Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Key study. Method according to OECD 421, GLP study. The NOAEL (based on toxicity) for both parent animals and first generation male/female Sprague Dawley rats was found to be 100 mg/kg bw; the NOAEL (based on reproductive ability) for parent male/female Sprague Dawley rats was found to be 40 mg/kg bw.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
August 30, - December 10, 2010.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Reason / purpose:
reference to other assay used for intermediate effect derivation
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Justification for study design:
SPECIFICATION OF STUDY DESIGN WITH JUSTIFICATIONS
- Premating exposure duration for parental (P0) animals: 2 weeks
- Basis for dose level selection: the dose of this study was set based on the results of the 28-day repeated dose toxicity test (See 'Cross-reference'): In this test, animals were dosed with 15, 100 or 500 mg/kg test item for 28 days. In the high dose group, 2 females died, and reduced locomotor activity, supression of body weight gain, increase in liver weight, thickening of the duodenal mucosa, etc. were observed in the remaining animals; no effects were observed at 100 mg/kg bw. In the range finding study, toxicity symptoms such as reduced locomotor activity were also observed in animals dosed with 125 and 250 mg/kg test item for 7 days. Based on the available information, the high dose of this study was set at 250 mg/kg bw, and the spacing factor was 2.5, so the medium dose was 100 mg/kg bw and the low dose 40 mg/kg bw.
- Route of administration: oral, gavage
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
- Purity: 99.1%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Refrigerated until use, storage under light shielding (measured temperature 3 to 7 ° C).
- Stability under test conditions: The stability of the test substance was confirmed at Hadano Laboratory before administration (August 30, 2010) and after the end of the administration period (December 10, 2010).
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl: CD (SD), SPF
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Atsugi Breeding Center (Japan).
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: (P) 10 wks.
- Weight at study initiation: (P) Males: 354.0 to 436.0 g; Females: 215.5 to 280.2 g.
- Housing: individually housed, 2 animals per cage at the time of mating.
- Diet: ad libitum feed (CE-2, CLEA Japan).
- Water: ad libitum tap water.
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 25ºC
- Humidity (%): 40 - 75%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12h light/dark cycle.

IN-LIFE DATES: From: September 1, 2010 (62 males and 73 females, arrival date).
Route of administration:
oral: gavage
Vehicle:
olive oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The medium was warmed at 40ºC or less, and the test item was weighed and added to the solution, at a concentration of 5% (w/v). Two further solutions were prepared at concentrations of 2 and 0.8 % (w/v). These solutions were stored in a refrigerated space (3 - 10ºC) and shielded from light, and used within 7 days after preparation (storage: from September 14th to October 30th, 2010). The doses were administered by gavage.

VEHICLE
- Justification for use and choice of vehicle (if other than water): the test item was found to be moisture sensitive; the stability of the test item in olive oil was confirmed by analysis.
- Concentration in vehicle: 5, 2 and 0.8% (w/v)
- Amount of vehicle (if gavage): 5 mL/kg
- Purity: according to Japanese Pharmacopoeia.
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: 2 weeks
- Proof of pregnancy: vaginal plug and sperm in vaginal smear, referred to as [day 0] of pregnancy.
- Further matings after two unsuccessful attempts: no.
- After successful mating each pregnant female was caged (how): females were individually housed in a plastic breeding cage for rats (350 w × 400 d × 180 h mm) from 18th day of pregnancy to 4th day of nursing, and a paper pulp chips (Pepper Clean, Japan SLC) were used as bedding. No anomalies in the breeding environment were observed during the period, the measured value of the temperature in the animal room was 23.5 to 24.0ºC, the measured value of humidity was 52.0 to 70.0%. In addition, it was confirmed that the analysis results of the feed, drinking water and bedding that were supplied were within the tolerance limits described in the standard operating procedure manual.
- Any other deviations from standard protocol: no.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test item content was measured by a validated analytical method after preparation of the dosing solution; the average content was found to be 90 - 110%, and the variation between measurements was within the acceptable range (average value ± 10%). No further details on analytical method provided (Annex D-1, D-2 not included).
Duration of treatment / exposure:
Males were treated from 2 weeks before mating until the day before necropsy (42 administrations in total). Females were treated from 2 weeks before mating until delivery.
Frequency of treatment:
daily.
Details on study schedule:
- Animals arrival date: September 1, 2010
- Quarantine end date: September 14, 2010
- Dosing prior to mating: 2 weeks
- Mating: up to 2 weeks
- Age at mating of the mated animals in the study: [12] weeks
- End of administration period: December 10, 2010
Dose / conc.:
40 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
250 mg/kg bw/day (nominal)
No. of animals per sex per dose:
13 male and 13 female per group.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: the dose of this study was set based on the results of the 28-day repeated dose toxicity test (See 'Cross-reference'): In this test, animals were dosed with 15, 100 or 500 mg/kg test item for 28 days. In the high dose group, 2 females died, and reduced locomotor activity, supression of body weight gain, increase in liver weight, thickening of the duodenal mucosa, etc. were observed in the remaining animals; no effects were observed at 100 mg/kg bw. In the range finding study, toxicity symptoms such as reduced locomotor activity were also observed in animals dosed with 125 and 250 mg/kg test item for 7 days. Based on the available information, the high dose of this study was set at 250 mg/kg bw, and the spacing factor was 2.5, so the medium dose was 100 mg/kg bw and the low dose 40 mg/kg bw.
Positive control:
Not required.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily before and after administration during administration and once daily during other rearing periods.

