Reaction mass of (2E)-Tridec-2-enenitrile and (2Z)-Tridec-2-enenitrile and (3E)-Tridec-3-enenitrile and (3Z)-Tridec-3-enenitrile

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

The NOAEL for reproductive toxicity was considered to be 200 mg/kg bw/day for males and females. The NOAEL for developmental toxicity was considered to be 200 mg/kg bw/day for pups.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-05-03 to 2016-09-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Remarks:

(Crl:CD(SD)), SPF

Details on species / strain selection:

Sprague-Dawley rats are commonly used in both the general systemic toxicity and reproductive and developmental toxicity studies with a large historical control data base. In addition, the rat is a required species in the regulatory guidelines.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Females nulliparous and non-pregnant: yes (not stated directly but females were determined to have normal estrous cycle)
- Age at study initiation: male: 11 weeks; female: 12 weeks
- Weight at study initiation: male: 368.9 – 479.3 g; female: 224.7 – 274.6 g
- Fasting period before study: no
- Housing: Animals per cage: 1 (during the quarantine-acclimation period), 1 (during the dosing period), 1 male and 1 female (during the mating period), 1 female and neonates (during the lactation period); Stainless wire mesh cages, 260W×350D×210H (mm) and Polycarbonate cage 260W×420D×180H (mm)
- Diet: ad libitum, Pelleted rodent chow (Teklad Certified Irradiated Global 18% Protein Rodent Diet 2918C)
- Water: ad libitum, tap water
- Acclimation period: 20 days

DETAILS OF FOOD AND WATER QUALITY:
The certificate of feed analysis was provided by the supplier and the results of feed analysis met the allowable standard of the test facility.
Public tap water was filtered and irradiated by ultraviolet light. Samples of drinking water are analyzed for specified microorganisms once a month and all environmental contaminants once a year according to the Regulation of Quality Criteria for Potable Water and Test. The results of water analysis met the allowable standard of the test facility.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.1 – 24.3
- Humidity (%): 48.0 – 69.7
- Air changes (per hr): 10 – 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The required amount of the test substance was weighed using an electronic balance (BP3100S, CP423S, Sartorius, Germany) and placed in a container. The test substance was mixed with a small amount of vehicle to dissolve using a magnetic stirrer, and then the vehicle was gradually added to yield the desired concentrations. The dosing formulations were stored in a refrigerator (3.9 – 5.4 °C). These dosing formulations were used within 7 days.

VEHICLE
- Justification for use and choice of vehicle: Through the preliminary solubility test to determine the solubility and dispersion characteristics of the test substance, corn oil was selected as the vehicle because the test substance was well dissolved in it.
- Concentration in vehicle: 0.2 and 200 mg/mL
- Amount of vehicle: 5 mL/kg
- Lot/batch no.: MKBV2080V

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 2 weeks
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: single
- Any other deviations from standard protocol: no

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analyses of the dosing formulations were conducted using a gas chromatography (GC-2010 series, Shimadzu Corp., Japan). Samples were taken three times from the middle of each dosing formulation prior to dosing and analyzed for verification of dose level concentration. The results of dose concentration analyses were determined to be 96.47 – 109.60 %. These results were within the acceptable limits (±15% of nominal values).

Duration of treatment / exposure:

Males of the main group were dosed for a total of 50 days (prior to mating for 2 weeks, during 2 weeks of mating and 22 days of post-mating). Males and females of the recovery groups were dosed for 50 days. Females of the main group were dosed for 2 weeks prior to mating until Postpartum Day 13.

Frequency of treatment:

once daily

Details on study schedule:

- F1 parental animals were not mated.

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Control

Dose / conc.:

20 mg/kg bw/day (nominal)

Remarks:

Low dose

Dose / conc.:

60 mg/kg bw/day (nominal)

Remarks:

Mid dose

Dose / conc.:

200 mg/kg bw/day (nominal)

Remarks:

High dose

No. of animals per sex per dose:

Main groups: 12
Recovery groups: 6

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: In previously conducted 2-week repeated oral dose range finding study all animals were dead or moribund at 1000 mg/kg bw/day. Furthermore salivation and an increase in the relative liver weight were noted in females at 300 mg/kg bw/day. Therefore, the high dose level was selected at 200 mg/kg bw/day. Then, the mid and low dose levels were selected at 60 and 20 mg/kg bw/day, respectively.
- Post-exposure recovery period in satellite groups: 2 weeks

