate gland, seminal vesicles with coagulating glands, testes, penis
Spermatogenic cycle: Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was examined.
Litter observations:
A parturition check was performed from Day 20 to Day 25 post coitum. Gestation periods were taken as the time between the day of successful mating (Day 0 post coitum) and the day of commencement of birth (i.e. first detected presence of offspring in the cage). The day that offspring were first detected in the cage was considered Day 0 post partum. As soon as possible after parturition was considered complete (Day 0 or 1 post partum), all pups (live and dead) were counted, sexed and only live pups were identified. Live pups were individually weighed on Days 1 and 4 post partum. Pups dying during the lactation period were weighed before the despatch to necropsy. Observations were performed once daily for all litters.
Postmortem examinations (parental animals):
Animals selected for blood collection were killed by exsanguination under isofluorane anaesthesia. Animals not selected for blood collection were killed by carbon dioxide asphyxiation.
Parental males: The males were killed after the mating of all females, after at least 28 days of treatment period.
Parental females: The females with live pups were killed on Day 4 post partum. The females showing no evidence of copulation were killed 25 days after the last day of the mating session.
The females which did not give birth 25 days after positive identification of mating were killed shortly after.
The clinical history of the animals was studied and a detailed post mortem examination was conducted (including examination of the external surface and orifices). Changes were noted, the requisite organs weighed and the required tissue samples preserved in fixative and processed for histopathological examination.

All females were examined also for the following:
– external and internal abnormalities;
– number of visible implantation sites (pregnant animals);
– number of corpora lutea (pregnant animals).
Uteri of females with no visible implantationswere immersed in a 20% solution of ammonium
sulphide to reveal evidence of implantation.

Organ weights: From all animals completing the scheduled test period, the following organs were dissected free of fat and weighed.
Adrenal glands,
Brain (cerebrum, cerebellum, medulla/pons),
Epididymides,
Heart,
Kidneys,
Liver,
Ovaries and oviducts,
Parathyroid glands,
Prostate gland,
Seminal vesicles with coagulating glands
Spleen,
Testes,
Thymus (where present),
Thyroid,
Uterus including cervix.
The ratios of organ weight to body weight were calculated for each animal.

Tissues fixed and preserved: Samples of all the following tissues (all parental animals) were fixed and preserved in 10% neutral buffered formalin (except eyes, optic nerves, testes and epididymides which were fixed inModified Davidson’s fluid and preserved in 70% ethyl alcohol):
Abnormalities,
Adrenal glands,
Bone marrow (from femur),
Bone marrow (from sternum),
Brain (cerebrum, cerebellum, medulla/pons),
Caecum,
Clitoral gland,
Colon,
Duodenum,
Epididymides,
Heart,
Ileum,
Jejunum (including Peyer’s patches),
Kidneys,
Liver,
Lungs (including mainstem bronchi),
Lymph nodes – cervical,
Lymph nodes – mesenteric,
Nasal cavity,*
Oesophagus*,
Ovaries and oviducts,
Parathyroid glands,
Pituitary gland,
Penis,
Prostate gland,
Rectum,
Sciatic nerve,
Seminal vesicles with coagulating glands,
Spinal column*,
Spinal cord (cervical, thoracic, lumbar),
Spleen,
Stomach (forestomach and glandular,)
Testes,
Thymus (where present)
Thyroid
Trachea
Urinary bladder
Uterus – cervix
Vagina
* = not examined as no signs of toxicity or target organ involvment were noted.

