Pentaerythritol, mixed esters with linear and branched fatty acids

Endpoint summary

Description of key information

Repeated dose toxicity (oral, WoE, 28d), rat: NOAEL ≥1000 mg/kg bw/day (m/f)

Repeated dose toxicity (inhalation, 90d), rat: NOAEC = 0.5 mg/L air (m/f) (RA from CAS 67762-53-2)

Repeated dose toxicity (dermal, 90d), rat: NOAEL ≥2000 mg/kg bw/day (m/f) (RA from CAS 67762-53-2)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Italy s.r.l., San Pietro al Natisone (UD), Italy.
- Age at study initiation: 6 to 7 weeks old
- Weight at study initiation: 176 to 200 g for males and 151 to 175 g for females
- Allocation: The rats were allocated to groups by computerised stratified randomisation to give approximately equal initial group mean body weights
- Housing: 5 of one sex to a cage, in clear polisulphone solid bottomed cages measuring 59.5⇥38⇥20 cm with a stainless steel mesh lid and floor (Techniplast Gazzada S.a.r.l., Buguggiate, Varese). Nesting material was provided inside suitable bedding bags and changed at least twice a week.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C ± 2 °C
- Humidity (%): 55 % ± 15 %
- Air changes (per hr): 15 to 20 air changes per hour and the rooms were lit by artificial light for 12 hours each day.

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

The test item was administered orally by gavage at a dose volume of 5 mL/kg body weight. Control animals received the vehicle alone at the same dose volume. The dose was administered to each animal on the basis of the most recently recorded body weight and the volume administered was recorded for each animal. During the gestation period, dose volumes were calculated according to individual body weight on Days 0, 7, 14 and 20 post coitum and on Day 1 post partum. Thereafter individual dose volumes remained constant.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analytical method was validated in the present study in the range from 20 to 200 mg/mL. Linearity, accuracy and precision were within the limits stated in RTC SOPs for suspension (r > 0.98; accuracy 90-110%; precision CV < 5%).
Samples of the formulations prepared on Day 1 and Last Week were analysed to check the concentration. Results of the analyses were within the acceptability limits stated in RTC SOPs for concentration of solutions (85-115%).

Duration of treatment / exposure:

Males: 5 weeks; females: 7 days a week, for 2 consecutive weeks prior to pairing and thereafter during pairing, gestation and post partum periods up to Day 3 post partum.

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
100 mg/kg/day
Basis:
actual ingested

Remarks:

Doses / Concentrations:
300 mg/kg/day
Basis:
actual ingested

Remarks:

Doses / Concentrations:
1000 mg/kg/day
Basis:
actual ingested

No. of animals per sex per dose:

Each group comprised 10 male and 10 female rats

Control animals:

yes, concurrent vehicle

Observations and examinations performed and frequency:

MORTALITY: all animals were checked early in each working day and again in the afternoon.

DETAILED CLINICAL OBSERVATIONS: Once before com- mencement of treatment (data not tabulated) and daily during treatment, each animal was observed and any clinical signs were recorded. Observations were performed at the same time interval each day, the interval was selected taking into consideration the presence of post-dose reactions.

NEUROBEHAVIOURAL EXAMINATION:
- FUNCTIONAL OBSERVATION BATTERY TESTS: Once before commencement of treatment and once a week thereafter, each animal was given a detailed clinical examination. Each animal was removed from the home cage and observed in an open arena.
- GRIP STRENGTH AND SENSORY REACTIVITY TO STIMULI: Once during the study, towards the end of treatment, 5 males and 5 females were randomly selected (computer generated random order) from each group for evaluation of sensory reactivity to stimuli of different modalities (e.g. auditory, visual and proprioceptive stimuli); an assessment of grip strength was also performed. For males the tests were performed on Day 30 of study and for females on Day 3 post partum.
- MOTOR ACTIVITY ASSESSMENT: Once during the study, towards the end of treatment, 5 males and 5 females were randomly selected from each group and the motor activity was measured (60 minutes) by an automated activity recording device. Measurements were performed using a computer generated random order. For males the test was performed on Day 30 of study and for females on Day 3 post partum.

BODY WEIGHT: Males were weighed on the day of allocation to treatment groups, on the day of treatment commencement, weekly thereafter and at termination. Females were weighed on the day of allocation to treatment groups, on the day of treatment commencement, weekly thereafter until positive identification of mating and on Days 0, 7, 14 and 20 post coitum. Dams were also weighed on Days 1 and 4 post partum.

FOOD CONSUMPTION: The weight of food consumed by each cage of males and females was recorded weekly during the pre-mating period starting from allocation. Individual food consumption for the females was measured on Days 7, 14 and 20 post coitum starting from Day 0 post coitum and on Day 4 post partum starting from Day 1 post partum.