Positive control:

No positive control was conducted.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once before the test and once a week throughout the dosing and recovery periods

BODY WEIGHT: Yes
- Time schedule for examinations: males of main group and males/females of recovery group: just prior to dosing on Day 1 (the first day of dosing), once a week throughout the dosing period and recovery period, the day prior to necropsy and on the day of necropsy (fasted body weights); females of main group: Body weights of females of the main group were recorded just prior to dosing on Day 1 (the first day of dosing), once a week throughout the dosing period, on Gestation Days 0, 7, 14 and 20, on Postpartum Days 0, 4 and 13, the day prior to necropsy and on the 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: No
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No
Individual food consumption was calculated by subtracting the amount of residual feed from the amount presented.

OTHERS:
OPHTHALMOSCOPIC EXAMINATION: Yes, as part of the detailed neurological examination

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At necropsy blood samples were collected from the abdominal aorta.
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes, approximately 18 hours
- How many animals: all
- Parameters checked in table [No.1] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At necropsy blood samples were collected from the abdominal aorta.
- Animals fasted: Yes, approximately 18 hours
- How many animals: all
- Parameters checked in table [No.2] were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: two days before necropsy
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Yes
- Parameters checked in table [No.3] were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes, cursory
- Time schedule for examinations: once before the test and once a week throughout the dosing and recovery periods
- Dose groups that were examined: all animals
- Battery of functions tested: motor activity, central and autonomic nervous system effects and behavior

NEUROBEHAVIOURAL EXAMINATION: Yes, detailed
- Time schedule for examinations: few days before necropsy
- Dose groups that were examined: six males and six females were randomly selected from the main groups in addition to all recovery animals
- Battery of functions tested: Pinna reflex, auditory (sound) reflex, corneal reflex, pupillary reflex, grip strength test, Rotarod test, spontaneous motor activity test

IMMUNOLOGY: Yes
- Time schedule for examinations: pups: on PND 4 and 13; adults: at termination
- How many animals: at least two pups per litter, all adult males and dams of the main group
- Dose groups that were examined: all
- Parameters examined: Total Thyroxine (T4) [ug/dL] Method: Chemiluminescent competitive immunoassay

Oestrous cyclicity (parental animals):

Observations of estrous cycle in females of the main group was conducted from dosing initiation day to confirmed copulation day and necropsy day. Smears of vaginal mucosa were prepared in the morning daily.

Sperm parameters (parental animals):

Parameters examined in P male parental generations:
testis weight, epididymis weight

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, anogenital distance (AGD), presence of nipples in male pups

GROSS EXAMINATION OF DEAD PUPS:
no, possible cause of death was not determined for pups born or found dead.

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: No observations conducted.

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:
ENDOCRINE FUNCTION: Yes
- Time schedule for examinations: pups: on PND 4
- How many animals: at least two pups per litter
- Dose groups that were examined: all
- Parameters examined: Total Thyroxine (T4) [ug/dL] Method: Chemiluminescent competitive immunoassay

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals of the main group were sacrificed on Day 51. All animals of the recovery groups were sacrificed two weeks after final dosing.
- Maternal animals: All surviving animals of the main group were sacrificed on Postpartum Day 14. All animals of the recovery groups were sacrificed two weeks after final dosing.

GROSS NECROPSY
- Complete gross postmortem examinations were conducted on all animals including the external surfaces and internal organs.

Organ weights:
The testis and epididymis of all adult males were weighed.
Six males and six females were randomly selected from the main group animals in addition to all recovery animals for necropsy. The organ weights from organs listed in Table No. 4 were determined.

HISTOPATHOLOGY: Yes. The testis, epididymis and eyes with optic nerves were fixed in Davidson’s fixative. All other tissues were preserved in 10% neutral buffered formalin. The thyroid from one male and one female pups per litter were preserved in 10% neutral buffered formalin on PND 13. Organs listed in Table No. 5 were prepared for histopathology. Examinations were conducted in six males and six females from the control, low, mid and high groups, for all gross, macroscopic lesions in all animals and in addition target organs noted at the high dose were examined for the recovery groups.