After dehydration and embedding in paraffin wax, sections of the tissues were cut at 5 micrometer thickness and stained with haematoxylin and eosin.
In addition, the testes and epididymides were cut at 2-3 micrometer thickness and stained with Periodic Acid Schiff (PAS). The morphological evaluation of theseminiferous epithelium (staging of spermatogenic cycle) was also performed.
The examination was restricted as detailed below:
1. Tissues specified above from 5 males and 5 females randomly selected (animals
evaluated for clinical pathology) in the control and high dose groups killed at term.
2. All abnormalities in all groups.
Postmortem examinations (offspring):
Pups that had completed the scheduled test period (Day 4 post partum) were euthanised by intraperitoneal injection of Sodium Thiopenthal.
All pups found dead in the cage were examined for external and internal abnormalities. All live pups sacrificed at termination (Day 4 post partum) were killed and examined for external abnormalities and sex confirmation by gonadal inspection.
Statistics:
Standard deviationswere calculated as appropriate. For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data.
The non-parametric Kruskal-Wallis analysis of variance (non-continuous variables) was used for the other parameters.
The criterion for statistical significance was p < 0.05. The mean values, standard deviations and statistical analysis were calculated from actual values in the computer without rounding off.
Reproductive indices:
Group mean values were calculated for all parameters. Data from non-pregnant females were excluded from group mean calculations as appropriate.
The following reproductive indices were calculated:
Males:
Copulation Index (%) = no. of animals mated x100/ no. of animals paired.
Fertility Index (%) = no. of males which induced pregnancy x100/ no. of males paired.

Females:
Copulation Index (%) = no. of animals mated x100/ no. of animals paired.
Fertility Index (%) = no. of pregnant females x100/ no. of females paired.

Offspring viability indices:
Males and females:
Pre-coital or Copulatory interval (mean time to mating) = Number of days between pairing and mating.

Females:
Pre-implantation loss was calculated as a percentage from the formula:
(no. of corpora lutea − no. of visible implantation/ no. of corpora lutea) x 100;
Pre-birth loss (post-implantation loss) was calculated as a percentage from the formula:
(no. of visible implantation − total litter size at birth/ no. of visible implantation) x 100;

Pup loss at birth was calculated as a percentage from the formula:
(Total litter size − live litter size at birth/ Total litter size) x 100;

Cumulative pup loss on Day 4 post partum was calculated as a percentage from the formula:
(Total litter size at birth − live litter size at Day 4/ Total litter size at birth) x 100;

Sex ratios were calculated at birth and on Day 4 post partum and were presented as the
percentage of males per litter.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Green staining of the tail was the main clinical sign recorded in all high dose animals and occasionally in the mid-dose group.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Some significant differences in body weight gain were recorded in high dose females before pairing (decreased and increased mean values on Days 8 and 15, respectively), compared to the control group.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption was comparable between groups throughout the study.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not specified
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No treatment-related changes were noted.
The sporadic lesions reported in control and treated animals were considered to be an expression of spontaneous and/or incidental pathology or are commonly seen in this species and age.

Spermatogenic cycle:
Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was noted.
Histopathological findings: neoplastic:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
The outcome of mating performance (the mean time between pairing to mating, copulatory and fertility indices) did not show differences related to treatment.

Details on results (P0)

Three non-pregnant females were recorded, 2 in the mid-dose group and 1 in the high dose group.
At necropsy, unilateral implantation was recorded in one mid-dose animal, although no presence of offspring was detected in the cage tray during parturition check. However, based on the red discarge increment in the body weight during gestation and the lower body weight at necropsy (Day 26 post coitum), it was considered that the animal was pregnant and gave birth. The number of females with live pups on Day 4 post partum was 10 in each of the control and low dose groups, 7 in the mid-dose group and 9 in the high dose group.
Green staining of the tail was the main clinical sign recorded in all high dose animals and occasionally in the mid-dose group.
In addition, tooth missing and emaciated appearance was recorded in one high dose female during gestation. Reduced body weight was indeed observed in this animal that then recovered and no other signs were recorded.
At observation of the cage tray, green staining was observed in the high dose group. This staining was explained by the colour of the test item which was likely excreted in the urine and faeces.

Observation of treated animals at removal from the cage and in an open arena did not reveal significant changes when compared to controls.
Motor activity, grip strength and sensory reaction to stimuli measurements recorded at the end of treatment did not show differences of toxicological relevance between control and treated groups of both sexes. The significant differences of lower grip strength in Group 2 or reduced landing food splay in high dose females (Group 4) were considered incidental.