CLINICAL PATHOLOGY INVESTIGATIONS: As a part of the sacrificial procedure, samples of blood were withdrawn under isofluorane anaesthesia from the abdominal vena cava from 5 males and 5 females (females with viable litters) randomly selected from each group, under condition of food deprivation. At the same time interval, individual overnight urine samples were also collected from the same male animals under the same conditions. Before starting urine collection, water bottles were removed from each cage and each animal received approximately 10 mL/kg of drinking water by gavage, in order to obtain urine samples suitable for analysis. After collection of the urine samples, the water bottles were supplied again to the animals.

HAEMATOLOGY: Haematocrit, Haemoglobin, Red blood cell count, Reticulocyte count, Mean red blood cell volume, Mean corpuscular haemoglobin, Mean corpuscular haemoglobin concentration, White blood cell count, Differential leucocyte count, Neutrophils, Lymphocites, Eosinophils, Basophils, Monocytes, Large unstained cells, Platelets

COAGULATION TEST: Prothrombin time

CLINICAL CHEMISTRY: Alkaline phosphatase, Alanine aminotransferase, Aspartate aminotransferase, Gamma-glutamyltransferase, Urea, Creatinine, Glucose, Triglycerides, Inorganic phosphorus, Total bilirubin, Total cholesterol, Bile acids, Total protein, Albumin, Globulin, A/G Ratio, Sodium, Potassium, Calcium, Chloride

URINALYSIS: Appearance, Volume, Specific gravity, pH, Protein, Glucose, Ketones, Bilirubin, Urobilinogen, Blood
The sediment, obtained from centrifugation at approximately 3000 rpm for 10 minutes, was examined microscopically for: Epithelial cells, Leucocytes, Erythrocytes, Crystals, Spermatozoa and precursors, Other abnormal components

Sacrifice and pathology:

Terminal studies:
- Euthanasia: Parental animals that had completed the scheduled test period were killed by exsanguination under isofluorane anaesthesia.
- Necropsy: The clinical history of the males and females of the parental generation was studied and a detailed post mortem examination was conducted (including examination of the external surface and orifices). Changes were noted, the requisite organs weighed and the required tissue samples preserved in fixative and processed for histopathological examination.

Statistics:

Standard deviations were calculated as appropriate. For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data. The non-parametric Kruskal-Wallis analysis of variance was used for the other parameters. Intergroup differences between the control and treated groups were assessed by the non-parametric version of the Williams test. The mean values, standard deviations and statistical analysis were calculated from actual values in the computer without rounding off.

Clinical signs:

no effects observed

Description (incidence and severity):

No mortality occurred throughout the study and no treatment-related signs were noted.

Mortality:

no mortality observed

Description (incidence):

No mortality occurred throughout the study and no treatment-related signs were noted.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption was unaffected by treatment in both sexes during the study.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Description (incidence and severity):

No remarkable changes were noted at post mortem examination in treated animals, when compared with controls.

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

HAEMATOLOGY: Lymphocytosis was recorded in a number of males dosed with 300 and 1000 mg/kg/day. Lymphocytes mean group values were 36% and 41%, respectively, above controls. The other statistically significant differences between control and treated animals (mean corpuscular haemoglobin concentration in males dosed with 100 mg/kg/day, erythrocytes in females of the same group and basophils in females dosed with 1000 mg/kg/day) were of minimal severity and/or not dose-related, therefore considered of no toxicological significance.
Coagulation: No changes were recorded in the coagulation parameters.

CLINICAL CHEMISTRY: Statistically significant differences between control and treated animals were observed, such as: decrease of triglycerides and chloride in males dosed with 300 mg/kg/day, decrease of albumin in those dosed with 100 mg/kg/day, increase of alkaline phosphatase and decrease of total protein in females treated with 100 mg/kg/day, decrease of globulin in those receiving 100 and 1000 mg/kg/day, increase of albumin/globulin ratio and decrease of creatinine in those treated with 1000 mg/kg/day. Due to the direction of changes and/or the absence of dose-relation, these were considered incidental.

URINALYSIS: Reduced diuresis was recorded in males dosed with 100 mg/kg/day. In addition, urinary haemoglobin, associated with the presence of erythrocytes in the urinary sediment, was recorded in one control animal and one male dosed with 100 mg/kg/day. Due to the absence of dose-relation, the above changes were considered unrelated to treatment.

NEUROBEHAVIOUR:
- Clinical observations (Functional Observation Battery Tests): Neurotoxicity assessment (removal of animals from the home cage and open arena)
Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test
item.
- Motor activity, grip strength and sensory reaction to stimuli: Motor activity and sensory reaction to stimuli measurements recorded at the end of treatment were comparable between control and treated groups in animals of both sexes. Variations recorded in mean grip strength in Group 4 males receiving 1000 mg/kg bw/day and mean landing foot splay in males receiving 300 and 1000 mg/kg bw/day were considered incidental, since they were observed in one sex only and without correlation with the dose.

HISTOPATHOLOGY: The histopathological changes reported in control and treated animals, such as mucosal erosion in the stomach, lymphoid depletion in the thymus, lymphoid hyperplasia in the spleen or malignant nephroblastoma, seen in a control male, were considered to be an expression of spontaneous and/or incidental pathology, commonly seen in this species and age under our experimental conditions.

