1-Propene, hydroformylation products, high-boiling

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD Guideline Study, GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

adopted 03 Oct 2008

GLP compliance:

yes (incl. QA statement)

Remarks:

Experimental Toxicology and Ecology BASF SE 67056 Ludwigshafen, Germany

Limit test:

no

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): Oxooel 740
- Physical state: liquid/ yellowish, clear
- Lot/batch No.: Tk251_20090716
- Storage condition of test material: refrigerator (KS), dry storage

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Sulzfeld, Germany
- Age at study initiation: 42±1 days (age when supplied: 34±1 days)
- Housing: 5 animals per cage
- Diet: Kliba maintenance diet mouse/rat “GLP”, meal, supplied by Provimi Kliba SA, Kaiseraugst, Switzerlandad libitum; ad libitum
- Water: drinking water (from water bottles), ad libitum
- Acclimation period: 8 days

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

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test substance was applied as a solution. To prepare this solution, the appropriate amount of test substance was weighed out depending on the desired concentration. Then, Olive Oil Ph. Eur was filled up to the desired volume, subsequently released manually. During
administration of the test substance, preparations were kept homogeneous by stirring with a
magnetic stirrer. The test-substance preparations were produced once a week.

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 20, 60, or 200 g/L
- Amount of vehicle (if gavage):
- Lot/batch no. (if required):
- Purity:

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Capillary gas chromatography with area-% and correction with the water content. Apparatus: Capillary gas chromatograph with split injector and flame ionization detector

Duration of treatment / exposure:

28 days

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300, 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: A range-finding study at dose levels of 0, 300 and 1000 mg/kg bw/d for 2 weeks was performed (experimental conduct in accordance with GLP, without GLP-status). The following test item-related adverse findings were noted:
1000 mg/kg bw/d: Salivation slight and moderate after treatment was observed in 3 males and 3 females.
300 mg/kg bw/d: No substance-related findings were observed.
The following dose levels were selected for the present study:
1000 mg/kg bw/d as high dose, 300 mg/kg bw/d as mid dose, 100 mg/kg bw/d as low dose
The dose levels for this study were selected in accordance with the requirements set forth in the test guidelines and based on the results of the preliminary range finding test.

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: A check for moribund and dead rats was made twice daily on working days and once daily on Saturdays, Sundays and public holidays. All animals were checked daily before as well as <1 hour and 3-4 hours after the administration for clinically abnormal signs. Abnormalities and changes were documented for each animal.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Detailed clinical observations were performed in all rats prior to the administration period and thereafter at weekly intervals.The findings were ranked according to the degree of severity, if applicable. The rats were transferred to a standard arena (50 × 37.5 cm with sides of 25 cm high). 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 and during the administration period on day 0 (start of the administration period) and thereafter at weekly intervals.

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily visual inspection

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: In the morning, blood was taken from the retroorbital venous plexus from fasted animals.
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked: Leukocyte count (WBC), Erythrocyte count (RBC), Hemoglobin (HGB), Hematocrit (HCT), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular haemoglobin concentration (MCHC), Platelet count (PLT), Differential blood count, Reticulocytes, Prothrombin time (HQT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: In the morning, blood was taken from the retroorbital venous plexus from fasted animals.
- Animals fasted: Yes
- How many animals: all rats
- Parameters checked: Alanine aminotransferase, aspartate aminotransferase, Alkaline phosphatase, γ-Glutamyltransferase, Sodium, Potassium, Chloride, Inorganic phosphate, Calcium, Urea, Creatinine, Glucose, Total bilirubin, Total protein, Albumin, Globulins, Triglycerides, Cholesterol, Magnesium, Bile acids

URINALYSIS: Yes
- Time schedule for collection of urine: For urinalysis the individual animals were
transferred to metabolism cages (withdrawal of food and water) and urine was collected
overnight.
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked: pH, Protein, Glucose, Ketones, Urobilinogen, Bilirubin, Blood, Specific gravity, Sediment, Color, turbidity, Volume

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: A Functional observational battery (FOB) and a measurement of motor activity (MA) was performed at the end of the administration (day 26/ d27) period starting at about 10:00 h
- Dose groups that were examined: all rats
Functional observation battery (FOB)
The FOB started with passive observations without disturbing the rats, followed by removal from the home cage, open field observations in a standard arena and sensorimotor tests as well as reflex tests. The findings were ranked according to the degree of severity, if applicable.