No differences in body weight were recorded between groups of both sexes throughout the study.
Some significant differences in body weight gain were recorded in high dose females before pairing (decreased and increased mean values on Days 8 and 15, respectively), compared to the control group.

Food consumption was also comparable between groups throughout the study.

Slight reticulocytosis was recorded in males dosed at 1000 mg/kg body weight/day (27% above controls), with no related changes in the other red blood cell parameters. In addition, leucocytosis was recorded in two high dose males. Leucocytes were approximately 47% above controls. The other statistically significant differences between control and treated males (mean corpuscular volume and mean corpuscular haemoglobin) were of minimal severity (up to 7%), therefore they were considered to be incidental findings.
Prothrombin time and activated thromboplastin time were slightly decreased in two males dosed at 250 mg/kg body weight/day. Due to the absence of dose-relation,
these changes were considered incidental.
Compared with mean control data, one high dose male (1000 mg/kg body weight/day) showed increase of alanine and aspartate aminotransferases (approximately 2.1 fold), glucose (1.6 fold) and bile acids (6.5 fold). Due to the minimal incidence, this change cannot be conclusively attributed to treatment. Increased glucose was also recorded in other males dosed at 62.5 and 1000 mg/kg body weight/day. Compared with mean control data, the increments were 1.3 and 1.4 fold, respectively.

No relevant changes were found in terminal body weight or organ weights between groups.
Significant reduction in relative weights to terminal body weight of prostate in the low and high dose groups were obtained. All these findings were not considered of toxicological importance.

Animals killed at termination did not show relevant macroscopic changes that could be treatment-related. The changes observed such as green staining of tail or green content of stomach were attributed to the colour of the test item, while enlarged liver in rats of both sexes or reduced thymus in control and treated females are suggested to be incidental and/or due to physiological pregnancy changes, often seen in this kind of study in some treated Sprague Dawley rats of the same age.

No treatment-related changes were noted at histopathological examination.
The sporadic lesions reported in control and treated animals were considered to be an expression of spontaneous and/or incidental pathology or are commonly seen in this species and age.
Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was noted.

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects

Target system / organ toxicity (P0)

Key result
Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No remarkable differences were noted in the clinical signs of pups sacrificed at term between groups. Similar findings were recorded which included pale appearance, cold to touch, apparently no food intake and small in size. These findings were considered unrelated to treatment. Left damaged hindlimb was recorded in one control pup. Missing and/or found dead pups were also recorded during the observation period in all groups.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
All organs autolysed in abdominal and/or thoracic cavities were noted at necropsy in the deceased pups of all groups without substantial differences.
At final sacrifice on Day 4 post partum, malrotated left hindlimb was recorded in one control pup (Dam no. 7) and no milk in the stomach was recorded in two other control pups (Dam nos. 7 and 17). One low dose pup of dam no. 29 showed dark area in the muzzle. All these findings were considered incidental. No abnormalities were recorded in the mid- and high dose groups.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Mean litter and pup weights were comparable between groups on Days 1 and 4 post partum.
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:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment-related findings were recorded at necropsy of deceased pups or those sacrificed at term.

All organs autolysed in abdominal and/or thoracic cavities were noted at necropsy in the deceased pups of all groups without substantial differences. One control pup of dam no. 7 showed malrotated left hindlimb. No milk in the stomach was also reported in the control group. One low dose pup of dam no. 29 showed dark area in the muzzle. All these findings were considered incidental. No abnormalities were recorded in the mid- and high dose groups.
Histopathological findings:
not examined

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Effect levels (F1)

Key result
Dose descriptor:
NOEL
Generation:
F1
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects

Target system / organ toxicity (F1)

Critical effects observed:
no

Overall reproductive toxicity

Key result
Reproductive effects observed:
no

Applicant's summary and conclusion

Conclusions:
In conclusion, no toxic effects of the tested substance were seen after repeated dose and on reproduction/ development in Sprague Dawley rats.
On the basis of the results obtained in this study, the NOAEL (No Observed Adverse Effect Level) for both general toxicity and reproduction/developmental toxicity was considered to be above 1000 mg/kg body weight/day for males and females of the parental and F1 generation.
Executive summary:

A combined repeat-dose/reproductive screening study was performed to obtain information on the possible toxic effects on Sprague Dawley rats of both sexes, after repeated dosing, as well as any effects of the test item on male and female reproductive performance, such as gonadal function, conception, parturition and early lactation. Recovery from any treatment-related effects was also investigated during a period of 2 weeks.