SPERMATOGENIC CYCLE: A detailed qualitative examination of the testes was performed in five control and high dose group males. Seminiferous tubules were evaluated with respect to their stage in the sperma- togenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was noted.

Key result

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:

body weight and weight gain

clinical signs

mortality

Remarks on result:

not determinable due to absence of adverse toxic effects

Critical effects observed:

not specified

Conclusions:

Based on the results of the present study, the NOAEL for general toxicity was considered to be the highest dose of 1000 mg/kg bw/day.

Executive summary:

The purpose of this study was to generate information on toxic effects on rats after repeated oral dosing of the test item. Experimental procedures were based on the OECD Guideline no 422 and the study was performed according to GLP.

Both male and female rats were treated for approximately 5 weeks. Three different doses were tested (100, 300 and 1000 mg/kg bw/day) and compared to a control group that received only the vehicle (corn oil).

No differences in body weights and food consumption were observed in treated animals compared to the control group.

No clinical signs were observed during the study. No adverse findings were recorded in clinical pathology investigations (haematology, clinical chemistry and urine analysis) apart for lymphocytosis in mid- and high dose groups in males. No relevant differences were recorded in the absolute and relative organ weights of treated animals. No treatment-related changes were noted at macroscopic and microscopic observations.

Based on the results the NOAEL (No Observed Adverse Effect Level) for general toxicity was considered to be the highest dose of 1000 mg/kg bw/day.

Reference 2

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

Nov - Dec 1992

Reason / purpose for cross-reference:

read-across source

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

Remarks:

Doses / Concentrations:
1000 ppm, 5000 ppm, 12500 ppm
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
112, 562, 1450 mg/kg bw/d
Basis:
other: actual ingested for males

Remarks:

Doses / Concentrations:
119, 586, 1613 mg/kg bw/d
Basis:
other: 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:

1 613 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No effects observed in female rats

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: corresponding to 12500 ppm

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises adequate, reliable (Klimisch score 2 due to read-across) and consistent studies from reference substances with similar structure and intrinsic properties. Read-across is justified based on common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to the endpoint discussion for further details).
The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

(limited parameters examined, no daily observation)

GLP compliance:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Taconic, Germantown, NY, USA
- Weight at study initiation: males: 379-388 g ; females: 234-239 g
- Housing: animals were housed in the exposure chambers (feed and water was removed during exposure)

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

whole body

Vehicle:

air

Remarks on MMAD:

MMAD / GSD: 1.0 µm/ approx. 1.8

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 1000 L stainless steel and glass exposure chambers; chambers contained catch pans between each of three leveles of cages.
- System of generating particulates/aerosols: The test material was aerosolized directly from the liquid by a modified Lakin nebulizer on each chamber. The test material was in a straight-walled glass flask and the barrels of the nebulizer were immersed under the level of the liquid in order to maximize the amount of material generated. The distance from the nebulizer to the walls of the flask was approx. 3 cm. After exiting the flask, the aerosol passed through a glass impactor to remove most of the larger particles. The remaining aerosol was mixed with the main air stream for each chamber before entering the chamber.
- Temperature and humidity in air chamber (by a Taylor wet/dry bulb hydrometer approx. every 30 min during each exposure): approx. 23 °C, 56 - 64%
- Air flow rate: approx. 300 L/min (mean chamber flow per group: 297, 308, 342, and 243 L/min, respectively)

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric sampling on glass fiber filters (3 times during each exposure); some filters were additionally analyzed by GC/MS
- Samples taken from breathing zone: yes
Nominal concentrations were determined as the loss of weight of fluid from the generator divided by total air flow through the chamber.

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

6 hours/day
5 days/week

Remarks:

Doses / Concentrations:
0.00 ± 0.00, 0.05 ± 0.01, 0.17 ± 0.01, and 0.56 ± 0.02 mg/L
Basis:
analytical conc.

Remarks:

Doses / Concentrations:
0.05, 0.15, and 0.5 mg/L
Basis:
nominal conc.

No. of animals per sex per dose:

15
(Additional 10 male rats per group were included for examination of pulmonary function tests and analysis of pulmonary hydroxyproline following exposure.)

