Registration Dossier

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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
developmental toxicity
Remarks:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
end of 2012 until spring 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
November 2012 - June 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
22nd March 1996
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories, B.V. Kreuzelweg 53 5961 NM Horst / Netherlands
- Age at study initiation: 11 weeks
- Weight at study initiation: Males: 260 to 390 g, Females: 175 to 240 g
- Fasting period before study: no
- Housing: individually, following exceptions: During overnight matings, male and female mating partners were housed together. Pregnant animals and their litters were housed together until PND 4 (end of lactation).
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: August 9th, 2011 To: October 6th, 2011
Route of administration:
oral: gavage
Vehicle:
corn oil
Remarks:
suspension
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test substance concentrations in the dose formulations ranged from 81.0% to 105.2% with reference to the nominal and were within the accepted range of ±20%.
Duration of treatment / exposure:
The duration of treatment covered a 2-week premating and a mating period in both sexes, approximately 1 week post-mating in males,
and the entire gestation period as well as 4 days of lactation and 2 weeks thereafter in females.
Frequency of treatment:
daily
Dose / conc.:
30 mg/kg bw/day (actual dose received)
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: range finder test 100, 300 and 1000 mg/kg bw
Positive control:
no
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: before the start of the administration period in order to randomize the animals. During the administration period body weight was determined on study day 0 (start of the administration period) and thereafter once a week at the same time of the day (in the morning).

FOOD CONSUMPTION
- weekly
- 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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: in the course of FOB
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: end of administration period
- Anaesthetic used for blood collection: Yes, anaesthetized using isoflurane (Isoba®, Essex GmbH, Munich, Germany)
- Animals fasted: Yes
- How many animals: 5/sex/dose

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: end of administration period
- Animals fasted: Yes
- How many animals: 5/sex/dose

URINALYSIS: Yes
- Time schedule for collection of urine: males: after mating, females: 1 day before end of administration period
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: male: postnatal day 0, female: 10d after gestation
- Dose groups that were examined: all
- Battery of functions tested: sensory activity / grip strength / motor activity
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
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
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

HISTOPATHOLOGY: Yes, control and high dose group, gross lesions in all animals
1. All gross lesions
2. Adrenal glands
3. Bone marrow (femur)
4. Brain
5. Cecum
6. Cervix
7. Coagulating glands
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. Oviducts
20. Peyer’s patches
21. Prostate
22. Rectum
23. Sciatic nerve
24. Seminal vesicles
25. Spinal cord (cervical, thoracic and lumbar cords)
26. Spleen
27. Stomach (forestomach and glandular stomach)
28. Testes
29. Thymus
30. Thyroid glands
31. Trachea
32. Urinary bladder
33. Uterus
34. Vagina
Other examinations:
Bile acids
Furthermore, an additional blood sample (0.5 mL) was be collected into serum tubes for possible future measurement of thyroid releasing hormone (TSH), and the thyroid hormones triiodothyronine (T3) and thyroxine (T4) (added by amendment if applicable). Serum samples are stored at -80 ± 10°C. Any samples remaining at finalization of the study report, were discarded.
Statistics:
Blood parameters:
For parameters with bidirectional changes:
Non-parametric one-way analysis using KRUSKAL-WALLIS test. 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 hypothesis of equal medians
For parameters with unidirectional changes:
Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians

Urinalysis parameters: WILCOXON-test (one-sided)

Food consumption: DUNNETT-test (two-sided)

fertility indices: FISHER'S EXACT test

Proportions of affected pups per litter with necropsy observations: WILCOXON-test

Weight parameters: KRUSKAL-WALLIS test
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
300 mg/kg bw/d:
- several clinical signs were noted over the whole treatment with earliest onset on the fourth day of treatment: bedding in mouth, breathing noises and labored breathing, ruffled fur, visible weight loss and generally weakened condition

100 mg/kg bw/d:
- isolated breathing noises recorded in two males and one female. Based on the low incidence and transient occurrence, these findings were not considered to be toxicologically relevant.

