Registration Dossier

Administrative data

Description of key information

OECD 422: NOAEL (males) = 100 mg/kg bw/d
OECD 408: NOAEL (males/females) = 300 mg/kg bw/d

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline 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: U.S. EPA, 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:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH (females) and Charles River, UK (males)
- Age at study initiation:10-12 weeks (males/females)
- Weight at study initiation: m (314.4-316.7); f (195.8-199.5)
- Fasting period before study: no data
- Housing:individually in type M III polycarbonate cages supplied by Becker & Co., Castrop Rauxel, Germany (floor area of about 800 cm2)
- Diet (e.g. ad libitum):ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 6-8 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C):20-24 °C
- Humidity (%):30-70%
- Air changes (per hr):10 air changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance was applied as a solution. To prepare the solution, the appropriate amount of test substance was weighed out depending on the desired concentration. Then the vehicle (propylene glycol) was filled up to the desired volume, subsequently mixed using a magnetic stirrer.
The test-substance solutions were prepared in such intervals that the stability was guaranteed.

VEHICLE
- Justification for use and choice of vehicle (if other than water): solubility
- Concentration in vehicle:

group dose conc. Male Female Male
0 0 0 5 10 10
1 100 2.0 5 10 10
2 300 6.0 5 10 10
3 1000 20.0 5 10 10

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses of the test-substance preparations
The analyses of the test substance preparations were carried out at the Analytical Chemistry Laboratory of Experimental Toxicology and Ecology of BASF SE as a part of the study.
The stability of the test substance in propylene glycol stored deep frozen over a period of 7 days followed by 4 hours at room temperature was demonstrated before the start of the study (Project No.: 01Y0471/09Y004).
Concentration control and homogeneity analyses of the test-substance preparations were performed in samples of all concentrations at the start and at the end of the administration period.

Food analysis
The food used in the study was assayed for chemical and microbiological contaminants.

Drinking water analysis
The drinking water is regularly assayed for chemical contaminants both by the municipal authorities of Frankenthal and by the Environmental Analytics Water/Steam Monitoring Department of BASF SE as well as for bacteria by a contract laboratory. The German “Trinkwasserverordnung” (Drinking Water Regulation) will serve as the guideline for maximum tolerable contaminants.

Bedding and enrichment analysis
The bedding and the enrichment are regularly assayed for specific contaminants (chlorinated hydrocarbons and heavy metals).
Duration of treatment / exposure:
females were treated for 49 days and males for 35
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
100, 300, 1000 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
10 m / 10 f
Control animals:
yes, concurrent vehicle
Details on study design:
Dose selection rationale: In a previously performed 14-days test study (Project No.: 10C0471/09069) ASA was administered by gavage to groups of 3 male and 3 female Wistar rats at dose levels of 0, 100, 300, 1000 mg/kg body weight/day (mg/kg bw/d). Propylene glycol swas the vehicle.
Positive control:
none
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: before and after treatment for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity. Abnormalities and changes were documented for each animal.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:before and after treatment. The littering and lactation behavior of the dams was generally evaluated in the mornings in combination with the daily clinical inspection of the dams.The day of littering was considered to be a 24-hour period from about 15.00 h of one day until about 15.00 h of the following day.

BODY WEIGHT: Yes
- Time schedule for examinations:once a week. 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.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Once a week (in a period of 7 days) 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 on PND 0 and 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).
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood:towards the end of the administration period (study days 35 (males) and 49 (females))
- Anaesthetic used for blood collection: No
- Animals fasted: No
- How many animals:
- Parameters checked in table [No.?] were examined.

CLINICAL CHEMISTRY: Yes / No / No data
- Time schedule for collection of blood:
- Animals fasted: Yes / No / No data
- How many animals: 5 randomly selected
- Parameters c Leukocyte count, Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, Mean corpuscular hemoglobin, Mean corpuscular hemoglobin concentration, Platelet count, Differential blood count, Reticulocytes were examined.

URINALYSIS: Yes
- Time schedule for collection of urine:towards the end of the administration period (study days 35 (males) and 49 (females))
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters: pH, Protein, Glucose, Ketones, Urobilinogen, Bilirubin, Blood, Specific gravity, Sediment, Color, turbidity, Volume were examined.