Control animals:

yes, concurrent no treatment

yes, sham-exposed

Details on study design:

- Dose selection rationale: The highest dose was expected to result in abnormal accumulation of test material in the lung and possible impairment of normal clearance mechanisms. The low dose is a factor of 10 above the TLV (treshold limit value) for mineral oil mistes, no untoward effects were expected at this level.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (except weekends)

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

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

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

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

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

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

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
Lung function: The animals were anaesthetized and pulmonray function tests were performed (deflation quasistatic pressure-volume cureved, functional residual capactiy, and maximal forced exhalation maneuver). After the pulmonray function tests, the lungs were removed and all lobes were weighed. Lobes were frozen for analysis of hydroxyproline content and analysis of test material remaining in the lung.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes; organ weights (adrenals, kidney, spleen, brain, liver, testes, epididymides, ovaries, thymus, heart, prostate, uterus, right middle lung lobe
HISTOPATHOLOGY: Yes (untreated and high-dose): adrenals, ovaries, sternum, pancreas, brain, salivary gland, eye, spleen heart, stomach, colon, testes, duodenum, thymus, kidneys, thyroid, liver, tracheobronchial lymph nodes, lung, nasal turbinates, thigh muscle, urinary bladder, sciatic nerve, and any gross lesions. Only the lungs and tracheobronchial lymph nodes of the untreated controls were processed. 10 males of group 1, 2 and 5 (untreated, sham-exposed, and high-dose) were evaluated for morphology, number of sperm and number of testicular spermatids.

Statistics:

ANOVA and Tukey´s multiple range test: body weighs, male reproductive endpoints, haematology, and serum chemistry
ANOVA and Duncan´s multiple range test: organ weights, pulmonary function, and pulmonary hydroxyproline

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

increased total weight of the lung lobes (high-dose)

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

mild macrophage accumulation in the lung (high-dose). Non adverse.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

mild macrophage accumulation in the lung (high-dose): Non adverse.

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No treatment-related clinical signs were and no mortalities observed.

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

HAEMATOLOGY
No treatment-related changes were observed.

CLINICAL CHEMISTRY
No treatment-related changes were observed.

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

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

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

OTHER FINDINGS
- Sperm morphology: No treatment-related effects were noted in sperm morophology or in sperm and spermatid counts.
- Lung function: There were no significant differences between any groups for any of the pulmonary function parameters. The only parameter affected by exposure was the total weight of the five lung lobes. Weight for the high-dose group was significantly greater than the other groups.

Dose descriptor:

NOAEC

Effect level:

0.5 mg/L air (analytical)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 2) from a reference substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to the endpoint discussion for further details).
The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

(limited parameters examined, no daily observation)

GLP compliance:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Taconic, Germantown, NY, USA
- Weight at study initiation: males: 379-388 g ; females: 234-239 g
- Housing: animals were housed in the exposure chambers (feed and water was removed during exposure)

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

whole body

Vehicle:

air

Remarks on MMAD:

MMAD / GSD: 1.0 µm/ approx. 1.8

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 1000 L stainless steel and glass exposure chambers; chambers contained catch pans between each of three leveles of cages.
- System of generating particulates/aerosols: The test material was aerosolized directly from the liquid by a modified Lakin nebulizer on each chamber. The test material was in a straight-walled glass flask and the barrels of the nebulizer were immersed under the level of the liquid in order to maximize the amount of material generated. The distance from the nebulizer to the walls of the flask was approx. 3 cm. After exiting the flask, the aerosol passed through a glass impactor to remove most of the larger particles. The remaining aerosol was mixed with the main air stream for each chamber before entering the chamber.
- Temperature and humidity in air chamber (by a Taylor wet/dry bulb hydrometer approx. every 30 min during each exposure): approx. 23 °C, 56 - 64%
- Air flow rate: approx. 300 L/min (mean chamber flow per group: 297, 308, 342, and 243 L/min, respectively)

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric sampling on glass fiber filters (3 times during each exposure); some filters were additionally analyzed by GC/MS
- Samples taken from breathing zone: yes
Nominal concentrations were determined as the loss of weight of fluid from the generator divided by total air flow through the chamber.

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

6 hours/day
5 days/week

Remarks:

Doses / Concentrations:
0.00 ± 0.00, 0.05 ± 0.01, 0.17 ± 0.01, and 0.56 ± 0.02 mg/L
Basis:
analytical conc.

Remarks:

Doses / Concentrations:
0.05, 0.15, and 0.5 mg/L
Basis:
nominal conc.

No. of animals per sex per dose:

15
(Additional 10 male rats per group were included for examination of pulmonary function tests and analysis of pulmonary hydroxyproline following exposure.)

Control animals:

yes, concurrent no treatment

yes, sham-exposed

Details on study design:

- Dose selection rationale: The highest dose was expected to result in abnormal accumulation of test material in the lung and possible impairment of normal clearance mechanisms. The low dose is a factor of 10 above the TLV (treshold limit value) for mineral oil mistes, no untoward effects were expected at this level.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (except weekends)

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

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

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

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

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

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

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
Lung function: The animals were anaesthetized and pulmonray function tests were performed (deflation quasistatic pressure-volume cureved, functional residual capactiy, and maximal forced exhalation maneuver). After the pulmonray function tests, the lungs were removed and all lobes were weighed. Lobes were frozen for analysis of hydroxyproline content and analysis of test material remaining in the lung.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes; organ weights (adrenals, kidney, spleen, brain, liver, testes, epididymides, ovaries, thymus, heart, prostate, uterus, right middle lung lobe
HISTOPATHOLOGY: Yes (untreated and high-dose): adrenals, ovaries, sternum, pancreas, brain, salivary gland, eye, spleen heart, stomach, colon, testes, duodenum, thymus, kidneys, thyroid, liver, tracheobronchial lymph nodes, lung, nasal turbinates, thigh muscle, urinary bladder, sciatic nerve, and any gross lesions. Only the lungs and tracheobronchial lymph nodes of the untreated controls were processed. 10 males of group 1, 2 and 5 (untreated, sham-exposed, and high-dose) were evaluated for morphology, number of sperm and number of testicular spermatids.

