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Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Oral: NOAEL (rat – general toxicity and reproductive/developmental toxicity): 200 mg/kg bw/day ; male/female, OECD TG 421, 2019

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04-06-2018 to 08-11-2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 870.3550
Version / remarks:
US EPA OPPTS 870.3550, Reproduction/Developmental Toxicity Screening Test, (2000)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: Not applicable.
- Specific activity: Not applicable.
- Locations of the label: Not applicable.
- Expiration date of radiochemical substance: Not applicable.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: (i) Prior to formulation: In refrigerator (2-8°C) protected from light container flushed with nitrogen ; (ii) Post-formulation: Dosing formulations were prepared weekly as a solution, formulated in daily portions and stored in the refrigerator protected from light and flushed with nitrogen. Test item dosing formulations were kept at room temperature until dosing. The dosing formulations were removed from the refrigerator and stirred at room temperature for at least 30 minutes before dosing. Dosing was done within 2 hours after the formulations were taken out of the refrigerator.
- Stability under test conditions: Stable.
- Solubility and stability of the test substance in the solvent/vehicle: During the course of the study at two occasions during the treatment phase, stability of the prepared formulation was determined at 2 hours at room temperature and at one occasion 7 days in a refrigerator under protection from light and flushed with nitrogen. Duplicate sets of each sample (approximately 500 mg) were sent to the analytical laboratory. Stability results were considered acceptable if the sample analysis results were within or equal to ±10% of the concentration determined by the initial analysis of each formulation. Analysis of Group 2 and Group 4 formulations after storage yielded a relative difference of ≤ 10%. The test item was considered stable in vehicle formulations during storage at room temperature protected from light for at least 2 hours and flushed with nitrogen and/or in a refrigerator (2-8°C) for 7 days, flushed with nitrogen.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not applicable.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Not applicable. Substance was a liquid fully soluble in vehicle. Dosing formulations were prepared weekly as a solution, formulated in daily portions and stored in the refrigerator protected from light and flushed with nitrogen. Test item dosing formulations were kept at room temperature until dosing. The dosing formulations were removed from the refrigerator and stirred at room temperature for at least 30 minutes before dosing.
- Preliminary purification step (if any): Not applicable.
- Final dilution of a dissolved solid, stock liquid or gel: Not applicable.
- Final preparation of a solid: Not applicable.

FORM AS APPLIED IN THE TEST (if different from that of starting material): Liquid formulations in vehicle.

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable): Not applicable.

OTHER SPECIFICS: Not applicable.
Species:
rat
Strain:
other: Crl: WI(Han)
Details on species / strain selection:
The species and strain was selected in accordance with the OECD TG 421 and the other relevant guidelines.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier (reported in the full study report)
- Females (if applicable) nulliparous and non-pregnant: Yes.
- Age at study initiation: (P) Males 10 – 11 weeks ; Females 13 – 14 weeks
- Weight at study initiation: (P) Males: 263 - 305 g; Females: 197 - 243 g
- Fasting period before study: No.
- Housing: On arrival and following the pretest (females only) and pre-mating period, animals were group housed (up to 5 animals of the same sex and same dosing group together) in polycarbonate cages (Macrolon MIV type). During the mating phase, males and females were cohabitated on a 1:1 basis in Macrolon plastic cages (MIII type). During the post-mating phase, males were housed in their home cage (Macrolon MIV type) group housed up to 5 per cage. Females were individually housed in Macrolon plastic cages (MIII type). During the lactation phase, females were housed in Macrolon plastic cages (MIII type). Cages contained appropriate and were equipped with water bottles.
- Use of restrainers for preventing ingestion (if dermal): Not applicable.
- Diet (e.g. ad libitum): Certified pelleted and powdered diet, ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Males: nine days before commencement of treatment. Females: 8 days before commencement of treatment.

DETAILS OF FOOD AND WATER QUALITY: Feed: Certified pelleted and powdered diet, ad libitum – batch numbers and certificates of analysis provided in the full study report. The diet, drinking water, bedding and environmental enrichment were considered not to contain any contaminant at a level that might have affected the purpose or integrity of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 – 24 (actual daily mean: 20 to 22)
- Humidity (%): 40 – 70 (actual daily mean: 48 to 74%, this deviation in the maximum was not considered to have affected the integrity of the study or clinical condition of the males/females)
- Air changes (per hr): > 10 per hour (no recirculation)
- Photoperiod (hrs dark / hrs light): 12 h light / 12 h dark

IN-LIFE DATES: From: 2018-07-26 To: 2018-09-24
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Remarks:
Polyethylene glycol 300 (or: PEG 300)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing formulations were prepared weekly as a solution, formulated in daily portions and stored in the refrigerator protected from light and flushed with nitrogen. The stability and homogeneity of the test item formulations were determined. The formulations were determined to be acceptably stable and homogenous.