NEUROBEHAVIOURAL EXAMINATION:No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes


Organ weights
The following weights were determined in all animals sacrificed on schedule:
1. Anesthetized animals
2. Adrenal glands
3. Brain
4. Cauda epididymis
5. Epididymides
6. Heart
7. Kidneys
8. Liver
9. Ovaries
10. Pituitary gland
11. Prostate
12. Seminal vesicles with coagulation glands
13. Spleen
14. Testes
15. Thymus
16. Thyroid glands (with parathyroid glands)
17. Uterus

Organ/tissue fixation
The following organs or tissues of parental animals were fixed in 4% formaldehyde solution or in modified Davidson’s solution:
1. All gross lesions
2. Adrenal glands
3. Aorta
4. Bone marrow (femur)
5. Brain
6. Cecum
7. Cervix
8. Coagulation glands
9. Colon
10. Duodenum
11. Epididymides (modified Davidson`s solution)
12. Esophagus
13. Eyes with optic nerve
14. Female mammary gland
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 (mesenteric and axillary lymph nodes)
24. Nose (nasal cavity)
25. Ovaries
26. Oviducts
27. Pancreas
28. Parathyroid glands
29. Pharynx
30. Pituitary gland
31. Prostate
32. Rectum
33. Salivary glands (mandibular and sublingual glands)
34. Seminal vesicle
35. Sciatic nerve
36. Skeletal muscle
37. Spinal cord (cervical, thoracic and lumbar cords)
38. Spleen
39. Sternum with marrow
40. Stomach (forestomach and glandular stomach)
41. Testes (modified Davidson’s solution)
42. Thymus
43. Thyroid glands
44. Trachea
45. Urinary bladder
46. Uterus
47. Vagina


HISTOPATHOLOGY: Yes
1. All gross lesions
2. Adrenal glands
3. Bone marrow (femur)
4. Brain
5. Cecum
6. Cervix
7. Coagulation gland
8. Colon
9. Duodenum
10. Epididymides
11. Heart
12. Ileum
13. Jejunum
14. Kidneys
15. Liver
16. Lung
17. Lymph nodes (mesenteric and axillary lymph nodes)
18. Ovaries
19. Peyer’s patches
20. Prostate
21. Rectum
22. Sciatic nerve
23. Seminal vesicle
24. Spleen
25. Spinal cord (cervical, thoracic and lumbar cords)
26. Stomach (forestomach and glandular stomach)
27. Testes
28. Thymus
29. Thyroid glands (with parathyroid glands)
30. Trachea
31. Urinary bladder
32. Uterus
33. Vagina

Other examinations:
CLINICAL PATHOLOGY
In the morning, blood was taken from the retroorbital venous plexus from fasted animals. The animals were anaesthetized using isoflurane. The blood sampling procedure and the subsequent analysis of the blood and serum samples were carried out in a randomized sequence. For urinalysis the individual animals were transferred to metabolism cages (withdrawal of food and water) and urine was collected overnight. The urine samples were evaluated in a randomized sequence
The assays of blood and serum parameters were performed under internal laboratory quality control conditions with commercial reference controls to assure reliable test results.
The results of the clinical pathology examinations are expressed in units of the International System (SI).
The examinations were carried out in 5 animals per test group and sex.
Statistics:
Statistics of the clinical examinations
DUNNETT-test , FISHER'S EXACT test, WILCOXON-test, KRUSKAL-WALLIS test

