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REACH

Monopentaerythritol tetraesters and dipentaerythritol hexaesters of 2-ethylhexanoic and n-valeric acids

EC number: 943-730-3 | CAS number: -

Life Cycle description
Uses advised against

Toxicological information

Endpoint summary

Administrative data

Description of key information

Oral (rat): NOAEL ≥ 1000 mg/kg bw/day

Read-across from structural analogue source substances Pentaerythritol tetraesters of n-decanoic, n-heptanoic, n-octanoic and n-valeric acids (CAS No. 68424-31-7), Dipentaerythritol hexaesters with fatty acids, C5 and C9iso (CAS No. 647028-25-9), Hexanoic acid, 2-ethyl-, 2,2-bis [ [(2-ethyl-1-oxohexyl)oxy] methyl] -1,3-propanediyl ester (CAS No. 7299-99-2), and Pentaerythritol ester of pentanoic acids, mixed esters with pentaerythritol, isopentanoic and isononanoic acid (CAS No. 146289-36-3)

Inhalation (similar to OECD 413, rat): NOAEC = 0.5 mg/L air

Read-across from structural analogue source substance Fatty acids, C5-9 tetraesters with pentaerythritol (CAS No. 67762-53-2)

Dermal (similar to OECD 411, rat): NOAEL ≥ 2000 mg/kg bw/day

Read-across from structural analogue source substance Fatty acids, C5-9 tetraesters with pentaerythritol (CAS No. 67762-53-2)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen all close all

Reference 1

Endpoint:: sub-chronic toxicity: oral
Type of information:: experimental study
Adequacy of study:: weight of evidence
Study period:: 17 Nov 1997 - 16 Mar 1998
Reliability:: 1 (reliable without restriction)
Rationale for reliability incl. deficiencies:: guideline study
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
Dose / conc.:: 100 mg/kg bw/day (actual dose received)
Dose / conc.:: 300 mg/kg bw/day (actual dose received)
Dose / conc.:: 1 000 mg/kg bw/day (actual dose received)
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
Critical effects observed:: no

Reference 2

Endpoint:: short-term repeated dose toxicity: oral
Type of information:: experimental study
Adequacy of study:: weight of evidence
Study period:: 19 Feb 1998 - 30 March 1999
Reliability:: 1 (reliable without restriction)
Rationale for reliability incl. deficiencies:: guideline study
Qualifier:: according to guideline
Guideline:: OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Version / remarks:: adopted in 1995
Qualifier:: according to guideline
Guideline:: EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
GLP compliance:: yes (incl. QA statement)
Remarks:: The Department of Health of the Government of the United Kingdom, UK
Species:: rat
Strain:: other: Sprague-Dawley Crl:CD®BR
Sex:: male/female
Details on test animals or test system and environmental conditions:: TEST ANIMALS
- Source: Charles River Limited, Margate, Kent, UK
- Age at study initiation: 5 to 6 weeks
- Weight at study initiation: 127 – 161 g (males) and 117 – 152 g (females)
- Housing: animals were housed in groups of 5 of the same sex per cage in polypropylene grid-floor cages
- Diet: Rat and Mouse SQC Expanded Diet No.1, pelleted, ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2
- Humidity (%): 55 ± 15
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:: oral: gavage
Vehicle:: arachis oil
Remarks:: BP
Details on oral exposure:: PREPARATION OF DOSING SOLUTIONS:
Formulations were prepared weekly and stored at approximately +4°C in the dark.

VEHICLE
- Concentration in vehicle: 75 mg/mL, 250 mg/mL, 500 mg/mL
- Amount of vehicle (if gavage): 2 mL/kg/day
Analytical verification of doses or concentrations:: yes
Details on analytical verification of doses or concentrations:: The concentration of test item in the test material formulations was determined by gas chromatography (GC) by means of an external standard technique. The prepared formulations were within ± 10% of the nominal concentration.
Duration of treatment / exposure:: 28 days
Frequency of treatment:: once daily, 7 days/week
Dose / conc.:: 150 mg/kg bw/day (actual dose received)
Dose / conc.:: 500 mg/kg bw/day (actual dose received)
Dose / conc.:: 1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:: 5
Control animals:: yes, concurrent vehicle
Details on study design:: - Dose selection rationale: dose levels were chosen based on the results of a foregoing range-finding study, in which animals were orally exposed to 150, 500 and 1000 mg/kg bw/day by gavage for 14 days (Jones, L.J., 2000). No adverse effects were observed during the study period in any animal. Therefore, 150, 500 and 1000 mg/kg bw/day were selected as the dose levels for the main study.
Observations and examinations performed and frequency:: DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: immediately before dosing and one and five hours after dosing during the working week and immediately before dosing and one hour after dosing at weekends.

BODY WEIGHT: Yes
- Time schedule for examinations: individual body weights were recorded on day 0 (the day before the start of treatment) and on days 7, 14, 21, and 28. Body weights were also recorded at terminal of the study

FOOD CONSUMPTION AND EFFICIENCIES: Yes
- Cage group mean weekly food consumption and weekly food efficiencies were evaluated.

WATER CONSUMPTION: Yes
- Water intake was observed daily, for each cage group, by visual inspection of the water bottles for any overt changes.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at end of the study (Day 28)
- Anaesthetic used for blood collection: No
- Animals fasted: No
- How many animals: all animals from each test and control group
- Parameters examined: haemoglobin, erythrocyte count, haematocrit, erythrocyte indices (mean corpuscular haemoglobin, mean corpuscular volume, mean corpuscular haemoglobin concentration (MCHC)), total leucocyte count, differential leucocyte count (neutrophils, lymphocytes, monocytes, eosinophils, basophils), platelet count (PLT), reticulocyte count.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at end of the study (Day 28)
- Animals fasted: No
- How many animals: all animals from each test and control group
- Parameters examined: urea, glucose, total protein, albumin, albumin/globulin ratio, calcium, inorganic phosphorus, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, creatinine, sodium, potassium, chloride, total cholesterol, total bilirubin

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: prior to the start of treatment and on days 2, 8, 15 and 23.
- Dose groups that were examined: all animals
- Battery of functions tested: sensory reactivity (grasp response, vocalisation, toe pinch, tail pinch, finger approach, touch escape, pupil reflex, startle reflex, blink reflex)/ grip strength (forelimb/hindlimb) / motor activity

BEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: prior to the start of treatment and on days 2, 8, 15 and 23.
- Dose groups that were examined: all animals
- Parameters observed: gait, tremors, twiches, convulsions, bizarre/abnormal/stereotypic behaviour, pilo-erection, exophthalmia, lachrymation, hyper/hypothermia, skin colour, respiration, palpebral closure, urination, defecation, transfer arousal, tail elevation.
Sacrifice and pathology:: GROSS PATHOLOGY: Yes. All animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.
HISTOPATHOLOGY: Yes. Adrenals, aorta (thoracic), bone and bone marrow (femur including stifle joint), bone and bone marrow (sternum), muscle (skeletal), oesophagus, ovaries, pancreas, pituitary, brain (including cerebrum, cerebellum and pons), caecum, colon duodenum, epididymides, eyes, gross lesions, heart, ileum, jejunum, kidneys, liver, lungs (with bronchi), lymph nodes (cervical and mesenteric), prostate, rectum, salivary glands (submaxillary), sciatic nerve, seminal vesicles, skin (hind limb), spinal cord (cervical), spleen, stomach, testes, thymus, thyroid/parathyroid, trachea, urinary bladder, uterus.
Other examinations:: Organ weights: adrenals, kidneys, testes, brain, liver, thymus, epididymes, ovaries, heart, spleen
Statistics:: Haematological, blood chemical, organ weight (absolute and relative to terminal body weight), weekly body weight gain and quantitative functional performance and sensory reactivity data were considered for dose response relationships by linear regression analysis followed by one way analysis of variance (ANOVA) including Levene’s test for homogeneity of variance. Where variances were shown to be homogeneous pair wise comparisons were conducted by means of Dunnett’s test. Where Levene’s test showed unequal variances the data were analysed using non-parametric methods: Kruskal-Wallis ANOVA and Mann Whitney “U” test.