Home cage observations
The rats were observed in their closed home cages; any disturbing activities (touching the cage or rack, noise) were avoided during these examinations in order not to influence the behavior of the rats. Attention was paid to:
1. posture
2. tremor
3. convulsions
4. abnormal movements
5. impairment of gait
6. other findings.

Open field observations
The rats were transferred to a standard arena (50 × 50 cm with sides of 25 cm high) and observed for at least 2 minutes. Following parameters were examined:
1. behavior when removed from cage
2. fur
3. skin
4. salivation
5. nose discharge
6. lacrimation
7. eyes/ pupil size
8. posture
9. palpebral closure
10. respiration
11. tremors
12. convulsions
13. abnormal movements
14. impairment of gait
15. activity/ arousal level
16. feces (number of fecal pellets/ appearance/ consistency) within two minutes
17. urine (appearance/ quantity) within two minutes
18. number of rearings within two minutes.

Sensorimotor tests/reflexes
The rats were removed from the open field and subjected to following sensorimotor or reflex tests:
1. approach response
2. touch response
3. vision ("visual placing response")
4. pupillary reflex
5. pinna reflex
6. audition ("startle response")
7. coordination of movements ("righting response")
8. behavior during "handling"
9. vocalization
10. pain perception ("tail pinch")
11. grip strength of forelimbs
12. grip strength of hindlimbs
13. landing foot-splay test
14. other findings.

Motor activity assessment (MA)
Motor activity (MA) was measured on the same day as FOB was performed. The examinations were performed using the TSE Labmaster System supplied by TSE Systems GmbH, Bad Homburg, Germany. For this purpose, the rats were placed in new clean polycarbonate cages for the time of measurement. Eight-teen beams were allocated per cage. The numbers of beam interrupts were counted over 12 intervals for 5 minutes in each case. The sequence at which the rats were placed in the cages was selected at random.
Motor activity measurements were carried out from 13:00 h onwards. On account of the measuring variant "staggered", the starting time varied by the time, which was needed to place the rats in the cages. For each rat, measurement was started individually when the 1st beam was interrupted and finished exactly 1 hour later. No food or water was offered to the rats during measurements. After the transfer of the last rat in each case, the room of measurement was darkened.
onal observational battery

OTHER: Hormones: Total triiodothyronine (T3), Total thyroxine (T4), Thyroid stimulating hormone (TSH)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Organ weights
Weight assessment was carried out on all animals sacrificed at scheduled dates. The following weights were determined:
1. Anesthetized animals
2. Adrenal glands
3. Brain
4. Epididymides
5. Heart
6. Kidneys
7. Liver
8. Ovaries
9. Prostate
10. Seminal vesicles with coagulation glands
11. Spleen
12. Testes
13. Thymus
14. Thyroid glands
15. Uterus with cervix

Organ/tissue fixation
The following organs or tissues 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. Colon
9. Duodenum
10. Epididymides
11. Esophagus
12. Extraorbital lacrimal glands
13. Eyes with optic nerve
14. Femur with knee joint
15. Heart
16. Ileum
17. Jejunum
18. Kidneys
19. Larynx
20. Liver
21. Lungs
22. Lymph nodes (mesenteric and axillary lymph nodes)
23. Mammary gland (males and females)
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 with coagulation glands
35. Sciatic nerve
36. Skeletal muscle
37. Skin
38. Spinal cord (cervical, thoracic and lumbar cords)
39. Spleen
40. Sternum with marrow
41. Stomach (forestomach and glandular stomach)
42. Testes (modified Davidson’s solution)
43. Thymus
44. Thyroid glands
45. Trachea
46. Urinary bladder
47. Uterus
48. Vagina
From the liver, each one slices of the Lobus dexter medialis and the Lobus sinster lateralis was fixed in Carnoy’s solution and embedded in paraplast.

Histopathology
Fixation was followed by histotechnical processing, examination by light microscopy and assessment of:
1. All gross lesions (all animals affected per test group)
2. Adrenal glands (all animals per test group in the control and the high dose group)
3. Bone marrow (femur) (all animals per test group in the control and the high dose group)
4. Brain (all animals per test group in the control and the high dose group)
5. Cecum (all animals per test group in the control and the high dose group)
6. Cervix (all animals per test group in the control and the high dose group)
7. Colon (all animals per test group in the control and the high dose group)
8. Duodenum (all animals per test group in the control and the high dose group)
9. Epididymides (all animals per test group in the control and the high dose group)
10. Eyes with optic nerve (all animals per test group in the control and the high dose group)
11. Heart (all animals per test group in the control and the high dose group)
12. Ileum (all animals per test group in the control and the high dose group)
13. Jejunum (with Peyer’s patches) (all animals per test group in the control and the high dose group)
14. Kidneys (all animals per test group in the control and the high dose group)
15. Liver (all animals per test group in all groups)
16. Lung (all animals per test group in the control and the high dose group)
17. Lymph nodes (mesenteric and axillary lymph nodes) (all animals per test group in the control and the high dose group)
18. Ovaries (with oviducts) (all animals per test group in the control and the high dose group)
19. Pituitary gland (all animals per test group in the control and the high dose group, all male animals in the low and mid dose group)
20. Prostate (all animals per test group in the control and the high dose group, all affected animals in the low and mid dose group)
21. Rectum (all animals per test group in the control and the high dose group)
22. Sciatic nerve (all animals per test group in the control and the high dose group)
23. Spinal cord (cervical, thoracic and lumbar cords) (all animals per test group in the control and the high dose group)
24. Spleen (all animals per test group in the control and the high dose group)
25. Seminal vesicle with coagulation glands (all animals per test group in the control and the high dose group)
26. Skeletal muscle (all animals per test group in the control and the high dose group)
27. Sternum with marrow (all animals per test group in the control and the high dose group)
28. Stomach (forestomach and glandular stomach) (all animals per test group in the control and the high dose group)
29. Testes (all animals per test group in the control and the high dose group, all affected animals in the low and mid dose group)
30. Thymus (all animals per test group in the control and the high dose group)
31. Thyroid glands (with parathyroid glands) (all animals per test group in the control and the high dose group)
32. Trachea (all animals per test group in the control and the high dose group)
33. Urinary bladder (all animals per test group in the control and the high dose group)
34. Uterus (all animals per test group in the control and the high dose group)
35. Vagina (all animals per test group in the control and the high dose group)

The immunorelevant organs and tissues were evaluated according to the following
parameters:
Thymus:
• Increased/decreased grade of cortico-medullar ratio (related only to area)
• Increase of starry sky cells
• Changes of cellular density in the cortex
• Changes of cellular density in the medulla
Spleen:
• Changes of the cellularity of PALS, lymphoid follicles, marginal zone, red pulp
• Altered cellular composition of follicles
• Altered number of germinal centers
Lymph nodes (mesenteric and axillary lymph nodes):
• Changes in the cellularity of follicles, interfollicular area, paracortical area, medulla
• Altered cellular composition of paracortex
• Altered number of germinal centers
• Hyperplasia of high endothelial venules
Peyer's patches (of the jejunum):
• Changes of the cellularity of follicles (including mantle zone and germinal centers)
• Changes of the cellularity of interfollicular area
Bone marrow:
• Changes of the cellularity
• Changes of the myeloid/erythroid ratio
Whenever the histopathologic evaluation of the immunorelevant organs and tissues did not reveal a morphologic alteration of these items and/or whenever no other pathologic finding was noted, these organs were diagnosed as "no abnormalities detected”.
Special attention was given for the synchrony of the morphology of the estrous cycle in ovaries, uterus, cervix, and vagina.

Statistics:

Clinical examinations
Means and standard deviations of each test group were calculated.
Body weight, body weight change: DUNNETT's test (two-sided).
Feces, rearing, grip strength forelimbs, grip strength hindlimbs, foot-splay test,motor activity: Non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided); WILCOXON test (two-sided).

Clinical pathology; pathology
Means, medians and standard deviations of each test group were calculated for several parameters.
Clinical pathology parameters, urine volume, urine specific gravity, pathology parameters: KRUSKAL-WALLIS test (two-sided); WILCOXON test (two-sided).
Urinalysis, except color, turbidity, volume and specific gravity: FISHER's exact test.

Results and discussion

Results of examinations

Details on results:

CLINICAL SIGNS AND MORTALITY: No rat died prematurely in the present study.
1000mg/kg bw/day: Slight and moderate salivation after treatment was observed in all male and four female rats from day 1 onwards on most of the days.
300 mg/kg bw/day: Slight salivation after treatment was observed in all male rats and one female rat from day 3 onwards on several days.
100 mg/kg bw/day: Slight salivation after treatment was observed in 1 male and 1 female rat on single days.
In all cases, salivation was reversible within at least 3 hours. From the temporary, short appearance immediately after dosing it is likely, that this finding was induced by a bad taste of the test substance or local affection of the upper digestive tract. This finding was not considered to be an adverse and toxicologically relevant effect.

BODY WEIGHT AND WEIGHT GAIN: No test substance-related changes on body weight parameters were observed.

FOOD CONSUMPTION: No test substance-related effects on food consumption were obtained.

WATER CONSUMPTION: No test substance-related findings were observed.

HAEMATOLOGY: No treatment-related changes among hematological parameters were measured.

CLINICAL CHEMISTRY: No treatment-related, adverse changes among clinical chemistry parameters were measured.

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

NEUROBEHAVIOUR: No test substance-related findings were observed in the FOB and MA.

ORGAN WEIGHTS: 1000 mg/kg bw/day: The significant testes weight decrease in males (72% of the control; p <= 0.01) was regarded as treatment-related and correlated with histopathological findings. A weak significant weight decrease was found in the prostate of the same male animals (68% of the control; p <= 0.05) Here, a treatment-related effect could not be ruled out.
No histopathological correlate was found for the adrenal glands weight increase (118% of the control; p <= 0.05) and the brain weight decrease (92% of the control; p <= 0.01)in females. Therefore, these findings were considered to be incidental.
All other mean absolute weight parameters did not show relevant differences when compared to the control group 0 and were considered to be within the normal range.

RELATIVE ORGAN WEIGHTS: 1000 mg/kg bw/day: The significant weight decrease observed in testes of males (75% of the control; p <= 0.01) correlated with histopathological findings and was considered to be treatment-related.
Although the relative prostate weight in the same animal group did not reveal a significant weight decrease (70% of the control), this change was regarded to be relevant. A significant liver weight increase in males (115% of the control, p <= 0.01) and females (110% of the control; p <= 0.05) occurred without histopathological correlate. However, due to the higher significant value in males than in females a treatment-related effect in the liver of males could not be completely ruled out. In females, the weight increase was rather regarded as incidental due to the low significant value.
All other mean relative weight parameters of treated animals did not show relevant
differences when compared to the control groups.

GROSS PATHOLOGY: 1000 mg/kg bw/day: Histopathological findings were observed in testes, epididymides, prostate and pituitary gland of male animals.

Testes and epididymides
A treatment-related diffuse tubular degeneration ranging from slight to severe was observedin the testes of all male animals. This finding was characterized by depletion of pachytene spermatocytes and round spermatids in all stages, leaving intact elongated spermatids and sperms in the tubules. In the epididymides of some of these animals, minimal to moderate amount of cellular debris within the lumen (3/5 male animals) was accompanied by oligospermia (2/5 male animals). Both were more pronounced in the caput. These findings were considered to be secondary to the testicular degeneration.

Prostate
A minimal reduction in the size of the prostate was observed in single control as well as treated animals of the low dose test group and the mid dose test group. However, animals in the 1000 mg/kg bw/d group revealed a slight increase in the incidence of these findings (4/5 male animals) that may reflect the absolute weight decrease of the prostate in this test group. Therefore, a treatment-related effect could not be ruled out.

Pituitary
A minimal to slight increase in the number of enlarged basophilic cells with cytoplasmic vacuoles was observed in the pars distalis of the pituitary gland in single control as well as treated animals of the low dose test group and the mid dose test group. However, animals in the 1000 mg/kg bw/d group revealed a slight increase in the incidence (4/5 male animals) of this finding. Therefore, a treatment-related effect could not be completely ruled out.

All other findings noted were either single observations or were biologically equally distributed between the control and the test substance-treated rats. All of them were considered to be incidental and/or spontaneous in origin.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: overall effects; no adverse effects at the highest dose tested

2

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: absolute and relative weight decrease in testes accompanied by histopathological changes affecting spermatogenesis

3

Dose descriptor:

LOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: absolute and relative weight decrease in testes accompanied by histopathological changes affecting spermatogenesis

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Under the conditions of the present study, the no observed adverse effect level (NOAEL) was 300 mg/kg bw/d for male and 1000 mg/kg bw/d for female Wistar rats.