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring were sacrificed at PND 4 or 13.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of external examinations.

HISTOPATHOLOGY / ORGAN WEIGTHS
The thyroid from one male and one female pups per litter were preserved in 10% neutral buffered formalin on PND 13.

Statistics:

The statistical analysis of this study was conducted using the SAS program (SAS® 9.3, SAS Institute Inc., U.S.A.).
For the data including body weights, food consumption, estrous cycle, mating period, gestation period, post-implantation loss, body weights and birth and survival rates of pups, AGD index, nipple number, thyroid hormone value, urine volume, organ weights, sensory reactivity and motor activity, the Bartlett test was conducted to test for homogeneity of variance (significance level: 0.05). One-way analysis of variance (ANOVA) test was employed on homogeneity, if significant (significance level: 0.05), followed by Dunnett’s t-test for multiple comparisons (significance levels: 0.05 and 0.01, two-tailed). Kruskal-Wallis test was employed on heterogeneity, and if significant (significance level: 0.05), Steel’s test was performed for multiple comparisons (significance levels: 0.05 and 0.01, two-tailed).
The data of mating index, fertility index and other data associated with gestation were analyzed utilizing Fisher’s exact test (significance levels: 0.05 and 0.01).
For the data of the recovery groups, Folded-F test was employed to test for homogeneity of variance (significance level: 0.05, two-tailed). Student t-test was employed for homogeneity, but if overruled, Aspin-Welch t-test was applied (significance levels: 0.05 and 0.01, two-tailed).

Reproductive indices:

Mating index, mating period, gestation period, male and female fertility index and gestation index were calculated.

Offspring viability indices:

Mean litter size, live birth index, viability index on postnatal day 0 and 4, sex ratio were calculated.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

In the main groups, salivation was sporadically observed in six males at 200 mg/kg bw/day from Day 22 to the end of dosing. Also, salivation was observed temporarily in one female at 20 mg/kg bw/day and 2 females at 200 mg/kg bw/day.
In the recovery groups, salivation was often observed in three males and three females at 200 mg/kg bw/day from Day 14 to the end of dosing.
Salivation observed in both sexes at 200 mg/kg bw/day was considered to be a test substance-related effect.
No clinical signs were observed in males and females of the main and recovery groups in the detailed examinations.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

In the main and recovery groups, all animals survived the duration of the study. There was no effect on mortality.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No test substance-related effects on body weight changes were noted in both sexes at 20, 60 and 200 mg/kg bw/day in the main and recovery groups.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

A statistically significant decrease in food consumption was noted in females at 20 mg/kg bw/day in the main group on Gestation Day 7. A statistically significant increase in food consumption was noted in females at 200 mg/kg bw/day in the recovery group on Day 50. However, these statistical significances were not considered to be a test substance-related change since the difference was of small magnitude and it was not related to the body weight changes.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Description (incidence and severity):

No test substance-related effects on pupillary reflex or corneal reflex were observed in animals of both sexes in the main and recovery groups when compared to the control group.

Haematological findings:

no effects observed

Description (incidence and severity):

No adverse effects were observed in any animal in the main and recovery groups.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No adverse effects were observed in any animal in the main and recovery groups.

Urinalysis findings:

no effects observed

Description (incidence and severity):

In males of the main and recovery groups, no effects were noted in any dosing group.

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

No test substance-related effects on auditory reflex or pinna reflex were observed in animals of both sexes in the main and recovery groups when compared to the control group.
In the main and recovery groups, there were no test substance-related effects in the grip strength test and spontaneous motor activity of both sexes when compared to the control group. A statistically significant increase was noted in rota rod test in males at 60 mg/kg/day when compared to control group. However, it was considered not to have a meaning of toxicological significance since it was without dose-dependency.
In the recovery groups, there were no test substance-related effects in the grip strength test, rota rod test and spontaneous motor activity when compared to the control group.

Immunological findings:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Microscopic changes observed in the stomach of both sexes were most likely to be test substance-related.
Epithelial hyperplasia/hyperkeratosis with inflammatory cell infiltration in the forestomach submucosa was noted in one female at 200 mg/kg bw/day. This finding corresponded to macroscopically observed polyp/thickening of the forestomach. Erosion on mucosa in the glandular stomach was observed in one male at 200 mg/kg bw/day and females at 60 and 200 mg/kg bw/day. This finding was in concordance with macroscopically observed focus on mucosa of the glandular stomach.
At the end of the recovery period, these findings were not observed in any animal.

