3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

screening for reproductive / developmental oral toxicity (OECD 421, rats): NOAEL = 200 mg/kg bw/day

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 Dec 2016 - 01 Aug 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

adopted Jul 2016

Deviations:

no

GLP compliance:

yes

Species:

rat

Strain:

other: Crl:CD(SD)

Details on species / strain selection:

The rat, for which the OECD Guideline 421 is designed, was selected for this study. In addition, reproductive and teratological background data for this strain of rats are well established and historical data are available.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc. (Atsugi Breeding Center)
- Age at study initiation: (P) Males: 11 weeks; Females: 10 weeks
- Weight at study initiation: (P) Males: 368 - 437 g; Females: 244 - 280 g
- Housing: Animals were individually housed from Day 0 to Day 18 of gestation. One animal/sex during co-housing and one dam with its offspring from Day 18 of gestation to the end of the lactation period were housed per cage, respectively. Stainless steel chip trays (changed at least twice weekly) and bedding material were used for stainless steel wire mesh cages. Bedding material was used for polycarbonate cages. Wood blocks for gnawing were placed in the cages during the housing period.
- Diet: NMF (pelleted feed) by Oriental Yeast Co., Ltd. (Chiba), ad libitum
- Water: Drinking water, ad libitum
- Acclimation period: At least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.5 - 23.2 (actual)
- Humidity (%): 52.0 - 70.8 (actual)
- Air changes (per hr): 15 - 17
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 10 Jan 2017 To: 08 Feb 2017 (males) and 02 - 10 Mar 2017 (females)

Route of administration:

oral: gavage

Vehicle:

other: olive oil containing 1.0% (w/v) Tween 80

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Dosing solutions were prepared weekly by dissolving 1.50, 2.56 and 7.50 mL of the test material in olive oil containing 1.0% (w/v) Tween 80 yielding a final concentration of 1, 5 and 40 mg/mL, respectively.

VEHICLE
- Concentration in vehicle: 1, 5 and 40 mg/mL, respectively
- Amount of vehicle: 5 mL/kg bw/d

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: A maximum of 14 days
- Proof of pregnancy: Sperm in vaginal smear or a vaginal plug in the vagina referred to as Day 0 of pregnancy
- After successful mating each pregnant female was caged: Individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Stability and homogeneity of the test article in the vehicle was investigated at the Chemicals Evaluation and Research Institute, Japan Hita Laboratory (Study Code No. X18-0838). It has been confirmed that 0.04 and 10.0% (w/v) of the test substance formulations in the vehicle (olive oil containing 1.0% (w/v) Tween 80) are homogeneous and stable for 8 days after preparation when stored in a cold dark place.
The verification of the dosing concentrations were analysed using gas chromatography at the first and at the last preparation of all dosing formulations at the Chemicals Evaluation and Research Institute, Japan Hita Laboratory. No peaks derived from the test substance were present in the chromatogram of the control formulation and the measured concentrations of the test substance dosing formulations were 91.6- 93.8% of the nominal concentrations.

Duration of treatment / exposure:

Males: From 14 days prior to mating, throughout the mating period, until the last day of the mating period.
Females: From 14 days prior to mating, throughout the mating and gestation periods, until Day 13 of lactation.

Frequency of treatment:

daily, 7 days/week

Dose / conc.:

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

Dose / conc.:

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

Dose / conc.:

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

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: In a previously conducted 28-day repeated dose oral toxicity study at the same testing facility (Study No. B11-0838) in rats at dose levels of 5, 25 and 200 mg/kg bw/day, findings noted in males were centrilobular lipid droplets in the hepatocytes and microgranuloma in the liver at 25 and 200 mg/kg bw/day and a large liver and increased liver weights (absolute and relative) at 200 mg/kg bw/day. However, no appreciable treatment-related toxicity was noted in any female. Based on these results as well as the dosing period of up to approximately 9 weeks for females in the current study, 200 mg/kg bw/day was selected as the high dose level, and 25 and 5 mg/kg bw/day were selected as the mid and low dose levels, respectively.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes, all parental animals. cage-side observations were conducted for each animal. When some abnormalities were suspected, such animals were removed from the cages and observed.
- Time schedule: Twice daily (pre-dosing and up to 1 hour post-dosing) during the dosing period and once daily (in the morning) during the other periods. Males were observed from Day 1 of dosing to the day of necropsy. Females were observed from Day 1 of dosing to Day 14 of lactation.

