Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and OECD 422 compliant study with well characterized material
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: EPA, Health Effects Test Guidelines; OPPTS 870.3650: Combined Repeated Dose Toxicity Study With the Reproduction/Developmental Toxicity Screening Test (Jul 2000)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
other: Wistar Rat, Crl:WI(Han)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Age at study initiation: (P) 10-12 weeks; (F1)
- Weight at study initiation: males: 304 - 338 g; females: 189 - 210 g
- Housing: The rats were housed individually in Makrolon type M III cages supplied by Becker & Co., Castrop-Rauxel, Germany (floor area of about 800 cm²). Pregnant animals and their litters were housed together until day of parturition (PND) 4 (end of lactation).
- Diet: ground Kliba maintenance diet mouse-rat “GLP”, meal, Provimi Kliba SA, Kaiseraugst, Switzerland, ad libitum
- Water: ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 08-18-2011 To: 09-16-2011
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
- The appropriate amount of test substance was weighed out depending on the desired concentration. Then, drinking water was filled up to the desired volume, subsequently released with a high speed homogenizer. During administration of the test substance, preparations were kept homogeneous by stirring with a magnetic stirrer. The test substance preparations were produced at least once a week.

- The administration volume was 10 mL/kg bw.

Details on mating procedure:
- Fourteen days after the beginning of treatment, males and females from the same test group were mated for a maximum of 2 weeks.
Throughout the mating period, each female animal was paired with a predetermined male animal from the same test group.
- M/F ratio per cage: 1:1
- Length of cohabitation: overnight
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy and the following day gestation day (GD) 1
- After successful mating each pregnant female was caged (how): Pregnant animals and their litters were housed together until PND 4 (end of lactation).
- The females were allowed to litter and rear their pups until day 4 after parturition.
On PND 4, all pups were sacrificed under Isoflurane anesthesia with CO₂ and examined.

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration control analyses of all concentrations revealed that the values were in the expected range of the target concentrations, i.e. were always in a range of about 95-101% of the nominal concentrations.
Duration of treatment / exposure:
The duration of treatment covered a 2-week pre-mating and mating period in both sexes, approximately 1 week post-mating in males, and the entire gestation period as well as 4 days of lactation in females followed by an additional treatment until one day before sacrifice.
Females: 52 days
Males: 31 days
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
100, 300 and 1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Parental animals: Observations and examinations:
see also repeated dose toxicity: keyBASF85R0060/11C032, 2012.Repeated dose toxicity, oral, rat.OECD422
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once daily for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity; a check for moribund and dead animals was made twice daily on working days and once daily on Saturdays, Sundays and public holidays.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior to the administration period and thereafter at weekly intervals. The findings were ranked according to the degree of severity, if applicable. Thereby, the following parameters were examined:
1. abnormal behavior during “handling”
2. fur
3. skin
4. posture
5. salivation
6. respiration
7. activity/arousal level
8. tremors
9. convulsions
10. abnormal movements
11. impairment of gait
12. lacrimation
13. palpebral closure
14. exophthalmus
15. feces (appearance/consistency)
16. urine
17. pupil size

BODY WEIGHT: Yes
- Time schedule for examinations: before the start of the administration period in order to randomize the animals; during the administration period on study day 0 (start of the administration period) and thereafter once a week at the same time of the day (in the morning).
The following exceptions are notable for the female animals:
• During the mating period the parental females were weighed on the day of positive evidence of sperm (GD 0) and on GD 7, 14 and 20.
• Females with litter were weighed on the day of parturition (PND 0) and on PND 4.
• Females without a litter and without positive evidence of sperm in the vaginal smear were weighed weekly.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Generally, food consumption was determined once a week for male and female parental animals, with the following exceptions:
• Food consumption was not determined during the mating period (male and female F0 animals).
• Food consumption of the F0 females with evidence of sperm was determined on GD 0, 7, 14 and 20.
• Food consumption of F0 females, which gave birth to a litter was determined for PND 4.
Food consumption was not determined in females without positive evidence of sperm (during the mating period of dams used in parallel) and females without litter (during the lactation period of dams used in parallel) and in males after the premating period.

Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
- Pup number and status at delivery
• Total number of pups and the number of liveborn and stillborn pups in each litter on the day of birth.
• Pups, which died before the first determination of their status on the day of birth, were defined as stillborn pups.
- Pup viability/mortality
• Check for dead or moribund pups twice daily on workdays and once on Saturdays, Sundays or public holidays.
• Number and percentage of dead pups on the day of birth (PND 0) and of pups dying between PND 1 - 4 (lactation period).
• Pups which died accidentally or were sacrificed due to maternal death were not included in these calculations.
• The number of live pups/litter was calculated on the day after birth, and on lactation day 4.
- Pup clinical observations
• The live pups were examined daily for clinical symptoms (including gross-morphological findings) during the clinical inspection of the dams. If pups showed particular findings, these were documented with the dam concerned.
- Pup body weight data
• The pups were weighed on the day after birth (PND 1) and on PND 4.
• “Runts” were defined on the basis of the body weights on PND 1. "Runts" are pups that weigh less than 75% of the mean weight of the respective control pups.
• Sex ratio at day 0 and day 4 after birth = number of live male or female pups on day 0 and 4/ number of live male and female pups on day 0 and 4 x 100
On the day of birth (PND 0) the sex of the pups was determined by observing the distance between the anus and the base of the genital tubercle. The sex of the pups was finally confirmed at necropsy.

GROSS EXAMINATION OF DEAD PUPS:
• All surviving pups (sacrificed on PND 4 under isoflurane anesthesia with CO2), all stillborn pups and those pups, which died ahead of schedule, were examined externally, eviscerated and their organs were assessed macroscopically.
All pups were discarded after their evaluation.
Postmortem examinations (parental animals):
see also repeated dose toxicity: keyBASF85R0060/11C032, 2012.Repeated dose toxicity, oral, rat.OECD422
SACRIFICE
- All animals were sacrificed by decapitation under Isoflurane anesthesia.

GROSS NECROPSY
- The exsanguinated animals were necropsied and assessed by gross pathology.

HISTOPATHOLOGY / ORGAN WEIGHTS
Weight parameters:
Weight assessment was carried out on all animals.
1. Anesthetized animals
2. Epididymides
3. Testes
The following weights were determined in 5 animals/sex and test group (females with litters, same animals as used for clinical pathology examinations):
1. Adrenal glands
2. Brain
3. Heart
4. Kidneys
5. Liver
6. Spleen
7. Thymus

- Organ / Tissue preservation
The following organs / tissues were preserved in neutral-buffered 4% formaldehyde or in modified Davidson’s solution:
1. Adrenal glands
2. All gross lesions
3. Aorta
4. Bone marrow (femur)
5. Brain
6. Cecum
7. Cervix
8. Coagulating glands
9. Colon
10. Duodenum
11. Eyes with optic nerve (modified Davidson’s solution)
12. Esophagus
13. Extraorbital lacrimal gland
14. Epididymides (modified Davidson’s solution)
15. Femur with knee joint
16. Heart
17. Ileum
18. Jejunum (with Peyer’s patches)
19. Kidneys
20. Larynx
21. Liver
22. Lungs
23. Lymph nodes (axillary and mesenteric)
24. Mammary gland (male and female)
25. Nose (nasal cavity)
26. Ovaries (modified Davidson’s solution)
27. Oviducts
28. Pancreas
29. Parathyroid glands
30. Pharynx
31. Pituitary gland
32. Prostate gland
33. Rectum
34. Salivary glands (mandibular and sublingual)
35. Sciatic nerve
36. Seminal vesicles
37. Skeletal muscle
38. Spinal cord (cervical, thoracic and lumbar cord)
39. Spleen
40. Sternum with marrow
41. Stomach (forestomach and glandular stomach)
42. Target organs
43. Testes (modified Davidson’s solution)
44. Thymus
45. Thyroid glands
46. Trachea
47. Urinary bladder
48. Uterus
49. Vagina

From the liver, each one slices of the lobus dexter medialis and the lobus sinister lateralis were fixed in Carnoy’s solution and embedded in paraplast.