Statistics of clinical pathology
FISHER'S EXACT test, KRUSKAL-WALLIS test

Statistics of pathology
KRUSKAL-WALLIS test, WILCOXON-test
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
No mortality, piloerection, poor general state in female animals during lactation, insufficient maternal care
Mortality:
mortality observed, treatment-related
Description (incidence):
No mortality, piloerection, poor general state in female animals during lactation, insufficient maternal care
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Increased platelet counts in rats of both sexes, increased WBC counts, absolute neutrophil, relative and absolute monocyte as well as relative and absolute LUC counts in males
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Reduced prothrombin time in males / Decreased total protein and albumin values in males / Increased cholesterol values in males / Increased ALT activities and bilirubin values in rats of both sexes / Increased SGGT activities in males
Urinalysis findings:
no effects observed
Description (incidence and severity):
No treatment-related changes among urinalyses parameters were measured
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Diffuse squamous cell hyperplasia in the forestomach of all males and females. Multifocal atypical bile duct hyperplasia in the liver of 8 males and of all females.
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
No animal died prematurely during the study period.In test group 3 (1000 mg/kg bw/d), salivation after treatment was observed in 6 males from study week 1 onwards and in 2 males in study weeks 3 and 4. Three male animals in test group 2 (300 mg/kg bw/d) showed salivation after treatment in study weeks 3 and 4. In females no salivation was observed during the premating period. One male of test group 0 (control) showed an eye injury in study week 4.
Salivation after treatment was observed in 3 to 4 female animals of test group 3 (1000 mg/kg bw/d) from GD 3 onwards. Salivation after treatment was also observed in 1 female animal of test group 2 (300 mg/kg bw/d) from GD 8 onwards.
Five female animals of test group 3 (1000 mg/kg bw/d), i.e. animal Nos. 132, 134, 137, 138 and 139, showed insufficient maternal care of pups. In two litters no more pups were alive on PND 1 (animal No. 139) and 2 (animal No. 138). Four of these animals also showed piloerection, i.e. animal Nos. 132, 134, 138 and 139. In addition urine smeared anogenital region was observed in female No. 139. Three dams of test group 3 (1000 mg/kg bw/d), i.e. animal Nos. 135, 136 and 139, showed salivation after treatment. Animal No. 124 of test group 2 (300 mg/kg bw/d) showed insufficient maternal care, piloerection and salivation after treatment. Poor general state, piloerection and insufficient maternal care of pups were observed in animal No. 111 of test group 1 (100 mg/kg bw/d).

BODY WEIGHT AND WEIGHT GAIN
No significant changes in body weight and body weight change values for male and female animals were observed during the entire study period.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
No significant changes in food consumption values for male and female animals were observed during the entire study period. Although not significantly decreased, food consumption in female animals of test group 3 (1000 mg/kg bw/d) was only 78% compared to test group 0 (control) during lactation.

FOOD EFFICIENCY
no data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study)
no data

OPHTHALMOSCOPIC EXAMINATION
no data

HAEMATOLOGY
In male rats of test group 3 (1000 mg/kg bw/d) total white blood (WBC) counts were increased which was mainly due to an increase of the absolute neutrophil counts. In addition, absolute and relative monocyte counts (relative counts not significant) as well as absolute and relative large unstained cell (LUC) counts were higher in males of test group 3 (1000 mg/kg bw/d) compared to controls. In females of test group 2 (300 mg/kg bw/d) relative monocyte counts were lower compared to controls, but this decrease was not dose-dependent, and it was not accompanied by any other alteration in the differential blood cell counts in these animals. Therefore, this change was regarded as incidental and not treatment-related.
In male and female rats of test group 3 (1000 mg/kg bw/d) platelet counts were increased (not significantly in females).
In males of test group 3 (1000 mg/kg bw/d) the prothrombin time (Hepato Quick’s test) was reduced compared to controls.