Statistics:

ANOVA and Tukey´s multiple range test: body weighs, male reproductive endpoints, haematology, and serum chemistry
ANOVA and Duncan´s multiple range test: organ weights, pulmonary function, and pulmonary hydroxyproline

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

increased total weight of the lung lobes (high-dose)

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

mild macrophage accumulation in the lung (high-dose). Non adverse.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

mild macrophage accumulation in the lung (high-dose): Non adverse.

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No treatment-related clinical signs were and no mortalities observed.

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

HAEMATOLOGY
No treatment-related changes were observed.

CLINICAL CHEMISTRY
No treatment-related changes were observed.

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

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

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

OTHER FINDINGS
- Sperm morphology: No treatment-related effects were noted in sperm morophology or in sperm and spermatid counts.
- Lung function: There were no significant differences between any groups for any of the pulmonary function parameters. The only parameter affected by exposure was the total weight of the five lung lobes. Weight for the high-dose group was significantly greater than the other groups.

Dose descriptor:

NOAEC

Effect level:

0.5 mg/L air (analytical)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 2) from a reference substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to the endpoint discussion for further details).
The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

09 Jul - 10 Oct 1986

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

(no data on test substance purity, only 2 dose groups, open application, limited parameters examined)

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Lakeview, NJ, USA
- Age at study initiation: approx. 7 weeks
- Housing: individually in hanging, stainless steel cages with wire bottoms and fronts
- Diet: Purina Certified Lab Chow ' 5002 in pellet form; ad libitum
- Water: tap water; ad libitum
- Acclimation period: at least 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22
- Humidity (%): 40-60
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

open

Vehicle:

unchanged (no vehicle)

Details on exposure:

TEST SITE
- Area of exposure: no data
- Type of wrap if used: no wrap used, open
- Time intervals for shavings or clipplings: 24 h before the first treatment; at least weekly afterwards
- Application site: back (shaved)

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

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

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes (collars), removal during the weekend

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

5 days per week (65 exposures), 24 hours/day, removal of substance on saturdays (once a week)

Remarks:

Doses / Concentrations:
800 and 2000 mg/kg bw/day
Basis:
nominal per unit body weight

No. of animals per sex per dose:

10
(5 additional animals of the control and the high dose group were included for dermal bioavailability experiments only.)

Control animals:

yes, concurrent no treatment

Details on study design:

The test substance was dispensed by volume from a syringe and left uncovered on the shaved skin. The rats were fitted with cardboard Elizabethan collars to minimze ingestion of the test material.
The controls were treated in the same manner except that no material was applied to their skin.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations included: appearance, behaviour, secretory function and discharges

DETAILED CLINICAL OBSERVATIONS: No

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

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

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

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

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

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

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

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

NEUROBEHAVIOURAL EXAMINATION: No

OTHER: sperm morphology: at termination
- Parameters: percentage normal sperm, abnormal heads, headless, tailless, and curled tail

Sacrifice and pathology:

GROSS PATHOLOGY: Yes; organ weights: kidneys, liver; only males: brain, spleen; only females: thyroids
HISTOPATHOLOGY: Yes (no further information available)

Statistics:

The level of statistical significance was P < 0.05.

Clinical signs:

no effects observed

Dermal irritation:

effects observed, treatment-related

Description (incidence and severity):

slight erythemal and flaking; slight epidermal hyperplasia and chronic inflammation (both treatment groups)

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

reduced body weight gain in males (800 mg/kg: 7% and 2000 mg/kg: 10%)

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No treatment-related effects were observed.

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

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

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

URINALYSIS
No additional data given on Urinalysis in study report.

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

GROSS PATHOLOGY
No abnormalities were detected.

HISTOPATHOLOGY:
No abnormalities were detected.

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

SKIN PENETRATION
Skin penetration values of 2 - 6% were obtained.
The results of the in vivo skin penetration study indicate that the 13-week treatment with the test substance does not increase the skin penetration of the test substance (only the value for females was statistically different from the penetration in untreated animals). The skin penetration of untreated rats was less than 2% and the mean value for treated rats was approx. 6%. The recovery of radioactivity was measuered in the urine and faeces as well as the remaining radioactivity in tissue samples.