HISTOPATHOLOGY: Yes
Organ samples / Methods-Scope of examinations / Test group
1. All gross lesions: Hematoxylin-eosin (H&E), all affected animals per group, all treatment groups
2. Adrenal glands: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
3. Bone marrow (femur): Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
4. Brain: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
5. Cecum: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
6. Cervix: Hematoxylin-eosin (H&E), all affected animals per group, control and high dose group
7. Coagulation glands: Hematoxylin-eosin (H&E), all affected animals per group, control and high dose group
8. Colon: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
9. Duodenum: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
10. Epididymides: Hematoxylin-eosin (H&E), all affected animals per group
11. Heart: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
12. Ileum: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
13. Jejunum: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
14. Kidneys: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
15. Liver: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group; low and mid dose group: embedded in paraplast all animals per group.
16. Lung: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
17. Lymph nodes (mesenteric and axillary lymph nodes): Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
18. Ovaries: Hematoxylin-eosin (H&E), control and high dose group, all animals per group
19. Oviducts: Hematoxylin-eosin (H&E), control and high dose group, all animals per group
20. Peyer’s patches Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
21. Prostate: Hematoxylin-eosin (H&E), control and high dose group, all animals per group
22. Rectum: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
23. Sciatic nerve: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
24. Seminal vesicles: Hematoxylin-eosin (H&E), control and high dose group, all animals per group
25. Spinal cord (cervical, thoracic and lumbar cords): Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
26. Spleen: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
27. Stomach (forestomach and glandular stomach): Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
28. Testes: Hematoxylin-eosin (H&E), control and high dose group, all animals per group
29. Thymus: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
30. Thyroid glands: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
31. Trachea: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
32. Urinary bladder: Hematoxylin-eosin (H&E), 5 animals per sex per group, control and high dose group
33. Uterus: Hematoxylin-eosin (H&E), control and high dose group, all animals per group
34. Vagina: Hematoxylin-eosin (H&E), control and high dose group, all animals per group
Postmortem examinations (offspring):
SACRIFICE
- All surviving pups were sacrificed on PND 4 under isoflurane anesthesia with CO2.

GROSS NECROPSY
- All surviving pups, all stillborn pups and those pups, which died ahead of schedule, were examined externally, eviscerated and their organs were assessed macroscopically.
All pups were discarded after their evaluation.

Statistics:
- Food consumption (parental animals), body weight and body weight change (parental animals and pups; for the pup weights, the litter means were used), number of mating days, duration of gestation, number of implantation sites, postimplantation loss and % postimplantation loss, number of pups delivered per litter: DUNNETT-test (two-sided);
- Male and female mating indices, male and female fertility indices, gestation index, females with liveborn pups, females with stillborn pups, females with all stillborn pups, live birth index, pups stillborn, pups died, pups cannibalized, pups sacrificed moribund, viability index, number of litters with affected pups at necropsy: Pairwise comparison of each dose group with the control group using FISHER'S EXACT test for the hypothesis of equal proportions;
- Proportions of affected pups per litter with necropsy observations: Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians.
- Feces, rearing, grip strength of forelimbs and hindlimbs, landing foot-splay test, motor activity, clinical pathology parameters (except for urine color and turbidity), weight parameters: Non-parametric one-way analysis using KRUSKALWALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the equal medians.
Reproductive indices:
For the males, mating and fertility indices were calculated for F1 litters according to the following formulas:
- Male mating index (%) = number of males with confirmed mating*/number of males placed with females x 100
*defined by a female with vaginal sperm or with implants in utero
- Male fertility index (%) = number of males proving their fertility*/ number of males placed with females x 100
* defined by a female with implants in utero
For the females, mating, fertility and gestation indices were calculated for F1 litters according to the following formulas:
- Female mating index (%) = number of females mated*/ number of females placed with males x 100
* defined as the number of females with vaginal sperm or with implants in utero
- Female fertility index (%) = number of females pregnant*/ number of females mated** x 100
* defined as the number of females with implants in utero
** defined as the number of females with vaginal sperm or with implants in utero
- Gestation index (%) = number of females with live pups on the day of birth/number of females pregnant* x 100
* defined as the number of females with implants in utero
- Live birth index (%) = number of liveborn pups at birth/total number of pups born x 100
- Post implantation loss (%) = number of implantations number of pups delivered / number of implantations x 100
Offspring viability indices:
Viability index (%) = number of live pups on day 4 after birth/number of live pups on the day of birth x 100
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
- One male of the low dose group (100 mg/kg bw/d) was sacrificed moribund on study day 21 with a poor general state. Due to the isolated occurrence and the lack of a dose response relationship this was assessed as being incidental.
- All animals of test groups 100, 300 and 1000 mg/kg bw/d showed orange discolored feces from study day 1 onwards.
- Also during gestation all animals of the low and high dose group (100 and 1000 mg/kg bw/d) and 9/10 animals of the mid dose group (300 mg/kg bw/d) showed orange discolored feces during gestation.
- One sperm-positive F0 female of the low dose group and one sperm-positive F0 female of the mid dose group did not deliver any F1 pups.
- 9/10 animals of the low dose group, 8/10 animals of the mid dose group and all animals of the high dose group (100, 300 and 1000 mg/kg bw/d) showed orange discolored feces during lactation period.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
- In females of the mid dose group (300 mg/kg bw/d) food consumption during gestation was significantly decreased from days 0 to 7 and from days 7 to 14. Due to the normal range of the mean of means in the entire gestation period and the lack of a dose response relationship this was assessed as being incidental.
- No changes of toxicological concern with regard to body weight parameters of male animals were observed during the entire study period.
- The maternal body weight on study day 0 of the lactation period was decreased, but due to the isolated occurrence and the lack of a dose response relationship this was assessed as being incidental.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
- Male mating index: For almost all F0 parental males, which were placed with females to generate F1 pups, mating was confirmed, except one animal of the mid dose group. Thus, the male mating index was 100% in the low and high dose group (0, 100 and 1000 mg/kg bw/d). The mating index in the mid dose group (300 mg/kg bw/d) was 90%.
- Male fertility index: Fertility was proven for most of the F0 parental males within the scheduled mating interval to produce F1 litter.
One male animal of the low dose group and 2 males of the mid dose group did not generate F1 pups.
- The male fertility index ranged between 80% and 100% (Control: 100%; low dose group: 89%; mid dose group: 80% and high dose group: 100%). This finding reflected the normal range of biological variation inherent in the strain of rats used for this study as all respective values were within the range of the historical control data.