BODY WEIGHT: Yes, the body weight was recorded for all parental animals.
- Time schedule for examinations: Body weight of males was recorded once weekly from Day 1 of dosing to the day of necropsy. Females were weighted once weekly from Day 1 of dosing to the day of copulation, Days 0, 7, 14 and 20 of gestation and Days 0, 4, 7, 11 and 13 of lactation.

FOOD CONSUMPTION: Yes, food consumption was determined for all males from Day 1 of dosing to the day of initiation of mating and for all females from Day 1 of dosing to Day 13 of lactation (excluding the mating period).
- Provided amounts of feed were recorded on Day 1 of dosing, provided and residual amounts were recorded on the subsequent body weight measurement days and residual amounts were recorded on the day of initiation of the mating period. During gestation, provided amounts of feed were recorded on Day 0 of gestation, provided and residual amounts were recorded on the subsequent body weight measurement days and residual amounts were recorded on Day 20 of gestation. During lactation, provided amounts of feed were recorded on Day 0 of lactation, provided and residual amounts were recorded on the subsequent body weight measurement days and residual amounts were recorded on Day 13 of lactation. Daily food consumption was calculated and reported as the food consumption from the day of feeding to the day of residual amount measurement.

CLINICAL BIOCHEMISTRY
- Thyroid hormone (T4) levels were determined in all dams and all males using a Rat Thyroid Hormone Magnetic Bead Panel. Blood samples were collected from the jugular vein of unanesthetized dams at necropsy.

OTHER:
Observations for delivery and nursing behavior of dams:
- Delivery: The presence/absence of abnormal delivery and live newborns was investigated at least twice daily.
- Duration of gestation: The number of gestational days on the day of completion of delivery was determined.
- Poor nursing during the lactation period was investigated twice daily (pre-dosing and up to 1 h post-dosing) during the dosing period and once (in the morning) on the day of necorpsy. Observations included scattering of offspring in the cages, total litter loss, cannibalization etc.

Oestrous cyclicity (parental animals):

Vaginal smears were taken using cotton swabs at nearly the same time every morning, dried, stained with Giemsa and examined by microscopy to determine the phases of the estrous cycle.
The number of days from the beginning of one estrous cycle to the next (from estrous to the next estrous) was counted and the mean length of the estrous cycle in days was calculated.

Sperm parameters (parental animals):

Mean number of cells in 1 seminiferous tubule and Sertoli cell ratio (%) were determined (please refer to Histopathology under "Postmortem examinations (parental animals)").

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- Maximum of 8 pups/litter (4 pups/sex/litter as nearly as possible); excess pups were killed and discarded. Culling was not conducted when the litter size was less than 8.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
Total number, sex and body weights of live newborns and the number of newborns with external morphology (including the oral cavity); total number of dead newborns; for the dead offspring, external macroscopic abnormalities (especially external reproductive organs) were observed and recorded anogenital distance (AGD), presence of nipples/areolas in male pups

CLINICAL BIOCHEMISTRY
Thyroid hormone (T4) levels were determined in 2 offspring/litter of the culled offspring at adjustment of litter size at 4 days of age using a Rat Thyroid Hormone Magnetic Bead Panel. Blood samples were collected from the jugular vein of unanesthetized of unanesthetized animals, basically from 1 offspring/sex/litter. If only 1 sex of culled offspring was available, blood sampling was conducted for 2 offspring of the same sex. If there were no culled offspring available, blood sampling was not conducted.

