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Diss Factsheets

Administrative data

Description of key information

The  available subchronic repeated dose oral toxicity studies resulted in NOAEL of 1000 mg/kg bw/day or greater.
The available subchronic repeated dose inhalation toxicity study results in NOAEC >= 0.5mg/L.
The available subchronic repeated dose dermal toxicity study results in NOAEL of > = 2000 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
17 Nov 1997 - 16 Mar 1998
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP guideline study (OECD). According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (Dec 2012)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
ToxLabs Prüflabor GmbH, Greppin, Germany
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH
- Age at study initiation: 32-38 d
- Weight at study initiation: 148.5 g (mean value males), 136.7 g (mean value females)
- Housing: one or two animals in cages (Makrolon Type 3)
- Diet: Altromin 1326, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 6-8 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23
- Humidity (%): 30-60
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
other: distilled water containing 1% Tween 80
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 1%
- Lot/batch no. (if required): S23350 739
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Duplicate 2 mL samples of each formulation were taken and stored in the frozen state until measurement
Duration of treatment / exposure:
90 d
Frequency of treatment:
once daily, 7 days/week
Remarks:
Doses / Concentrations:
100, 300, 1000 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
10 (control, test and satellite groups)
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for selecting satellite groups: 10 animals each from the high dose and the vehicle group were used to investigate reversibility of possible effects
- Post-exposure recovery period in satellite groups: 28 d
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes, autonomic activity, presence of clonic or tonic movements, stereotypies, bizarre behavior
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes, changes in skin, fur, eyes, mucous membranes, gait, posture: response to handling; occurrence of secretions and excretions
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly from the start to the end of the study

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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: prior to the administration and at the end of the study
- Dose groups that were examined: All (surviving) animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Just prior to killing at the end of the study
- Anesthetic used for blood collection: Yes (Ether)
- Animals fasted: Yes, over night
- How many animals: All animals
- Parameters examined: erythrocyte count, hemoglobin concentration, packed cell volume, platelet count, total leukocyte count, leukocyte differential count, prothrombine time, fibrinogen concentration

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Just prior to killing at the end of the study (including the satellite groups)
- Animals fasted: Yes
- How many animals: all animals
- Parameters examined: alkaline aminotransferase, alkaline phosphatase, aspartate aminotransferase, bilirubin, blood urea nitrogen, calcium, creatinine, fasting glucose, phosphorus, total cholesterol, total protein, albumin, chloride, potassium, sodium

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Prior to administration, at monthly intervals and in the last week of dosing and in the last week of the recovery period.
- Dose groups that were examined: all
- Battery of functions tested: sensory activity (auditory, visual and proprioceptive stimuli) / grip strength / motor activity
Sacrifice and pathology:
GROSS PATHOLOGY: cranial, thoracic and abdominal cavities were opened and examined macroscopically
HISTOPATHOLOGY: adrenals, aorta, brain (3 sections), epididymides, eye, femur, heart, kidney, liver, lungs (incl. mainstem bronchi), mesenteric lymph node, muscle incl. sciatic nerve, oesophagus, ovaries, pancreas, pituitary, prostate, seminal vesicle, skin incl. mammary glands, small and large intestine (including peyer´s patches), spinal chord (3 levels), spleen, sternum with bone marrow, stomach, submandibular lymph node, testes, thymus, thyroid (incl. parathyroids), trachea, urinary bladder, uterus
Other examinations:
Organ weights of adrenals, brain, epididymides, testes, heart, kidneys, liver, ovaries, spleen, testes and thymus
Statistics:
Body weights, food consumption: Welch t-test

Haematology, coagulation, clinical biochemistry and absolute and relative organ weights: Dunnett´s test

Differential leukocyte count: Mean, range and standard deviation
Clinical signs:
no effects observed
Mortality:
no mortality observed
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 examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
high dose group (female): fibrinogen concentration of plasma and creatinine content increased, high dose group (male/female): alkaline phosphatase increased, middle and high dose groups (male/female): sreum urea nitrogen increased, all non-adverse
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
increased kidney weights for high dose males and females, non-adverse
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
intracellular fat and low-grade fatty degenerations of hepatocytes in all male animals, female control, middle dose and high dose groups, non-adverse
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
Two animals died shortly after administration due to incorrect gavage shown by lungs filled with blood. None of the animals showed any alterations of their general state of well-being and behaviour at any observation period (few observations were made substance independent and for a short period of time).

BODY WEIGHT AND WEIGHT GAIN
Not affected by the test compound.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Not dose-dependently influenced.

OPHTHALMOSCOPIC EXAMINATION
No alterations.

HAEMATOLOGY
Not influenced.

CLINICAL CHEMISTRY
The fibrinogen concentration of the plasma was increased in the female animals of the high dose group, this was no longer apparent at the end of the treatment-free period.
The activity of alkaline phosphatase of the serum significantly increased in the high dose group, males and femals. This indicates damage to liver cells and/or an increased function rate. This finding was no longer apparent at the end of the treatment-free period.
The serum urea nitrogen was significantly increased in the middle and high dose group of the males and in the high dose group of female animals. The creatinine content was significantly increased in all male and in the high dose group of the female animals. The phosphorus content was significantly increased dose-dependently in all female animals and the sodium content was dose-dependently decreased in the male animals, significantly in the animals of the middle and high dose groups. These effects were no longer apparent at the end of the treatment-free period.