DIET PREPARATION
- Rate of preparation of diet (frequency): Not applicable.
- Mixing appropriate amounts with (Type of food): Not applicable.
- Storage temperature of food: Not applicable.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Applicant assessment indicates: PEG 300 was considered as appropriate based on test item solubility and use in preceding studies.
- Concentration in vehicle: Dosing formulations were prepared freshly or weekly as a solution. For Days 1 and 2 of dosing, freshly prepared formulations were supplied for dosing (they were kept at room temperature until use). All subsequent dosing formulations were prepared beforehand in daily portions and stored in the refrigerator until the day of use. Based on stability data, the formulations were stored in the refrigerator for a maximum of 7 days. After removal from the refrigerator, the formulations were acclimatised at room temperature and stirred for at least 30 minutes before dosing. Dosing was done within 2 hours after the formulations were prepared or taken out of the refrigerator with continuous stirring prior to use. Concentrations in vehicle were as follows: 0 (control) mg/kg/bw [or 0 mg/mL], 60 mg/kg/bw [or 12 mg/mL], 200 mg/kg/bw [or 40 mg/mL], 600 mg/kg/bw [or 120 mg/mL]. The test item concentrations for each group are indicated in table 1. Dose-formulations were analysed during the study and were reported as with ± 10% applied limits.
- Amount of vehicle (if gavage): Treatment volume was 5 mL/kg for control (negative, untreated group) and all treatment groups with applicable test item concentrations per group.
- Purity: Information on supplier is provided in the full study report.
- Other: Dose-formulations were analysed during the study and were reported as with ± 10% applied limits. With the exception: The formulation of Group 4 was not homogeneous (i.e. coefficient of variation of 15%). A comprehensive evaluation of the cause of the deviation of the Group 4 formulation from target concentration and homogeneity revealed the following: (1) the analysis was performed in samples taken from freshly prepared formulations, (2) A correct amount of test item en vehicle were used to prepare to formulation and (3) A large difference between each of the duplicate samples at all sampling heights was observed.
Based on these findings, it was considered that the low accuracy at the high concentration was probably related to incomplete dissolution of the test item in the vehicle in freshly prepared formulations. Therefore, sampling and subsequent analysis of another Group 4 formulation that had been stored for at least 24 hours in the refrigerator was performed. The concentrations analysed in the Group 4 formulation that had been stored in the refrigerator were in agreement with target concentrations (i.e. mean accuracies between 90% and 110% and coefficient of variation of 10%). Fresh formulations were used for dosing the animals on Day 1 and 2 of treatment only. On all other dosing occasions, formulations that had been stored for one or more days in the refrigerator were used. Therefore, it was considered that dosing formulations containing incompletely dissolved test item might only have been used on the first 2 days of dosing. It was considered that the impact on the study was minimal.
Details on mating procedure:
M/F ratio per cage: 1:1 within the same treatment groups.
- Length of cohabitation: up to 2 weeks
- Proof of pregnancy: Confirmed by evidence of sperm in the vaginal lavage or by the appearance of an intravaginal copulatory plug. Day 0 post-coitum was when positive evidence of mating was detected (daily checks).
- Further matings after unsuccessful attempts: No.
- After successful mating each pregnant female was caged (how): Females were individually housed in Macrolon plastic cages (MIII type)
- Any other deviations from standard protocol: No
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
- The homogeneity and stability was confirmed once during the study to assess accuracy, homogeneity and stability over 2 hours at room temperature and 7 days in a refrigerator. Chemical analysis for Group 4 (600 mg/kg bw/day) were performed twice to assess accuracy, homogeneity and stability over 2 hours at room temperature.
- The analysis was performed in a validated analytical procedure. This consisted of UPLC-UV quantitative analysis with external calibration within a dedicated formulation analysis report reference within the full study report. Five calibration solutions in the concentration range of 2.0 – 40 mg/L were prepared from two stock solutions. The end solution of the calibration solutions was 50/50 (v/v) acetonitrile/water. Analysis was conducted in duplicate per concentration. Separately, approximately 500 mg vehicle was accurately weighed and spiked with the test item at a target concentration of 10.7 or 107 mg/g. The QC samples were prepared in volumetric flasks of 20 mL. The flasks were filled up to mark with acetonitrile. The solutions were 100-fold further diluted with 50/50 (v/v) acetonitrile/water to obtain concentrations within the calibration range. These were then subjected to analysis by UPLC-UV. The analytical method was validated (details available within the full study report). With LOQ = 10.7 mg/g in vehicle; coefficient of correlation (r) = > 0.99 and repeatability (n=10) of CoV < 6%. Accuracy and precision was confirmed in the range 85 – 115%. Mean procedural recovery was 98% (n=5) at 10.7 mg/g and 101% (n=5) at 108 mg/g. Samples were determined to be adequately stable.
- Mean concentrations of dose-formulations analysed during the study were within ± 10% applied limits confirming accurate test item/vehicle formulation.
- Other: The formulation of Group 2 was homogeneous (i.e. coefficient of variation ≤ 10%). The formulation of Group 4 was not homogeneous (i.e. coefficient of variation of 15%). Therefore a second set of Group 4 was measured. This second set fell within the criterion of ≤ 10%. For further information see ‘details on exposure’.
Duration of treatment / exposure:
Males: minimum of 14 days prior to mating and during the mating period and up to termination/necropsy (ca. 29 days)
Females: 14 days prior to mating (with the objective to cover at least two complete estrous cycles), the variable time to conception, the duration of pregnancy and at least 14-16 days after delivery, up to and including the day before scheduled necropsy (50 to 56 days ; females that failed to deliver or had total litter lost were treated for 42 to 44 days).
Frequency of treatment:
Daily; at approximately the same time each day.
F1 generation were not dosed.
Details on study schedule:
- Age at study initiation: (P) Males 10 – 11 weeks ; Females 13 – 14 weeks
- Age at mating of the mated animals in the study: F0 generation: Males: ca. 12 – 13 weeks ; Females: 15 - 16 weeks. (i.e. after two weeks treatment).
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Group 1 - Vehicle Control Group
Dose / conc.:
60 mg/kg bw/day (nominal)
Remarks:
Group 2 - Low Treatment Group
Dose / conc.:
200 mg/kg bw/day (nominal)
Remarks:
Group 3 - Intermediate Treatment Group
Dose / conc.:
600 mg/kg bw/day (nominal)
Remarks:
Group 4 - High Treatment Group
No. of animals per sex per dose:
10 per sex per dose (10 male / 10 female)
F1 generation were not dosed.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were based on the results of a previously conducted: (i) 14-day range finding study by dietary route which determined unpalatability (decreased food consumption) that dietary route of exposure was not practicable ; (ii) 5-day oral gavage range finder and (iii) 28-day toxicity study by the oral gavage route (report numbers attached to and cited in the full study report). The dose levels selected for the study were the same as used in the previous study.
- Rationale for animal assignment (if not random): Randomly assigned
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily (and during acclimatisation period, at least once daily).

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: F0: At least once daily ; during dosing period: after dosing

BODY WEIGHT: Yes
- Time schedule for examinations: F0 males: before dosing, day 2, 3, 4, 5, and 8. Weekly thereafter plus day of necropsy ; F0 females: before dosing, day 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation PND 1, 4, 7, 10 and 13 plus day of necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Not applicable.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not applicable.
- Other: Food consumption was monitored during the study: F0 males: day 1, 5, 8 and then quantitatively weekly ; except for males and females during mating and/or females without evidence of mating. F0 females post-mating: day 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation PND 1, 4, 7, and 13. Mean daily consumption per animal (g/animal/day) was calculated for each phase.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not applicable.
- Time schedule for examinations: Not applicable.

OTHER:
1, See ‘FOOD CONSUMPTION’ field.
2. THYROID HORMONE ANALYSIS: Yes
- Time schedule: F0 males/females: on day of necropsy ; F1: 2 per litter PND 4 and PND 14 to 16.
- Assessment: T4 assessment ; T3/TSH assessment if treatment related changes were noted.
Oestrous cyclicity (parental animals):
Estrous cycles were evaluated by examining the vaginal cytology of samples obtained by vaginal lavage.

- Daily performed for all F0 females beginning 14 days prior to treatment (pretest), first 14 days of treatment, during mating until evidence of copulation.
- Daily performance for those females with no evidence of copulation until termination of mating period.
- Final vaginal lavage taken on day of necropsy (except for females that indicated spontaneous mortality).
Sperm parameters (parental animals):
Parameters examined in F0 male parental generations:
testis weight, epididymis weight and histology/histopathology

For the testes of all males of Groups 1 and 4, a detailed qualitative histopathology examination was made, taking into account the tubular stages of the spermatogenic cycle. The epididymis was also subject to histology.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: No.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: clinical observations, litter size, sex ratio, survival indices, ano-genital distance, body weight change, presence of nipple/areolae count in male offspring , macroscopic pathology / abnormalities. Histopathology. Thyroid Hormone Analysis.

GROSS EXAMINATION OF DEAD PUPS:
Yes, where required or if applicable.

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: No.

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: No.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: F0 Males: minimum after 28 days treatment.
- Maternal animals: F0 females: (post-natal day) PND 14 to 16 ; F0 females failing to deliver: minimum Day 25 post-coitum ; F0 females with total litter loss: within 24 hours of last pup death or missing

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera. With special attention to reproductive organs.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues were prepared for microscopic examination and weighed, respectively. For: Cervix, Epididymis, Gland, coagulation , Gland, mammary , Gland, parathyroid , Gland, pituitary, Gland, prostate, Gland, seminal vesicle , Gland, thyroid , Gross lesions/masses, Ovaries, Testes, Uterus and Vagina
The organ weights were taken for: Epididymis, Gland, coagulation (together with seminal vesicles), Gland, parathyroid (together with thyroid), Gland, prostate , Gland, seminal vesicle, Gland, thyroid, Testes
For bilateral organs, left and right organs were weighed together, unless specified above. Requisite organs were weighed at scheduled intervals.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at typically either PND > 4 and < 7 (two pups per litter) and/or remaining: PND 14 to 16.
- In addition to blood collection the F1 offspring were subjected to postmortem examinations (macroscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera. Particular attention was paid to the external genitalia and reproductive organs. Thyroids were retained from one male and one female offspring in each litter on PND 14 to 16.

HISTOPATHOLOGY / ORGAN WEIGHTS
The thyroid tissues were prepared for microscopic examination from one male and one female offspring in each litter. Organ weights were not performed for F1 males or F1 females.
Statistics:
All statistical tests were conducted at the 5% significance level. All pairwise comparisons were conducted using two sided tests and were reported at the 1% or 5% levels.
The following pairwise comparisons were made:
Group 2 vs. Group 1
Group 3 vs. Group 1
Group 4 vs. Group 1
This was subject to parmetric, non-parametric statistical analysis using: Dunnetts-test and/or Steel-test as applicable.
Fishers exact test was used to compare all groups.
Reproductive indices:
Mating index (%) ; Precoital time ; Fertility index (%) ; Gestation index (%) ; Duration of gestation ; Number of Implantation Sites
Offspring viability indices:
Parturition/Maternal Care ; Post-implantation survival index (%) ; Viability index (%) ; Live Birth index (%) ; Percent live males at first litter check (%) ; Percent live females at first live litter check (%) ; Viability Index (%) , Lactation Index (%) and Sex Ratio
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: piloerection (1 female week 1 to 3 ; 7 females week 5 to 7), yellow discolouration of urine (majority of males and females ; in week 1 (males) and 2 (females)). Where piloerection was noted, it was observed for periods of approximately 1-12 days.