- Female mating index: The female mating index calculated after the mating period for F1 litter was 100% for the control and the low and high dose group (100 and 1000 mg/kg bw/d) and 90% for the mid dose group (300 mg/kg bw/d). The mean duration until sperm detected (GD 0) was 3.6, 3.6, 3.2 and 1.9 days in the control and the treatment groups 1, 2 and 3.

- Female fertility index: All sperm positive rats delivered pups with the exception of one female of the low dose group and two females of the mid dose group, which did not become pregnant. The female fertility index varied between 89% and 100% (Control: 100%; low dose group: 90%; mid dose group: 89% and high dose group: 100%). Three female animals, which delivered no pups had no implantation sites.

- The mean duration of gestation was 21.9 days in the control group and in the low and high dose group (100 and 1000 mg/kg bw/d). The duration in the mid dose group (300 mg/kg bw/d) was significantly increased (22.4 days) but was without a dose-response relationship or occurred in single rats only, these observations were considered to have been incidental.

- Gestation index: The gestation index was 100% in all test groups.

- Live birth indices: The rate of liveborn pups was not affected by the test substance, as indicated by live birth indices of 99% (Control, low and high dose group) and 100% (mid dose group). Single stillborn pups were only seen in the control and the low and high dose group. Thereby, the live birth index was in the range of the rat strain used (Historical Control Data).

- Postimplantation loss: Implantation, postimplantation loss and delivery were not affected by the treatment, since neither the mean number of implantation sites nor the average litter size showed any statistically significant differences between the groups.

- Fertility: The number of males with no offsprings and of not pregnant females is given in the following tables:
Male animals:
Test group (mg/kg bw/day): 0 / 100 / 300 / 1000
Number of animals: 10 / 9 / 10 / 10
Number of animals with no offspring: 0 / 1 / 2 / 0

Female animals:
Test group (mg/kg bw/day): 0 / 100 / 300 / 1000
Number of animals: 10 / 10 / 10 / 10
Number of animals not pregnant: 0 / 1 / 2 / 0

- All females that were not pregnant and all male mating partners did not show gross lesions in reproduction relevant organs.

ORGAN WEIGHTS (PARENTAL ANIMALS)
- Absolute weights
When compared to the control group, the mean absolute kidney weight was significantly altered in the mid dose group (300 mg/kg bw/d): 94% (p ≤ 0.05).

- Relative Organ Weights
When compared to control group, none of the mean relative weights determined was significantly altered.
The reduction in kidney weight in females of the mid dose group (300 mg/kg bw/d) is regarded to be incidental due to a missing altered relative organ weight, a missing dose response relationship and missing histopathologic findings that could explain the weight reduction.
All other mean weight parameters did not show significant differences when compared to the control groups.