The haematology variable basophils was not analysed since consistently greater than 30% of the data were recorded as the same value.

Probability values (p) are presented as follows:

p< 0.001***
p < 0.01**
p< 0.05*
p ≥0.05 (not significant)
Clinical signs:: no effects observed
Mortality:: no mortality observed
Body weight and weight changes:: effects observed, treatment-related
Description (incidence and severity):: 1000 and 500 mg/kg/day: statistically significant reduction in body weight in males during week 1 (non adverse)
Food consumption and compound intake (if feeding study):: not examined
Food efficiency:: no effects observed
Water consumption and compound intake (if drinking water study):: no effects observed
Ophthalmological findings:: not examined
Haematological findings:: effects observed, treatment-related
Description (incidence and severity):: 500 and 1000 mg/kg/day: statistical significant increase in platelet count and reduction in mean corpuscular haemoglobin concentration respectively (non adverse)
Clinical biochemistry findings:: no effects observed
Urinalysis findings:: not examined
Behaviour (functional findings):: effects observed, treatment-related
Description (incidence and severity):: 1000 mg/kg/day: increase in total and asymptotic motor activity in males. Females from the same treatment group showed a decrease in asymptotic activity (non adverse)
Organ weight findings including organ / body weight ratios:: effects observed, treatment-related
Description (incidence and severity):: 500 mg/kg/day: statistically significant reduction in absolute epididymides weight (non adverse)
Gross pathological findings:: effects observed, treatment-related
Description (incidence and severity):: 150 mg/kg/day (males and females) and 1000 mg/kg/day (one female): dark foci on the lungs (non adverse)
Histopathological findings: non-neoplastic:: effects observed, treatment-related
Description (incidence and severity):: 1000 mg/kg/day: 3 males showed globular accumulations of eosinophilic material in the proximal tubular epithelium (non adverse)
Histopathological findings: neoplastic:: not examined
Details on results:: CLINICAL SIGNS AND MORTALITY
No mortality occurred during the study period. No clinical signs of toxicity were observed in test or control animals throughout the study. However, noisy respiration was observed in one treated male animal (1000 mg/kg/day group) and one treated female animal (500 mg/kg/day group) one hour after dosing on day 2. These isolated and transient observations showed no convincing dose-relationship and were considered to be of no toxicological importance.
One treated female (1000 mg/kg/day) showed fur loss from day 21 onwards and a control female developed a scab on the head from day 25. Since such findings are occasionally reported in group housed rats, they are considered to be incidental.

BODY WEIGHT AND WEIGHT GAIN
No adverse effect on body weight was noted. Nevertheless a statistically significant reduction in body weight gain was apparent for 1000 and 500 mg/kg/day males during week 1 of the study. The dose relationship was not credible and the intergroup differences were considered to be a result of slightly higher than usual control group body weights gains.

HAEMATOLOGY
No treatment-related changed in the haematological parameters were measured.
Males treated with 1000 mg/kg/day showed a statistically significant reduction (p< 0.05) in mean corpuscular haemoglobin concentration when compared to controls. Under these conditions and in absence of any other haematological changes, the intergroup difference was considered to be accidental.
A statistically significant (p < 0.05) increase in platelet count was detected but this was confined to 500 mg/kg/day males and was considered to be incidental (see Table 2 under "Any other information on results incl. tables").

FUNCTIONAL PERFORMANCE TESTS
Males treated with 1000 mg/kg/day showed a statistically significant increase in total and asymptotic motor activity while females from the same treatment group displayed a statistically significant decrease in asymptotic activity. In the absence of any other evidence to suggest neurotoxicity or other toxicologically significant effects of treatment, these isolated intergroup differences were considered to be accidental and of no toxicological significance (see Table 3 under "Any other information on results incl. tables").

ORGAN WEIGHTS
No treatment-related effect on body weight was noted. Males treated with 500 mg/kg/day showed a statistically significant reduction in absolute epididymides weight when compared with control but, in the absence of a convincing dose-response relationship, the intergroup difference was considered to be incidental and of no toxicological importance (see Table 4 under "Any other information on results incl. tables").

GROSS PATHOLOGY
Necropsy revealed no substance-related findings. One male and female animal treated with 150 mg/kg/day and a female animal treated with 1000 mg/kg/day displayed dark foci on the lungs. These findings showed non dose-related response and where considered to be of non toxicological importance.

HISTOPATHOLOGY: NON-NEOPLASTIC
Three males treated with 1000 mg/kg/day demonstrated globular accumulations of eosinophilic material in the proximal tubular epithelium which should be regarded as a possible effect of treatment. The authors considered this finding consistent with the appearance of hydrocarbon nephropathy, which results from the excessive accumulation of α2-microglobulin in renal proximal tubular epithelium of adult male rats, which does not represent a hazard to human health.
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
Critical effects observed:: no

Reference 3

Endpoint:: short-term repeated dose toxicity: oral
Type of information:: experimental study
Adequacy of study:: weight of evidence
Study period:: Nov - Dec 1992
Reliability:: 1 (reliable without restriction)
Rationale for reliability incl. deficiencies:: guideline study
Qualifier:: equivalent or similar to guideline
Guideline:: OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:: no
GLP compliance:: yes
Limit test:: no
Species:: rat
Strain:: other: Alpk:APfSD
Sex:: male/female
Details on test animals or test system and environmental conditions:: TEST ANIMALS
- Source: Zeneca Pharmaceuticals, Alderly Park, Macclesfield, Cheshire, UK
- Age at study initiation: 28 d
- Weight at study initiation: 148.45 g (males); 122.6 g (females)
- Housing: sexes separately, five per cage, cages had measurements of 26.5 x 50.0 x 20.0 cm and were constructed of stainless steel mesh with one solid side.
- Diet: CT1 diet; Special Diets Services Limited, Witham, Essex, UK
- Water: tap water, ad libitum
- Acclimation period: approx. 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 45-65 (71 at one occasion)
- Air changes (per hr): 25-30
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: November 1992 To: December 1992
Route of administration:: oral: feed
Vehicle:: other: in diet
Details on oral exposure:: PREPARATION OF DOSING SOLUTIONS: All diet preparations were based on CT1 diet (Special Diets Services Limited, Witham, Essex, UK). They were prepared by grinding the appropriate amount of test substance with 1 kg of milled CT1 diet. This premix was then added to 14 kg of diet and mixed thoroughly with a Pharma Blender Model PMA 100S (T K Filder).

DIET PREPARATION
- Rate of preparation of diet (frequency): 15 kg batches
- Mixing appropriate amounts with (Type of food): CT1 diet (Special Diets Services Limited, Witham, Essex, UK)
- Storage temperature of food: - 20°C, stored at room temperature for usage up to 14 days
Analytical verification of doses or concentrations:: yes
Details on analytical verification of doses or concentrations:: Chemical stability was determined for diet preparations over a period of 5 weeks following storage at room temperatureT or at -20°C.
Samples were extracted by chemical shaking with ethyl acetate. The supernatant was diluted with ethyl acetate to give solutions containing appropriate concentrations of the test substance. Extracts were analysed by gas chromatography using flame ionisation detection. The extract concentration was calculated by reference to data from a standard containing a known concentration.
Duration of treatment / exposure:: daily
Frequency of treatment:: 28 d
Dose / conc.:: 1 000 ppm
Remarks:: nominal in diet
Dose / conc.:: 5 000 ppm
Remarks:: nominal in diet
Dose / conc.:: 12 500 ppm
Remarks:: nominal in diet
Dose / conc.:: 112 mg/kg bw/day (actual dose received)
Remarks:: actual ingested for males
Dose / conc.:: 562 mg/kg bw/day (actual dose received)
Remarks:: actual ingested for males
Dose / conc.:: 1 450 mg/kg bw/day (actual dose received)
Remarks:: actual ingested for males
Dose / conc.:: 119 mg/kg bw/day (actual dose received)
Remarks:: actual ingested for females
Dose / conc.:: 586 mg/kg bw/day (actual dose received)
Remarks:: actual ingested for females
Dose / conc.:: 1 613 mg/kg bw/day (actual dose received)
Remarks:: actual ingested for females
No. of animals per sex per dose:: 5
Control animals:: yes
Details on study design:: - Dose selection rationale: Based on results of preliminary feeding studies
Observations and examinations performed and frequency:: CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked: changes in clinical condition and behaviour and significant changes were recorded.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: on Days 8, 15, 22, 29
- observations included, but were not limited to the assessment of autonomic function (e.g. lacrimation, salivation, piloerection, exophthalmus, urination, defecation, pupillary function, ptosis); description, incidence and severity of any convulsions, tremors, abnormal motor function, alteration in respiration, reactivity to stimuli, changes in the level of arousal, sensorimotor responses

BODY WEIGHT: Yes, measurement in replicate order immediately before feeding and at the same day once a week until termination.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as mg food/kg body weight: Yes, on a weekly basis
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

HAEMATOLOGY: Yes
At termination, all rats were bled by cardiac puncture and samples were collected. Parameters determined: Hemoglobin, red cell count, haematocrit, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, platelet count, white blood cell count, neutrophil count, lymphocyte count, monocyte count, eosinophil count, prothrombin time and kaolin-cephalin time

CLINICAL CHEMISTRY: Yes
At termination, all rats were bled by cardiac puncture and samples were collected. Parameters determined: Albumin, total protein, cholesterol, triglycerides, urea, creatinine, glucose, total bilirubin and alkaline phosphatase, plasma gamma-glutamyl transferase, plasma alanine aminotransferase, plasma aspartate aminotransferase, plasma creatine kinase, plasma sodium, plasma potassium, plasma chloride, plasma calcium and plasma phosphorus

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: on Days 8, 15, 22, 29
- Dose groups that were examined: All
- Battery of functions tested: sensory activity / grip strength
Sacrifice and pathology:: GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes (adrenals, aorta, bladder, bone and bone marrow (femur), brain, caecum, colon, cervical lymph node, cervix, colon, duodenum, epididymis, eye and harderian gland, heart, ileum, jejunum, kidney, liver, lungs, mammary gland, mesenteric lymph node, nasal passages, oesophagus, oral cavity, ovaries, pancreas, parathyroid glad, pituary gland, prostate gland, rectum, salivary glands, sciatic nerve, seminal vesicles, skin, spinal chord, spleen, sternum, stomach, testes, thymus, thyroid gland, trachea, uterus, voluntary muscle)
Statistics:: Bodyweights were considered by analysis of covariance on initial body weight, separately for males and females.
Time to tail flick and fore and hindlimb grip strength at weeks 2, 3, 4 and 5 were considered by analysis of variance, separately for both sexes.
Haematological and clinical blood parameters were considered by analysis of variance.
Organ weights were considered by analysis of variance and covariance on final body weight separately for both sexes.
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
Haematological findings:: effects observed, treatment-related
Description (incidence and severity):: reduction in haemoglobin and haematocrit (males), reductions in haemoglobin, haematocrit and in white blood cell count (females).
Clinical biochemistry findings:: effects observed, treatment-related
Description (incidence and severity):: minor reductions in plasma cholesterol, triglyceride, total protein levels and plasma alanine transferase activities in males
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 (males), slightly increased kidney weights (females), increased liver weights (males/females) in 5000 and 12500 ppm groups
Gross pathological findings:: no effects observed
Histopathological findings: non-neoplastic:: effects observed, treatment-related
Description (incidence and severity):: kidney: increased tubular hyaline droplet formation, tubular basophilia, granular cast formation (males). liver: minimal hepatocyte hypertrophy in 4/5 male rats
Histopathological findings: neoplastic:: no effects observed
Details on results:: DIET ANALYSIS:
All diets prepared were found to be within 4% of the target concentration. The homogeneity of the test material in the diet, determined at 1000 and 12500 ppm inclusion levels was within 2 % of the overall mean concentration for both levels. Chemical stability of the test material, assessed at the 1000 and 12500 ppm inclusion levels stored at room temperature or at -20 °C was satisfactory over the period of use.
Dose rates (based on nominal dietary levels) were highest at the start of the study and declined rapidly during the period of rapid growth to week 4.

MORTALITY
There were no mortalities.

FUNCTIONAL OBSERVATION BATTERY
A slightly reduced splay reflex was observed in one female of the 1000 ppm group (on days 29 and 30), in one male of the 5000 ppm group (on day 29) and in one male of the 12.500 ppm group (on day 29). As isolated observations, these were considered to be incidental.
There were no differences in time to tail flick in either sex which could be attributed to treatment. The statistically significant increase in time to response observed on day 22 for males (5000 ppm) and day 8 for females (1000 ppm) were considered to be incidental to treatment in the absence of similar changes at higher dose levels.
There was no evidence of any treatment related effects on forelimb or hindlimb grip strength.
Any other statistically significant changes were considered spurious and unrelated to treatment with the test material.

BODY WEIGHT AND WEIGHT GAIN
There were no statistically significant effects on body weight and all final bodyweights were within 3% of the respective controls, after adjusting for initial weight differences.

FOOD CONSUMPTION
Food consumption in all treated groups remained similar to, or exceeded that, of the respective control group throughout the study.

HAEMATOLOGY
There were statistically significant reductions in haemoglobin and haematocrit at 12.500 ppm in male rats. Statistically significant reductions in haemoglobin and haematocrit were seen in females at 1000 and 5000 ppm and in white blood cell count at 1000 ppm. In the absence of a coherent dose-response relationship, these differences were considered incidental to treatment.
Any other statistically significant changes were considered spurious and unrelated to treatment with the test material.

CLINICAL CHEMISTRY
There were minor reductions in plasma cholesterol, triglyceride and total protein levels and plasma alanine transferase activities in males at 12500 ppm compared to controls. Any other statistically significant changes were considered spurious and unrelated to treatment with the test material.
ORGAN WEIGHTS
Kidney weights adjusted for body weight were statistically significant increased in males at 5000 and 12500 ppm. All the females in the treatment groups had slightly raised kidney weights compared to control, but none achieved statistical significance, and there was no evidence of a coherent dose response relationship.
Liver weights adjusted for body weight were statistically significant increased in both sexes at 12500 ppm and in males at 5000 ppm.
Any other statistically significant changes were considered spurious and unrelated to treatment with the test material.

PATHOLOGY:
Macroscopic findings:
No treatment-related macroscopic findings were apparent at the end of the study.
HISTOPATHOLOGY:
Microscopic findings:
Treatment related findings were present in the kidney of male rats from all dose groups. In the 5000 and 12500 ppm dose group these comprised increased tubular hyaline droplet formation and tubular basophilia in all animals, and granular cast formation in four of the 5000 ppm animals and all of the 12500 ppm animals; the latter occurring at the cortico-medullary injection. In the 1000 ppm group, increased renal hyaline droplet formation and/or tubular basophilia were seen, but not granular cast formation.
In the liver, there was minimal hepatocyte hypertrophy in 4/5 male rats in the 12500 ppm group.

The increased kidney weights and microscopic findings of renal tubular basophilia, granular cast formation and increased hyaline droplet formation present in male rats at 5000 and 12.500 ppm are clearly treatment related. These findings are consistent with the well characterized light hydrocarbon nephropathy described for male rats, following to a variety of chemicals including light hydrocarbons such as unleaded gasoline and trimethyl pentane. The characteristics include an increased accumulation of hyaline droplets in male rat kidneys, the main constituent of which is alpha 2µ-globulin (Alden et al. Adv. Modern Environ Toxicol 7: 107-120 (1984); Stonard et al. Renal Heterogeneity and Target Cell Toxicity. Bach PH and Lock EA Eds, John Wiley and Sons (1985)). It is widely accepted that this phenomenon is specific to male rat and as such appears to have no relevance for man (Swenberg et al. Toxicol and App. Pharmacol. 97: 35-46 (1989)).
Dose descriptor:: NOAEL
Effect level:: 1 450 mg/kg bw/day (actual dose received)
Based on:: test mat.
Sex:: male
Basis for effect level:: other: no effects observed in male rats
Dose descriptor:: NOAEL
Effect level:: 1 613 mg/kg bw/day (actual dose received)
Based on:: test mat.
Sex:: female
Basis for effect level:: other: no effects observed in female rats
Critical effects observed:: no

Reference 4

Endpoint:: short-term repeated dose toxicity: oral
Remarks:: combined repeated dose and reproduction / developmental screening
Type of information:: experimental study
Adequacy of study:: weight of evidence
Study period:: 22 Apr - 17 Jun 2004
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)
Deviations:: no
GLP compliance:: yes
Limit test:: no
Species:: rat
Strain:: Crj: CD(SD)
Sex:: male/female
Details on test animals or test system and environmental conditions:: TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc., Yokohama, Japan
- Age at study initiation: 10 weeks
- Weight at study initiation: males: 330.8 - 420.4 g (mean 374.0 g), females: 207.2 - 245.0 g (mean 228.3 g)
- Fasting period before study: no fasting period
- Housing: steel cage with wire floor
- Diet: CE-2 from CLEA Japan, Inc., Tokyo, Japan, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.0 - 24.5
- Humidity (%): 49.0 - 67.0
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:: oral: gavage
Vehicle:: corn oil
Details on oral exposure:: PREPARATION OF DOSING SOLUTIONS:
A 50% w/v solution was prepared by diluting test substance in vehicle. This solution was diluted serially with vehicle to prepare 5 and 15% w/v solutions. Gavage solution was prepared within 8 days before administration, since the stability was verified for 8 days. Prepared samples were stored at room temperature in the dark.

VEHICLE
- Justification for use and choice of vehicle: test substance is insoluble in water, but not in corn oil
- Lot/batch no. (if required): V2P1825
Analytical verification of doses or concentrations:: yes
Details on analytical verification of doses or concentrations:: 0.5 mL of each prepared solution was mixed with hexane to be confirmed by GC. The resulting analytical concenrations were 107 - 110% of the nominal concentrations
Duration of treatment / exposure:: Males: 42 days
Females: from Day 14 before mating to Day 4 of lactation
Females (satellite group): 42 days

Recovery period :
- Males: 14 days
- Females (satellite): 14 days
Frequency of treatment:: once daily
Dose / conc.:: 100 mg/kg bw/day (actual dose received)
Dose / conc.:: 300 mg/kg bw/day (actual dose received)
Dose / conc.:: 1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:: Control: 7 males, 100 mg/kg: 12 males, 300 mg/kg: 12 males, 1000 mg/kg: 7 males;
12 females per dose;
control and 1000 mg/kg for satellite group; 5 males and 5 females per dose
Control animals:: yes, concurrent vehicle
Details on study design:: - Dose selection rationale: The dose were based on the results of a preliminary test, where groups of male and female rats received doses of 100, 300 and 1000 mg/kg bw, respectively. No effects on general condition, body weight gain nor organ weights of liver, kidney or spleen was observed in any of the treated animals. Based on this, the maximum dose for the study was set to 1000 mg/kg bw, while the medium and low dose were set to 300 and 100 mg/kg bw, respectively.
- Post-exposure recovery period in satellite groups: 14 days
Observations and examinations performed and frequency:: CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day during treatment period and once a day during recovery period

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:
Males: Day 0, 7, 14, 21, 28, 35 and 42 of treatment
Males in satellite groups: Day 0, 7, 14, 21, 28, 35, 42 of treatment, Day 7 and 14 of recovery period
Females: Day 0, 7, 14, 21, 28, 35, 42 of treatment and once from Day 0 of lactation to Day 4 of lactation
Females in satellite groups: Day 0, 7, 14, 21, 28, 35, 42 of treatment, Day 7 and 14 of recovery period

BODY WEIGHT: Yes
- Time schedule for examinations:
Males: Day 1, 7, 14, 21, 28, 35, 42 of administration and before sacrifice
Males and females in satellite groups: Day 1, 7, 14, 21, 28, 35 , 42 of administration, Day 1, 7, 14 of recovery period and before sacrifice
Females: Day 1, 7, 14, 21 of treatment, Day 0, 7, 14, 20 of pregnancy, Day 0 and 4 of lactation, and before sacrifice