Salivation (seen in control, 60, 200 and 600 mg/kg bw/day) was considered not toxicologically relevant. Taking into account nature and severity and time of occurrence (after dosing) and was considered related to administration of the test item.

Other incidental findings: scabs, alopecia, red staining, exophthalmos, lethargy, hunched posture, laboured respiration, rales and a lean posture. These findings were considered as within the normal background range for the species/strain and therefore of no toxicological relevance.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
No mortality occurred during the study period that was considered to be related to treatment with the test item.

At 600 mg/kg bw/day dose level: one female: was for a large part cannibalized (including reproductive organs). The thyroid glands did not show any microscopic changes. Clinical signs noted for this female were rales of low severity in the days before its death and slight body weight loss was observed for this female from day 2 to 4 of treatment.

At 60 mg/kg bw/day dose level: one female. No relevant clinical signs (salivation was noted on Day 2 and 3) were observed and body weights and food consumption were in normal range. There were no macroscopic findings except advanced autolysis and the examined thyroid gland and ovaries, uterus, cervix and vagina did not have abnormal microscopic findings.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: Males: statistically significantly reduced body weight gain was observed from Day 5 of treatment onwards, resulting in approximately 7% lower body weights at termination. The body weights of these males were statistically significantly lower on days 22 and 29 of treatment when compared to controls. ; Females: Minimal body weight loss (maximally 2%) was observed in all groups, including controls, over the four days of treatment. On Day 5, the females of the groups had returned to their start weight, except for those of the high dose group. A further body weight loss was noted in these latter females, with a maximum loss of approximately 4% on Day 8. At start of mating (Day 15 of treatment), they had returned to their start weight, but their body weight were still (approximately 3%) lower compared to controls. At start of mating (Day 15 of treatment), they had returned to their start weight, but their body weight were still (approximately 3%) lower compared to controls. Although body weight gain during post coitum was comparable to controls, the difference in body weights after delivery had increased to slightly more than 10%, being lower in high dose females than in controls. During the progress of lactation, the body weight gain in females at 600 mg/kg became higher than that in controls, achieving a level of statistical significance on Day 13 of lactation. This resulted in similar body weights in females of all groups at termination.

At 200 mg/kg bw/day dose level: Males: no noted effect ; Females: Minimal body weight loss (maximally 2%) was observed in all groups, including controls, over the four days of treatment. On Day 5, the females had returned to their start weight.

At 60 mg/kg bw/day dose level: Same as 200 mg/kg bw/day dose level.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: males/females: a slight or markedly reduced treatment related reduction in absolute food consumption was observed over days 1-8, respectively.

Further statistically significant differences in food consumption for females were considered incidental and not related to treatment.
Food efficiency:
not examined
Description (incidence and severity):
See 'Food consumption'
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No effects were observed.
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
See: THYROID HORMONE ANALYSIS
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):
There were no treatment related changes seen in histopathology related to treatment.

All recorded microscopic findings were within the normal range of background gross observations or the species/strain and therefore of no toxicological relevance.

There were no morphological findings in the reproductive organs of either sex which could be attributed to the test item and stage aware evaluation of the testes did not show any indication of abnormal spermatogenesis.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
There were no treatment related changes seen in histopathology related to treatment.

All recorded microscopic findings were within the normal range of background gross observations or the species/strain and therefore of no toxicological relevance.
Other effects:
no effects observed
Description (incidence and severity):
THYROID HORMONE ANALYSIS
1. Serum levels of T4 in F0-males were dose dependently lower by 20%, 38% and 44% at 60, 200 and 600 mg/kg, respectively. All values remained within the historical control range.
2. Serum levels of T4 in F0-females were dose dependently lower by 19%, 23% and 33% at 60, 200 and 600 mg/kg, respectively. All values remained within the historical control range.

Changes in T4 levels were possibly related to the changes in the livers of male and female rats (as previously observed) at similar dose levels in the previously performed 28-day toxicity study (cited within the full study report).

There was no correlates to the Thyroid in gross pathology, organ weights or histopathology. Consequently, T3 and TSH were not further analysed.
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
Length and regularity of the estrous cycle were considered not to have been affected by treatment.

During the pre-mating period, irregular cycling was observed in two females (no.58, at 60 mg/kg and no.74 at 600 mg/kg) and another female (no.76 at 600 mg/kg) being acyclic. In addition, in two females (no.60 at 60 mg/kg and no.72 at 600 mg/kg) it was not possible to determine the regularity of their estrous cycles. Based on the absence of a clear dose response relationship and the fact that also these females became pregnant and delivered offspring, these findings were considered unrelated to treatment and of no toxicological significance.

Gestation length and gestation index were unaffected by treatment.
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
There were no morphological findings in the reproductive organs of either sex which could be attributed to the test item and stage aware evaluation of the testes did not show any indication of abnormal spermatogenesis.
Reproductive performance:
no effects observed
Description (incidence and severity):
There was not effect on fertility index or mating index (mating performance) related to treatment. Precoital time and Number of implantation sites was unaffected by treatment.

Fertility indices were 100, 90, 100 and 100% for the control, 60, 200 and 600 mg/kg groups, respectively.

All females showed evidence of mating.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
body weight and weight gain
Critical effects observed:
no
Clinical signs:
no effects observed
Description (incidence and severity):
There were no treatment related clinical signs considered related to treatment.

For 4 out of 5 pups found dead at first litter check at 60 and 200 mg/kg, no milk was found in the stomach. For all pups of litter no. 76 (600 mg/kg), no milk in the stomach was found on PND 2 and for 8/9 pups, cold to touch was noted. Other findings included blue spots on the head or abdomen and scabs on the snout.

The nature and incidence of these and other clinical signs remained within the range considered normal for pups of this age, and were therefore not considered to be toxicologically relevant.
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
There was no treatment related mortality associated with treatment within F1 males or females.

For 4 out of 5 pups found dead at first litter check at 60 and 200 mg/kg, no milk was found in the stomach. For all pups of litter no. 76 (600 mg/kg), no milk in the stomach was found on PND 2 and for 8/9 pups, cold to touch was noted. Other findings included blue spots on the head or abdomen and scabs on the snout.

The nature and incidence of these and other clinical signs remained within the range considered normal for pups of this age, and were therefore not considered to be toxicologically relevant.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: Body weights of pups were lower from postnatal Day (PND) 1 in males, females and for males and females combined. Body weights were approximately 9-13% lower on PND 1 and approximately 13-18% lower on the following days up to PND 13 when compared with the concurrent controls.

All remaining recorded macroscopic findings were within the normal range of background gross observations or the species/strain and therefore of no toxicological relevance.
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:
no effects observed
Description (incidence and severity):
See: THYROID HORMONE ANALYSIS
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no changes seen macroscopically in offspring considered related to treatment.

Anogenital distance (absolute and normalized for body weight) in male and females was considered not to be affected by treatment.
Treatment had no effect on areola/nipple retention. No males at PND 13 had nipples.
Histopathological findings:
no effects observed
Description (incidence and severity):
No macroscopic findings were notedthat were considered to be related to treatment.
Other effects:
no effects observed
Description (incidence and severity):
THYROID HORMONE ANALYSIS
Serum T4 levels in PND 14-16 males and females were considered not to be affected by treatment.

In the related gross pathology/histopathology:

The thyroids of the F1 offspring were considered unaffected by treament.