GROSS PATHOLOGY (PARENTAL ANIMALS)
- The following relevant gross findings were observed (incidence is given in the table below):
Gross finding / Male animals (0 / 100 / 300 / 1000 mg/kg bw/day) // Female animals (0 / 100 / 300 / 1000 mg/kg bw/day)
Glandular stomach, discoloration of contents / 0 / 6 / 5 / 6 // 0 / 8 / 9 / 10
Jejunum, discoloration of contents / 0 / 1 / 0 / 0 // 0 / 4 / 4 / 9
Cecum, discoloration of contents / 0 / 5 / 10 / 9 // 0 / 0 / 0 / 0
Colon, discoloration of contents / 0 / 0 / 0 / 1 // 0 / 5 / 9 / 10
These findings are regarded to be treatment related.
All other gross lesions noted were single observations and they were regarded to have developed spontaneously and unrelated to compound and treatment.

- One male animal of the low dose group (100 mg/kg bw/d) had to be sacrificed. It revealed enlargement of several organs and lymph nodes which was in line with the histopathologic finding “Lymphoma malignant”. It was a single case and not regarded to be treatment related but spontaneous in origin.

HISTOPATHOLOGY (PARENTAL ANIMALS)
- The discoloration of the content of some parts of the digestive tract could not be observed in histopathology in the high dose group (1000 mg/kg bw/d). Probably the discoloration could not be detected because the content was completely or mostly lost during processing of the tissue.
The discoloration of the content in the digestive tract was regarded to be a consequence to the incorporation of the test substance which is of orange color. Therefore, the gross findings in the remaining animals and animals of the low dose group (100 mg/kg bw) and mid dose group (300 mg/kg bw/d) were not investigated in addition.

- Fertility: All organs of the reproduction tract did not show any relevant finding. All investigated not pregnant females as well as the remaining male mating partners did not show relevant histopathological findings that could explain infertility.

All other findings noted were either single observations or they were biologically equally distributed between control and treatment group. All of them were considered to be incidental or spontaneous in origin and without any relation to treatment.



Dose descriptor:
NOAEL
Remarks:
reproductive performance and fertility
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: NOAEL corresponding to the highest dose tested.
VIABILITY (OFFSPRING)
F1 generation litter/pups
• Pup number and status at delivery
- The mean number of delivered F1 pups per dam was evenly distributed about the groups.
- The single stillborn pup in the control group, low dose group (100 mg/kg bw/d) and high dose group (1000 mg/kg bw/d) were incidental and in the normal range of biological variation inherent in the strain of rats used for this study.
• Pup viability/mortality
- The viability index indicating pup mortality during lactation (PND 0 - 4) varied between 99% (control group, low and mid dose group) and 100% (high dose group) and was in the normal range of biological variation inherent in the strain of rats used for this study.
• Sex ratio
- The sex distribution and sex ratios of live F1 pups on the day of birth and PND 4 did not show substantial differences between the control and the test substance-treated groups. Slight differences were regarded to be spontaneous in nature.

CLINICAL SIGNS (OFFSPRING)
- The surviving F1 pups of any test group did not show adverse clinical signs up to scheduled sacrifice on PND 4.

BODY WEIGHT (OFFSPRING)
- Mean pup body weights/pup body weight changes of all pups in all test groups were comparable to the concurrent control values except in males of the mid dose group (300 mg/kg bw/d). These male pups showed slightly increased body weight values on PND 1 and 4 compared to control group, this was assessed as being incidental.
- In the low and mid dose group (100 and 300 mg/kg bw/d) one female runt and in the high dose group (1000 mg/kg bw/d) two in female and one male runts were seen.
These values were within the range of the biological variation inherent in the strain of rats used for this study.

GROSS PATHOLOGY (OFFSPRING)
- A few F1 pups showed spontaneous findings at gross necropsy, such as post mortem autolysis. These findings occurred without any relation to dosing and/or can be found in the historical control data at comparable or even higher incidences.
Thus, all these findings were not considered to be associated to the test substance.

Reproductive effects observed:
not specified
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Study duration:
subacute
Species:
rat
Quality of whole database:
Valid without restriction
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

There is a reliable study available to assess the potential of the substance for repeated dose toxicity after oral dosing.