Postmortem examinations (parental animals):

SACRIFICE AND GROSS NECROPSY: After completion of blood collection from the jugular vein, the males and females were anesthetized by inhalation of isoflurane. The abdomen was incised and the animals were euthanized by exsanguination from the posterior vena cava and abdominal aorta for complete macroscopic postmortem examinations.
Males:
- 1 day after the last day of the mating period for all surviving males.
- Preservation of the specimens: Liver, seminal vesicles (including coagulating glands), prostate (ventral lobe), thyroids, organs/tissues with gross lesions and the ear with ID No. were fixed and preserved in 10 vol% neutral buffered formalin. The testes and epididymides were fixed and preserved in Bouin’s fixative.

Females:
- On Day 14 of lactation for all surviving dams, on the day on which a total litter loss occurred and promptly after being found dead, respectively.
- Preservation of the specimens: Liver, ovaries, uterus and mammary glands for all surviving dams; ovaries, uterus, mammary glands and organs/tissues with gross lesions for dams with total litter loss; brain, pituitary, thymus, lungs, heart, liver, spleen, adrenals, kidneys, ovaries, uterus, mammary glands and offspring for dead dams

HISTOPATHOLOGY / ORGAN WEIGHTS
Males:
- The following organs were weighed: Liver, testes (separately for the right and left), epididymides (separately for the right and left), the levator ani plus bulbocavernosus muscle complex, Cowper’s gland, glans penis and body weights on the day of necropsy (for calculation of organ-to-terminal body weight ratios).
- Histopathology: The testes and epididymides from the control and high dose groups were routinely processed and stained with hematoxylin and eosin (HE). All histopathological specimens were examined by light microscopy. For the testes, the number of cells (Sertoli cells, spermatogonium, pachytene spermatocytes and spermatids) in stages VII and VIII were counted for 3 seminiferous tubules/animal. In addition, the number of Leydig cells/clump was counted for 3 cell clumps/animal. Then, the mean number of cells and Sertoli cell ratio were calculated:
Mean number of cells in 1 seminiferous tubule = Total number of each cell type / Number of target seminiferous tubules
Sertoli cell ratio (%) = 100 × Mean number of each cell type* / Mean number of Sertoli cells
* Cells other than Sertoli cells in the seminiferous tubules

Females:
- The following organ was weighed: Liver and body weights on the day of necropsy (for calculation of organ-to-terminal body weight ratios).
- Histopathology: The ovaries from the control and high dose groups were routinely processed and stained with hematoxylin and eosin (HE). All histopathological specimens were examined by light microscopy.

Postmortem examinations (offspring):

SACRIFICE: The F1 offspring was sacrificed at 14 days of age. The animals were anesthetized by inhalation of isoflurane. The abdomen was incised and the animals were euthanized by exsanguination from the posterior vena cava and abdominal aorta for complete macroscopic postmortem examinations. For the animals used for determination of T4 levels, these procedures were conducted after blood sampling.

Statistics:

Information on statistical analysis is given under "any other information on materials and methods incl. tables"

Reproductive indices:

- Copulation index = (Number of pairs with successful copulation / Number of pairs mated) × 100
- Fertility index = (Number of pregnant females / Number of pairs with successful copulation) × 100
- Gestation index = (Number of dams with live newborns / Number of pregnant dams) × 100

Offspring viability indices:

- Duration of gestation
- Delivery index = (Number of newborns / Number of implantation sites) × 100
- Birth index = (Number of live newborns / Number of implantation sites) × 100
- Sex ratio of live newborns
- Incidence of external malformations of live newborns
- Viability index on Day 4 of lactation = (Number of live offspring on Day 4 of lactation / Number of live newborns) × 100
- Viability index on Day 14 of lactation = (Number of live offspring on Day 14 of lactation / Number of live offspring after adjustment of litter size at 4 days of age) × 100

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Females:
200 mg/kg bw/day: 1/10 dams showed emaciation attributable to decreased food consumption on Days 4 to 6 of lactation.
25 mg/kg bw/day: 1/2 dams with total litter loss showed a decrease in movement on Day 2 of lactation.