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

The estrus cycle lengths (day) of females in the 0 (control), 20, 60 and 200 mg/kg bw/day dosing groups were 4.0, 4.1, 4.1 and 4.0 days, respectively. There were no statistically significant differences in any dosing group. The estrous cycle of females on the day of necropsy was diestrus except one female at 200 mg/kg bw/day which had no pups.

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Description (incidence and severity):

- Mating: In the control groups and 20, 60 and 200 mg/kg bw/day dose groups, the mating periods were 1.7, 2.7, 1.5 and 1.8 days, the mating index was 100%, the gestation periods were 22.1, 22.0, 22.3 and 22.3 days, and the fertility indices of animals of both sexes were 100.0%, respectively. There were no statistically significant differences in any dose group.
- Delivery observation: In the control groups and 20, 60 and 200 mg/kg bw/day dose groups, the gestation index was 100%, the post-implantation loss rates were 5.3, 6.8, 12.6 and 9.6%, the live birth indices were 94.7, 93.2, 87.4 and 90.4%, the mean litter sizes were 13.7, 14.3, 13.0 and 13.0, the viability indices on Postnatal Day (PND) 0 were 98.7, 99.3, 97.6 and 98.4%, and the viability indices on PND 4 were 99.0, 98.8, 99.4 and 90.0%, respectively. There were no statistically significant differences in any dose group. In addition, normal delivery was observed in all animals of the control groups and 20, 60 and 200 mg/kg bw/day dose groups, and the sex ratios on PND 0 were 1.2, 1.1, 1.3 and 1.4, respectively.

Key result

Dose descriptor:

NOAEL

Effect level:

200 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

reproductive function (oestrous cycle)

reproductive function (sperm measures)

reproductive performance

Key result

Dose descriptor:

NOAEL

Effect level:

60 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: systemic toxicity

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

60 mg/kg bw/day (nominal)

System:

gastrointestinal tract

Organ:

stomach

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

not specified

Clinical signs:

not examined

Dermal irritation (if dermal study):

not examined

Mortality:

not examined

Body weight and weight changes:

not examined

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

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

not examined

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test substance-related changes.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

In the control groups and 20, 60 and 200 mg/kg bw/day dose groups, the viability indices on Postnatal Day (PND) 0 were 98.7, 99.3, 97.6 and 98.4%, and the viability indices on PND 4 were 99.0, 98.8, 99.4 and 90.0%. There were no statistically significant differences in any dose group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no test substance-related changes in body weights of pups at 20, 60 and 200 mg/kg bw/day.

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:

no effects observed

Description (incidence and severity):

There were no test substance-related changes in external examinations of pups at 20, 60 and 200 mg/kg bw/day.

Histopathological findings:

not examined

Other effects:

no effects observed

Description (incidence and severity):

There were no test substance-related changes in AGD index of pups at 20, 60 and 200 mg/kg bw/day.
Nipple numbers were 0 in male pups at 0, 20, 60 and 200 mg/kg bw/day on PND 12. There was no nipple retention in male pups at 20, 60 and 200 mg/kg bw/day on PND 12.

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

no effects observed

Description (incidence and severity):

There were no test substance-related changes in thyroid hormone (T4) level of F1 pups at 20, 60 and 200 mg/kg bw/day.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

200 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: developmental toxicity

Clinical signs:

not examined

Dermal irritation (if dermal study):

not examined

Mortality / viability:

not examined

Body weight and weight changes:

not examined

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:

not examined

Histopathological findings:

not examined

Other effects:

not examined

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

Key result

Reproductive effects observed:

no

Conclusions:

The NOAEL for reproductive toxicity was considered to be 200 mg/kg bw/day for males and females. The NOAEL for developmental toxicity was considered to be 200 mg/kg bw/day for pups.