2. Offspring Ano-Genital Distance and Male Nipple Counts
Ano-genital distances for male or female offspring were unaffected by treatment. Treatment had no effect on areola/nipple retention. No males at PND 13 had nipples.
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Reproductive Toxicity:
At up to 600 mg/kg bw/day: no treatment-related changes were noted in any of the reproductive parameters investigated in this study (i.e. mating and fertility indices, precoital time, number of implantations, estrous cycle, spermatogenic profiling, and histopathological examination of reproductive organs).
Developmental Toxicity:
At 600 mg/kg bw/day: A treatment related effect on body weights of pups was observed. As at Day 1 of lactation females had a lower body weight, this may be related to the observed lower body weights of pups. Since the body weights of the F1 females remained lower than controls to normal body weights up to Day 13, this finding was considered adverse. No effects were observed at 200 mg/kg bw/day or lower.
Key result
Dose descriptor:
NOAEL
Remarks:
reproduction toxicity
Generation:
F1
Effect level:
>= 600 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to adverse toxic effects at highest dose / concentration tested
Remarks:
No reproduction toxicity was observed up to the highest dose level tested
Dose descriptor:
NOAEL
Remarks:
developmental toxicity
Generation:
F1
Effect level:
>= 200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
Remarks on result:
other:
Remarks:
treatment related effect on body weights was observed at 600 mg/kg
Critical effects observed:
no
Reproductive effects observed:
no
Conclusions:
Under the conditions of this study, the no-observed-adverse-effect level (NOAEL) for systemic toxicity was considered to be 200 mg/kg bw/day due to clinical signs and reduced body weight gain. For reproductive toxicity the NOAEL for males and females is 600 mg/kg bw/day. For developmental toxicity the NOAEL for males and females is 200 mg/kg bw/day due to reduced body weight gain.
Executive summary:

The study was performed according the requirements of OECD TG 421 guideline under GLP conditions. The purpose of this study was to assess general systemic toxic potential in rats, including a screening test for reproductive / development effects and assessment of endocrine disruptor relevant endpoints, with administration of the test item by oral gavage administration for a least four weeks in Crl:WI(Han) rats. Three groups, each comprising ten male and ten female rats, received test item at doses of 60, 200 or 600 mg/kg/day. A control group of ten males and ten females was similarly dosed with vehicle alone (PEG 300), at the same dose volume as the treated groups (5 mL/Kg). The following parameters and end points were evaluated in this study: mortality/ moribundity, clinical signs, body weight and food consumption, estrous cycle determination, measurement of thyroid hormone T4 (F0-males and females), gross necropsy findings, organ weights and histopathologic examinations. In addition, the following reproduction/developmental parameters were determined: mating and fertility indices, precoital time, number of implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights, sex, anogenital distance, areola/nipple retention and macroscopy, measurement of thyroid hormone T4 (PND 14-16 pups)). Test formulations prepared were considered homogeneous at the concentrations tested and analysis of the accuracy revealed acceptable levels. Test formulations prepared were considered stable, for at least 2 hours at room temperature. Test formulations for Group 2 were also considered stable for 7 days in the refrigerator. The test item doses were there therefore considered based on nominal concentrations.The mortality observed in this study (one mortality at 60 and 600 mg/kg bw/day, respectively) was considered unrelated to treatment, due to a lack of a dose response and replacements to the two decedents indicated comparable effects to treatment to survivors.Within clinical signs: piloerection was noted in females at dose 600 mg/kg and occurred in one female from week 1-3 of treatment and in several females from approximately Week 5 until Week 7 of treatment. Yellow discolouration of the urine was observed in both males and females at dose 600 mg/kg from Week 1 (males) and Week 2 (females) of treatment but was speculated as being related to higher bilirubin concentrations in plasma as found in a previously performed 28-day toxicity study. Lower body weight gain was observed for males at 600 mg/kg during the treatment period, resulting in approximately 7% lower body weights at termination. In females at 600 mg/kg, a higher body weight loss was observed over the first week compared to that in the other groups (maximally 4% versus 2%). Lower body weights were noted in high dose females at start of mating and directly after delivery. Any effect on body weights during the post coitum were difficult to evaluate as they are also affected by the growth of the fetuses. During lactation a higher body weight gain was observed in the high dose females, resulting in similar body weights in females of all groups at termination. Absolute and relative food consumption were treatment relatedly lower in males and females at 600 mg/kg during days 1-8 of treatment. As food consumption recovered to normal levels in the following days, this lower food consumption was considered non-adverse. Serum levels of T4 in F0-males and females were dose dependently reduced at all tested dose levels up to 600 mg/kg. Although all values remained within the historical control range.Changes in T4 levels were possibly related to the changes in the livers of male and female rats (as previously observed) at similar dose levels in the previously performed 28-day toxicity study. There was no correlates to the Thyroid in gross pathology, organ weights or histopathology. Consequently, T3 and TSH were not further analysed. No reproduction toxicity was observed up to the highest dose level tested. Within developmental toxicity, a treatment related effect on body weights of pups was observed at 600 mg/kg bw/day. As at Day 1 of lactation females had a lower body weight, this may be related to the observed lower body weights of pups. Since the body weights of the pups remained lower than controls up to Day 13, this finding was considered adverse. Under the conditions of this study, the no-observed-adverse-effect level (NOAEL) for systemic toxicity was considered to be 200 mg/kg bw/day due to clinical signs and reduced body weight gain. For reproductive toxicity the NOAEL for males and females is 600 mg/kg bw/day. For developmental toxicity the NOAEL for males and females is 200 mg/kg bw/day due to reduced body weight gain.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
600 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The key study is GLP compliant and of a high quality (Klimisch 1); The available information as a whole meets the tonnage driven information requirements of REACH.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Key study : Reproductive/developmental screening study – Oral: OECD TG 421, 2019 : The study was performed according the requirements of OECD TG 421 guideline under GLP conditions. The purpose of this study was to assess general systemic toxic potential in rats, including a screening test for reproductive / development effects and assessment of endocrine disruptor relevant endpoints, with administration of the test item by oral gavage administration for a least four weeks in Crl:WI(Han) rats. Three groups, each comprising ten male and ten female rats, received test item at doses of 60, 200 or 600 mg/kg/day. A control group of ten males and ten females was similarly dosed with vehicle alone (PEG 300), at the same dose volume as the treated groups (5 mL/Kg). The following parameters and end points were evaluated in this study: mortality/ moribundity, clinical signs, body weight and food consumption, estrous cycle determination, measurement of thyroid hormone T4 (F0-males and females), gross necropsy findings, organ weights and histopathologic examinations. In addition, the following reproduction/developmental parameters were determined: mating and fertility indices, precoital time, number of implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights, sex, anogenital distance, areola/nipple retention and macroscopy, measurement of thyroid hormone T4 (PND 14-16 pups)). Test formulations prepared were considered homogeneous at the concentrations tested and analysis of the accuracy revealed acceptable levels. Test formulations prepared were considered stable, for at least 2 hours at room temperature. Test formulations for Group 2 were also considered stable for 7 days in the refrigerator. The test item doses were there therefore considered based on nominal concentrations.The mortality observed in this study (one mortality at 60 and 600 mg/kg bw/day, respectively) was considered unrelated to treatment, due to a lack of a dose response and replacements to the two decedents indicated comparable effects to treatment to survivors.Within clinical signs: piloerection was noted in females at dose 600 mg/kg and occurred in one female from week 1-3 of treatment and in several females from approximately Week 5 until Week 7 of treatment. Yellow discolouration of the urine was observed in both males and females at dose 600 mg/kg from Week 1 (males) and Week 2 (females) of treatment but was speculated as being related to higher bilirubin concentrations in plasma as found in a previously performed 28-day toxicity study. Lower body weight gain was observed for males at 600 mg/kg during the treatment period, resulting in approximately 7% lower body weights at termination. In females at 600 mg/kg, a higher body weight loss was observed over the first week compared to that in the other groups (maximally 4% versus 2%). Lower body weights were noted in high dose females at start of mating and directly after delivery. Any effect on body weights during the post coitum were difficult to evaluate as they are also affected by the growth of the fetuses. During lactation a higher body weight gain was observed in the high dose females, resulting in similar body weights in females of all groups at termination. Absolute and relative food consumption were treatment relatedly lower in males and females at 600 mg/kg during days 1-8 of treatment. As food consumption recovered to normal levels in the following days, this lower food consumption was considered non-adverse. Serum levels of T4 in F0-males and females were dose dependently reduced at all tested dose levels up to 600 mg/kg. Although all values remained within the historical control range.Changes in T4 levels were possibly related to the changes in the livers of male and female rats (as previously observed) at similar dose levels in the previously performed 28-day toxicity study. There was no correlates to the Thyroid in gross pathology, organ weights or histopathology. Consequently, T3 and TSH were not further analysed. No reproduction toxicity was observed up to the highest dose level tested. Within developmental toxicity, a treatment related effect on body weights of pups was observed at 600 mg/kg bw/day. As at Day 1 of lactation females had a lower body weight, this may be related to the observed lower body weights of pups. Since the body weights of the pups remained lower than controls up to Day 13, this finding was considered adverse. Under the conditions of this study, the no-observed-adverse-effect level (NOAEL) for systemic toxicity was considered to be 200 mg/kg bw/day due to clinical signs and reduced body weight gain. For reproductive toxicity the NOAEL for males and females is 600 mg/kg bw/day. For developmental toxicity the NOAEL for males and females is 200 mg/kg bw/day due to reduced body weight gain.