CLINICAL CHEMISTRY
In male and female rats of test groups 2 and 3 (300 and 1000 mg/kg bw/d) the alanine aminotransferase (ALT) activity was increased. The ALT values in rats of test group 2 (300 mg/kg bw/d) were within (females) or at the border of (males) the historical control range (ALT males: 0.46-0.94 µkat/L; ALT females: 0.42-0.73 µkat/L, PART III, Supplement). In males of this test group marginal higher ALT activities were the only altered parameter. In addition to the ALT activity increase in females of test group 2 (300 mg/kg bw/d), significantly higher total bilirubin values were measured, but these values were also in the historical control range (bilirubin: 2.30-3.46 µmol/L, PART III, Supplement). Therefore, the ALT activity increases in rats of both sexes of test group 2 (300 mg/kg bw/d) as well as the higher bilirubin values in females of the same test group were regarded as incidental and not treatment-related (ECETOC Technical Report No. 85, 2002). In 2 males of test group 3 (1000 mg/kg bw/d; Nos. 32 and 35) the -glutamyltransferase (SGGT) activity was increased.
In males of test group 3 (1000 mg/kg bw/d) albumin and total protein values (not significant) were decreased, whereas cholesterol values in males of the same test group were increased.
The bilirubin values in females of test group 3 (1000 mg/kg bw/d) were higher compared to controls and at least 1 male rat (No. 35) in the same test group did also have a high total bilirubin level.
In females of test groups 1 and 2 (100 and 300 mg/kg bw/d) lower triglyceride levels were measured compared to controls, but this decrease was not dose-dependent and, therefore, it was regarded as incidental rather than treatment-related.

URINALYSIS
No treatment-related changes among urinalyses parameters were measured.

NEUROBEHAVIOUR
no data

ORGAN WEIGHTS
All mean absolute weight parameters in test groups 1-3 (100, 300 and 1000 mg/kg bw/d) did not show significant differences when compared to test group 0 (control).
When compared to test group 0 (control; set to 100%), the mean relative weights of following organs were significantly increased: kidney, testes (male); liver (female)
All other mean relative weight parameters did not show significant differences when compared to test group 0 (control).

The increase of the relative kidney weight in males of test group 1 (100 mg/kg bw/d) was regarded to be incidental as no dose-response relationship occurred. The increased relative liver weight in females of test group 2 (300 mg/kg bw/d) was also considered to be incidental because the mean absolute weight was not significantly increased and no histopathological correlate was present.
For the increase of the relative testes weight in males of test group 3 (1000 mg/kg bw/d) there were no histopathological correlates. Furthermore, the absolute weight of testes did not show significant differences. Therefore, the increased testes weight was related to the slightly but not significantly decreased terminal body weight (-5%) in these males.

GROSS PATHOLOGY
Forestomach: Three males of test group 2 (300 mg/kg bw/d) and 6 males of test group 3 (1000 mg/kg bw/d) showed a yellow-white deposition on the forestomach epithelium. A thickening of the forestomach wall was observed in all males of test group 3 (1000 mg/kg bw/d).
Duodenum: The duodenal wall was slightly thickened in 7 males of test group 3 (1000 mg/kg bw/d).
All other gross lesions occurred singly and were considered to be incidental.

HISTOPATHOLOGY: NON-NEOPLASTIC
Forestomach: A diffuse squamous cell hyperplasia was observed in the forestomach of 1 control female and of 1 female in test group 1 (100 mg/kg bw/d). In test groups 2 (300 mg/kg bw/d) and 3 (1000 mg/kg bw/d), incidence and severity of diffuse squamous cell hyperplasia were increased dose-related. The squamous cell hyperplasia was characterized by an increased number of epithelial cells showing small exophytic structures into the lumen or small endophytic finger-like projections. The hyperplasia was accompanied by an infiltration of some lymphoid cells and granulocytes. The hyperplasia correlated with the thickening of the forestomach wall that was grossly diagnosed in all males of test group 3 (1000 mg/kg bw/d). The increased occurrence of squamous cell hyperplasia was considered to be treatment-related.
The squamous cell hyperplasia occurred mostly in combination with hyperkeratosis. The macroscopically diagnosed yellow-white deposition on the forestomach epithelium corresponded histopathologically with a diffuse orthokeratotic hyperkeratosis, characterized by an increased thickening of the superficial non-nucleated keratin layer. The incidence of hyperkeratosis was increased treatment-related in males of test group 2 (300 mg/kg bw/d) as well as in males and females of test group 3 (1000 mg/kg bw/d).
The occurrence of the minimal squamous cell hyperplasia in the forestomach of 1 control female and 1 female of test group 1 (100 mg/kg bw/d) as well as the occurrence of hyperkeratosis in 1 control female, in 2 females of test group 1 (100 mg/kg bw/d) and in 2 females in test group 2 (300 mg/kg bw/d) were considered to be incidental.