These signs were considered not to be treatment-related since 1) they were limited to one dam in each group; 2) no treatment-related effects on food consumption were noted at 200 mg/kg bw/day and 3) the sign observed in the 25 mg/kg bw/day group was also observed in the dam that died in the control group.

Please refer to table 1 and 2 under "any other information on results incl. tables".

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, non-treatment-related

Description (incidence):

Females:
One dam in the control group died after exhibiting a decrease in movement, soiled fur and scattering of all offspring in the cage on Day 1 of lactation. Since death occurred 1 day after delivery, the cause of death was judged to be a deteriorated physical condition due to poor recovery from delivery. Gross pathology of this animal revealed a small thymus and discoloration of the spleen (please refer to "Gross pathological findings").

Please refer to table 2 under "any other information on results incl. tables".

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Females:
During the lactation period, statistically significant decreases in body weight gain were noted in the 5 mg/kg bw/day group on Days 11 and 13 of lactation as compared to the control group. However, these decreases were not dose-related and were therefore considered to be incidental changes unrelated to treatment with the test substance.

Food consumption and compound intake (if feeding study):

no effects observed

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:

effects observed, treatment-related

Description (incidence and severity):

Males:
A significant decrease in thyroid hormone (T4) levels was noted in the 200 mg/kg bw/day group when compared with the control group.

Please refer to table 1 and 2 under "any other information on results incl. tables".

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

effects observed, non-treatment-related

Description (incidence and severity):

25 mg/kg bw/d: 1 pair failed to mate
Scattering of all offspring in the cage was observed in 2 dams of the 200 mg/kg bw/day group and in 1 dam of the 25 mg/kg bw/day group on the day of and/or 1 day after delivery, all of which were assumed to be due to the poor nursing behavior of the dams. Of these dams, total litter loss occurred for 1 dam of the 200 mg/kg bw/day and for 1 dam of the 25 mg/kg bw/day group on Day 2 of lactation (please refer to table 2 under "any other information on results incl. tables").

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

organ weights and organ / body weight ratios

gross pathology

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No adverse effects observed at the highest tested dose

Key result

Dose descriptor:

NOAEL

Remarks:

parental reproductive function

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed at the highest tested dose

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

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

System:

hepatobiliary

Organ:

liver

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

No milk in the stomach of offspring was observed for 3 high-dosed dams and subnormal body surface temperature of offspring was also observed in 1 of these dams on Day 1 of lactation. Both signs were observed in 1 mid-dosed dam on the day of delivery and/or Day 1 of lactation (please refer to table 3 under "any other information on results incl. tables").
These signs were assumed to be due to poor nursing behavior of the dams.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

Total litter loss occurred on Day 2 of lactation in 1 dam each in the 200 and 25 mg/kg bw/d groups. Death of some offspring/litter occurred in 5, 3, 0 and 4 dams in the control and 5, 25 and 200 mg/kg bw/d groups, respectively. The incidence of death was not dose-related and therefore the deaths were considered to be unrelated to treatment with the test substance (please refer to table 3 under "any other information on results incl. tables").

Body weight and weight changes:

no effects observed

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

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

200 mg/kg bw/d: In 1 female offspring discoloration of the spleen, dilation and retention of contents in the stomach and a greenish brown focus in the liver was observed. Since no gross pathological lesions were observed in any other offspring in this group, the effects were considered to be incidental lesions unrelated to treatment with the test substance (please refer to table 3 under "any other information on results incl. tables").

Histopathological findings:

not examined

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

Key result

Dose descriptor:

NOAEL

Remarks:

developmental toxicity

Generation:

F1

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed at the highest tested dose.