Effects on developmental toxicity

Description of key information

Oral: NOAEL (rat – general toxicity and reproductive/developmental toxicity): 200 mg/kg bw/day ; male/female, OECD TG 421, 2019

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04-06-2018 to 08-11-2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 870.3550
Version / remarks:
US EPA OPPTS 870.3550, Reproduction/Developmental Toxicity Screening Test, (2000)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: Not applicable.
- Specific activity: Not applicable.
- Locations of the label: Not applicable.
- Expiration date of radiochemical substance: Not applicable.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: (i) Prior to formulation: In refrigerator (2-8°C) protected from light container flushed with nitrogen ; (ii) Post-formulation: Dosing formulations were prepared weekly as a solution, formulated in daily portions and stored in the refrigerator protected from light and flushed with nitrogen. Test item dosing formulations were kept at room temperature until dosing. The dosing formulations were removed from the refrigerator and stirred at room temperature for at least 30 minutes before dosing. Dosing was done within 2 hours after the formulations were taken out of the refrigerator.
- Stability under test conditions: Stable.
- Solubility and stability of the test substance in the solvent/vehicle: During the course of the study at two occasion during the treatment phase, stability of the prepared formulation was determined at 2 hours at room temperature and at one occasion 7 days in a refrigerator under protection from light and flushed with nitrogen. Duplicate sets of each sample (approximately 500 mg) were sent to the analytical laboratory. Stability results were considered acceptable if the sample analysis results were within or equal to ±10% of the concentration determined by the initial analysis of each formulation. Analysis of Group 2 and Group 4 formulations after storage yielded a relative difference of ≤ 10%. The test item was considered stable in vehicle formulations during storage at room temperature protected from light for at least 2 hours and flushed with nitrogen and/or in a refrigerator (2-8°C) for 7 days, flushed with nitrogen.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not applicable.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Not applicable. Substance was a liquid fully soluble in vehicle. Dosing formulations were prepared weekly as a solution, formulated in daily portions and stored in the refrigerator protected from light and flushed with nitrogen. Test item dosing formulations were kept at room temperature until dosing. The dosing formulations were removed from the refrigerator and stirred at room temperature for at least 30 minutes before dosing.
- Preliminary purification step (if any): Not applicable.
- Final dilution of a dissolved solid, stock liquid or gel: Not applicable.
- Final preparation of a solid: Not applicable.

FORM AS APPLIED IN THE TEST (if different from that of starting material): Liquid formulations in vehicle.

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable): Not applicable.

OTHER SPECIFICS: Not applicable.
Species:
rat
Strain:
other: Crl: WI(Han)
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier (reported in the full study report)
- Females (if applicable) nulliparous and non-pregnant: Yes.
- Age at study initiation: (P) Males 10 – 11 weeks ; Females 13 – 14 weeks
- Weight at study initiation: (P) Males: 263 - 305 g; Females: 197 - 243 g
- Fasting period before study: No.
- Housing: On arrival and following the pretest (females only) and pre-mating period, animals were group housed (up to 5 animals of the same sex and same dosing group together) in polycarbonate cages (Macrolon MIV type). During the mating phase, males and females were cohabitated on a 1:1 basis in Macrolon plastic cages (MIII type). During the post-mating phase, males were housed in their home cage (Macrolon MIV type) group housed up to 5 per cage. Females were individually housed in Macrolon plastic cages (MIII type). During the lactation phase, females were housed in Macrolon plastic cages (MIII type). Cages contained appropriate and were equipped with water bottles.
- Use of restrainers for preventing ingestion (if dermal): Not applicable.
- Diet (e.g. ad libitum): Certified pelleted and powdered diet, ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Males: nine days before commencement of treatment. Females: 8 days before commencement of treatment.

DETAILS OF FOOD AND WATER QUALITY: Feed: Certified pelleted and powdered diet, ad libitum – batch numbers and certificates of analysis provided in the full study report. The diet, drinking water, bedding and environmental enrichment were considered not to contain any contaminant at a level that might have affected the purpose or integrity of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 – 24 (actual daily mean: 20 to 22)
- Humidity (%): 40 – 70 (actual daily mean: 48 to 74%, this deviation in the maximum was not considered to have affected the integrity of the study or clinical condition of the males/females)
- Air changes (per hr): > 10 per hour (no recirculation)
- Photoperiod (hrs dark / hrs light): 12 h light / 12 h dark

IN-LIFE DATES: From: 2018-07-26 To: 2018-09-24
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Remarks:
Polyethylene glycol 300 (or: PEG 300)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing formulations were prepared weekly as a solution, formulated in daily portions and stored in the refrigerator protected from light and flushed with nitrogen. The stability and homogeneity of the test item formulations were determined. The formulations were determined to be acceptably stable and homogenous.