NEUROBEHAVIOUR
No changes in grip strength, motor activity and sensory response.

ORGAN WEIGHTS
Absolute and relative kidney weights were increased in all male animals in the high dose group which was still present after the recovery period. Absolute and relative liver weights were increased in both sexes but this was no longer apparent after the recovery period in females. Other significant differences seem to be incidental.

GROSS PATHOLOGY
No substance-dependent changes.

HISTOPATHOLOGY: NON-NEOPLASTIC
Intracellular lipid droplets in hepatocytes of the female animals in the high and mid dose group (5-90% of the observed area) with cell lesions were clearly caused dose-dependently by the test article. There was no special localization of the changes of hepatocytes in the liver lobules. In most cases only low grade intracellular lipopexia occurred in the male animals.
Stomach: Oesophagal part and cardia with multilayered squamous epithelium, leukocyte infiltration in the submucosa and fibrous repair, fibrotic regions in the submucosa of the glandular stomach
Lungs: Atelectactic and emphysemic areas
Thymus: Partial substitution of the parenchyma by fibrinogenesis
Skin: Benign fibrous proliferation
Sciatic nerve: Thickening of the perineurium and thickening of the adventitia of the vessels


Dose descriptor:
NOAEL
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 observed in the highest dose level.
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 2 due to read across) study from reference substance with similar structure and intrinsic properties. Read-across is justified based on common origin, common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to endpoint discussion for further details). The selected studies are thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (limited parameters examined, no daily observation, no data on test substance purity)
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)
Deviations:
yes
Remarks:
(limited parameters examined, no daily observation)
GLP compliance:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Taconic, Germantown, NY, USA
- Weight at study initiation: males: 379-388 g ; females: 234-239 g
- Housing: animals were housed in the exposure chambers (feed and water was removed during exposure)
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
whole body
Vehicle:
air
Remarks on MMAD:
MMAD / GSD: 1.0 µm/ approx. 1.8
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 1000 L stainless steel and glass exposure chambers; chambers contained catch pans between each of three leveles of cages.
- System of generating particulates/aerosols: The test material was aerosolized directly from the liquid by a modified Lakin nebulizer on each chamber. The test material was in a straight-walled glass flask and the barrels of the nebulizer were immersed under the level of the liquid in order to maximize the amount of material generated. The distance from the nebulizer to the walls of the flask was approx. 3 cm. After exiting the flask, the aerosol passed through a glass impactor to remove most of the larger particles. The remaining aerosol was mixed with the main air stream for each chamber before entering the chamber.
- Temperature and humidity in air chamber (by a Taylor wet/dry bulb hydrometer approx. every 30 min during each exposure): approx. 23 °C, 56 - 64%
- Air flow rate: approx. 300 L/min (mean chamber flow per group: 297, 308, 342, and 243 L/min, respectively)

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric sampling on glass fiber filters (3 times during each exposure); some filters were additionally analyzed by GC/MS
- Samples taken from breathing zone: yes
Nominal concentrations were determined as the loss of weight of fluid from the generator divided by total air flow through the chamber.
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
6 hours/day
5 days/week
Remarks:
Doses / Concentrations:
0.00 ± 0.00, 0.05 ± 0.01, 0.17 ± 0.01, and 0.56 ± 0.02 mg/L
Basis:
analytical conc.
Remarks:
Doses / Concentrations:
0.05, 0.15, and 0.5 mg/L
Basis:
nominal conc.
No. of animals per sex per dose:
15
(Additional 10 male rats per group were included for examination of pulmonary function tests and analysis of pulmonary hydroxyproline following exposure.)
Control animals:
yes, concurrent no treatment
yes, sham-exposed
Details on study design:
- Dose selection rationale: The highest dose was expected to result in abnormal accumulation of test material in the lung and possible impairment of normal clearance mechanisms. The low dose is a factor of 10 above the TLV (treshold limit value) for mineral oil mistes, no untoward effects were expected at this level.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (except weekends)

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: 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: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: Yes
- How many animals: all core animals
- Parameters examined: complete blood count (CBC) (white blood cell count (WBC), red blood cell count (RBC), hemoglobin (Hb), hematocrit (Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and platelets) and differential count.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: Yes
- How many animals: all core animals
- Parameters examined: glucose, urea nitrogen, total protein, albumin (A), globulin (G), A/G, sorbitol dehydrogenase, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase, total bilirubin, creatinine colesterol, triglycerides, uric acid, Cl, Ca, Na, K, and P