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Day 43 (on day after last treatment) for males, Day 15 of recovery period for satellite animals of both sexes, Day 4 of lactation for dams, Day 26 of pregnancy for non delivered dams
- Anesthetic used for blood collection: Yes (sodium pentobarbital)
- Animals fasted: Yes (18 - 24 hours)
- How many animals: 5 in each group
- Parameters checked: erythrocytes count (RBC), hemoglobin, hematocrit, Mean Cell Volume (MCV), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Con. (MCHC), leykocyte count (WBC), differential leukocyte count, platelet, Prothrombin time (PT), Activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Day 43 (on day after last treatment) for males, Day 15 of recovery period for satellite animals of both sex, Day 4 of lactation for dams, Day 26 of pregnancy for non delivered dams
- Anaesthetic used for blood collection: Yes (pentobarbital sodium)
- Animals fasted: Yes (18 - 24 hours)
- Parameters checked: total protein, albumin, A/G, blood urea nitrogen, creatinine, glucose, total cholesterol, triglyceride, Alkaline phosphatase (ALP), Alanine transaminase (ALT (GPT)), Aspartate transaminase (AST (GOT)), γ-GTP, total bilirubin, inorganic phosphorous, calcium, Na, K, Cl

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at the end of dosing period and at the end of recovery period
- Dose groups that were examined: all group
- Battery of functions tested: pupilary reflex, eyelid reflex, visual placing, withdrawal reflex, Payer's reaction, startle reaction, righting reflex
Sacrifice and pathology:: Termination:
- Males: day 43 of treatment and day 15 of recovery
- Females: day 5 of lactation
- Females (satellite): day 15 of recovery
- Offspring: 4 days after birth

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes
brain, the pituitary gland, spinal cord, heart, lung and bronchus, liver, kidney, thymus, spleen, adrenal gland, thyroid gland, mandibular lymph node, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, testis, epididymis, prostate, seminal vesicles including coagulation glands, ovary, uterus, vagina, mesenteric lymph node, sciatic nerve, urinary bladder, bone marrow of femur
Other examinations:: Estrus cycle, reproductive performance, observation of pups
Statistics:: Fisher test, Mann-Whitney-U-Test, Student's t-test, Aspin-Welch test, Bartlett test, Kruskal-Wallis test, Dunnett test
Clinical signs:: effects observed, treatment-related
Description (incidence and severity):: incidental effects which were not treatment-related (non adverse)
Mortality:: mortality observed, treatment-related
Description (incidence):: incidental effects which were not treatment-related (non adverse)
Body weight and weight changes:: effects observed, treatment-related
Description (incidence and severity):: 100 mg/kg bw: reduced body weight in males (non adverse)
Food consumption and compound intake (if feeding study):: effects observed, treatment-related
Description (incidence and severity):: 1000 mg/kg bw: significantly more food consumption during days 29 - 30 by satellite group (non adverse)
Food efficiency:: not examined
Water consumption and compound intake (if drinking water study):: not examined
Ophthalmological findings:: not examined
Haematological findings:: effects observed, treatment-related
Description (incidence and severity):: 300 mg/kg bw: increased hematocrit and hemoglobin in females (non adverse)
Clinical biochemistry findings:: effects observed, treatment-related
Description (incidence and severity):: 1000 mg/kg bw: increase of blood urea nitrogen in females at the end of recovery period (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):: 1000 mg/kg bw: increase in relative liver weight in females (non adverse)
Gross pathological findings:: effects observed, treatment-related
Description (incidence and severity):: one tumor in the femur (non adverse)
Histopathological findings: non-neoplastic:: effects observed, treatment-related
Description (incidence and severity):: all changes were found distributed over all groups, thus they were regarded as spontaneous incidences (non adverse)
Histopathological findings: neoplastic:: no effects observed
Description (incidence and severity):: one tumor in the femur (non adverse)
Details on results:: CLINICAL SIGNS AND MORTALITY
No mortality was observed in all groups.
An emaciation was observed at day 40 of treatment (Day 1 of lactation) in the female control group, but this was recovered after one day. Loss of the upper incisor was observed at day 25 - 32 in a female of the 300 mg/kg group but this was considered due to physical shock and were not affected by administration of the test substance.

BODY WEIGHT AND WEIGHT GAIN
Males of the 100 mg/kg group showed body weight loss compared to the control group, but this was not statistically significant. There was no difference in female groups and satellite groups compared with control groups.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Females of the satellite 1000 mg/kg group showed significantly more food consumption during days 29 - 30 compared to the control group. However, this change was considered as not compound-related as no difference in other groups was found, both within the treatment period as well as in the recovery period.

HAEMATOLOGY
At the end of administration, hematocrit and hemoglobin were significantly increased in females of the 100 mg/kg group. Prothrombin time was prolonged in females of the 300 mg/kg group. These changes were not considered as compound-related changes, since a dose dependency was not observed. No other significant differences were observed.

CLINICAL CHEMISTRY
At the end of dosing period, significant differences between all treatment groups and the control groups were not found.
Only concentration of blood urea nitrogen was increased in females of the 1000 mg/kg group at the end of recovery period. This change was not considered as compound-related change, since no corresponding abnormalities were observed in the kidney.

NEUROBEHAVIOUR
No abnormalities were observed.

ORGAN WEIGHTS
At the end of dosing period, a significantly increase of absolute weight of brain was found in females (300 mg/kg), but the change of relative weight was not significant. No significant differences in males between treatment group and the control group were observed.
At the end of recovery period, a significantly increase of relative liver weight in females of the 1000 mg/kg group was noted, while no differences in males were found. The change was not considered as compound-related, since no corresponding abnormalities were observed in the livers.

GROSS PATHOLOGY
At the end of dosing period, no abnormalities were observed. At the end of recovery period, a tumor in right hind femur was observed in one male of the control group.

HISTOPATHOLOGY: NON-NEOPLASTIC
At the end of dosing period, localized atrophy of the seminiferous tubules were observed in three males of the 100 mg/kg group and in one male of the 300 mg/kg group. Three of these animals had cell debris in lumen in epidedymis. Livers in males of the 1000 mg/kg and control groups exhibited fatty change in hepatocyte and microgranuloma. In all control group animals and three animals of 1000 mg/kg, eosinophilic bodies were found in cortex of the kidney, while slightly basophilic tubules were found in kidney cortex of two animals in the control group and in four animals of the 1000 mg/kg group. Extramedullary hematopoiesis and deposit of brown pigment was found in the spleen of all animals of the control and the 1000 mg/kg group. In addition, focal degeneration/fibrosis in the myocardium, accumulation of focal foam cell in the adveolus and mineralization of the arterial wall in lung, as well as lymphocytes and neutrophil cellular infiltration in prostate were observed both in control groups and 1000 mg/kg groups. However, these change were not significantly different but accidental.
In one female of the 1000 mg/kg group, follicular cyst, increased arterial follicle and decreased corpus luteum was found in ovary at the end of dosing period. In liver, fatty change in periportal hepatocytes and microgranuloma was observed in all animals of control and 1000 mg/kg groups, respectively. Basophilic tubule in the cortex of the kidney was observed in one animal at 1000 mg/ kg and slight mineralization was found in one control animal and one animal dosed with 1000 mg/kg, respectively. In the spleen, extramedullary hematopoiesis and brown deposit were found in all animals of the control and the 1000 mg/kg group. In addition, focal degeneration/fibrosis in myocardium, mineralization on arterial wall of lung and atrophy in thymus were also found in both control and 1000 mg/kg. However, these change were not significantly different but accidental.
At the end of recovery period, intramembranous ossification and periosteum proliferation were found in one male in control. These histopathological findings are due to fracture of the right femur. In one female of 1000 mg/kg, follicular cyst was found.
Dose descriptor:: NOAEL
Effect level:: >= 1 000 mg/kg bw/day (nominal)
Based on:: test mat.
Sex:: male/female
Basis for effect level:: other: no adverse effects observed
Critical effects observed:: no