The test item, formulated in sesame oil, was administered orally, by gavage, at the dosages of 62.5, 250 and 1000 mg/kg body weight/day.

Reproductive/Developmental toxicity

Thein vivoparameters did not show signs of toxicological importance. Green staining of the tail was mainly recorded in high dose animals receiving 1000 mg/kg body weight/day and occasionally in the mid-dose group (250 mg/kg body weight/day), and was related to the colour of the test item (dark green textile dye). In addition, green staining in the cage tray was recorded likely due to the compound excreted with urine and/or faeces.

Furthermore, neurotoxicity assessment including functional tests such as hindlimb landing foot splay, sensory reactivity tostimuliincluding grip strength and motor activity did not show differences of toxicological relevance between control and treated groups in the animals of both sexes.

There were no relevant effects on body weight or food consumption. The significant differences of body weight gain recorded before pairing in females of the high dose group on Day 8 (decrease) and on Day 15 (increase) of study, respectively, were considered to represent biological variability and hence not to be of toxicological importance.

No findings of toxicological relevance were recorded at haematological and biochemistry investigations.

Concerning the reproductive parameters, no relevant differences were found in terms of mating performance including the pre-coital interval (number of days paired to sperm positive day) and the copulatory evidence (the positive identification of mating i.e. the presence of sperm and/or copulation plugin situor in the cage). The resulting copulatory index, calculated as the percentage of animals mated respect to those paired, was comparable between groups.

No significant differences were found in terms of number of implantation sites and/or corpora lutea between the groups. All dams had comparable length of gestation and gave birth on Day 22post coitumas mean value Furthermore, sex ratio calculated as the percentage of males in the litter did not differ between the groups.

No effects were seen onbodyweight of pups at birth and on Day 4post partumand no abnormalities were detected in pups sacrificed at term. Similar findings were found in decedent pups of all groups.

The macroscopic observation of organs did not show relevant findings. The changes observed such as green staining of tail or green content of stomach were attributed to the colour of the test item, while enlarged liver in rats of both sexes or reduced thymus in control and treated females are suggested to be incidental and/or due to physiological pregnancy changes, often seen in this kind of study in some treated Sprague Dawley rats of the same age. No remarkable alterations in the organ weights were found that could be considered treatment-related.

No treatment-related changes were noted at histopathological examination including the evaluation of the spermatogenic cycle.

Repeated dose - Recovery groups

Thein vivoparameters did not reveal signs of toxicity. No signs of toxicological importance were recorded at daily clinical observation including the neurotoxicity assessment. Green staining of the tail was also recorded in the animals dosed at 1000 mg/kg body weight/day during treatment and recovery phases as well as green staining in the cage tray was recorded during the treatment phase. Body weight and food consumption did not significantly differ between groups during treatment and recovery phases.

At clinical pathology investigation, changes recorded during the dosing phase showed reversibility. Other significant differences of slight leucopenia in males or increased mean corpuscular haemoglobin concentration in females were considered incidental findings of no toxicological importance.

No treatment-related changes were recorded at macroscopic examination, including organ weights.

Histopathological evaluation was not conducted as no adverse findings were noted in main group animals.

In conclusion, no toxic effects of the tested substance were seen after repeated dose and on reproduction/development in Sprague Dawley rats.

On the basis of the results obtained in this study, the NOAEL (No Observed Adverse Effect Level) for both general toxicity and reproduction/developmental toxicity was considered to be 1000 mg/kg body weight/day for males and females of the parental and Fl generation.