30 mg/kg bw/d:
- ruffled fur was intermittently noted in one male and in one female. Based on the low incidence and transient occurrence, these findings were not considered to be toxicologically relevant.
Mortality:
mortality observed, treatment-related
Description (incidence):
Three males and one female treated at 300 mg/kg bw/day did not survive until the scheduled necropsy.
- One male and one female were killed for humane reasons after 28 days of treatment. Marked body weight loss, ruffled fur, labored breathing and breathing noises along with a generally weakened condition were observed.
- Two males died spontaneously on day 25 of the pairing period and day 1 of the after pairing period, respectively. Body weight loss, breathing
noises with increasing severity towards the end of pre-pairing and throughout pairing until death were oberved.
- For all decedent animals dark red lungs and/or incompletely collapsed lungs were noted during necropsy.
Although the distribution of the deaths suggested a relationship to treatment the presence of bronchopneumonia, bronchiolitis/peribronchiolitis and/or fluid in the lungs was considered to provide strong evidence that death or moribund condition had, in all of the decedents, resulted from gavage error.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, a slight treatment-related reduction in body weights and body weight gain were noted in males and females.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, a slight treatment-related reduction in food consumption was noted in males and females.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, 3-fold higher alanine aminotransferase and slightly higher aspartate aminotransferase levels in males and approximately 2-fold higher alanine aminotransferase levels in females were recorded (not reaching statistical significance and still within the range of historical control data). These higher values deemed related to treatment, also in view of the changes in liver weights noted in males.
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):
In males at 300 mg/kg bw/day, statistically significantly lower absolute liver weights and their ratios to brain were recorded when compared with controls.
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no macroscopical findings that were deemed related to treatment with the test item.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Histopathological changes considered to be associated with the administration of the test item were present in the mesenteric lymph nodes of females given 30 mg/kg bw/day and both sexes given 100 or 300 mg/kg bw/day.
Histopathological findings: neoplastic:
no effects observed
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
Critical effects observed:
no
Conclusions:
Based on these results, the NOAEL (No Observed Adverse Effect Level) for general toxicity was considered to be 100 mg/kg body weight/day based on clinical signs, reduced body weights and
food consumption, and increased serum ALAT- and ASAT activities at 300 mg/kg bw/day.
Reason / purpose for cross-reference:
reference to same study
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
end of 2012 until spring of 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories, B.V. Kreuzelweg 53 5961 NM Horst / Netherlands
- Age at study initiation: 11 weeks
- Weight at study initiation: males: 260-390 g, females: 175-240 g
- Fasting period before study: no
- Housing: individually, following exceptions: During overnight matings, male and female mating partners were housed together. Pregnant animals and their litters were housed together until PND 4 (end of lactation).
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/-3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: August 9th, 2011 To: October 6th, 2011
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: maximum 2 weeks
- Proof of pregnancy: copulation plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): pregnant animals and litter together
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test substance concentrations in the dose formulations ranged from 81.0% to 105.2% with reference to the nominal and were within the accepted range of ±20%.
Duration of treatment / exposure:
The duration of treatment covered a 2-week premating and a mating period in both sexes, approximately 1 week post-mating in males,
and the entire gestation period as well as 4 days of lactation and 2 weeks thereafter in females.
Frequency of treatment:
daily
Details on study schedule:
- Age at mating of the mated animals in the study: 13-14 weeks

Dose / conc.:
30 mg/kg bw/day (actual dose received)
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: range finder test 300 and 1000 mg/kg bw/d
Positive control:
no
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: before the start of the administration period in order to randomize the animals. During the administration period body weight was determined on study day 0 (start of the administration period) and thereafter once a week at the same time of the day (in the morning).