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: Male animals were weighed on the day 1 (administration start date), 7, 14, 21, 28, 35, 42 days and on the autopsy day. Female animals were weighed on days 1, 7 and 14 of administration, pregnancy 0 (mating confirmation date), 7, 14, 20 days, day of delivery, 4 days after and on the autopsy day. If any animals died, they were weighed at the time of death.

FOOD CONSUMPTION AND COMPOUND INTAKE (not a feeding study):
- Male animals were measured for food intake on days 1 to 2, 7 to 8, 14 to 15, 29 to 30, 35 to 36, and 41 to 42.
- Female animals were measured for food intake on days 1 to 2, 7 to 8, 14 to 15 of administration; days 0 to 1, 7 to 8, 14 to 15, 20 to 21 of gestation and days 3 to 4 of rearing.
Oestrous cyclicity (parental animals):
For each group, vaginal smear specimens were prepared every day after the start of administration following the sexual cycle observation up to the grouping day, and the sex cycle was observed until the copulation was confirmed. In addition, the average number of days from estrous period to estrous period was calculated for each individual.
Sperm parameters (parental animals):
Parameters examined in [P] male parental generations:
testis weight, epididymis weight, sperm count in testes, sperm count in epididymides.
Litter observations:
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.

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities.

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: Not specified.

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: Not specified.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals as soon as possible after the last litters were produced.
- Maternal animals: All surviving animals after the last litter of each generation was weaned (day 4).

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
- In males: liver, duodenum, prostate, seminal vesicle (including coagulated gland), mammary gland.
- In females: liver, duodenum, ovary, uterus, vagina, mammary gland. If the female or all babies died, also: heart, brain, kidney, lung. In females that had copulated but did not deliver or that died before delivery, liver and ovarian weights were measured.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated were prepared for microscopic examination and weighed:
- In males, histopathological examination was performed on the liver, duodenum, testis, epididymis (0.1 M phosphate buffered 10% formalin solution).
- In females, histopathological examination was performed on the liver, duodenum and ovaries. In females that had copulated but did not deliver or that died before delivery, the number of corpus luteum was counted under a stereoscopic microscope and the implantation rate [(number of implantation / number of pregnant corpus luteum) X 100,%] was determined. In females that died or when all babies died, histopathological examinations also included: heart, brain, kidney, lung.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at [4] days of age.
- These animals were subjected to postmortem examinations as follows: all pups were subject to macroscopic examination, and any organ presenting abnormalities was fixed in 0.1 M phosphate buffered 10% formalin solution and stored.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
Statistics:
Fisher's exact test of significance (5%) was carried out on the frequency, the mating rate, the conception rate and the morphological abnormality frequency of the babies.From the histopathological findings of the treated groups, the graded pathological tissue findings were determined by Marm-Whitney's U test and the total value of positive groups was determined by Fisher's one-sided probability test. Significant difference test with the group was conducted (significance: 5%).
For the other data, a value obtained for each individual or an average value for each litter was taken. First, the uniformity of variance of each group was tested (significance: 5%) by Bartlett's method. When the variance was uniform, a one-way analysis of variance (significance level: 5%) was performed, and when significance was found between groups, multiple comparisons were performed by the Dmrnett method (significance Level: 5%). Finally, Kruskal-Wallis's rank test (significance level: 5%) was performed when the variance was 0 in any group and when the variance was not uniform, significance was observed between the groups In the case of multiple comparison by Dunnett type test method (significance level: 5%).
Reproductive indices:
- Copulation index = [Number of copulated pairs / number of mated pairs (%)]
- Fertility index = [number of fertile males / copulation index x 100 (%)]
- Implantation index = [Number of implantation scars / number of corpora lutea x 100 (%)]
Offspring viability indices:
- Gestation index = [Number of dams with live offspring / number of pregnant dams x 100 (%)]
- Sex ratio = [Number of male offspring / (number of male + female offspring)]
- Delivery index = [Number of offspring at birth / number of implantation scars x 100 (%)]
- Birth index = [Number of live offspring at birth / number of implantation scars x 100 (%)]
- Live birth index = [Number of live offspring at birth / number of offspring at birth x 100 (%)]
- Viability index = [Number of live offspring 21 days after birth / number of live offspring after culling x 100 (%)]
Clinical signs:
no effects observed
Mortality:
mortality observed, treatment-related
Description (incidence):
Females: one animal in the 100 mg/kg group died at perinatal stage, five in the 250 mg/kg group (table 2).
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In the 250 mg/kg administration group, suppression of increase in body weight was observed in both males and females. No significant differences with the control were observed at any other dose.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In the 250 mg/kg administration group, female food intake decreased. No effects were observed at 100 mg/kg or less.
Food efficiency:
not specified
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):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
The liver weight of the 250 mg / kg administration group showed an increasing trend in both males and females. There was no significant difference between the treated group and the control group in absolute weight and relative weight of testis, epididymis, prostate (vaginal), seminal vesicle and ovary.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
- Males: miniaturisation of the testis at 100 and 250 mg/kg; thickening of the intestinal wall of the duodenum.
- Females: In the 250 mg/kg group, thickening of the intestinal wall of the duodenum was seen in 1 animal, darkening of the lungs in 3 animals, and emphysema in 1 animal, spleen miniaturisation in all animals, dark spots in the stomach mucosa in 4 animals, and red urine in 2 animals. In females that died during pregnancy or parturition, the fetuses remained in the uterus.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Histopathological examination of the liver revealed bile duct expansion and diffuse Kupffer cell proliferation around the Gleason sheath in males and females at 250 mg / kg administration group. On histopathological examination of the duodenum, thickening of the mucosa was observed in males and females at the dose of 250 mg / kg. Degeneration / necrosis of the proximal renal tubular epithelium in the kidney cortex was observed in females whose whole litter died by nursing day 4.
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed
Reproductive function: oestrous cycle:
effects observed, non-treatment-related
Description (incidence and severity):
The female sex cycle, mating rate and conception rate, number of corpus luteums of pregnant animals, implantation number, implantation rate and pregnancy period were not affected by administration of test substance.
The estrous regression days in the 250 mg/kg group increased, but it had been observed before administration that the same group's estrus regression days tended to be extended.
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
testis, epididymis, prostate and seminal vesicle weights were not affected by the administration of the test substance.
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
Failure of labor condition was observed in the group administered 100 and 250 mg / kg, and the birth rate decreased: the number of births and the number of babies born decreased, the live birth rate and the birth rate decreased. No effects on delivery rate, neonatal survival rate or sex ratio.
Key result
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
mortality
body weight and weight gain
histopathology: non-neoplastic
Key result
Dose descriptor:
NOAEL
Effect level:
40 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive performance
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
There was no effect of administration of the test substance on the sex ratio, the form and weight of the newborn infant.
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
The birth rate, fertility rate and neonatal survival rate decreased in the 250 mg/kg administration group.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
There was no effect of administration of the test substance on the sex ratio, the form and weight of the newborn infant.
Food efficiency:
not specified
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
Other effects:
no effects observed
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
Key result
Reproductive effects observed:
yes
Lowest effective dose / conc.:
100 mg/kg bw/day (nominal)
Treatment related:
yes
Relation to other toxic effects:
reproductive effects occurring together with other toxic effects, but not as a secondary non-specific consequence of other toxic effects

Table 2. Mortality in parental animals.

Animal number

Dose

(mg/kg)

Death period

Internal uterine findings

at necropsy

F03002

100

During labor

8 fetus remains

F04001

250

Nursing 3rd

No abnormality

F04004

250

Pregnancy 23rd

15 fetus remains

F04006

250

During labor

2 fetus remains

F04008

250

During labor

10 fetus remains

F04009

250

During labor

1 fetus remains

Table 3. Body weights of male rats.

Group

 

Olive oil (vehicle)

DCC

40mg/kg

DCC

100mg/kg

DCC

250mg/kg

Number of males

 

13

13

13

13

Days of administration

1

397.4

±

15.0

402.5

±

14.7

397.0

±

10.8

400.3

±

14.7

 

7

429.3

±

19.5

427.2

±

22.0

421.2

±

14.8

420.7

±

16.6

 

14

459.0

±

24.9

455.4

±

24.8

449.9

±

20.0

445.0

±

20.3

 

21

481.7

±

24.1

479.8

±

26.1

470.2

±

22.0

460.9

±

23.6

 

28

510.7

±

26.4

502.4

±

26.5

494.1

±

24.1

485.5

±

29.5

 

35

533.7

±

30.6

526.6

±

26.3

516.2

±

28.0

506.9

±

32.2

 

42

554.0

±

31.8

547.2

±

30.3

535.6

±

31.0

525.5

±

35.5

Each value shows mean (g)±S.D.

Significantly different from the control group (*: P<0.05, **: P<0.01).

Table 4. Body weights of female rats.  

Group

 

Olive oil (vehicle)

DCC 40 mg/kg

DCC 100 mg/kg

DCC 250 mg/kg

Number of females

 

13

13

13

13

Days of administration

 

 

 

 

 

 

1

247.3 ± 11.9

248.1 ± 9.4

246.1 ± 12.2

250.2 ± 11.6

 

7

258.6 ± 12.3

258.9 ± 10.3

259.7 ± 12.0

257.5 ± 13.9

 

14

270.9 ± 15.9

270.9 ± 15.9

266.1 ± 16.7

260.7 ± 16.2

Number of dams

 

13

12

12

12

Days of pregnancy

0

280.9 ± 12.6

279.7 ± 14.4

276.9 ± 19.4

267.5 ± 15.3

 

7

314.9 ± 17.1

309.7 ± 17.0

308.1 ± 21.4

293.9 ± 17.5 *

 

14

351.6 ± 17.8

342.1 ± 18.4

347.7 ± 25.8

326.4 ± 18.5 **

 

20

432.6 ± 25.8

415.0 ± 20.8

423.5 ± 30.0

392.3 ± 22.9 **

Number of dams

 

13

12

11

10

Days of lactation

0

323.8 ± 24.3

315.8 ± 34.0

307.7 ± 34.2

291.5 ± 36.1

 

4

339.6 ± 26.7 (12)

335.3 ± 20.8 (10)

331.2 ± 26.9 (10)

325.2 ± 13.4 (4)

Each value shows mean (g) ± S.D. Significantly different from the control group (*: P<0.05, **: P<0.01). Figures in parentheses indicate number of dams.