DIET PREPARATION
- Rate of preparation of diet (frequency): Not applicable.
- Mixing appropriate amounts with (Type of food): Not applicable.
- Storage temperature of food: Not applicable.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Applicant assessment indicates: PEG 300 was considered as appropriate based on test item solubility and use in preceding studies.
- Concentration in vehicle: Dosing formulations were prepared freshly or weekly as a solution. For Days 1 and 2 of dosing, freshly prepared formulations were supplied for dosing (they were kept at room temperature until use). All subsequent dosing formulations were prepared beforehand in daily portions and stored in the refrigerator until the day of use. Based on stability data, the formulations were stored in the refrigerator for a maximum of 7 days. After removal from the refrigerator, the formulations were acclimatised at room temperature and stirred for at least 30 minutes before dosing. Dosing was done within 2 hours after the formulations were prepared or taken out of the refrigerator with continuous stirring prior to use. Concentrations in vehicle were as follows: 0 (control) mg/kg/bw [or 0 mg/mL], 60 mg/kg/bw [or 12 mg/mL], 200 mg/kg/bw [or 40 mg/mL], 600 mg/kg/bw [or 120 mg/mL]. The test item concentrations for each group are indicated in table 1. Dose-formulations were analysed during the study and were reported as with ± 10% applied limits.
- Amount of vehicle (if gavage): Treatment volume was 5 mL/kg for control (negative, untreated group) and all treatment groups with applicable test item concentrations per group.
- Purity: Information on supplier is provided in the full study report.
- Other: Dose-formulations were analysed during the study and were reported as with ± 10% applied limits. With the exception: The formulation of Group 4 was not homogeneous (i.e. coefficient of variation of 15%). A comprehensive evaluation of the cause of the deviation of the Group 4 formulation from target concentration and homogeneity revealed the following: (1) the analysis was performed in samples taken from freshly prepared formulations, (2) A correct amount of test item en vehicle were used to prepare to formulation and (3) A large difference between each of the duplicate samples at all sampling heights was observed.
Based on these findings, it was considered that the low accuracy at the high concentration was probably related to incomplete dissolution of the test item in the vehicle in freshly prepared formulations. Therefore, sampling and subsequent analysis of another Group 4 formulation that had been stored for at least 24 hours in the refrigerator was performed. The concentrations analysed in the Group 4 formulation that had been stored in the refrigerator were in agreement with target concentrations (i.e. mean accuracies between 90% and 110% and coefficient of variation of 10%). Fresh formulations were used for dosing the animals on Day 1 and 2 of treatment only. On all other dosing occasions, formulations that had been stored for one or more days in the refrigerator were used. Therefore, it was considered that dosing formulations containing incompletely dissolved test item might only have been used on the first 2 days of dosing. It was considered that the impact on the study was minimal.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
- The homogeneity and stability was confirmed once during the study to assess accuracy, homogeneity and stability over 2 hours at room temperature and 7 days in a refrigerator. Chemical analysis for Group 4 (600 mg/kg bw/day) were performed twice to assess accuracy, homogeneity and stability over 2 hours at room temperature.
- The analysis was performed in a validated analytical procedure. This consisted of UPLC-UV quantitative analysis with external calibration within a dedicated formulation analysis report reference within the full study report. Five calibration solutions in the concentration range of 2.0 – 40 mg/L were prepared from two stock solutions. The end solution of the calibration solutions was 50/50 (v/v) acetonitrile/water. Analysis was conducted in duplicate per concentration. Separately, approximately 500 mg vehicle was accurately weighed and spiked with the test item at a target concentration of 10.7 or 107 mg/g. The QC samples were prepared in volumetric flasks of 20 mL. The flasks were filled up to mark with acetonitrile. The solutions were 100-fold further diluted with 50/50 (v/v) acetonitrile/water to obtain concentrations within the calibration range. These were then subjected to analysis by UPLC-UV. The analytical method was validated (details available within the full study report). With LOQ = 10.7 mg/g in vehicle; coefficient of correlation (r) = > 0.99 and repeatability (n=10) of CoV < 6%. Accuracy and precision was confirmed in the range 85 – 115%. Mean procedural recovery was 98% (n=5) at 10.7 mg/g and 101% (n=5) at 108 mg/g. Samples were determined to be adequately stable.
- Mean concentrations of dose-formulations analysed during the study were within ± 10% applied limits confirming accurate test item/vehicle formulation.
- Other: The formulation of Group 2 was homogeneous (i.e. coefficient of variation ≤ 10%). The formulation of Group 4 was not homogeneous (i.e. coefficient of variation of 15%). Therefore a second set of Group 4 was measured. This second set fell within the criterion of ≤ 10%. For further information see ‘details on exposure’.
Details on mating procedure:
M/F ratio per cage: 1:1 within the same treatment groups.
- Length of cohabitation: up to 2 weeks
- Proof of pregnancy: Confirmed by evidence of sperm in the vaginal lavage or by the appearance of an intravaginal copulatory plug. Day 0 post-coitum was when positive evidence of mating was detected (daily checks).
- Further matings after unsuccessful attempts: No.
- After successful mating each pregnant female was caged (how): Females were individually housed in Macrolon plastic cages (MIII type)
- Any other deviations from standard protocol: No
Duration of treatment / exposure:
Males: minimum of 14 days prior to mating and during the mating period and up to termination/necropsy (ca. 29 days)
Females: 14 days prior to mating (with the objective to cover at least two complete estrous cycles), the variable time to conception, the duration of pregnancy and at least 14-16 days after delivery, up to and including the day before scheduled necropsy (50 to 56 days ; females that failed to deliver or had total litter lost were treated for 42 to 44 days).
Frequency of treatment:
Daily; at approximately the same time each day.
F1 generation were not dosed.
Duration of test:
- Age at study initiation: (P) Males 10 – 11 weeks ; Females 13 – 14 weeks
- Age at mating of the mated animals in the study: F0 generation: Males: ca. 12 – 13 weeks ; Females: 15 - 16 weeks. (i.e. after two weeks treatment).
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Group 1 - Vehicle Control Group
Dose / conc.:
60 mg/kg bw/day (nominal)
Remarks:
Group 2 - Low Treatment Group
Dose / conc.:
200 mg/kg bw/day (nominal)
Remarks:
Group 3 - Intermediate Treatment Group
Dose / conc.:
600 mg/kg bw/day (nominal)
Remarks:
Group 4 - High Treatment Group
No. of animals per sex per dose:
10 per sex per dose (10 male / 10 female)
F1 generation were not dosed.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were based on the results of a previously conducted 28-day toxicity study (Report number attached to and cited in the full study report). The dose levels selected for the study were the same as used in the previous study.
- Rationale for animal assignment (if not random): Randomly assigned
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily (and during acclimatisation period, at least once daily).

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: F0: At least once daily ; during dosing period: after dosing

BODY WEIGHT: Yes
- Time schedule for examinations: F0 males: before dosing, day 2, 3, 4, 5, and 8. Weekly thereafter plus day of necropsy ; F0 females: before dosing, day 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation PND 1, 4, 7, 10 and 13 plus day of necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Not applicable.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not applicable.
- Other: Food consumption was monitored during the study: F0 males: day 1, 5, 8 and then quantitatively weekly ; except for males and females during mating and/or females without evidence of mating. F0 females post-mating: day 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation PND 1, 4, 7, and 13. Mean daily consumption per animal (g/animal/day) was calculated for each phase.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not applicable.
- Time schedule for examinations: Not applicable.

OTHER:
1, See ‘FOOD CONSUMPTION’ field.
2. THYROID HORMONE ANALYSIS: Yes
- Time schedule: F0 males/females: on day of necropsy ; F1: 2 per litter PND 4 and PND 14 to 16.
- Assessment: T4 assessment ; T3/TSH assessment if treatment related changes were noted.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes (if no visibile implantation sites)
- Number of implantations: Yes
- Number of early resorptions: No
- Number of late resorptions: No
Fetal examinations:
- External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [all per litter]
- Skeletal examinations: Yes: [all per litter]
- Head examinations: Yes: [all per litter]
Statistics:
All statistical tests were conducted at the 5% significance level. All pairwise comparisons were conducted using two sided tests and were reported at the 1% or 5% levels.
The following pairwise comparisons were made:
Group 2 vs. Group 1
Group 3 vs. Group 1
Group 4 vs. Group 1
This was subject to parmetric, non-parametric statistical analysis using: Dunnetts-test and/or Steel-test as applicable.
Fishers exact test was used to compare all groups.

The following data types were analyzed at each timepoint separately:
Body weight, using absolute weights and gains over appropriate study periods
Food consumption, over appropriate study periods during gestation and lactation
Litter size, survival indices and sex ratio
Ano-genital distance
Organ weights, both absolute and adjusted for terminal body weight

Comparisons were performed:
Group 1 vs 2, 3 and 4

Typical statistical analysis included:
parametric analysis was performed if Bartlett's test for variance homogeneity (Bartlett 1937)
For all other comparisons the F1 approximate test was applied. Which included:
parametric monotonic trend test, such as Williams’ test (Williams 1971, 1972)
If the F1 approximate test was significant, Dunnett's test (Dunnett 1955, 1964) was performed instead

Additional tests, included Kruskal-Wallis’ test (Kruskal and Wallis 1952, 1953), Wilcoxon rank sum tests (Wilcoxon 1945) and/or Shirley's test (Shirley 1977) and Steel's test (Steel 1959), as appropriate.

For litter size and survival indices, Fisher’s exact tests (Fisher 1973).

Sex ratio were analyzed equivalent to Lipsitz (Lipsitz 1991). Each treated group was compared to control using a Wald chi-square test.

For organ weight data, analysis of covariance was performed using terminal body weight as covariate (Angervall and Carlstrom, 1963), unless non-parametric methods were applied.
Indices:
Mating index (%) ; Precoital time ; Fertility index (%) ; Gestation index (%) ; Duration of gestation ; Number of Implantation Sites ; Parturition/Maternal Care ; Post-implantation survival index (%) ; Viability index (%) ; Live Birth index (%) ; Percent live males at first litter check (%) ; Percent live females at first live litter check (%) ; Viability Index (%) , Lactation Index (%) and Sex Ratio
Historical control data:
Historical control data was available.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: piloerection (1 female week 1 to 3 ; 7 females week 5 to 7), yellow discolouration of urine (majority of males and females ; in week 1 (males) and 2 (females)). Where piloerection was noted, it was observed for periods of approximately 1-12 days.

Salivation (seen in control, 60, 200 and 600 mg/kg bw/day) was considered not toxicologically relevant. Taking into account nature and severity and time of occurrence (after dosing) and was considered related to administration of the test item.