Dose descriptor:

NOAEL

Effect level:

>= 2 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: highest dose tested

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 2) from a reference substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to the endpoint discussion for further details).
The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

09 Jul - 10 Oct 1986

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

(no data on test substance purity, only 2 dose groups, open application, limited parameters examined)

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Lakeview, NJ, USA
- Age at study initiation: approx. 7 weeks
- Housing: individually in hanging, stainless steel cages with wire bottoms and fronts
- Diet: Purina Certified Lab Chow ' 5002 in pellet form; ad libitum
- Water: tap water; ad libitum
- Acclimation period: at least 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22
- Humidity (%): 40-60
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

open

Vehicle:

unchanged (no vehicle)

Details on exposure:

TEST SITE
- Area of exposure: no data
- Type of wrap if used: no wrap used, open
- Time intervals for shavings or clipplings: 24 h before the first treatment; at least weekly afterwards
- Application site: back (shaved)

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

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

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes (collars), removal during the weekend

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

5 days per week (65 exposures), 24 hours/day, removal of substance on saturdays (once a week)

Remarks:

Doses / Concentrations:
800 and 2000 mg/kg bw/day
Basis:
nominal per unit body weight

No. of animals per sex per dose:

10
(5 additional animals of the control and the high dose group were included for dermal bioavailability experiments only.)

Control animals:

yes, concurrent no treatment

Details on study design:

The test substance was dispensed by volume from a syringe and left uncovered on the shaved skin. The rats were fitted with cardboard Elizabethan collars to minimze ingestion of the test material.
The controls were treated in the same manner except that no material was applied to their skin.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations included: appearance, behaviour, secretory function and discharges

DETAILED CLINICAL OBSERVATIONS: No

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

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

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

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

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

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

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

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

NEUROBEHAVIOURAL EXAMINATION: No

OTHER: sperm morphology: at termination
- Parameters: percentage normal sperm, abnormal heads, headless, tailless, and curled tail

Sacrifice and pathology:

GROSS PATHOLOGY: Yes; organ weights: kidneys, liver; only males: brain, spleen; only females: thyroids
HISTOPATHOLOGY: Yes (no further information available)

Statistics:

The level of statistical significance was P < 0.05.

Clinical signs:

no effects observed

Dermal irritation:

effects observed, treatment-related

Description (incidence and severity):

slight erythemal and flaking; slight epidermal hyperplasia and chronic inflammation (both treatment groups)

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

reduced body weight gain in males (800 mg/kg: 7% and 2000 mg/kg: 10%)

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No treatment-related effects were observed.

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

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

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

URINALYSIS
No additional data given on Urinalysis in study report.

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

GROSS PATHOLOGY
No abnormalities were detected.

HISTOPATHOLOGY:
No abnormalities were detected.

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

SKIN PENETRATION
Skin penetration values of 2 - 6% were obtained.
The results of the in vivo skin penetration study indicate that the 13-week treatment with the test substance does not increase the skin penetration of the test substance (only the value for females was statistically different from the penetration in untreated animals). The skin penetration of untreated rats was less than 2% and the mean value for treated rats was approx. 6%. The recovery of radioactivity was measuered in the urine and faeces as well as the remaining radioactivity in tissue samples.

Dose descriptor:

NOAEL

Effect level:

>= 2 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: highest dose tested

Critical effects observed:

not specified

Endpoint conclusion

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 2) from a reference substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to the endpoint discussion for further details).
The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Additional information

Justification for analogue read-across

There are no data on the repeated dose toxicity of Pentaerythritol, mixed esters with linear and branched fatty acids. The assessment was therefore based on studies conducted with analogue substances as part of a read across approach, which is in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. 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 analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

Repeated dose toxicity, oral, subacute

CAS 68424-31-7

A 28-day study was conducted with Fatty acids, C5-10, esters with pentraerythritol according to OECD guideline 407 and under GLP conditions (Brammer, 1993). The test substance was administered in concentrations of 1000 ppm, 5000 ppm and 12500 ppm (corresponding to 112, 562 and 1450 mg/kg bw/day for male and 119, 586 and 1613 mg/kg bw/day for female rats) to 5 Alpk:APfSD rats/sex/dose for 28 consecutive days. Control animals received the plain diet. There was no mortality and no toxicologically relevant clinical signs were observed during the study period. No significant differences in body weight, body weight gain and food consumption between the control group and treatment groups were noted. 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. These effect were considered incidental as they were only observed in one sex and not dose-related. There were minor reductions in plasma cholesterol, triglyceride and total protein levels and plasma alanine transferase activities in males at 12500 ppm compared to control males. As these effects were not dose-related and only observed in one sex they are not considered to be toxicologically relevant. The relative kidney weights significantly increased in males at 5000 and 12500 ppm, which is considered to be related to the histopathological changes observed. The relative liver weights were significantly increased in both sexes at 12500 ppm and in males at 5000 ppm. This is considered to be an adaptive response to the high doses of fatty ester. 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 there were only isolated observations, these were considered to be incidental. 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. 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 of increased tubular hyaline droplet formation and tubular basophilia in all animals, and granular cast formation in 4/5 of the 5000 ppm animals and 5/5 of the 12500 ppm animals. In the 1000 ppm group, increased renal hyaline droplet formation and/or tubular basophilia were seen, but not granular cast formation. Hyaline droplet formation (the main constituent of which is alpha-2µ-globulin) in the kidney of male rats is widely accepted to be species- and sex specific, and as such is considered to have no relevance to man. In the liver, there was minimal hepatocyte hypertrophy in 4/5 male rats in the 12500 ppm group, which is considered to be an adaptive response to the high intake of fatty ester. Based on the results of the study, the NOAEL was considered to be≥12500 ppm (equivalent to 1450 mg/kg bw/day for male rats and 1613 mg/kg bw/day for female rats).