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
Lung function: The animals were anaesthetized and pulmonray function tests were performed (deflation quasistatic pressure-volume cureved, functional residual capactiy, and maximal forced exhalation maneuver). After the pulmonray function tests, the lungs were removed and all lobes were weighed. Lobes were frozen for analysis of hydroxyproline content and analysis of test material remaining in the lung.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes; organ weights (adrenals, kidney, spleen, brain, liver, testes, epididymides, ovaries, thymus, heart, prostate, uterus, right middle lung lobe
HISTOPATHOLOGY: Yes (untreated and high-dose): adrenals, ovaries, sternum, pancreas, brain, salivary gland, eye, spleen heart, stomach, colon, testes, duodenum, thymus, kidneys, thyroid, liver, tracheobronchial lymph nodes, lung, nasal turbinates, thigh muscle, urinary bladder, sciatic nerve, and any gross lesions. Only the lungs and tracheobronchial lymph nodes of the untreated controls were processed. 10 males of group 1, 2 and 5 (untreated, sham-exposed, and high-dose) were evaluated for morphology, number of sperm and number of testicular spermatids.
Statistics:
ANOVA and Tukey´s multiple range test: body weighs, male reproductive endpoints, haematology, and serum chemistry
ANOVA and Duncan´s multiple range test: organ weights, pulmonary function, and pulmonary hydroxyproline
Clinical signs:
no effects observed
Mortality:
no mortality observed
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:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
increased total weight of the lung lobes (high-dose)
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
mild macrophage accumulation in the lung (high-dose). Non adverse.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
mild macrophage accumulation in the lung (high-dose): Non adverse.
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
No treatment-related clinical signs were and no mortalities observed.

BODY WEIGHT AND WEIGHT GAIN
Increased body weights were observed in treated males. The difference compared to control was statistically significant, but as no clear dose-response was seen and the difference was lower than 7%, it was not considered to be of toxicological relevance.

HAEMATOLOGY
No treatment-related changes were observed.

CLINICAL CHEMISTRY
No treatment-related changes were observed.

ORGAN WEIGHTS
The lungs had a minimal increase in weight after exposure to 0.50 mg/L. Other organ weights were not affected by exposure to the test substance.

GROSS PATHOLOGY
The number of macrophages in the pulmonary alveoli increased slightly. This increase was small considering the high (500 mg/nr) aerosol concentration.

HISTOPATHOLOGY: NON-NEOPLASTIC
Microscopic examination of the lungs of animals in the high-dose group revealed one to two plump macrophages with sparse cytoplasmic vacuoles in less than 1.0% of the aveoli (in controls less than 0.1% would be expected).

OTHER FINDINGS
- Sperm morphology: No treatment-related effects were noted in sperm morophology or in sperm and spermatid counts.
- Lung function: There were no significant differences between any groups for any of the pulmonary function parameters. The only parameter affected by exposure was the total weight of the five lung lobes. Weight for the high-dose group was significantly greater than the other groups.
Dose descriptor:
NOAEC
Effect level:
0.5 mg/L air (analytical)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 2 due to read across) study from reference substance with similar structure and intrinsic properties. Read-across is justified based on common origin, common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to endpoint discussion for further details). The selected studies are thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Repeated dose toxicity: inhalation - local effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (limited parameters examined, no daily observation, no data on test substance purity)
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)
Deviations:
yes
Remarks:
(limited parameters examined, no daily observation)
GLP compliance:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Taconic, Germantown, NY, USA
- Weight at study initiation: males: 379-388 g ; females: 234-239 g
- Housing: animals were housed in the exposure chambers (feed and water was removed during exposure)
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
whole body
Vehicle:
air
Remarks on MMAD:
MMAD / GSD: 1.0 µm/ approx. 1.8
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 1000 L stainless steel and glass exposure chambers; chambers contained catch pans between each of three leveles of cages.
- System of generating particulates/aerosols: The test material was aerosolized directly from the liquid by a modified Lakin nebulizer on each chamber. The test material was in a straight-walled glass flask and the barrels of the nebulizer were immersed under the level of the liquid in order to maximize the amount of material generated. The distance from the nebulizer to the walls of the flask was approx. 3 cm. After exiting the flask, the aerosol passed through a glass impactor to remove most of the larger particles. The remaining aerosol was mixed with the main air stream for each chamber before entering the chamber.
- Temperature and humidity in air chamber (by a Taylor wet/dry bulb hydrometer approx. every 30 min during each exposure): approx. 23 °C, 56 - 64%
- Air flow rate: approx. 300 L/min (mean chamber flow per group: 297, 308, 342, and 243 L/min, respectively)

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric sampling on glass fiber filters (3 times during each exposure); some filters were additionally analyzed by GC/MS
- Samples taken from breathing zone: yes
Nominal concentrations were determined as the loss of weight of fluid from the generator divided by total air flow through the chamber.
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
6 hours/day
5 days/week
Remarks:
Doses / Concentrations:
0.00 ± 0.00, 0.05 ± 0.01, 0.17 ± 0.01, and 0.56 ± 0.02 mg/L
Basis:
analytical conc.
Remarks:
Doses / Concentrations:
0.05, 0.15, and 0.5 mg/L
Basis:
nominal conc.
No. of animals per sex per dose:
15
(Additional 10 male rats per group were included for examination of pulmonary function tests and analysis of pulmonary hydroxyproline following exposure.)
Control animals:
yes, concurrent no treatment
yes, sham-exposed
Details on study design:
- Dose selection rationale: The highest dose was expected to result in abnormal accumulation of test material in the lung and possible impairment of normal clearance mechanisms. The low dose is a factor of 10 above the TLV (treshold limit value) for mineral oil mistes, no untoward effects were expected at this level.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (except weekends)

