Fatty acids, C9, hexaesters with dipentaerythritol

Endpoint summary

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:

weight of evidence

Study period:

Nov - Dec 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Read across study

Justification for type of information:

Based on its composition and physico chemical properties, fatty acids, C5-10, esters with pentaerythritol is considered to be a suitable read across substance to address the repeated dose toxicity endpoint of the test substance, fatty acids, C9, hexaesters with dipentaerythritol.

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)).

Key result

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

Key result

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

Key result

Critical effects observed:

no

Conclusions:

Under the study conditions, NOAEL values of 1450 and 1613 mg/kg/day could be identified for male and female rats, respectively.

Executive summary:

A study was conducted to determine the toxicity of the read across substance, fatty acids, C5-10, esters with pentaerythritol, to rats after repeated oral administration according to OECD Guideline 407, in compliance with GLP. The test substance was administered for 28 d to groups of 5 animals per sex and dose at concentrations of 1000, 5000 and 12500 ppm, equivalent to 112, 562 and 1450 mg/kg bw/day for male and 119, 586 and 1613 mg/kg bw/day for female rats, respectively. There were no toxicologically significant effects on bodyweight, food consumption and clinical condition up to and including the highest dose level. Changes in some clinical chemistry and red blood cell-related parameters were observed in male rats at 12500 ppm but these were minor and considered to be of no toxicological significance. There were no clinical sings indicative of neurological changes in the brains of the 12500 ppm group. A minimal hepatocyte hypertrophy, present in males in the 12500 ppm group, is 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. Under the study conditions, NOAEL values of 1450 and 1613 mg/kg/day could be identified for male and female rats, respectively (Zeneca, 1993).

Reference 2

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

19 Feb 1998 - 30 March 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No ophthalmoscopic examination and no urinalysis and no data on analytical purity. Read across study.

Justification for type of information:

Based on its composition and physico chemical properties, fatty acids, C5-9, hexaesters with dipentaerythritol is considered to be a suitable read across substance to address the repeated dose toxicity endpoint of the test substance, fatty acids, C9, hexaesters with dipentaerythritol.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 1995

Deviations:

yes

Remarks:

no ophthalmoscopic examination and no urinalysis and no data on analytical purity

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Deviations:

yes

Remarks:

no ophthalmoscopic examination and no urinalysis and no data on analytical purity

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, 250 and 500 mg/mL
- Amount of vehicle (if gavage): 2 mL/kg bw

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

Remarks:

Doses / Concentrations:
150, 500 and 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: 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 1 to 5 h after dosing during the working week and immediately before dosing and 1 h 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 termination 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 the end of the study (Day 28)
- 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: Day 28
- 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 Day 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 Day 2, 8, 15 and 23
- Dose groups that were examined: all animals
- Parameters observed: gait, tremors, twitches, convulsions, bizarre/abnormal/stereotypic behaviour, pilo-erection, exophthalmia, lachrymation, hyper/hypothermia, skin colour, respiration, palpebral closure, urination, defecation, transfer arousal, tail elevation

OTHER
ORGAN WEIGHTS: adrenals, kidneys, testes, brain, liver, thymus, epididymis, ovaries, heart, spleen

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.

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 bw/day: reduction in body weight in males during Week 1 (non adverse)

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):

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

500 and 1000 mg/kg bw/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 bw/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 bw/day: statistically significant reduction in absolute epididymides weight (non adverse)

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

150 mg/kg bw/day (one male and one female) and 1000 mg/kg bw/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 bw/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 bw/day) and one treated female animal (500 mg/kg bw/day) one h 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 bw/day) showed fur loss from Day 21 onwards and one 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. A statistically significant reduction in body weight gain was apparent for males in the high- and mid dose-group during Week 1 of the study (see Table 1 under "Any other information on results incl. tables"). The dose relationship was not credible and the intergroup differences were considered to be a result of slightly higher than usual control group body weight gains.

HAEMATOLOGY
No treatment-related changes in the haematological parameters were measured.
Males treated with 1000 mg/kg bw/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 bw/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 bw/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 effects on organ weight were noted. Males treated with 500 mg/kg bw/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 bw/day and a female animal treated with 1000 mg/kg bw/day showed dark foci on the lungs. These findings showed no dose-related response and where considered to be of no toxicological importance.

HISTOPATHOLOGY: NON-NEOPLASTIC
Three males treated with 1000 mg/kg bw/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.

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:

other: overall effects, NOAEL corresponding to the highest dose tested

Key result

Critical effects observed:

no

Table 1. Group mean weekly body weight gains (males).

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

*: significant different from control group p < 0.05

**: significant different from control group p < 0.01

Table 2. Group mean haematological values (males).

Dose Level	MCHC (g/d)	PLT (10^9/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

*: significant different from control group p < 0.05

Table 3. Group mean functional performance test values.

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*

*: significant different from control group p < 0.05

**: significant different from control group p < 0.01

Table 4. Group mean absolute organ weights.

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

**: significant different from control group p < 0.01

Conclusions:

Under the study conditions, the NOAEL in rats was 1000 mg/kg bw/day, the highest dose tested.