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Table 1: Summary of results in parental males

Daily Dose (mg/kg/day)		0	5	25	200
No. Animals		10	10	10	10
No. Died or Sacrificed Moribund		0	0	0	0
Copulation Index (%)		100.0	100.0	90.0	100.0
Fertility Index (%)		100.0	100.0	100.0	100.0
Gross Pathology: Liver, Large		0	0	0	7
Liver Weights (mean)	Absolute (g)	15.9	16.9	17.4	23.4**
	Relative (%)	3.304	3.519	3.669 *	4.932**
Thyroid Hormone (T4) Level (ng/mL, mean)		541.0	529.8	506.3	474.8**

* p < 0.05; ** p < 0.01

Table 2: Summary of results in parental females

Daily Dose (mg/kg/day)		0	5	25	200
No. Animals		10	10	10	10
No. Died or Sacrificed Moribund		1 (L1)	0	0	0
No. Dams that Delivered Live Newborns		10	10	9	9
No. Total Litter Loss		0	0	1 (L2)	1 (L2)
Until Copulation
Estrous Cycle (mean)		4.2	4.2	4.1	4.2
Copulation Index (%)		100.0	100.0	90.0	100.0
Fertility Index (%)		100.0	100.0	100.0	100.0
Pre-coital Period (mean)		2.1	3.4	2.8	2.7
Gestation Period
Duration of Gestation (mean)		22.0	22.0	22.1	22.0
Implantation Sites (mean)		16.4	16.1	16.4	16.0
Gestation Index (%)		100.0	100.0	100.0	100.0
Delivery Index (%)		96.4	88.9	95.1	93.9
Abnormal Parturition		−	−	−	−
Birth Index (%)		89.3	87.1	84.5	91.2
Lactation Period
Clinical Observation: Decrease in Movement		1 a	0	1 b	0
Clinical Observation: Soiled Fur		1 a	0	0	0
Clinical Observation: Emaciation		0	0	0	1
Scattering of all Offspring in the Cage		1 a	0	1 b	2 c
Gross Pathology: Thymus, Small		1 a	0	1 b	1 b
Gross Pathology: Spleen, Discoloration, pale		1 a	0	0	0
Gross Pathology: Spleen, Small		0	0	1 b	0
Liver Weights (mean)	Absolute (g)	15.7	16.9	16.9	17.4
	Relative (%)	4.130	4.402	4.410	4.672**
Thyroid Hormone (T4) Level (ng/mL, mean)		374.2	379.2	362.4	382.6

** p < 0.01

L: Lactatiopn period

Table 3: Sumamry of results in offpring (F1)

No. Litters		10	10	9	9
Mean No. Newborn/Litter		15.8	14.2	15.7	15.0
Mean No. Live Newborn/Litter		14.6	13.9	13.9	14.6
Sex Ratio (Male/Total)		0.50	0.47	0.37	0.55
External Malformation		0	0	0	0
No. Dead Newborn		12	3	16	4
Clinical Observation	No milk in the stomach	1	0	1	3
Clinical Observation	Subnormal body surface temperature	1	0	1	1
Clinical Observation	Death of some offspring	5	3	0	4
Viability Index on Day 4 of Lactation (0-4, %)		86.9	98.0	88.9	74.1
Thyroid Hormone (T4) Level (ng/mL, mean)		113.5	130.8	130.3	131.2
Viability Index on Day 14 of Lactation (4 - 14, %)		100.0	100.0	100.0	100.0
Gross Pathology	Spleen: Discoloration, pale	0	0	0	1
Gross Pathology	Stomach: Dilation	0	0	0	1
Gross Pathology	Stomach: Retention, contents	0	0	0	1
Gross Pathology	Liver: Focus, greenish brown	0	0	0	1
Thyroid Hormone (T4) Level (ng/mL, mean) 14 days of age		327.2	332.7	328.3	323.0

Conclusions:

Based on the results of the study, the test substance had no effect on reproductive performance.