FOOD CONSUMPTION
- weekly
- 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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: in the course of FOB
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: end of administration period
- Anaesthetic used for blood collection: Yes, anaesthetized using isoflurane (Isoba®, Essex GmbH, Munich, Germany)
- Animals fasted: Yes
- How many animals: 5/sex/dose

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: end of administration period
- Animals fasted: Yes
- How many animals: 5/sex/dose

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: male: postnatal day 0, female: 10d after gestation
- Dose groups that were examined: all
- Battery of functions tested: sensory activity / grip strength / motor activity
Sperm parameters (parental animals):
stages of spermatogenesis were examined in histopathology
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality up to day 4, presence of gross anomalies

GROSS EXAMINATION OF DEAD PUPS:
yes, all stillborn pups and those pups, which died ahead of schedule, were examined externally, eviscerated and their organs were assessed macroscopically
Postmortem examinations (parental animals):
GROSS PATHOLOGY: Yes
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
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

HISTOPATHOLOGY
ORGAN WEIGHTS: Adrenal glands, Brain, Heart, Kidneys, Liver, Spleen, Thymus

HISTOPATHOLOGY: Yes, control and high dose group, gross lesions in all animals
1. All gross lesions
2. Adrenal glands
3. Bone marrow (femur)
4. Brain
5. Cecum
6. Cervix
7. Coagulating glands
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. Oviducts
20. Peyer’s patches
21. Prostate
22. Rectum
23. Sciatic nerve
24. Seminal vesicles
25. Spinal cord (cervical, thoracic and lumbar cords)
26. Spleen
27. Stomach (forestomach and glandular stomach)
28. Testes
29. Thymus
30. Thyroid glands
31. Trachea
32. Urinary bladder
33. Uterus
34. Vagina
Postmortem examinations (offspring):
SACRIFICE
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows: All pups delivered from the F0 parents were examined as soon as possible on the day of birth

GROSS NECROPSY
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.
Statistics:
- Means and standard deviations of various data were calculated and included in the report.
• The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables were assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data were not assumed to follow a normal distribution.
• Fisher's exact-test was applied if the variables could be dichotomized without loss of information.
Reproductive indices:
Male reproduction data:
- Male mating index
- Male fertility index

Female reproduction and delivery data:
- Female mating index
- Female fertility index
- Gestation index
- Live birth index
- Post implantation loss
Offspring viability indices:
Pup number and status at delivery
Pup viability/mortality
Sex ratio
Pup body weight data
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
300 mg/kg bw/d:
- several clinical signs were noted over the whole treatment with earliest onset on the fourth day of treatment: bedding in mouth, breathing noises and labored breathing, ruffled fur, visible weight loss and generally weakened condition

100 mg/kg bw/d:
- isolated breathing noises recorded in two males and one female. Based on the low incidence and transient occurrence, these findings were not considered to be toxicologically relevant.