Executive summary:

A study was conducted to determine the toxicity of the read across substance,fatty acids, C5-9, hexaesters with dipentaerythritol,to rats after repeated oral administration according to OECD Guideline 407, in compliance with GLP. Groups of 5 male and 5 female Sprague Dawley rats were administered 150, 500 and 1000 mg/kg bw/day of the test substance in arachis oil for 28 d by gavage. A concurrent negative control group receiving the vehicle arachis oil only. No clinical signs or mortality occurred in relation to the treatment in any animal. Isolated and transient observations such as noisy respiration in 2 animals and fur loss in one animal were observed without dose-relationship and therefore considered to be of no toxicological importance. No adverse effect on clinical chemistry parameters and food and water consumption was noted. There was no adverse effect on bodyweight. A reduction in bodyweight gain in the high- and mid-dose group in Week 1 was considered to be a result of slightly higher than usual control group bodyweight gains. No treatment-related changes in the haematological parameters were measured. However, a reduction in mean corpuscular haemoglobin concentration in the male high-dose group and an increase in platelet count in the male mid-dose group were apparent. In the absence of any other haematological changes, these differences were considered to be accidental. The functional performance tests showed isolated intergroup differences which were considered to be accidental and of no toxicological relevance. No treatment-related effects on organ weights were noted. Males in the mid-dose group showed a reduction in absolute epididymides weight. In the absence of a dose-response relationship, this intergroup difference was considered to be incidental and of no toxicological significance. Necropsy revealed no treatment-related findings. One male and one female animal in the low- dose group and a female in the high-dose group showed dark foci on the lungs. These findings showed no dose-related response and where considered to be of no toxicological importance. Three males of the high-dose group demonstrated globular accumulations of eosinophilic substance in the proximal tubular epithelium. The author 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. No other histopathological changes were observed including effects on epididymides, testes, uterus and ovaries in any animal. Under the study conditions, the NOAEL for male and female rats was 1000 mg/kg bw/day (Jones, 2000).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Adequate

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Study 1:

A study was conducted to determine the toxicity of the read across substance, fatty acids, C5-9, hexaesters with dipentaerythritol, to rats after repeated oral administration according to OECD Guideline 407, in compliance with GLP. Groups of 5 male and 5 female Sprague Dawley rats were administered 150, 500 and 1000 mg/kg bw/day of the test substance in arachis oil for 28 d by gavage. A concurrent negative control group receiving the vehicle arachis oil only. No clinical signs or mortality occurred in relation to the treatment in any animal. Isolated and transient observations such as noisy respiration in 2 animals and fur loss in one animal were observed without dose-relationship and therefore considered to be of no toxicological importance. No adverse effect on clinical chemistry parameters and food and water consumption was noted. There was no adverse effect on bodyweight. A reduction in bodyweight gain in the high- and mid-dose group in Week 1 was considered to be a result of slightly higher than usual control group bodyweight gains. No treatment-related changes in the haematological parameters were measured. However, a reduction in mean corpuscular haemoglobin concentration in the male high-dose group and an increase in platelet count in the male mid-dose group were apparent. In the absence of any other haematological changes, these differences were considered to be accidental. The functional performance tests showed isolated intergroup differences which were considered to be accidental and of no toxicological relevance. No treatment-related effects on organ weights were noted. Males in the mid-dose group showed a reduction in absolute epididymides weight. In the absence of a dose-response relationship, this intergroup difference was considered to be incidental and of no toxicological significance. Necropsy revealed no treatment-related findings. One male and one female animal in the low- dose group and a female in the high-dose group showed dark foci on the lungs. These findings showed no dose-related response and where considered to be of no toxicological importance. Three males of the high-dose group demonstrated globular accumulations of eosinophilic substance in the proximal tubular epithelium. The author 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. No other histopathological changes were observed including effects on epididymides, testes, uterus and ovaries in any animal. Under the study conditions, the NOAEL for male and female rats was 1000 mg/kg bw/day (Jones, 2000).

 

Study 2:

A study was conducted to determine the toxicity of the read across substance, fatty acids, C5-10, esters with pentaerythritol, to rats after repeated oral administration according to OECD Guideline 407, in compliance with GLP. The test substance was administered for 28 d to groups of 5 animals per sex and dose at concentrations of 1000, 5000 and 12500 ppm, equivalent to 112, 562 and 1450 mg/kg bw/day for male and 119, 586 and 1613 mg/kg bw/day for female rats, respectively. There were no toxicologically significant effects on bodyweight, food consumption and clinical condition up to and including the highest dose level. Changes in some clinical chemistry and red blood cell-related parameters were observed in male rats at 12500 ppm but these were minor and considered to be of no toxicological significance. There were no clinical sings indicative of neurological changes in the brains of the 12500 ppm group. A minimal hepatocyte hypertrophy, present in males in the 12500 ppm group, is 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. Under the study conditions, NOAEL values of 1450 and 1613 mg/kg/day could be identified for male and female rats, respectively (Zeneca, 1993).

Justification for classification or non-classification

Based on the results of studies with appropriate read across substances, the test substance does not require classification for repeated dose toxicity according to EU CLP (EC 1272/2008) criteria.