Oral

A subacute study was conducted according to OECD 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) and GLP (BASF 2012; reliability score 1). The test substance was administered orally via gavage to groups of 10 male and 10 female Wistar rats (97.7% a.i. purity) at dose levels of 100, 300 and 1000 mg/kg bw. Drinking water served as vehicle. The duration of treatment covered a 2-week pre-mating and mating period in both sexes, approximately 1 week post-mating in males, and the entire gestation period as well as 4 days of lactation in females followed by an additional treatment until one day before sacrifice. After 2 weeks of premating treatment the F0 animals were mated to produce F1 generation pups.

 

A detailed clinical observation was performed in all animals before initial test substance administration and thereafter at weekly intervals. Food consumption and body weight was determined. Clinicochemical and hematological examinations, urinalyses as well as functional observational battery and motor activity examinations were performed towards the end of the administration period. All parental animals were assessed by gross pathology. Weights of selected organs were recorded and a histopathological examination was performed.

The pups were sexed and examined for macroscopically changes, body weight was determined and viability recorded. At necropsy on day of parturition (PND) 4, all pups were sacrificed and examined macroscopically for external and visceral findings.

 

No changes of toxicological concern with regard to food consumption and body weight parameters of male and female animals were observed during the entire study period. In females of the mid dose group food consumption during gestation was significantly decreased in the first two weeks. Due to the lack of a dose response relationship the reduced food consumption was assessed as being incidental.

Regarding clinical examinations, test substance-related, adverse findings were not observed in male or female parental animals of test groups 100, 300 and 1000 mg/kg bw/d during the entire study period. Due to the isolated occurrence and the lack of a dose response relationship, the incident of one male, sacrificed moribund on day 21 was assessed as being incidental. All animals showed orange discolored feces. This finding was substance related due to the orange color of the dye stuff. Clinical pathology revealed no treatment-related effects up to a dose of the compound of 1000 mg/kg bw/d. Macroscopic discoloration of the content of the digestive tract in numerous animals was regarded to be a consequence to the incorporation of the test substance which is of orange color and therefore treatment related but not adverse in nature. The significant reduction in absolute kidney weight in females of the mid dose group is regarded to be incidental due to a missing altered relative organ weight, a missing dose response relationship and missing histopathologic findings that could explain the weight reduction. All other mean weight parameters did not show significant differences when compared to the control groups.

Regarding reproductive performance and reproductive indices no test substance-related, adverse findings were noted. No delivery of pups and no implantation sites were observed in sperm-positive females of the low (1 animal) and mid dose (2 animals) group, respectively. Nevertheless, all organs of the reproduction tract did not show any relevant finding and all investigated not pregnant females as well as the remaining male mating partners did not show relevant histopathological findings that could explain infertility.

Concerning F1 pups, including litter data, viability, sex ration, clinical examinations and gross findings no test substance-related, adverse findings were noted.

 

 

CONCLUSION

Under the conditions of this reproduction/developmental toxicity screening test the NOAEL (no observed adverse effect level) for ) for reproductive performance and fertility was 1000 mg/kg bw/d for the F0 parental rats.

The NOAEL for developmental toxicity in the F1 progeny was found to be 1000 mg/kg bw/d.

The NOAEL for general, systemic toxicity was 1000 mg/kg bw/d based on the clinical, the clinical pathological and histopathological findings.

 

 


Short description of key information:
oral
Reproduction/developmental toxicity screening test: NOAEL for reproductive performance and fertility (F0 parental rats): 1000 mg/kg bw /d (BASF 2012), OECD 422, GLP, rat, gavage

dermal
no data

inhalation
no data

No substance-related effects were observed after repeated administration of high doses to animals.

Effects on developmental toxicity

Description of key information
oral
Reproduction/developmental toxicity screening test: NOAEL for developmental toxicity (F1 progeny): 1000 mg/kg bw /d (BASF 2012), OECD 422, GLP, rat, gavage
dermal
no data
inhalation
no data
No substance-related effects were observed after repeated administration of high doses to animals.

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available screening study is considered reliable and suitable for classification purposes under Directive 67/548/EEC. As a result the substance is not considered to be classified for reproduction toxicity under Directive 67/548/EEC as amended for the 31st time in Directive2009/2/EG.

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

The available screening study is reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is not considered to be classified for reproduction toxicity under Regulation (EC) No 1272/2008, as amended for the second time in Directive (EC 286/2011