Other incidental findings: scabs, alopecia, red staining, exophthalmos, lethargy, hunched posture, laboured respiration, rales and a lean posture. These findings were considered as within the normal background range for the species/strain and therefore of no toxicological relevance.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
No mortality occurred during the study period that was considered to be related to treatment with the test item.

At 600 mg/kg bw/day dose level: one female: was for a large part cannibalized (including reproductive organs). The thyroid glands did not show any microscopic changes. Clinical signs noted for this female were rales of low severity in the days before its death and slight body weight loss was observed for this female from day 2 to 4 of treatment.

At 60 mg/kg bw/day dose level: one female. No relevant clinical signs (salivation was noted on Day 2 and 3) were observed and body weights and food consumption were in normal range. There were no macroscopic findings except advanced autolysis and the examined thyroid gland and ovaries, uterus, cervix and vagina did not have abnormal microscopic findings.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: Males: statistically significantly reduced body weight gain was observed from Day 5 of treatment onwards, resulting in approximately 7% lower body weights at termination. The body weights of these males were statistically significantly lower on days 22 and 29 of treatment when compared to controls. ; Females: Minimal body weight loss (maximally 2%) was observed in all groups, including controls, over the four days of treatment. On Day 5, the females of the groups had returned to their start weight, except for those of the high dose group. A further body weight loss was noted in these latter females, with a maximum loss of approximately 4% on Day 8. At start of mating (Day 15 of treatment), they had returned to their start weight, but their body weight were still (approximately 3%) lower compared to controls. At start of mating (Day 15 of treatment), they had returned to their start weight, but their body weight were still (approximately 3%) lower compared to controls. Although body weight gain during post coitum was comparable to controls, the difference in body weights after delivery had increased to slightly more than 10%, being lower in high dose females than in controls. During the progress of lactation, the body weight gain in females at 600 mg/kg became higher than that in controls, achieving a level of statistical significance on Day 13 of lactation. This resulted in similar body weights in females of all groups at termination.

At 200 mg/kg bw/day dose level: Males: no noted effect ; Females: Minimal body weight loss (maximally 2%) was observed in all groups, including controls, over the four days of treatment. On Day 5, the females had returned to their start weight.

At 60 mg/kg bw/day dose level: Same as 200 mg/kg bw/day dose level.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: males/females: a slight or markedly reduced treatment related reduction in absolute food consumption was observed over days 1-8, respectively.

Further statistically significant differences in food consumption for females were considered incidental and not related to treatment.
Food efficiency:
not examined
Description (incidence and severity):
See 'Food consumption'
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No effects were observed.
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
See: THYROID HORMONE ANALYSIS
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no test item-related alterations in organ weights.

All organ weight differences observed, including those that reached statistical significance, were in line with a decreased body weight and unrelated to treatment.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: males: test item-related macroscopic findings in the liver (red-brown discoloration) in 4/10 males. Without microscopic correlation.

All remaining recorded macroscopic findings were within the normal range of background gross observations or the species/strain and therefore of no toxicological relevance.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no treatment related changes seen in histopathology related to treatment.

All recorded microscopic findings were within the normal range of background gross observations or the species/strain and therefore of no toxicological relevance.

There were no morphological findings in the reproductive organs of either sex which could be attributed to the test item and stage aware evaluation of the testes did not show any indication of abnormal spermatogenesis.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
There were no treatment related changes seen in histopathology related to treatment.

All recorded microscopic findings were within the normal range of background gross observations or the species/strain and therefore of no toxicological relevance.
Other effects:
no effects observed
Description (incidence and severity):
THYROID HORMONE ANALYSIS
1. Serum levels of T4 in F0-males were dose dependently lower by 20%, 38% and 44% at 60, 200 and 600 mg/kg, respectively. All values remained within the historical control range.
2. Serum levels of T4 in F0-females were dose dependently lower by 19%, 23% and 33% at 60, 200 and 600 mg/kg, respectively. All values remained within the historical control range.

Changes in T4 levels were possibly related to the changes in the livers of male and female rats (as previously observed) at similar dose levels in the previously performed 28-day toxicity study (cited within the full study report).

There was no correlates to the Thyroid in gross pathology, organ weights or histopathology. Consequently, T3 and TSH were not further analysed.
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
The total number of offspring born compared to the total number of uterine implantations was considered not to be affected by treatment.
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
Gestation index and duration of gestation were considered not to be affected by treatment
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
There was no treatment related effect to estrous cyclicity, pre-coital interval, fertility and mating performance by treatment. Litter size, offspring survival was not affected by treatment.

Within viability index: at 600 mg/kg bw/day there was some evidence in one litter at day 1 to 4 of poor maternal care.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 200 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
clinical signs
Abnormalities:
no effects observed
Description (incidence and severity):
No treatment related Adverse Effects observed up to the maximum dose level
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 600 mg/kg bw/day dose level: Body weights of pups were lower from postnatal Day (PND) 1 in males, females and for males and females combined. Body weights were approximately 9-13% lower on PND 1 and approximately 13-18% lower on the following days up to PND 13 when compared with the concurrent controls.

All remaining recorded macroscopic findings were within the normal range of background gross observations or the species/strain and therefore of no toxicological relevance.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
Live birth index (number of live offspring on PND 1 as percentage of total number of offspring born) was considered not to be affected by treatment.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
Sex ratio was considered not to be affected by treatment.

Sex ratios were 42/58, 49/51, 54/46, 47/53 for the control, 60, 200 and 600 mg/kg groups, respectively.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
The total number of offspring born compared to the total number of uterine implantations was considered not to be affected by treatment.

Litter size was considered not to be affected by treatment.

Changes in postnatal survival:
no effects observed
Description (incidence and severity):
The number of live offspring on Day 4 before termination compared to the number of offspring on Day 1 was considered not to be affected by treatment.

The mortality among pups up to Day 4 occurred in one control litter and three litters at 600 mg/kg. The distribution of the mortality over the dose groups did not indicate a relation to treatment. However, the relatively high number of dead pups in the high dose group was due to the death of nine pups in one litter. There was some evidence of poor maternal care.
External malformations:
no effects observed
Description (incidence and severity):
Ano-genital distances for male or female offspring were unaffected by treatment. A check was performed to assess for the presence or absence of nipple/areolae for the male offspring; no nipples were found.
Skeletal malformations:
no effects observed
Description (incidence and severity):
Macroscopic examination of offspring did not reveal any findings attributable to treatment.
Visceral malformations:
no effects observed
Description (incidence and severity):
Macroscopic examination of offspring did not reveal any findings attributable to treatment.
Other effects:
no effects observed
Description (incidence and severity):
THYROID HORMONE ANALYSIS
Serum T4 levels in PND 14-16 males and females were considered not to be affected by treatment.

In the related gross pathology/histopathology:

The thyroids of the F1 offspring were considered unaffected by treament.
Key result
Dose descriptor:
NOAEL
Remarks:
reproduction toxicity
Effect level:
>= 600 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to adverse toxic effects at highest dose / concentration tested
Dose descriptor:
NOAEL
Remarks:
developmental toxicity
Effect level:
>= 200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
fetal/pup body weight changes
Remarks on result:
other: treatment related effect on body weights was observed at 600 mg/kg
Abnormalities:
no effects observed
Developmental effects observed:
yes
Lowest effective dose / conc.:
600 mg/kg bw/day (nominal)
Treatment related:
yes
Relation to maternal toxicity:
developmental effects as a secondary non-specific consequence of maternal toxicity effects
Dose response relationship:
yes
Relevant for humans:
no
Conclusions:
Under the conditions of this study, the no-observed-adverse-effect level (NOAEL) for systemic toxicity was considered to be 200 mg/kg bw/day due to clinical signs and reduced body weight gain. For reproductive toxicity the NOAEL for males and females is 600 mg/kg bw/day. For developmental toxicity the NOAEL for males and females is 200 mg/kg bw/day due to reduced body weight gain.
Executive summary:

The study was performed according the requirements of OECD TG 421 guideline under GLP conditions. The purpose of this study was to assess general systemic toxic potential in rats, including a screening test for reproductive / development effects and assessment of endocrine disruptor relevant endpoints, with administration of the test item by oral gavage administration for a least four weeks in Crl:WI(Han) rats. Three groups, each comprising ten male and ten female rats, received test item at doses of 60, 200 or 600 mg/kg/day. A control group of ten males and ten females was similarly dosed with vehicle alone (PEG 300), at the same dose volume as the treated groups (5 mL/Kg). The following parameters and end points were evaluated in this study: mortality/ moribundity, clinical signs, body weight and food consumption, estrous cycle determination, measurement of thyroid hormone T4 (F0-males and females), gross necropsy findings, organ weights and histopathologic examinations. In addition, the following reproduction/developmental parameters were determined: mating and fertility indices, precoital time, number of implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights, sex, anogenital distance, areola/nipple retention and macroscopy, measurement of thyroid hormone T4 (PND 14-16 pups)). Test formulations prepared were considered homogeneous at the concentrations tested and analysis of the accuracy revealed acceptable levels. Test formulations prepared were considered stable, for at least 2 hours at room temperature. Test formulations for Group 2 were also considered stable for 7 days in the refrigerator. The test item doses were there therefore considered based on nominal concentrations.The mortality observed in this study (one mortality at 60 and 600 mg/kg bw/day, respectively) was considered unrelated to treatment, due to a lack of a dose response and replacements to the two decedents indicated comparable effects to treatment to survivors.Within clinical signs: piloerection was noted in females at dose 600 mg/kg and occurred in one female from week 1-3 of treatment and in several females from approximately Week 5 until Week 7 of treatment. Yellow discolouration of the urine was observed in both males and females at dose 600 mg/kg from Week 1 (males) and Week 2 (females) of treatment but was speculated as being related to higher bilirubin concentrations in plasma as found in a previously performed 28-day toxicity study. Lower body weight gain was observed for males at 600 mg/kg during the treatment period, resulting in approximately 7% lower body weights at termination. In females at 600 mg/kg, a higher body weight loss was observed over the first week compared to that in the other groups (maximally 4% versus 2%). Lower body weights were noted in high dose females at start of mating and directly after delivery. Any effect on body weights during the post coitum were difficult to evaluate as they are also affected by the growth of the fetuses. During lactation a higher body weight gain was observed in the high dose females, resulting in similar body weights in females of all groups at termination. Absolute and relative food consumption were treatment relatedly lower in males and females at 600 mg/kg during days 1-8 of treatment. As food consumption recovered to normal levels in the following days, this lower food consumption was considered non-adverse. Serum levels of T4 in F0-males and females were dose dependently reduced at all tested dose levels up to 600 mg/kg. Although all values remained within the historical control range.Changes in T4 levels were possibly related to the changes in the livers of male and female rats (as previously observed) at similar dose levels in the previously performed 28-day toxicity study. There was no correlates to the Thyroid in gross pathology, organ weights or histopathology. Consequently, T3 and TSH were not further analysed. No reproduction toxicity was observed up to the highest dose level tested. Within developmental toxicity, a treatment related effect on body weights of pups was observed at 600 mg/kg bw/day. As at Day 1 of lactation females had a lower body weight, this may be related to the observed lower body weights of pups. Since the body weights of the pups remained lower than controls up to Day 13, this finding was considered adverse. Under the conditions of this study, the no-observed-adverse-effect level (NOAEL) for systemic toxicity was considered to be 200 mg/kg bw/day due to clinical signs and reduced body weight gain. For reproductive toxicity the NOAEL for males and females is 600 mg/kg bw/day. For developmental toxicity the NOAEL for males and females is 200 mg/kg bw/day due to reduced body weight gain.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The key study is GLP compliant and of a high quality (Klimisch 1); The available information as a whole meets the tonnage driven information requirements of REACH.

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

Key Study : Reproductive/developmental screening study – Oral: OECD TG 421, 2019 : The study was performed according the requirements of OECD TG 421 guideline under GLP conditions. The purpose of this study was to assess general systemic toxic potential in rats, including a screening test for reproductive / development effects and assessment of endocrine disruptor relevant endpoints, with administration of the test item by oral gavage administration for a least four weeks in Crl:WI(Han) rats. Three groups, each comprising ten male and ten female rats, received test item at doses of 60, 200 or 600 mg/kg/day. A control group of ten males and ten females was similarly dosed with vehicle alone (PEG 300), at the same dose volume as the treated groups (5 mL/Kg). The following parameters and end points were evaluated in this study: mortality/ moribundity, clinical signs, body weight and food consumption, estrous cycle determination, measurement of thyroid hormone T4 (F0-males and females), gross necropsy findings, organ weights and histopathologic examinations.In addition, the following reproduction/developmental parameters were determined: mating and fertility indices, precoital time, number of implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights, sex, anogenital distance, areola/nipple retention and macroscopy, measurement of thyroid hormone T4 (PND 14-16 pups)). Test formulations prepared were considered homogeneous at the concentrations tested and analysis of the accuracy revealed acceptable levels. Test formulations prepared were considered stable, for at least 2 hours at room temperature. Test formulations for Group 2 were also considered stable for 7 days in the refrigerator. The test item doses were there therefore considered based on nominal concentrations.The mortality observed in this study (one mortality at 60 and 600 mg/kg bw/day, respectively) was considered unrelated to treatment, due to a lack of a dose response and replacements to the two decedents indicated comparable effects to treatment to survivors.Within clinical signs: piloerection was noted in females at dose 600 mg/kg and occurred in one female from week 1-3 of treatment and in several females from approximately Week 5 until Week 7 of treatment. Yellow discolouration of the urine was observed in both males and females at dose 600 mg/kg from Week 1 (males) and Week 2 (females) of treatment but was speculated as being related to higher bilirubin concentrations in plasma as found in a previously performed 28-day toxicity study. Lower body weight gain was observed for males at 600 mg/kg during the treatment period, resulting in approximately 7% lower body weights at termination. In females at 600 mg/kg, a higher body weight loss was observed over the first week compared to that in the other groups (maximally 4% versus 2%). Lower body weights were noted in high dose females at start of mating and directly after delivery. Any effect on body weights during the post coitum were difficult to evaluate as they are also affected by the growth of the fetuses. During lactation a higher body weight gain was observed in the high dose females, resulting in similar body weights in females of all groups at termination. Absolute and relative food consumption were treatment relatedly lower in males and females at 600 mg/kg during days 1-8 of treatment. As food consumption recovered to normal levels in the following days, this lower food consumption was considered non-adverse. Serum levels of T4 in F0-males and females were dose dependently reduced at all tested dose levels up to 600 mg/kg. Although all values remained within the historical control range.Changes in T4 levels were possibly related to the changes in the livers of male and female rats (as previously observed) at similar dose levels in the previously performed 28-day toxicity study. There was no correlates to the Thyroid in gross pathology, organ weights or histopathology. Consequently, T3 and TSH were not further analysed. No reproduction toxicity was observed up to the highest dose level tested. Within developmental toxicity, a treatment related effect on body weights of pups was observed at 600 mg/kg bw/day. As at Day 1 of lactation females had a lower body weight, this may be related to the observed lower body weights of pups. Since the body weights of the pups remained lower than controls up to Day 13, this finding was considered adverse. Under the conditions of this study, the no-observed-adverse-effect level (NOAEL) for systemic toxicity was considered to be 200 mg/kg bw/day due to clinical signs and reduced body weight gain. For reproductive toxicity the NOAEL for males and females is 600 mg/kg bw/day. For developmental toxicity the NOAEL for males and females is 200 mg/kg bw/day due to reduced body weight gain.

Justification for classification or non-classification

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for reproductive/developmental toxicity.

Within developmental toxicity: the reduced bodyweight gains of F1 males/females at 600 mg/kg bw/day was considered to be a non-specific consequence of F0 maternal toxicity in females at 600 mg/kg bw/day. No effects were observed at non-maternally toxic dose levels (NOAEL 200 mg/kg bw/day).

 

References:

1. ECHA Guidance on Application on the CLP Criteria, (v5.0, July 2017)

Additional information

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