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 available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.7, of Regulation (EC) No 1907/2006.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

The registered substance was tested in a screening study for reproductive and developmental toxicity according to OECD guideline 421 and in compliance with GLP (Key, 2017). The test substance was administered orally to 10 Sprague Dawley rats/sex/group at 5, 25 and 200 mg/kg/day to evaluate the potential adverse effects of the test substance on mating, fertility, pregnant/lactating female rats and offspring. The dosing periods set for males were from 14 days prior to mating, throughout the mating period, until the last day of the mating period and for females were from 14 days prior to mating, throughout the mating and gestation periods, until Day 13 of lactation. Animals in the control group received the vehicle, olive oil containing 1.0 % (w/v) Tween 80, in the same manner for comparison. In parental animals (P0), large liver in males and increase in the absolute and relative weights of the liver in males and females were noted at 200 mg/kg bw/day. No treatment-related effects were observed in the clinical observations, body weights or food consumption. Regarding the reproductive function of parental animals (P0), no treatment-related effects were noted on the estrous cycle, copulation index, fertility index, delivery, duration of gestation, gestation index, number of implantation sites, delivery index, birth index, nursing behavior or histopathology of reproductive organs. In the subsequent generation (F1), no treatment-related effects were noted in the clinical observations, body weights, number of newborns, number of live newborns, sex ratio, external morphology, viability index on Day 4 or 14 of lactation, anogenital distance, number of nipples/areolas or thyroid hormone (T4) levels. Based on these results, the no observed adverse effect levels (NOAELs) were considered to be 25 mg/kg bw/day for parental males and 200 mg/kg bw/day for parental females, parental reproductive function and offspring under the conditions of this study.

Effects on developmental toxicity

Description of key information

screening for reproductive / developmental toxicity oral (OECD 421, rats): NOAEL = 200 mg/kg bw/day

Effect on developmental toxicity: 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 available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.7, of Regulation (EC) No 1907/2006.

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

The registered substance was tested in a screening study for reproductive and developmental toxicity according to OECD guideline 421 and in compliance with GLP (Key, 2017). The test substance was administered orally to 10 Sprague Dawley rats/sex/group at 5, 25 and 200 mg/kg/day to evaluate the potential adverse effects of the test substance on mating, fertility, pregnant/lactating female rats and offspring. The dosing periods set for males were from 14 days prior to mating, throughout the mating period, until the last day of the mating period and for females were from 14 days prior to mating, throughout the mating and gestation periods, until Day 13 of lactation. Animals in the control group received the vehicle, olive oil containing 1.0 % (w/v) Tween 80, in the same manner for comparison. In parental animals (P0), large liver in males and increase in the absolute and relative weights of the liver in males and females were noted at 200 mg/kg bw/day. No treatment-related effects were observed in the clinical observations, body weights or food consumption. Regarding the reproductive function of parental animals (P0), no treatment-related effects were noted on the estrous cycle, copulation index, fertility index, delivery, duration of gestation, gestation index, number of implantation sites, delivery index, birth index, nursing behavior or histopathology of reproductive organs. In the subsequent generation (F1), no treatment-related effects were noted in the clinical observations, body weights, number of newborns, number of live newborns, sex ratio, external morphology, viability index on Day 4 or 14 of lactation, anogenital distance, number of nipples/areolas or thyroid hormone (T4) levels. Based on these results, the no observed adverse effect levels (NOAELs) were considered to be 25 mg/kg bw/day for parental males and 200 mg/kg bw/day for parental females, parental reproductive function and offspring under the conditions of this study.

Justification for classification or non-classification

The available data on toxicity to reproduction with the test substance do not meet the criteria for classification according to Regulation (EC) No 1272/2008, and are therefore conclusive but not sufficient for classification.