Endpoint conclusion

Endpoint conclusion:: no adverse effect observed
Dose descriptor:: NOAEL
: 1 000 mg/kg bw/day
Study duration:: subchronic
Species:: rat
Quality of whole database:: The available information comprises adequate and reliable (Klimisch score 1) studies from various source substances with similar structures and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected studies are thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 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:: experimental study
Adequacy of study:: key study
Reliability:: 2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:: comparable to guideline study with acceptable restrictions
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
Dose / conc.:: 0 mg/L air (analytical)
Dose / conc.:: 0.05 mg/L air (analytical)
Dose / conc.:: 0.17 mg/L air (analytical)
Dose / conc.:: 0.56 mg/L air (analytical)
Dose / conc.:: 0.05 mg/L air (nominal)
Dose / conc.:: 0.15 mg/L air (nominal)
Dose / conc.:: 0.5 mg/L air (nominal)
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 and no mortalities were 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:: 0.56 mg/L air (analytical)
Based on:: test mat.
Sex:: male/female
Basis for effect level:: other: no adverse effects observed
Critical effects observed:: no

Endpoint conclusion

Endpoint conclusion:: no adverse effect observed
Dose descriptor:: NOAEC
: 500 mg/m³
Study duration:: subchronic
Species:: rat
Quality of whole database:: The available information comprises an adequate and reliable (Klimisch score 2) study from a source substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected study is thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 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:: experimental study
Adequacy of study:: key study
Reliability:: 2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:: comparable to guideline study with acceptable restrictions
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
Dose / conc.:: 0 mg/L air (analytical)
Dose / conc.:: 0.05 mg/L air (analytical)
Dose / conc.:: 0.17 mg/L air (analytical)
Dose / conc.:: 0.56 mg/L air (analytical)
Dose / conc.:: 0.05 mg/L air (nominal)
Dose / conc.:: 0.15 mg/L air (nominal)
Dose / conc.:: 0.5 mg/L air (nominal)
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 and no mortalities were 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:: 0.56 mg/L air (analytical)
Based on:: test mat.
Sex:: male/female
Basis for effect level:: other: no adverse effects observed
Critical effects observed:: no

Endpoint conclusion

Endpoint conclusion:: no adverse effect observed
Dose descriptor:: NOAEC
: 500 mg/m³
Study duration:: subchronic
Species:: rat
Quality of whole database:: The available information comprises an adequate and reliable (Klimisch score 2) study from a source substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected study is thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 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:: experimental study
Adequacy of study:: key study
Study period:: 09 Jul - 10 Oct 1986
Reliability:: 2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:: comparable to guideline study with acceptable restrictions
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)
Dose / conc.:: 800 mg/kg bw/day (nominal)
Dose / conc.:: 2 000 mg/kg bw/day (nominal)
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: no adverse effects observed
Critical effects observed:: no

Endpoint conclusion

Endpoint conclusion:: no adverse effect observed
Dose descriptor:: NOAEL
: 2 000 mg/kg bw/day
Study duration:: subchronic
Species:: rat
Quality of whole database:: The available information comprises an adequate and reliable (Klimisch score 2) study from a source substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected study is thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 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:: experimental study
Adequacy of study:: key study
Study period:: 09 Jul - 10 Oct 1986
Reliability:: 2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:: comparable to guideline study with acceptable restrictions
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)
Dose / conc.:: 800 mg/kg bw/day (nominal)
Dose / conc.:: 2 000 mg/kg bw/day (nominal)
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: no adverse effects observed
Critical effects observed:: no

Endpoint conclusion

Endpoint conclusion:: adverse effect observed
Dose descriptor:: NOAEL
: 800 mg/cm²
Study duration:: subchronic
Species:: rat
Quality of whole database:: The available information comprises an adequate and reliable (Klimisch score 2) study from a source substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected study is thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 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

The read-across from analogue source substances approach comprises aliphatic esters of poly-functional alcohols containing two to six reactive hydroxyl groups and one to six fatty acid chains. The analogue approach contains mono constituent, multi-constituent and UVCB substances with fatty acid carbon chain lengths ranging from C5 - C28, which are mainly saturated but also mono unsaturated C16 and C18, polyunsaturated C18, branched C5 and C9, branched C14 - C22 building mono-, di-, tri-, and tetra esters with an alcohol (i.e. the polyol).

The available data allows for an accurate hazard and risk assessment of the target substance and the read-across concept is applied for the assessment of environmental fate and environmental and human health hazards. Thus, where applicable, environmental and human health effects are predicted from adequate and reliable data for source substances within the group by interpolation to the target substance applying the group concept in accordance with Annex XI, Item 1.5, of Regulation (EC) No. 1907/2006. In particular, for each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read-across, with due regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across.

A detailed justification for the grouping of chemicals and read-across is provided in the technical dossier (see IUCLID Section 7.1 and 13).

Oral repeated dose toxicity

There are several short-term and one sub-chronic repeated dose toxicity studies available for structural analogue source substances investigating the toxicity after oral exposure. All studies are accounted for by means of a Weight-of-Evidence approach.

In the first short-term (28 day) repeated dose toxicity study Fatty acids, C5-10, esters with pentaerythritol (CAS No. 68424-31-7) was investigated according to OECD Guideline 407 and under GLP conditions (WoE, RA-A, 68424-31-7, 1993). The test substance was administered in the diet 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.

The short-term (28-day) repeated dose toxicity of Dipentaerythritol hexaesters with fatty acids, C5 and C9iso (CAS No. 647028-25-9) was assessed in a study performed according to OECD Guideline 407 and in compliance with GLP provisions (WoE, RA-A, 647028-25-9, 2000a). Groups of 5 male and 5 female Crl:CD BR rats were dosed once daily, 7 day/week, with 150, 500 and 1000 mg/kg bw/day of the test substance by oral gavage for 28 consecutive days. The animals of a control group received only the vehicle arachis oil. No mortality occurred during the study period and no clinical signs of toxicity were observed in test or control animals throughout the study. Minor effects observed, e.g. fur loss in one female of the highest dose group, were considered incidental. No adverse effect on body weight was noted. Nevertheless a statistically significant reduction in body weight gain was apparent for 1000 and 500 mg/kg/day males during week 1 of the study. The dose relationship was not credible and the intergroup differences were considered to be a result of slightly higher than usual control group body weights gains. No treatment-related changes in the hematological parameters were measured. No treatment-related effect on organ weights was noted. Males treated with 500 mg/kg/day showed a statistically significant reduction in absolute epididymides weight when compared with control animals but, in the absence of a convincing dose-response relationship, the intergroup difference was considered to be incidental and of no toxicological importance. Necropsy revealed no substance-related findings. Three males treated with 1000 mg/kg/day demonstrated globular accumulations of eosinophilic material in the proximal tubular epithelium which could be regarded as a possible effect of treatment. The authors considered this finding consistent with the appearance of hydrocarbon nephropathy, which results from the excessive accumulation of α2-microglobulin in renal proximal tubular epithelium of adult male rats, which does not represent a hazard to human health. Taken all observations in consideration, the oral NOAEL value was therefore determined to be ≥ 1000 mg/kg bw/day, corresponding to the highest dose tested.

A combined repeated dose and reproduction / developmental toxicity screening study according to OECD Guideline 422 and under GLP conditions (MHWL, 2005b) was performed with Hexanoic acid, 2-ethyl-, 2,2-bis [ [(2-ethyl-1-oxohexyl)oxy] methyl] -1,3-propanediyl ester (CAS No. 7299-99-2). Doses of 100, 300 and 1000 mg/kg bw/day were administered to male and female Crj: CD(SD) rats once daily by oral gavage. All males and the females of the satellite group were dosed for 42 days while females of the dose groups were treated from Day 14 before mating to Day 4 of lactation. A 14-day recovery period for males and the females of the satellite group followed the dosing period. Animals of the control group received only the vehicle corn oil. No mortality was observed in all groups. Males of the 100 mg/kg group showed body weight loss compared to the control group, but this was not statistically significant. There was no difference in female groups and satellite groups compared with control groups. At the end of administration, hematocrit and hemoglobin were significantly increased in females of the 100 mg/kg group. Prothrombin time was prolonged in females of the 300 mg/kg group. These changes were not considered as compound-related changes, since a dose dependency was not observed. At the end of dosing period, significant differences between all treatment groups and the control groups were not found with regard to clinical chemistry parameters. No statistically significant or treatment-related effects were observed with respect to organ weights after the treatment period. The histopathological examination revealed only minor observations which were all considered to be accidental and of no toxicological relevance. Therefore, the oral NOAEL was established at ≥ 1000 mg/kg bw/day for male and female rats in this study.

In addition to the short-term repeated dose studies referred to above, a sub-chronic (90-day) oral toxicity study with Pentaerythritol ester of pentanoic acids and isononanoic acid (CAS No. 146289-36-3) was performed according to OECD Guideline 408 and under GLP conditions (Emery, 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. Therefore, a 90-day oral NOAEL of ≥ 1000 mg/kg bw/day was found for the test substance in male and female rats.

In conclusion, the available studies clearly indicate a very low level of repeated dose toxicity after oral exposure either in the diet (WoE, RA-A, 64842-31-7, 1993) or via gavage (WoE, RA-A, 647028-25-9, 2000a; MHWL, 2005b; Emery, 1998). Since no toxicologically relevant findings were detected in any of the studies and hence all NOAELs determined correspond to the highest doses tested, a NOAEL of ≥ 1000 mg/kg bw/day is used for risk assessment and C&L purposes of the target substance.

Inhalation repeated dose toxicity

A relevant key study investigating the effects of repeated inhalation exposure to a structural analogue substance is available.

A 90-day subchronic inhalation toxicity study was performed with Sprague-Dawley rats with Fatty acids, C5-9, tetraesters with pentaerythritol (CAS No. 67762-53-2) comparable to OECD guideline 413 (Exxon, 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 air. The respective controls (15 animals per sex and dose) inhaled clean air under the same conditions. Animals were observed for clinical sings, 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 hydroxy proline following exposure. No substance-related adverse effects were observed for body weight, body weight gain, mortality, clinical biochemistry and hematological 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 air.

Dermal repeated dose toxicity

Regarding repeated dose toxicity after dermal exposure, one relevant key study of a structural analogue source substance is available.

A 90-day dermal toxicity study with Fatty acids, C5-9, tetraesters with pentaerythritol (CAS No. 67762-53-2) was performed comparable to OECD Guideline 411 (Exxon, 1988a). 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.

Conclusion on repeated dose toxicity

Several reliable studies performed with analogue source substances are available investigating the repeated dose toxicity after the oral, inhalation and dermal route of exposure. All available data indicate a very low level of repeated dose toxicity for the analogue source substances as all NOAEL and NOAEC values determined correspond to the highest doses tested. Thus, no hazard for oral, inhalation and dermal repeated dose toxicity is identified for the target substance Monopentaerythritol tetraesters and dipentaerythritol hexaesters of 2-ethylhexanoic and n-valeric acids.

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 information on intrinsic properties of substances may be generated by means other than tests, e.g. using information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI states that “substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or ‘category’ of substances. This avoids the need to test every substance for every endpoint". Since the read-across concept is applied to the target substance Monopentaerythritol tetraesters and dipentaerythritol hexaesters of 2-ethylhexanoic and n-valeric acids, data gaps can be filled by interpolation from representative structural analogue source substances to avoid unnecessary animal testing.

The read-across concept is also used to derive the classification of the target substance taking the properties of the source substances into account. Based on the read-across concept, all available data on repeated dose toxicity via the oral, inhalation and dermal route of exposure do not meet the classification criteria according to Regulation (EC) No. 1272/2008 (CLP) and are therefore conclusive but not sufficient for classification.

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Dose Level	Increase in bodyweight (g) during Week 1 2 3 4
Group 1 0 mg/kg bw	62 ± 5	53 ± 4	51 ± 8	35 ± 5
Group 2 150 mg/kg bw	56 ± 5	54 ± 4	53 ± 8	38 ± 6
Group 3 500 mg/kg bw	50 ± 6**	51 ± 3	47 ± 9	38 ± 10
Group 4 1000 mg/kg bw	53 ± 5*	51 ± 5	51 ± 9	35 ± 6

Dose Level	MCHC (g/d)	PLT (10⁹/L)
Group 1 0 mg/kg bw	34.6 ± 0.4	792 ± 67
Group 2 150 mg/kg bw	34.3 ± 0.5	928 ± 115
Group 3 500 mg/kg bw	34.2 ± 0.3	944 ± 68*
Group 4 1000 mg/kg bw	33.7 ± 0.7*	920 ± 76

Dose Level (mg/kg bw/day)	Motor activity overall/ Number of animals (% mobile)	Motor activity final 20% of trial/ Number of animals (% mobile)	Motor activity final 20% of trial/ Number of animals (% activity)
control males	5.0 ± 3.8	0.0 ± 0.0	15.1± 25.4
control females	15.4 ± 6.9	9.2± 6.9	49.5± 6.9
150 males	5.0 ± 4.3	0.4± 0.9	21.1± 43.7
150 females	21.5 ± 4.6	14.9± 11.5	49.4± 28.7
500 males	5.8 ± 6.1	0.4± 0.5	11.8± 13.0
500 females	21.7 ± 7.5	47.2 ± 27.3	47.2± 27.3
1000 males	18.3 ± 5.9**	9.9 ± 8.5*	55.4± 32.2
1000 females	17.2 ± 6.0	0.3± 0.3	5.1± 7.6*

Dose Level	Epididymides (g)
Group 1 0 mg/kg bw	1.2251 ± 0.0555
Group 2 150 mg/kg bw	1.0521 ± 0.1711
Group 3 500 mg/kg bw	0.9431 ± 1.031**
Group 4 1000 mg/kg bw	1.0538 ± 0.1125

	Dosage (mg/kg bw)
Days of treatment	0	100	300	1000
1	374.6 ± 24.0	373.6 ± 18.0	374.1 ± 26.2	373.7 ± 21.3
7	409.6 ± 29.0	405.4 ± 23.9	407.0 ± 31.9	404.4 ± 26.6
14	441.4 ± 32.0	431.4 ± 27.4	439.4 ± 38.2	437.5 ± 31.9
21	463.1 ± 34.7	450.4 ± 30.5	467.1 ± 44.7	461.1 ± 27.9
28	489.7± 37.1	474.8 ± 34.5	495.7 ± 53.5	493.4 ± 32.1
35	513.2 ± 40.9	493.6 ± 39.3	519.3 ± 58.2	514.5 ± 35.4
42	534.4 ± 46.4	511.8 ± 41.7	544.3 ± 62.4	540.3 ± 37.8
Days of recovery
1	545.3 ± 55.9			544.0 ± 34.9
7	562.3 ± 59.9			570.0 ± 30.7
14	580.4 ± 57.1			594.3 ± 31.6

	Dosage (mg/kg bw)
	Females		Males
	0	1000	0	1000
Number of animals	5	5	5	5
Terminal body weight	539.1 ± 54.2	548.2 ± 30.6	281.6 ± 0.10	280.5 ± 18.4
Brain	2.06 ± 0.06	2.02 ± 0.08	1.89 ± .0.10	1.90 ± 0.04
Thymus / mg	226.7 ± 23.7	269.8 ± 620.7	245.4 ± 43.9	261.9 ± 50.7
Heart	1.49 ± 0.10	1.50 ± 0.15	0.93 ± 0.12	0.91 ± 0.05
Liver	14.80 ± 2.06	16.21 ± 2.32	7.58 ± 0.44	7.95 ± 0.64
Kidneys	3.45 ± 0.11	3.49 ± 0.33	1.92 ± 0.16	1.97 ± 0.25
Spleen	0.73 ± 0.18	0.88 ± 0.14	0.48 ± 0.03	0.52 ± 0.05
Adrenal galnd / mg	61.5 ± 11.8	67.9 ± 10.2	66.8 ± 6.4	65.6 ± 3.1
Testes	3.74 ± 0.33	3.41 ± 0.12
Epididymides	1.32 ± 0.07	1.30 ± 0.01

	Dosage (mg/kg bw)
Days of treatment	0	100	300	1000
1	228.4 ± 8.4	228.4 ± 5.4	227.7 ± 10.1	227.9 ± 9.8
7	237.6 ± 9.4	240.3 ± 7.9	237.6 ± 11.2	238.7 ± 12.9
14	247.5 ± 13.2	252.2 ± 11.1	249.3 ± 13.5	246.2 ± 14.9
Days of pregnancy
0	256.2 ± 12.2	260.3 ± 7.6	261.6 ± 17.5	259.0 ± 20.6
7	295.0 ± 16.1	298.5 ± 12.1	296.0 ± 23.7	299.3 ± 19.3
14	331.4 ± 17.4	334.5 ± 13.0	334.7 ± 26.4	332.8 ± 20.2
20	404.5 ± 24.0	406.8 ± 19.1	411.2 ± 31.0	401.9 ± 23.3
Days of lactation
0	292.1 ± 27.3	300.1 ± 24.7	297.6 ± 36.9	282.6 ± 29.1
4	327.9 ± 20.5	323.1 ± 14.8	325.0 ± 27.0	311.3 ± 22.6

	Dosage (mg/kg bw)
Days of treatment	0	1000
1	227.9 ± 5.9	230.8 ± 5.0
7	241.9 ± 4.5	242.5 ± 6.2
14	254.4 ± 3.6	253.8 ± 9.1
21	261.0 ± 11.5	261.8 ± 9.5
28	274.7 ± 12.0	273.4 ± 12.9
35	282.9 ± 13.3	278.4 ± 10.6
42	287.5 ± 11.2	286.9 ± 14.5
Days of recovery
1	290.1 ± 11.6	284.7 ± 17.4
7	297.2 ± 13.0	293.4 ± 17.0
14	307.1 ± 12.2	305.1 ± 20.3

	Dosage (mg/kg bw)
	0	100	300	1000
Number of animals	5	5	5	5
Terminal body weight	494.9 ± 36.3	466.3 ± 33.8	521.9 ± 61.8	489.6 ± 38.4
Brain	2.00 ± 0.03	1.98 ± 0.06	2.04 ± 0.10	2.06 ± 0.11
Thymus / mg	342.7 ± 99.4	330.9 ± 69.1	344.3 ± 79.6	350.3 ± 36.6
Heart	1.42 ± 0.10	1.43 ± 0.22	1.49 ± 0.11	1.51 ± 0.11
Liver	13.94 ± 1.18	12.64 ± 1.45	14.75 ± 2.39	13.90 ± 1.91
Kidneys	3.12 ± 0.19	3.23 ± 0.31	3.35 ± 0.12	3.36 ± 0.27
Spleen	0.77 ± 0.19	0.69 ± 0.06	0.85 ± 0.10	0.87 ± 0.10
Adrenal galnd / mg	58.9 ± 3.6	55.7 ± 2.7	64.4 ± 14.7	64.6 ± 13.4
Testes	3.23 ± 0.15	3.24 ± 0.25	3.44 ± 0.47	3.33 ± 0.47
Epididymides	1.19 ± 0.10	1.18 ± 0.12	1.28 ± 0.08	1.26 ± 0.14

	Dosage (mg/kg bw)
	0	100	300	1000
Number of animals	5	5	5	5
Terminal body weight	282.8 ± 23.5	295.2 ± 14.9	297.1 ± 24.8	279.5 ± 17.4
Brain	1.81 ± 0.06	1.87 ± 0.07	1.93 ± 0.06**	1.84 ± 0.04
Thymus / mg	182.8 ± 122.0	201.1 ± 44.5	205.3 ± 94.8	198.9 ± 70.1
Heart	0.91 ± 0.15	0.93 ± 0.08	0.99 ± 0.18	0.91 ± 0.05
Liver	9.75 ± 1.20	10.84 ± 1.46	10.30 ± 0.94	9.95 ± 1.55
Kidneys	1.94 ± 0.29	1.99 ± 0.23	2.03 ± 0.20	1.88 ± 0.21
Spleen	0.59 ± 0.10	0.62 ± 0.02	0.69 ± 0.16	0.60 ± 0.13
Adrenal galnd /mg	65.9 ± 7.0	76.1 ± 14.6	68.4 ± 7.8	75.0 ± 6.4

Registration Dossier

Registration Dossier

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Diss Factsheets

Monopentaerythritol tetraesters and dipentaerythritol hexaesters of 2-ethylhexanoic and n-valeric acids

Endpoint summary

Administrative data

Description of key information

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

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Reference 4

Endpoint conclusion

Repeated dose toxicity: inhalation - systemic effects

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Reference

Endpoint conclusion

Repeated dose toxicity: inhalation - local effects

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Endpoint conclusion

Repeated dose toxicity: dermal - systemic effects

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Reference

Endpoint conclusion

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint conclusion

Additional information

Justification for classification or non-classification

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