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: 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: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: Yes
- How many animals: all core animals
- Parameters examined: complete blood count (CBC) (white blood cell count (WBC), red blood cell count (RBC), hemoglobin (Hb), hematocrit (Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and platelets) and differential count.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: Yes
- How many animals: all core animals
- Parameters examined: glucose, urea nitrogen, total protein, albumin (A), globulin (G), A/G, sorbitol dehydrogenase, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase, total bilirubin, creatinine colesterol, triglycerides, uric acid, Cl, Ca, Na, K, and P

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
Lung function: The animals were anaesthetized and pulmonray function tests were performed (deflation quasistatic pressure-volume cureved, functional residual capactiy, and maximal forced exhalation maneuver). After the pulmonray function tests, the lungs were removed and all lobes were weighed. Lobes were frozen for analysis of hydroxyproline content and analysis of test material remaining in the lung.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes; organ weights (adrenals, kidney, spleen, brain, liver, testes, epididymides, ovaries, thymus, heart, prostate, uterus, right middle lung lobe
HISTOPATHOLOGY: Yes (untreated and high-dose): adrenals, ovaries, sternum, pancreas, brain, salivary gland, eye, spleen heart, stomach, colon, testes, duodenum, thymus, kidneys, thyroid, liver, tracheobronchial lymph nodes, lung, nasal turbinates, thigh muscle, urinary bladder, sciatic nerve, and any gross lesions. Only the lungs and tracheobronchial lymph nodes of the untreated controls were processed. 10 males of group 1, 2 and 5 (untreated, sham-exposed, and high-dose) were evaluated for morphology, number of sperm and number of testicular spermatids.
Statistics:
ANOVA and Tukey´s multiple range test: body weighs, male reproductive endpoints, haematology, and serum chemistry
ANOVA and Duncan´s multiple range test: organ weights, pulmonary function, and pulmonary hydroxyproline
Clinical signs:
no effects observed
Mortality:
no mortality observed
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:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
increased total weight of the lung lobes (high-dose)
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
mild macrophage accumulation in the lung (high-dose). Non adverse.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
mild macrophage accumulation in the lung (high-dose): Non adverse.
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
No treatment-related clinical signs were and no mortalities observed.

BODY WEIGHT AND WEIGHT GAIN
Increased body weights were observed in treated males. The difference compared to control was statistically significant, but as no clear dose-response was seen and the difference was lower than 7%, it was not considered to be of toxicological relevance.

HAEMATOLOGY
No treatment-related changes were observed.

CLINICAL CHEMISTRY
No treatment-related changes were observed.

ORGAN WEIGHTS
The lungs had a minimal increase in weight after exposure to 0.50 mg/L. Other organ weights were not affected by exposure to the test substance.

GROSS PATHOLOGY
The number of macrophages in the pulmonary alveoli increased slightly. This increase was small considering the high (500 mg/nr) aerosol concentration.

HISTOPATHOLOGY: NON-NEOPLASTIC
Microscopic examination of the lungs of animals in the high-dose group revealed one to two plump macrophages with sparse cytoplasmic vacuoles in less than 1.0% of the aveoli (in controls less than 0.1% would be expected).

OTHER FINDINGS
- Sperm morphology: No treatment-related effects were noted in sperm morophology or in sperm and spermatid counts.
- Lung function: There were no significant differences between any groups for any of the pulmonary function parameters. The only parameter affected by exposure was the total weight of the five lung lobes. Weight for the high-dose group was significantly greater than the other groups.
Dose descriptor:
NOAEC
Effect level:
0.5 mg/L air (analytical)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 2 due to read across) study from reference substance with similar structure and intrinsic properties. Read-across is justified based on common origin, common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to endpoint discussion for further details). The selected studies are thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: dermal
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
09 Jul - 10 Oct 1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (no data on test substance purity; only 2 dose groups, open application, limited parameters examined)
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Deviations:
yes
Remarks:
(no data on test substance purity, only 2 dose groups, open application, limited parameters examined)
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, Lakeview, NJ, USA
- Age at study initiation: approx. 7 weeks
- Housing: individually in hanging, stainless steel cages with wire bottoms and fronts
- Diet: Purina Certified Lab Chow ' 5002 in pellet form; ad libitum
- Water: tap water; ad libitum
- Acclimation period: at least 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22
- Humidity (%): 40-60
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
open
Vehicle:
unchanged (no vehicle)
Details on exposure:
TEST SITE
- Area of exposure: no data
- Type of wrap if used: no wrap used, open
- Time intervals for shavings or clipplings: 24 h before the first treatment; at least weekly afterwards
- Application site: back (shaved)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing; wiping off with a gauze pads every saturday (applications on working days)