 

Table 5. Organ weights of male rats

Group

 

Olive oil (vehicle)

DCC 40 mg/kg

DCC 100 mg/kg

DCC 250 mg/kg

Number of males

 

13

13

13

13

Body weight

(g)

528.4 ± 33.5

521.6 ± 28.4

507.6 ± 28.9

491.7 ± 33.8

Liver

(mg)

14917.5 ± 1803.5

14677.0 ± 1145.0

15266.9 ± 2121.6

15699.2 ± 1366.2

 

(mg/g)

28.160 ± 2.004

28.149 ± 1.780

29.991 ± 2.914

31.930 ± 1.807**

Testes

(mg)

3257.5 ± 563.1

3441.9 ± 243.3

3293.5 ± 419.8

3387.7 ± 532.2

 

(mg/g)

6.209 ± 1.230

6.618 ± 0.603

6.512 ± 0.912

6.922 ± 1.228

Epididymides

(mg)

1192.3 ± 194.7

1301.6 ± 102.0

1198.8 ± 88.7

1266.9 ± 158.8

 

(mg/g)

2.268 ± 0.406

2.501 ± 0.223

2.368 ± 0.207

2.588 ± 0.377

Prostate, ventral

(mg)

626.5 ± 101.8

637.6 ± 162.5

638.5 ± 130.1

650.3 ± 104.6

 

(mg/g)

1.194 ± 0.229

1.226 ± 0.313

1.256 ± 0.246

1.331 ± 0.245

Seminal vesicles

(mg)

1907.7 ± 430.4

1669.7 ± 338.9

1837.3 ± 216.5

1752.7 ± 126.9

 

(mg/g)

3.614 ± 0.781

3.205 ± 0.638

3.617 ± 0.336

3.570 ± 0.214

Each value shows mean±S.D. Significantly different from the control group (*: P<0.05, **: P<0.01).

Table 6. Organ weights of female rats

Group

 

Olive oil (vehicle)

DCC 40 mg/kg

DCC 100 mg/kg

DCC 250 mg/kg

Number of females

 

12

10

10

4

Body weight

(g)

307.7±25.5

306.0±20.4

297.5±23.7

289.6±20.6

Liver

(mg)

10146.4±740.0

10223.8±555.2

15266.9±2121.6

15699.2±1366.2

 

(mg/g)

33.120±3.014

33.501±2.342

34.760±2.501

35.956±4.321

Ovaries

(mg)

105.5±9.7

102.9±11.6

109.0±12.9

97.6±9.8

 

(mg/g)

0.344±0.039

0.336±0. 032

0.367±0.044

0.338±0.033

Table 7. Reproductive performance.

Group

Olive oil (vehicle)

DCC 40 mg/kg

DCC 100 mg/kg

DCC 250 mg/kg

Number of pairs

13

13

13

13

Number of copulated pairs

13

13

13

13

Copulation index

100

100

100

100

Number of fertile males

13

12

12

12

Fertility index

100

92.3

92.3

92.3

Pairing days until copulation

2.6±1.3 (13)

3.2±1.0 (13)

2.8±1.2 (13)

2.6± 1.0(13)

Table 8. Reproduction/developmental toxicity.

Group

Olive oil (vehicle)

DCC 40 m/kg

DCC100mg/kg

DCC250mg/kg

Number of dams

13

12

11

 

8

 

Gestation length (days)

Mean ± S.D. per dam

Number of corpora lutea

22.5 ± 0.5

22.3 ± 0.5

22.5 ± 0.5

 

22.5 ± 0.5

 

Total

216

187

202

 

193

 

Mean ± S.D. per dam

Number of implantation scars

16.6 ± 2,3

15.6 ± 1.4

16.8 ± 1.8

(12)

16.1 ± 1.9

(12)

Total

198

173

190

 

175

 

Mean ± S.D. per dam

15.2 ± 2.0

14.4 ± 1.4

15.8 ± 1.9

(12)

14.6 ± 5.3

(12)

Implantation index (%)

92.2 ± 8.7

92.7 ± 7.1

94.3 ± 7.6

(12)

90.7 ± 19.4

(12)

Gestation index (%)

100.0

100.0

91.7

 

66.7

 

Number of offspring at birth

 

 

 

 

 

 

Total

184

164

161

 

101

 

Mean ± S.D. per dam

Number of live offspring at birth

14.2 ± 2.8

13.7 ± 1.4

14.6 ± 1.4

 

12.6 ± 4.3

 

Male

87

69

78

 

39

 