30 mg/kg bw/d:
- ruffled fur was intermittently noted in one male and in one female. Based on the low incidence and transient occurrence, these findings were not considered to be toxicologically relevant.
Mortality:
mortality observed, treatment-related
Description (incidence):
Three males and one female treated at 300 mg/kg bw/day did not survive until the scheduled necropsy.
- One male and one female were killed for humane reasons after 28 days of treatment. Marked body weight loss, ruffled fur, labored breathing and breathing noises along with a generally weakened condition were observed.
- Two males died spontaneously on day 25 of the pairing period and day 1 of the after pairing period, respectively. Body weight loss, breathing
noises with increasing severity towards the end of pre-pairing and throughout pairing until death were oberved.
- For all decedent animals dark red lungs and/or incompletely collapsed lungs were noted during necropsy.
Although the distribution of the deaths suggested a relationship to treatment the presence of bronchopneumonia, bronchiolitis/peribronchiolitis and/or fluid in the lungs was considered to provide strong evidence that death or moribund condition had, in all of the decedents, resulted from gavage error.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, a slight treatment-related reduction in body weights and body weight gain were noted in males and females.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, a slight treatment-related reduction in food consumption was noted in males and females.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, 3-fold higher alanine aminotransferase and slightly higher aspartate aminotransferase levels in males and approximately 2-fold higher alanine aminotransferase levels in females were recorded (not reaching statistical significance and still within the range of historical
control data). These higher values deemed related to treatment, also in view of the changes in liver weights noted in males.
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
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Histopathological changes considered to be associated with the administration of the test item were present in the mesenteric lymph nodes of females given 30 mg/kg bw/day and both sexes given 100 or 300 mg/kg bw/day.
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Dose descriptor:
NOEL
Remarks:
fertility
Effect level:
300 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 up to and including the highest tested dose
Dose descriptor:
NOAEL
Remarks:
general toxicity
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
body weight and weight gain
food consumption and compound intake
clinical biochemistry
Critical effects observed:
no
Clinical signs:
no effects observed
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
The overall mean number of postnatal loss per dam was 0.0, 1.2, 0.3 and 0.2 at the dose level of 0, 30, 100 and 300 mg/kg bw/day.
Incidentally at 30 mg/kg bw/day, postnatal loss was statistically significantly higher than controls due to the loss of 13 pups in one litter (litter no. 140). This litter initially was quite large
with 16 pups, of which 12 were missing on day 2 post partum and one on day 3 post partum.
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Anogenital distance (AGD):
not examined
Nipple retention in male pups:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
One pup at 30 mg/kg bw/day had a sore on the right side at planned necropsy. At 300 mg/kg bw/day, two pups from one litter had necrosis of the tail, along with a reduction in size in one pup.
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Dose descriptor:
NOEL
Generation:
F1
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed up to and including the highest tested dose
Critical effects observed:
no
Reproductive effects observed:
no
Conclusions:
The NOEL (No Observed Effect Level) for reproduction and development was considered to be 300 mg/kg bw/day.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 422
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Fatty acids, tall-oil, reaction products with pentaethylenehexamine
EC Number:
288-927-9
EC Name:
Fatty acids, tall-oil, reaction products with pentaethylenehexamine
Cas Number:
85940-40-5
Molecular formula:
Not applicable (UVCB substance)
IUPAC Name:
Fatty acids, tall-oil, reaction products with pentaethylenehexamine
Test material form:
liquid: viscous

Test animals

Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories, B.V. Kreuzelweg 53 5961 NM Horst / Netherlands
- Age at study initiation: 11 weeks
- Weight at study initiation: males 260-390g females: 175-240 g
- Fasting period before study: no
- Housing: individually, following exceptions: During overnight matings, male and female mating partners were housed together. Pregnant animals and their litters were housed together until PND 4 (end of lactation).
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20+/- 3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: August 9th, 2011 To: October 6th, 2011

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test substance concentrations in the dose formulations ranged from 81.0% to 105.2% with reference to the nominal and were within the accepted range of ±20%.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: maximum 2 weeks
- Proof of pregnancy: copulation plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): pregnant animals and litter together

Duration of treatment / exposure:
The duration of treatment covered a 2-week premating and a mating period in both sexes, approximately 1 week post-mating in males,
and the entire gestation period as well as 4 days of lactation and 2 weeks thereafter in females.
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Dose / conc.:
30 mg/kg bw/day (actual dose received)
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: range finder test 300 and 1000 mg/kg bw, strong liver effects (ALT, AST, albumin) and decreased body weight at 1000 mg/kg bw

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: before the start of the administration period in order to randomize the animals. During the administration period body weight was determined on study day 0 (start of the administration period) and thereafter once a week at the same time of the day (in the morning).

FOOD CONSUMPTION
- weekly
- 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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: in the course of FOB
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: end of administration period
- Anaesthetic used for blood collection: Yes, anaesthetized using isoflurane (Isoba®, Essex GmbH, Munich, Germany)
- Animals fasted: Yes
- How many animals: 5/sex/dose

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: end of administration period
- Animals fasted: Yes
- How many animals: 5/sex/dose

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: male: postnatal day 0, female: 10d after gestation
- Dose groups that were examined: all
- Battery of functions tested: sensory activity / grip strength / motor activity
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: No
- Number of late resorptions: No
Fetal examinations:
- External examinations: Yes, all pubs
- Soft tissue examinations: necropsy observations macroscopically
- Skeletal examinations: No
- Head examinations: No
Other examinations:
- Sex ratio
- Pup body weight data
- Pup clinical observations
Statistics:
• Means and standard deviations of various data were calculated and included in the report.
• The Dunnett-test (many to one t-test) based on a pooled variance estimate would be applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test) would be applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test wouldl be applied if the variables could be dichotomized without loss of information.
Indices:
Pup number and status at delivery
Pup viability/mortality
Sex ratio
Pup body weight data

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
300 mg/kg bw/d:
- several clinical signs were noted over the whole treatment with earliest onset on the fourth day of treatment: bedding in mouth, breathing noises and labored breathing, ruffled fur, visible weight loss and generally weakened condition

100 mg/kg bw/d:
- isolated breathing noises recorded in one female. Based on the low incidence and transient occurrence, these findings were not considered to be toxicologically relevant.

30 mg/kg bw/d:
- ruffled fur was intermittently noted in one female. Based on the low incidence and transient occurrence, these findings were not considered to be toxicologically relevant.
Mortality:
mortality observed, treatment-related
Description (incidence):
- One female was killed for humane reasons after 28 days of treatment. Marked body weight loss, ruffled fur, labored breathing and breathing noises along with a generally weakened condition were observed. A dark red lung and/or incompletely collapsed lung was noted during necropsy.
The observed effects were considered to provide strong evidence that the moribund condition had resulted from a gavage error.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, a slight treatment-related reduction in body weights and body weight gain was noted.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, a slight treatment-related reduction in food consumption was noted.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
At 300 mg/kg bw/day, approximately 2-fold higher alanine aminotransferase levels in females were recorded (not reaching statistical significance and still within the range of historical
control data).
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Histopathological changes considered to be associated with the administration of the test item were present in the mesenteric lymph nodes of females given 30 mg/kg bw/day, 100 and 300 mg/kg bw/day.
Histopathological findings: neoplastic:
no effects observed

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
not examined
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
clinical signs
food consumption and compound intake

Results (fetuses)

Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
not examined
Visceral malformations:
no effects observed

Effect levels (fetuses)

Dose descriptor:
NOEL
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed up to and including the highest tested dose

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Developmental effects observed:
no

Any other information on results incl. tables

Table 1: Summary of reproductive performance

 

Group

1

2

3

4

 

 

(mg/kg/day)

(0)

(30)

(100)

(300)

 

Female numbers

49 - 60

61 – 72

134 - 140

73 – 84

141 - 147

85 – 96

148 - 157

Number of females paired

12

19

19

22

Number of females mated (A)

12

19

19

21

Number of females not pregnant (B)

2

7

8

8

Number of pregnant females which did not give birth to living pups (C)

 

1

 

0

 

0

 

2

Number of females with living pups at first litter check

 

9

 

12

 

11

 

11

Number of females which reared their pups until day 4post partum

 

9

 

12

 

11

 

11

(A) Female no. 91 did not mate during either of the two paring periods

(B) Female nos. 50, 51, 61, 62, 65, 66, 68, 69, 71, 73 to 77, 143, 144, 147, 85, 86, 89, 92, 94, 96, 155 and 156 were

not pregnant

(C) Female nos. 52, 90 (implantation sites only) and 151 (killed in extremis on day 11post coitum)

Applicant's summary and conclusion

Conclusions:
The NOEL (No Observed Effect Level) for reproduction and development was considered to be 300 mg/kg bw/day.