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): no data
- Concentration (if solution): undiluted
- Constant volume or concentration used: no

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes (collars), removal during the weekend
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
5 days per week (65 exposures), 24 hours/day, removal of substance on saturdays (once a week)
Remarks:
Doses / Concentrations:
800 and 2000 mg/kg bw/day
Basis:
nominal per unit body weight
No. of animals per sex per dose:
10
(5 additional animals of the control and the high dose group were included for dermal bioavailability experiments only.)
Control animals:
yes, concurrent no treatment
Details on study design:
The test substance was dispensed by volume from a syringe and left uncovered on the shaved skin. The rats were fitted with cardboard Elizabethan collars to minimze ingestion of the test material.
The controls were treated in the same manner except that no material was applied to their skin.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations included: appearance, behaviour, secretory function and discharges

DETAILED CLINICAL OBSERVATIONS: No

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: weekly
- Parameters evaluated: erythema and edema according to Draize, chronic deterioration: flaking, thickening, stiffening, cracking and slouthing

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: 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: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: No data
- How many animals: all animals
- Parameters examined: red blood cells, white blood cells, and platelets

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: No data
- How many animals: all animals
- Parameters examined: glucose, urea nitrogen, alanine aminotransferase, albumin, phosphorus; only females: lactate dehydrogenase, aspartate aminotransferase

URINALYSIS: Yes
- Time schedule for collection of urine: weeks 5 and 13
- Metabolism cages used for collection of urine: No
- Parameters examined: pH, bilirubin, specific gravity, urobilinogen, blood, protein, glucose, ketones

NEUROBEHAVIOURAL EXAMINATION: No

OTHER: sperm morphology: at termination
- Parameters: percentage normal sperm, abnormal heads, headless, tailless, and curled tail
Sacrifice and pathology:
GROSS PATHOLOGY: Yes; organ weights: kidneys, liver; only males: brain, spleen; only females: thyroids
HISTOPATHOLOGY: Yes (no further information available)
Statistics:
The level of statistical significance was P < 0.05.
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
slight erythemal and flaking; slight epidermal hyperplasia and chronic inflammation (both treatment groups)
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
reduced body weight gain in males (800 mg/kg: 7% and 2000 mg/kg: 10%)
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:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
No treatment-related effects were observed.

BODY WEIGHT AND WEIGHT GAIN
Treated males gained slightly less weight than the controls (800 mg/kg bw: 7%, 2000 mg/kg bw: 10%). As the difference is low, the decrease in body weight was not interpreted as a sign for systemic toxicity.

HAEMATOLOGY
No adverse effects on any haematologic parameters measured were observed.

CLINICAL CHEMISTRY
A few of the serum paramters of the high-dose animals were statistically different from the controls, but the differences were small, not consistent between males and females, and did not present any pattern suggestive of effects on any specific organ (no corresponding histological findings). Thus, the effects were considered not to be of toxicological relevance.
- high-dose males (compared to controls): glucose: -14%, albumin: -3%, and phoshorus: +16%
- high-dose females (compared to controls): lactate dehydrogenase: +45% (low-dose: +22%), and aspartate aminotransferase: +22%

URINALYSIS
No additional data given on Urinalysis in study report.

ORGAN WEIGHTS
Increased thyroid weight in the low-dose (+ 25%) females and decreased spleen weight (- 10%) in the low-dose males were not considered to be toxicologically relevant, as these effects were not observed in the high-dose groups.

GROSS PATHOLOGY
No abnormalities were detected.

HISTOPATHOLOGY:
No abnormalities were detected.

OTHER FINDINGS
- Sperm morphology: No effects on sperm morphology were detected.
- Local effects: Slight erythema and flaking of the skin were observed in the treated groups during the dosing phase. Microscopic examination of the skin indicated trace to slight epidermal hyperplasia and chronic inflammation of the superficial dermis.

SKIN PENETRATION
Skin penetration values of 2 - 6% were obtained.
The results of the in vivo skin penetration study indicate that the 13-week treatment with the test substance does not increase the skin penetration of the test substance (only the value for females was statistically different from the penetration in untreated animals). The skin penetration of untreated rats was less than 2% and the mean value for treated rats was approx. 6%. The recovery of radioactivity was measuered in the urine and faeces as well as the remaining radioactivity in tissue samples.
Dose descriptor:
NOAEL
Effect level:
>= 2 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: highest dose tested
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 2 due to read across) study from reference substance with similar structure and intrinsic properties. Read-across is justified based on common origin, common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to endpoint discussion for further details). The selected studies are thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Repeated dose toxicity: dermal - local effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: dermal
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
09 Jul - 10 Oct 1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (no data on test substance purity; only 2 dose groups, open application, limited parameters examined)
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Deviations:
yes
Remarks:
(no data on test substance purity, only 2 dose groups, open application, limited parameters examined)
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, Lakeview, NJ, USA
- Age at study initiation: approx. 7 weeks
- Housing: individually in hanging, stainless steel cages with wire bottoms and fronts
- Diet: Purina Certified Lab Chow ' 5002 in pellet form; ad libitum
- Water: tap water; ad libitum
- Acclimation period: at least 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22
- Humidity (%): 40-60
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
open
Vehicle:
unchanged (no vehicle)
Details on exposure:
TEST SITE
- Area of exposure: no data
- Type of wrap if used: no wrap used, open
- Time intervals for shavings or clipplings: 24 h before the first treatment; at least weekly afterwards
- Application site: back (shaved)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing; wiping off with a gauze pads every saturday (applications on working days)

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): no data
- Concentration (if solution): undiluted
- Constant volume or concentration used: no

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes (collars), removal during the weekend
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
5 days per week (65 exposures), 24 hours/day, removal of substance on saturdays (once a week)
Remarks:
Doses / Concentrations:
800 and 2000 mg/kg bw/day
Basis:
nominal per unit body weight
No. of animals per sex per dose:
10
(5 additional animals of the control and the high dose group were included for dermal bioavailability experiments only.)
Control animals:
yes, concurrent no treatment
Details on study design:
The test substance was dispensed by volume from a syringe and left uncovered on the shaved skin. The rats were fitted with cardboard Elizabethan collars to minimze ingestion of the test material.
The controls were treated in the same manner except that no material was applied to their skin.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations included: appearance, behaviour, secretory function and discharges

DETAILED CLINICAL OBSERVATIONS: No

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: weekly
- Parameters evaluated: erythema and edema according to Draize, chronic deterioration: flaking, thickening, stiffening, cracking and slouthing

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: 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: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: No data
- How many animals: all animals
- Parameters examined: red blood cells, white blood cells, and platelets

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: No data
- How many animals: all animals
- Parameters examined: glucose, urea nitrogen, alanine aminotransferase, albumin, phosphorus; only females: lactate dehydrogenase, aspartate aminotransferase

URINALYSIS: Yes
- Time schedule for collection of urine: weeks 5 and 13
- Metabolism cages used for collection of urine: No
- Parameters examined: pH, bilirubin, specific gravity, urobilinogen, blood, protein, glucose, ketones

NEUROBEHAVIOURAL EXAMINATION: No

OTHER: sperm morphology: at termination
- Parameters: percentage normal sperm, abnormal heads, headless, tailless, and curled tail
Sacrifice and pathology:
GROSS PATHOLOGY: Yes; organ weights: kidneys, liver; only males: brain, spleen; only females: thyroids
HISTOPATHOLOGY: Yes (no further information available)
Statistics:
The level of statistical significance was P < 0.05.
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
slight erythemal and flaking; slight epidermal hyperplasia and chronic inflammation (both treatment groups)
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
reduced body weight gain in males (800 mg/kg: 7% and 2000 mg/kg: 10%)
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:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
No treatment-related effects were observed.

BODY WEIGHT AND WEIGHT GAIN
Treated males gained slightly less weight than the controls (800 mg/kg bw: 7%, 2000 mg/kg bw: 10%). As the difference is low, the decrease in body weight was not interpreted as a sign for systemic toxicity.

HAEMATOLOGY
No adverse effects on any haematologic parameters measured were observed.

CLINICAL CHEMISTRY
A few of the serum paramters of the high-dose animals were statistically different from the controls, but the differences were small, not consistent between males and females, and did not present any pattern suggestive of effects on any specific organ (no corresponding histological findings). Thus, the effects were considered not to be of toxicological relevance.
- high-dose males (compared to controls): glucose: -14%, albumin: -3%, and phoshorus: +16%
- high-dose females (compared to controls): lactate dehydrogenase: +45% (low-dose: +22%), and aspartate aminotransferase: +22%

URINALYSIS
No additional data given on Urinalysis in study report.

ORGAN WEIGHTS
Increased thyroid weight in the low-dose (+ 25%) females and decreased spleen weight (- 10%) in the low-dose males were not considered to be toxicologically relevant, as these effects were not observed in the high-dose groups.

GROSS PATHOLOGY
No abnormalities were detected.

HISTOPATHOLOGY:
No abnormalities were detected.

OTHER FINDINGS
- Sperm morphology: No effects on sperm morphology were detected.
- Local effects: Slight erythema and flaking of the skin were observed in the treated groups during the dosing phase. Microscopic examination of the skin indicated trace to slight epidermal hyperplasia and chronic inflammation of the superficial dermis.

SKIN PENETRATION
Skin penetration values of 2 - 6% were obtained.
The results of the in vivo skin penetration study indicate that the 13-week treatment with the test substance does not increase the skin penetration of the test substance (only the value for females was statistically different from the penetration in untreated animals). The skin penetration of untreated rats was less than 2% and the mean value for treated rats was approx. 6%. The recovery of radioactivity was measuered in the urine and faeces as well as the remaining radioactivity in tissue samples.
Dose descriptor:
NOAEL
Effect level:
>= 2 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: highest dose tested
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 2 due to read across) study from reference substance with similar structure and intrinsic properties. Read-across is justified based on common origin, common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to endpoint discussion for further details). The selected studies are thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Additional information

Justification for grouping of substances and read-across

There are no data available for the repeated dose toxicity of 2,2-bis[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate (CAS 25811-35-2). In order to fulfil the standard information requirements set out in Annex VIII-IX, 8.6 in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from structurally related substances was conducted. In accordance with Article 13 (1) of Regulation (EC) No 1907/2006 “information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set put in Annex XI are met”. In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across).

Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006, whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity, the substance listed below are selected as reference substances for hazard assessment.

Overview for repeated dose toxicity

CAS

oral

NOAEL, rat (sex) mg/kg bw/day

Inhalation

NOAEC, rat (sex) mg/L air

Dermal

NOAEL, rat (sex) mg/kg bw/day

25811-35-2

Target substance

RA: CAS 189200 -42 -8 (28d)

RA : CAS 68424-31-7 (28-day)

RA: CAS 146289-36-3 (90-day)

RA: CAS 67762-53-2

RA: CAS 67762-53-2

68424-31-7

NOAEL(f)=1613 mg/kg/day (28-day)

NOAEL(m)=1450 mg/kg/day (28-day)

--

--

189200-42-8

NOAEL (m; f) >= 1000 mg/kg bw/day (90-day)

--

--

146289-36-3

NOAEL (m; f) >= 1000 mg/kg bw/day (90-day)

--

--

67762-53-2

--

0.5 mg/L air (analytical)

NOAEL (m; f) >= 2000 mg/kg bw/day (90-day)

The above mentioned substances are considered to be similar on the basis of structural similarity resulting in similar properties and/or activities. The available endpoint information is used to predict the same endpoints for 2,2-bis[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate (CAS 25811-35-2). A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

 

Subacute Oral Repeated Rose Toxicity

A 28 day study with Pentaerythritol tetraesters of n-decanoic, n-heptanoic, n-octanoic and n-valeric acids (CAS 68424 -31 -7)

was conducted according to OECD Guideline 407 and under GLP conditions (Brammer, 1993). The test substance was administered in concentrations of 1000 ppm, 5000 ppm and 12500 ppm (corresponding to 112, 562 and 1450 mg/kg bw/day for male and 119, 586 and 1613 mg/kg bw/day for female rats) to 5 Alpk:APfSD rats per sex and dose for 28 consecutive days. Control animals (5 per sex and dose) received the plain diet. There were no toxicologically significant effects on body weight, food consumption and clinical condition and mortality up to and including the highest dose level. Changes in clinical chemistry and red cell-related parameters were observed in male rats at 12500 ppm, but these were minor and considered not to be of toxicological significance. A minimal hepatocyte hypertrophy present in males of the 12500 ppm group was observed and considered to be evidence of an adaptive response. Microscopic examination of the kidneys from male animals from all dose groups revealed an increase in hyaline droplet formation (the main constituent of which is alpha-2µ-globulin) and tubular basophilia. This phenomenon is widely accepted to be specific to the male rat and as such is considered to have no relevance to man. A NOAEL of 1450 and 1613 mg/kg/d could be identified for male and female rats, respectively.

Subchronic Oral Repeated Dose Toxicity

A 90-day oral feeding toxicity study with Pentaerythritol ester of pentanoic acids and isononanoic acid (CAS 146289-36-3) was performed according to OECD Guideline 408 and under GLP conditions (Müller, 1998). Groups of 10 male and 10 female Wistar rats were exposed to the substance at 100, 300 and 1000 mg/kg bw/day by gavage daily, 7 days/week for 90 days. Satellite control and high dose groups containing 10 male and female animals each were observed for additional 28 days. Control animals (10 per sex and dose) received the concurrent vehicle, distilled water containing 1% Tween 80. Observations and examinations of the animals included clinical signs, body weight, food consumption, haematology, clinical chemistry, organ weights, neurobehaviour, gross necropsy and histopathology. The daily oral administration of the test substance was tolerated without any adverse effects up to 1000 mg/kg bw/day. No mortality was observed except for two animals that died shortly after administration due to incorrect gavage. Absolute and relative kidney weights were increased in all male animals in the high dose group which was still present after the recovery period. However, histopathology revealed no adverse effects in the kidney. Absolute and relative liver weights were increased in both sexes but this was no longer apparent after the recovery period in females. Other significant differences seem to be incidental. The activity of alkaline phosphatase of the serum significantly increased in the high dose group, males and females. This indicates damage to liver cells and/or an increased function rate. This finding was no longer apparent at the end of the treatment-free period. Except for the increased kidney weights and liver weight in the males, all changes (e.g. clinical chemical changes) were no longer apparent at the end of the treatment-free period. The increase in kidney weights in all male animals could be correlated to the formation of hyaline droplets a phenomenon widely accepted to be specific to the male rat and as such considered to have no relevance to man, a 90-day oral NOAEL of 1000 mg/kg bw/day was found for Pentaerythritol ester of pentanoic acids and isononanoic acid in male and female rats.

Conclusion for Repeated Dose Toxicity - Oral

In summary, 28-day studies conducted with structural related substances of 2,2-bis[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate (CAS 25811-35-2) showed no signs of overt toxicity. However, in both studies formation of an increased amount of hyaline droplets in the proximal cortical tubular epithelium of the kidney was observed. The 90-day study with Pentaerythritol ester of pentanoic acids and isononanoic acid (CAS 146289-36-3) did not show any sign of overt toxicity. However, increased kidney and liver weights in the male animals was observed.

In conclusion, since the effects observed are not considered to be systemic and relevant for humans, the NOAEL was found to exceed 1000 mg/kg bw for all substances based on the result from the 28 and 90-day studies.

Repeated Dose Toxicity - Inhalation

With regard to the endpoint repeated dose toxicity inhalation, one study is available.

Subchronic Inhalation Repeated Dose Toxicity

A 90-day subchronic inhalation toxicity study was performed with Sprague-Dawley rats with Fatty acids, C5-9, tetraesters with pentaerythritol (CAS 67762-53-2) comparable to OECD guideline 413 (Dulbey, 1992). 15 males and 15 females per group were whole body exposed to the test substance aerosol for 6 hours/day, 5 days/week at concentrations of 0.05, 0.15 and 0.5 mg/L. The respective controls (15 animals per sex and dose) inhaled clean air under the same conditions. Animals were observed for clinical signs, body weight, haematology, clinical chemistry, organ weights, gross necropsy and histopathological examinations. 10 additional male animals were included in every group for examination of pulmonary function tests and pulmonary hydroxyproline following exposure. No substance-related adverse effects were observed for body weight, body weight gain, mortality, clinical biochemistry and haematological parameters. The lungs of the high dose animals had a minimal increase in weight which correlated with slightly increased numbers of macrophages in the pulmonary alveoli. Thus, the NOAEC were found to be 0.5 mg/L.

Conclusion for Repeated Dose Toxicity - inhalation

For repeated dose inhalation toxicity, one study is available which was considered for assessment of of 2,2-bis[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate (CAS 25811-35-2) by analogue based read-across. The 90-day sub-chronic inhalation toxicity study showed a minimal effect on lungs including increased weight and slightly increased numbers of macrophages in pulmonary alveoli. Therefore, the NOAEC for repeated inhalation toxicity was found to be 0.5 mg/L for the 2,2-bis[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate (CAS 25811-35-2).

Repeated Dose Toxicity – dermal

A 90-day dermal toxicity study with Fatty acids, C5-9, tetraesters with pentaerythritol (CAS 67762-53-2) was performed comparable to OECD Guideline 411 (Cruzan, 1988). Groups of 10 male and female Sprague-Dawley rats were once daily (5 days/week, 24 hours/day) exposed to the substance at 800 and 2000 mg/kg bw for 90 days (65 applications in total). Application to the skin was done open without coverage. Animals were observed for clinical signs, body weight, dermal irritation, haematology, clinical chemistry, urinalysis, organ weights, gross necropsy and histopathological examinations.

Overall there were no adverse effects found after dermal application of the test substance for 90 days on the parameters investigated. Treated males gained less body weight than control animals. Since the effect was low and no dose-relation was observed, it was not considered to be due to systemic toxicity. Some serum parameters of the high dose group animals were significant different to the control, but since the differences were small and they did not present any pattern suggestive of effects on a specific organ, they were considered not to be of toxicological relevance. Both treated groups exhibited minimal erythema and flaking of the skin during the dosing phase. At microscopic examination it was identified as very minor epidermal hyperplasia and chronic inflammation of the superficial dermis. Since no effects of systemic toxicity were identified up to the highest dose tested, the 90 day dermal NOAEL was found to be 2000 mg/kg bw/day for Fatty acids, C5-9, tetraesters with pentaerythritol in Sprague-Dawley rats.

Conclusion for Repeated Dose Toxicity – Dermal

For repeated dose dermal toxicity, one study is available which was considered for assessment of 2,2-bis[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate (CAS 25811-35-2) by analogue based read-across. The 90-day sub-chronic dermal toxicity study showed no adverse effect. Therefore, the NOAEL for repeated inhalation toxicity was found to be 2000 mg/kg bw/day of 2,2-bis[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate (CAS 25811-35-2).


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Hazard assessment is conducted by means of read-across from structural analogue. The available study is adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substances and overall quality assessment (refer to the endpoint discussion for further details).

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
Hazard assessment is conducted by means of read-across from structural analogue. The available study is adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substances and overall quality assessment (refer to the endpoint discussion for further details).

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
Hazard assessment is conducted by means of read-across from structural analogue. The available study is adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substances and overall quality assessment (refer to the endpoint discussion for further details).

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
Hazard assessment is conducted by means of read-across from structural analogue. The available study is adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substances and overall quality assessment (refer to the endpoint discussion for further details).

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
Hazard assessment is conducted by means of read-across from structural analogue. The available study is adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substances and overall quality assessment (refer to the endpoint discussion for further details).

Justification for classification or non-classification

The available data on structural related substance for oral repeated dose toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.