Executive summary:

An oral repeated dose toxicity study was conducted according to OECD 422 in rats. The test substance was administered via gavage to groups of 12 male and 12 female rats at concentrations of 0, 20, 60 and 200 mg/kg bw/d. Males of the main group were dosed once daily for a total of 50 days (prior to mating for 2 weeks, during 2 weeks of mating and 22 days of post-mating), and females of the main group were dosed once daily for two weeks prior to mating, throughout gestation and for thirteen days after delivery. Also, males and females of the recovery groups were dosed for 50 days. All males of the main group and all animals of the recovery groups survived the duration of the study. Salivation was observed in males and females at 200 mg/kg bw/day in the main and recovery groups. No test substance-related adverse effects were noted in the results of detailed clinical signs, body weights, food consumption, estrous cycle, sensory function, motor activity, urinalysis, hematology, clinical chemistry, thyroid hormone (T4) of adult males and organ weights of animals of both sexes in the test substance-dosed groups. Focus of the glandular stomach was observed in one male at 200mg/kg bw/day and females at 60 and 200 mg/kg bw/day. Polyp/thickening of the forestomach mucosa was observed in one female at 200 mg/kg bw/day. Erosion on mucosa in the glandular stomach was observed in one male at 200 mg/kg bw/day and females at 60 and 200 mg/kg bw/day. Epithelial hyperplasia/hyperkeratosis with inflammatory cell infiltration in the forestomach submucosa was noted in one female at 200 mg/kg bw/day. No test substance-related adverse effects were noted in the results of the mating period, mating index, gestation period, male and female fertility indexes, gestation index, post-implantation loss rates, live birth index, mean litter size, external examination of pups, body weights of pups, sex ratio of pups and viability index of Postnatal Days 0 and 4. No test substance-related effects were noted in the results of anogenital distance (AGD) index of pups, nipple retention of male pups and thyroid hormone (T4) of pups. The test substance had no endocrine disrupting potential under the condition of this study. Based on the results of this study, the NOAEL for reproductive toxicity was considered to be 200 mg/kg bw/day for males and females. The NOAEL for developmental toxicity was considered to be 200 mg/kg bw/day for pups.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

200 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The study was conducted according to guideline and GLP.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

An oral repeated dose toxicity study was conducted according to OECD 422 in rats. The test substance was administered via gavage to groups of 12 male and 12 female rats at concentrations of 0, 20, 60 and 200 mg/kg bw/d. Males of the main group were dosed once daily for a total of 50 days (prior to mating for 2 weeks, during 2 weeks of mating and 22 days of post-mating), and females of the main group were dosed once daily for two weeks prior to mating, throughout gestation and for thirteen days after delivery. Also, males and females of the recovery groups were dosed for 50 days. All males of the main group and all animals of the recovery groups survived the duration of the study. Salivation was observed in males and females at 200 mg/kg bw/day in the main and recovery groups. No test substance-related adverse effects were noted in the results of detailed clinical signs, body weights, food consumption, estrous cycle, sensory function, motor activity, urinalysis, hematology, clinical chemistry, thyroid hormone (T4) of adult males and organ weights of animals of both sexes in the test substance-dosed groups. Focus of the glandular stomach was observed in one male at 200mg/kg bw/day and females at 60 and 200 mg/kg bw/day. Polyp/thickening of the forestomach mucosa was observed in one female at 200 mg/kg bw/day. Erosion on mucosa in the glandular stomach was observed in one male at 200 mg/kg bw/day and females at 60 and 200 mg/kg bw/day. Epithelial hyperplasia/hyperkeratosis with inflammatory cell infiltration in the forestomach submucosa was noted in one female at 200 mg/kg bw/day. No test substance-related adverse effects were noted in the results of the mating period, mating index, gestation period, male and female fertility indexes, gestation index, post-implantation loss rates, live birth index, mean litter size, external examination of pups, body weights of pups, sex ratio of pups and viability index of Postnatal Days 0 and 4. No test substance-related effects were noted in the results of anogenital distance (AGD) index of pups, nipple retention of male pups and thyroid hormone (T4) of pups. The test substance had no endocrine disrupting potential under the condition of this study. Based on the results of this study, the NOAEL for reproductive toxicity was considered to be 200 mg/kg bw/day for males and females. The NOAEL for developmental toxicity was considered to be 200 mg/kg bw/day for pups.

Effects on developmental toxicity

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

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

The available experimental test data is reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on reproductive toxicity, the substance is considered to be not classified, according to Regulation (EC) No 1272/2008 (CLP), as amended for the ninth time in Regulation (EU) No 2016/1179.