Liver: A multifocal atypical bile duct hyperplasia was observed in 2 males of test group 2 (300 mg/kg bw/d) as well as in 8 males and all females of test group 3 (1000 mg/kg bw/d). This finding was characterized by a multifocal proliferation of bile ducts in the portal region. The bile duct epithelium seemed slightly atypical. It was single layered, often cuboidal and slightly basophilic. Some mitotic figures and necrotic cells were present within the epithelium. The hyperplasia was associated with periductular fibrosis and periductular lymphohistiocytic infiltrates.
Furthermore, a minimal centrilobular, hepatocellular hypertrophy was observed in 8 males and 7 females of test group 3 (1000 mg/kg bw/d).

The occurrence of multifocal atypical bile duct hyperplasia and of centrilobular, hepatocellular hypertrophy was considered to be treatment-related.

Thyroid glands: A minimal or slight follicular hypertrophy/hyperplasia was observed in 1 control male, in 2 males of test group 1 (100 mg/kg bw/d), in one male of test group 2 (300 mg/kg bw/d), and in 4 males of test group 3 (1000 mg/kg bw/d). In affected animals the number of small follicles was slightly increased or the follicular epithelium was higher, varying in size from cuboidal cells to columnar cells.
For the slightly increased incidence of hypertrophy/hyperplasia in males of test group 3 (1000 mg/kg bw/d), a treatment related effect could not be ruled out.

Duodenum: For the macroscopically described thickening of duodenal wall in 7 males of test group 3 (1000 mg/kg bw/d), there was no histopathological correlate.

All other findings occurred either individually or were biologically equally distributed over the control group and the treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

Dose descriptor:
NOAEL
Remarks:
systemic effects
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: haematology; gross pathology; histopathology;
Dose descriptor:
NOAEL
Remarks:
fertility
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects on fertility observed
Dose descriptor:
dose level:
Effect level:
>= 300 - <= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: See remarks
Remarks on result:
other: Diffuse squamous cell hyperplasia in the forestomach of males and females and multifocal atypical bile duct hyperplasia in the liver of males
Dose descriptor:
dose level:
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: See remarks
Remarks on result:
other: See remarks in basis for effect level
Remarks:
Increased platelet counts in rats of both sexes Increased ALT activities and bilirubin values in rats of both sexes Increased SGGT activities in males Increased WBC counts, absolute neutrophil, relative and absolute monocyte as well as relative and absolute LUC counts in males Reduced prothrombin time in males Decreased total protein and albumin values in males Increased cholesterol values in males
Critical effects observed:
not specified

Functional observational battery

Home cage observations:

No test substance-related or spontaneous findings were observed in male and female animals of all test groups during the home cage observation.

  

Open field observations:

One female animal of test group 2 (300 mg/kg bw/d;animal No. 128) showed piloerection. All other male and female animals of all test groups did not reveal any test substance-related findings during home cage observation.

 

Sensorimotor tests/reflexes:

There were no test substance-related findings in male and female animals of all test groups. Any deviations from "zero values" were equally distributed between test substance-treated groups and controls or occurred in single animals only. Therefore,these observations were considered as being incidental.

 

Quantitative Parameters:

No test substance-related impaired parameters were observed in male and female animals of all test group

 Motor activity measurement

There were no significant deviations concerning the overall motor activity (summation of all intervals) in the male and female animals of all test groups in comparison to the concurrent control group.

Regarding single intervals, in males of test group 1 (100 mg/kg bw/d) one isolated significantly increased value was measured at interval 10. This finding was considered as being incidental since the overall motor activity was not changed and no findings were observed for female animals.

Conclusions:
The sults of this OECD 422 study are, that the no observed adverse effect level (NOAEL) for systemic toxicity in F0 males was 100 mg/kg bw/d due to pathological findings at 300 mg/kg bw/d and above including increased blood values and squamous cell hyperplasia in the forestomach of all animals. However, the NOAEL for systemic toxicity in F0 females could not be established, although the dose of 100 mg/kg bw/d was without pathological findings. This was due tothenoted insufficent maternal care at all dose levels correlates with increased pup mortality. The cause for the insufficent maternal care is presently undecided as the effect may be due to systemic maternal toxicity or due to an effect on the pups via lactation. The NOAEL for fertility in male and female Wistar rats was 1000 mg/kg bw/d.
Executive summary:

In a study performed according to OECD 422 and GLP, the test substance was administered orally by gavage to groups of 10 male and 10 female Wistar rats (F0 parental animals) at dose levels of 0 mg/kg body weight/day (mg/kg bw/d; vehicle control; test group 0), 100 mg/kg bw/d (test group 1), 300 mg/kg bw/d (test group 2) and 1000 mg/kg bw/d in propylene glycol, respectively. As results, the no observed adverse effect level (NOAEL) for systemic toxicity in F0 males was 100 mg/kg bw/d due to pathological findings at 300 mg/kg bw/d and above including increased blood values and squamous cell hyperplasia in the forestomach of all animals. However, the NOAEL for systemic toxicity in F0 females could not be established, although the dose of 100 mg/kg bw/d was without pathological findings. This was due to the noted insufficent maternal care at all dose levels correlates with increased pup mortality. The cause for the insufficent maternal care is presently undecided as the effect may be due to systemic maternal toxicity or due to an effect on the pups via lactation. The NOAEL for fertility in male and female Wistar rats was 1000 mg/kg bw/d.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
100 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
OECD 422 and OECD 408 study performed according GLP avaialable.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

In a study performed according to OECD 422 and GLP, the test substance was administered orally by gavage to groups of 10 male and 10 female Wistar rats (F0 parental animals) at dose levels of 0 mg/kg body weight/day (mg/kg bw/d; vehicle control; test group 0), 100 mg/kg bw/d (test group 1), 300 mg/kg bw/d (test group 2) and 1000 mg/kg bw/d in propylene glycol, respectively. As results, the no observed adverse effect level (NOAEL) for systemic toxicity in F0 males was 100 mg/kg bw/d due to pathological findings at 300 mg/kg bw/d and above including increased blood values and squamous cell hyperplasia in the forestomach of all animals. However, the NOAEL for systemic toxicity in F0 females could not be established, although the dose of 100 mg/kg bw/d was without pathological findings. This was due tothenoted insufficent maternal care at all dose levels correlates with increased pup mortality. The cause for the insufficent maternal care is presently undecided as the effect may be due to systemic maternal toxicity or due to an effect on the pups via lactation. The NOAEL for fertility in male and female Wistar rats was 1000 mg/kg bw/d.

Long-term toxicity of ASA was evaluated in a study in accordance with OECD 408, where groups of ten male and ten female Wistar Han™:RccHan™:WIST strain rats, were treated for ninety consecutive days, at dose levels of 0, 10, 100 and 300 mg/kg bw/day ASA in Arachis oil, respectively. Clinical signs, functional observations, body weight change, dietary intake and water consumption were monitored during the study. Hematology and blood chemistry were evaluated for all animals at the end of the study. Ophthalmoscopic examination was also performed on control group and high dose animals. All animals were subjected to gross necropsy examination and histopathological evaluation of selected tissues from high dose and control animals was performed. As result, the NOEL was considered to be 10 mg/kg bw/day for males and 100 mg/kg bw/day for females. The noted lower body weight gains in males treated with 300 mg/kg bw/day were considered insufficient in magnitude to represent an adverse effect of treatment. The increased liver weights in animals of either sex treated with 300 mg/kg bw/day and in males treated with 100 mg/kg bw/day, accompanied by associated increases in hepatic enzymes, in the absence of microscopic changes in the liver do not represent an adverse effect of treatment. The effects were considered to represent an adaptive response to treatment so that 300 mg/kg bw/day was regarded as the NOAEL for animals of either sex.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
OECD guideline study according to GLP with lowest observed effect level

Justification for classification or non-classification

There are conclusive and sufficient data for classification of with regard to repeated dose toxicity.

The substance is not classified for this endpoint in accordance to Directive 67/548/EEC or to the CLP Regulation (EC) No 1272/2008.