Female

92

90

73

 

39

 

Total

179

159

151

 

78

 

Mean ± S.D. per dam

Sex ratio

13.8 ± 2.6

13.3 ± 1.4

13.7 ± 2.1

 

9.8 ± 5.7

 

Mean±S.D. per dam

Number of dead offspring

0.49 ± 0.06

0.43 ± 0.10

0.51 ± 0.12

 

0.40 ± 0.28

 

Total

5

5

10

 

23

 

Mean ±S.D. per dam

Delivery index

0.4 ± 0.9

0.4 ± 0.7

1.9 ± 3.9

 

2.9 ± 4.5

 

Mean%±S.D. per dam

Birth index

92.1 ± 8.9

94.8 ± 5.5

95.3 ± 6.6

 

90.6 ± 12.4

 

Mean% ±S.D. per dam

Live birth index

89.8 ± 9.5

910 ± 5.6

81.9 ± 28.3

 

72.2 ± 35.0

 

Mean% ±S.D. per dam

97.6 ± 5.4

97.1 ± 4.6

93.9 ± 12.1

 

77.6 ± 33.2

 

Number of offspring on day 4

 

 

 

 

 

 

Male

78

58

72

 

28

 

Female

Sex ratio

83

74

63

 

17

 

Mean ±S.D. per dam

Viability index

0.49 ± 0.06 (12)

0.44 ± 0.09 (10)

0.53 ± 0.11

(10)

0.62 ± 0.09

(4)

Mean% ±S.D. per dam

90.9 ± 27.4

83.3 ± 38.9

86.2 ± 31.7

 

57.1 ± 53.5

(7)

Number of external abnormalities 0

0

0

0

 

0

 

Mean% ± S.D. per dam

0

0

0

 

0

 


Conclusions:
The NOAEL (based on toxicity) for both parent animals and first generation male/female Sprague Dawley rats was found to be 100 mg/kg bw; the NOAEL (based on reproductive ability) for parent male/female Sprague Dawley rats was found to be 40 mg/kg bw.
Executive summary:

A Reproduction / Developmental Toxicity Screening Test was conducted on the test item, according to OECD Guideline 421 (GLP study). 13 male and 13 female Sprague Dawley rats per group were exposed to 0 (control), 40, 100 or 250 mg/kg bw of test item by oral gavage, based on the results of a 28 -day repeated dose toxicity test performed previously. Males were dosed up to the day of conception and females were dosed throughout the study. Observations included clinical observations, body weight measurements, food intake, ostreus cycles, mating procedures, behavioural studies, evaluation of reproduction, weight of organs, and histopathological examinations. Toxicity signs were observed at the highest dose (lower body weight, enlarged liver, thickening of duodenum walls), and effects on reproduction were observed at the mid and high doses (lower birth rate, maternal deaths). Based on the available information, the NOAEL for general toxicity was set at 100 mg/kg bw, both for P0 and F1 generations, and the NOAEL for reproductive toxicity was set at 40 mg/kg bw.

Effect on fertility: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
40 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The available key study has a Klimisch score of 2.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

A Reproduction / Developmental Toxicity Screening Test was conducted on the test item, according to OECD Guideline 421 (GLP study). 13 male and 13 female Sprague Dawley rats per group were exposed to 0 (control), 40, 100 or 250 mg/kg bw of test item by oral gavage, based on the results of a 28 -day repeated dose toxicity test performed previously. Males were dosed up to the day of conception and females were dosed throughout the study.Observations included clinical observations, body weight measurements, food intake, ostreus cycles, mating procedures, behavioural studies, evaluation of reproduction, weight of organs, and histopathological examinations. Toxicity signs were observed at the highest dose (lower body weight, enlarged liver, thickening of duodenum walls), and effects on reproduction were observed at the mid and high doses (lower birth rate, maternal deaths). Based on the available information, the NOAEL for general toxicity was set at 100 mg/kg bw, both for P0 and F1 generations, and the NOAEL for reproductive toxicity was set at 40 mg/kg bw.

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

Based on the available information, the substance is not classified for reproductive toxicity according to CLP Regulation (EC) no. 1272/2008.