 

CAS 7299-99-2

A combined repeated dose toxicity and reproduction/developmental toxicity screening study (according to OECD guideline 422 and in compliance with GLP) was performed with Hexanoic acid, 2-ethyl-, 2,2-bis [ [(2-ethyl-1-oxohexyl)oxy] methyl] -1,3-propanediyl ester (Ohta, 2005). Rats were administered 0, 100, 300 and 1000 mg/kg bw/day of the test substance once daily for 42 days (males) and up to 54 days (females) via gavage. All groups had 12 females, while there were 7 males in the control, low-dose and high-dose groups, and 12 males in the mid-dose group. The application started two weeks before mating on test day one and ended on the day of or one day before sacrifice. Day of sacrifice was on test day 42 for the male rats and on lactation day 4 or shortly thereafter for the female rats. A satellite group of 5 rats/sex/dose with a 14-day recovery period was included for the control and high-dose group. There was no mortality during the study period. No toxicologically relevant clinical signs were observed. The body weight and body weight gain was comparable between the control and treatment groups. The food consumption in high-dose satellite females was significantly increased, but is not considered to be treatment-related as no effect on body weight was noted. At the end of recovery period, a significant increase in 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, as no corresponding biochemical or histopathological changes were observed. The statistically significant differences in haematological parameters between control and treated animals (increase in hematocrit and hemoglobin levels in females of the 100 mg/kg group, increased prothrombin time in females of the 300 mg/kg group) were of low magnitude and/or not dose-related, and therefore considered incidental. The concentration of blood urea nitrogen was increased in satellite females of the 1000 mg/kg group at the end of recovery period. As it was only observed in one sex, and due to the absence of relevant histopathological findings, this change is not considered to be toxicologically relevant. There was no significant difference between control and treatment groups during the observational and neurological screenings. The macroscopic inspection at autopsy and subsequent histopathological examination did not show any toxicologically relevant changes. The NOAEL for systemic toxicity was considered to be≥1000 mg/kg bw/day.

 

CAS 126-57-8

A combined repeated dose toxicity and reproduction/developmental toxicity screening study (according to OECD guideline 422 and in compliance with GLP) 2-ethyl-2-[[(1-oxononyl)oxy]methyl]propane-1,3-diyl dinonan-1-oate (CAS RN 126-57-8) (Salvador, 2015). Both male and female rats were treated for approximately 5 weeks. Three different doses were tested (100, 300 and 1000 mg/kg bw/day) and compared to a control group that received only the vehicle (corn oil). No differences in body weights and food consumption were observed in treated animals compared to the control group. No clinical signs were observed during the study. No adverse findings were recorded in clinical pathology investigations (haematology, clinical chemistry and urine analysis) apart for lymphocytosis in mid- and high dose groups in males. No relevant differences were recorded in the absolute and relative organ weights of treated animals. No treatment-related changes were noted at macroscopic and microscopic observations. Based on the results the NOAEL (No Observed Adverse Effect Level) for general toxicity was considered to be the highest dose of 1000 mg/kg bw/day.

 

Repeated dose toxicity, inhalation, subchronic

CAS 67762-53-2

A 90-day subchronic inhalation toxicity study was performed with Fatty acids, C5-9, tetraesters with pentaerythritol, following a protocol similar to OECD guideline 413 (Dulbey, 1992). 15 rats/sex/dose were exposed whole-body to the test substance aerosol for 6 hours/day, 5 days/week at concentrations of 0.05, 0.15 and 0.5 mg/L (0.05, 0.16 and 0.56 mg/L nominal concentration). The sham-control group (15 animals/sex) was treated with clean air under the same conditions, while a second control group remained untreated. 10 additional male animals were included in every group for pulmonary function tests (deflation quasistatic pressure-volume cureved, functional residual capactiy, and maximal forced exhalation manoeuvre) and histopathological examination (analysis of hydroxyproline content and analysis of test material remaining in the lung). There was no mortality during the study period. No toxicologically relevant clinical signs were observed. The body weight of treated animals was increased significantly compared with the control group. As the difference was less than 10% this is considered to be an incidental occurrence. The body weight and body weight gain of females was comparable between the control and treatment groups. No substance-related adverse effects were observed clinical biochemistry and haematological parameters. The lungs of the high-dose animals had a minimal increase in weight which correlated with slightly increased numbers of macrophages in the pulmonary alveoli. 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 alveoli (in controls less than 0.1% would be expected). However, as the changes observed in the lungs were minor, they were not considered to be toxicologically relevant. No treatment-related effects were noted in sperm morphology or in sperm and spermatid counts. There were no significant differences between any groups for any of the pulmonary function parameters. The total weight of the five lung lobes of the high-dose group was significantly higher than for the sham-exposed controls and other exposed groups, but not greater than the untreated controls. This effect is not seen as toxicologically relevant. Therefore, the NOAEC was considered to be≥0.56 mg/L (nominal concentration).

 

Repeated dose toxicity, dermal, subchronic

CAS 67762-53-2

A 90-day dermal toxicity study with Fatty acids, C5-9, tetraesters with pentaerythritol was performed following a protocol similar to OECD guideline 411 (Cruzan, 1988). 10 Sprague-Dawley rats/sex/dose were exposed to 800 and 2000 mg/kg bw/day of the test substance via open application for 90 days (5 days/week). The application was continuous (24 hours/day), and the application site was washed clean once per week. The animals wore collars to reduce the possibility of ingestion through grooming. 5 additional rats/sex were included in the control and high-dose group to assess the dermal bioavailability of the test substance. The penetration rate was measured as recovery of radioactivity in the urine and faeces as well as the remaining radioactivity in tissue samples. There was no mortality during the study period. No toxicologically relevant clinical signs were observed. The body weight gain was 7% lower in the low-dose group, and 10% lower in the high-dose group, compared with the control males. As the difference was relatively low, the decrease in body weight gain was not considered to be toxicologically relevant. The body weight and body weight gain of females was comparable between the control and treatment groups. The aspartate aminotransferase level in high-dose females was significantly increased. The increased activity of hepatic enzymes indicates an adaptive increase in hepatic metabolism caused by the treatment, but is not considered to be toxicologically relevant as no other effects were noted in the liver. Other changes observed in clinical chemistry parameters in the low-dose group only or in one sex only are considered to be incidental. No toxicologically relevant changes were seen on organ weights. No effects on sperm morphology were observed. Both treated groups showed minimal erythema and flaking of the skin at the application site. Microscopic examination showed very minor epidermal hyperplasia and chronic inflammation of the superficial dermis. The macroscopic inspection at autopsy and subsequent histopathological examination of remaining organs and tissues did not reveal any treatment-related changes.

 

The results of the in vivo skin penetration study performed as part of the repeated dose toxicity study indicate that the 90-day treatment did not increase the skin penetration of the test substance significantly overall, as 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 rats treated for 90 days was approximately 6%. As no effects of systemic toxicity were identified up to the highest dose tested, the 90 day dermal NOAEL was found to be≥2000 mg/kg bw/day for rats.

 

Overall conclusion for repeated dose toxicity

The data for the source substances showed that no effects were observed up to and including the recommended limit values in studies conducted via the oral, inhalation and dermal routes. Therefore, as the available data did not identify any hazard for repeated dose toxicity, Pentaerythritol, mixed esters with linear and branched fatty acids is not expected to be hazardous following repeated exposure.

 

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:

Hazard assessment is conducted by means of read-across from a structural analogue. The selected study is the most adequate and reliable study based on the identified similarities in structure and intrinsic properties between the source and target substance and overall assessment of quality, duration and dose descriptor level (refer to the endpoint discussion for further details).

 

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:

Hazard assessment is conducted by means of read-across from a structural analogue. The selected study is the most adequate and reliable study based on the identified similarities in structure and intrinsic properties between the source and target substance and overall assessment of quality, duration and dose descriptor level (refer to the endpoint discussion for further details).

 

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:

Hazard assessment is conducted by means of read-across from a structural analogue. The selected study is the most adequate and reliable study based on the identified similarities in structure and intrinsic properties between the source and target substance and overall assessment of quality, duration and dose descriptor level (refer to the endpoint discussion for further details).

 

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:

Hazard assessment is conducted by means of read-across from a structural analogue. The selected study is the most adequate and reliable study based on the identified similarities in structure and intrinsic properties between the source and target substance and overall assessment of quality, duration and dose descriptor level (refer to the endpoint discussion for further details).

 

Justification for selection of repeated dose toxicity dermal - local effects endpoint:

Hazard assessment is conducted by means of read-across from a structural analogue. The selected study is the most adequate and reliable study based on the identified similarities in structure and intrinsic properties between the source and target substance and overall assessment of quality, duration and dose descriptor level (refer to the endpoint discussion for further details).

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 "General Requirements for Generation of Information on Intrinsic Properties of substances", information on intrinsic properties of substances may be generated by means other than tests e.g. from information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI, "General rules for adaptation of this standard testing regime set out in Annexes VII to X” 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 analogue concept is applied to Pentaerythritol, mixed esters with linear and branched fatty acids, data will be generated from data for reference source substance(s) to avoid unnecessary animal testing. Additionally, once the analogue read-across concept is applied, substances will be classified and labelled on this basis.

Therefore, based on the analogue read-across approach, the available data on repeated dose toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification