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Description of key information

Oral (OECD 407, GLP, rat): NOAEL ≥ 1000 mg/kg bw/day

Read-across from structural analogue source substances Dipentaerythritol hexaesters with fatty acids, C5 and C9iso (CAS No. 647028-25-9), Pentaerythritol tetraesters of n-decanoic, n-heptanoic, n-octanoic and n-valeric acids (CAS No. 68424-31-7), and Fatty acids, C8-10 mixed esters with dipenaterythritol, isooctanoic acid, pentaerythritol and tripentaerythritol (CAS No. 189200-42-8)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Guideline study with acceptable restrictions (Lack of data on test substance. Not all required organs were weighed and examined during necropsy (only liver, kidneys, adrenals, brain and testes/ovaries) and histopathology (only liver, kidneys, adrenals, heart, spleen, lung, testes/ovaries and gross lesions))
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
(not all required organs were examined during necropsy and histopathology, positive control was included)
GLP compliance:
yes
Species:
rat
Strain:
other: Crl:CD BR
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Stone Ridge, NY, USA
- Age at study initiation: approximately 7 weeks
- Weight at study initiation: 216.6 - 251.2 g (males); 161.9 - 187.8 g (females)
- Assigned to test groups randomly: yes, by a computer-generated body weight sorting program
- Housing: the animals were housed individually in suspended stainless steel and wire mesh cages with absorbent paper below the cages.
- Diet: certified Rodent Diet # 5002 (pellets) (PMI Feeds Inc.), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 13 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 40 - 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
From: 28 Sep 1994
To: 27 Oct 1994
Route of administration:
oral: gavage
Vehicle:
other: polyethylene glycol (PEG 400)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test substance was diluted in the vehicle to a concentration of 2% (w/v).

VEHICLE
- Justification for use and choice of vehicle (if other than water): the test substance was soluble in PEG 400 at the concentrations required for this study.
- Concentration in vehicle: 2% (w/v)
- Amount of vehicle (if gavage): 5 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability, uniformity and concentration of the test substance and the positive control substance in their vehicles were analysed. The analysis for the stability and uniformity were initiated prior to or concomitant with the initiation of dosing. Concentration analysis were performed for the mixtures of weeks 1 and 4.
A solution of 2% (w/v) test substance in PEG 400 was stable for at least 8 d at room temperature. The relative standard deviation of a solution of 2% of the test substance in PEG 400 was 0.732. The analysis of the concentration indicated that all solutions were within 9% of the nominal concentrations for weeks 1 and 4.
Duration of treatment / exposure:
28 d
Frequency of treatment:
once daily, 7 d/week
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
500 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Positive control:
acrylamide
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily monday through friday, once daily on weekends and holidays

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: on the day of initiation of dosing (Day 0), on Days 7, 14, 21, 27 and on the day of sacrifice (Day 28)

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

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Day 28
- Anaesthetic used for blood collection: es, methoxyflurane
- Animals fasted: Yes
- How many animals: all surviving animals
- Parameters observed: hematocrit, hemoglobin, erythrocyte count, leukocyte count (total and differential), platelet count, reticulocyte count, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration; to estimate the clotting potential prothrombin time and thromboplastin time were observed

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Day 28
- Animals fasted: Yes
- How many animals: all surviving animals
- Parameters observed: albumin, urea nitrogen, calcium, creatinine, electrolytes (Na+, CI-, K+), glucose, phosphorus, gamma glutamyl transpeptidase, serum alanine aminotransferase, serum aspartate aminotransferase, serum alkaline phosphatase, total protein, total bilirubin, cholesterol and triglycerides

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: prior to initiation of dosing, on Day 8 and on the last day of dosing (both at least 1 h after dosing)
- Dose groups that were examined: all surviving animals
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, including organ weights of liver, kidneys, adrenals, brain and testes/ovaries
HISTOPATHOLOGY: Yes, histopathological examination of liver, kidneys, adrenals, spleen, heart, lungs, testes/ovaries and gross lesions from the animals of the vehicle and positive control groups and the high dose group; histopathological examination of liver, kidneys, lungs and gross lesions from the animals of the low and mid dose groups.
Statistics:
The statistical evaluation of equality of means was done by an appropriate one way analysis of variance (ANOVA) and a test for ordered response in the dose groups. To determine if the dose groups have equal variance, Bartlett's Test was performed. If the variances were equal, the testing was done using parametric methods, otherwise nonparametric techniques were used.
For the parametric procedures, a standard one way ANOVA using the F distribution to assess significance was used. If significant differences among the means were indicated, Dunnett's Test was used to determine which treatment groups differed significantly from control. In addition to the ANOVA, a standard regression analysis for linear response in the dose groups was performed. The regression also tested for linear lack of fit in the model.
For the nonparametric procedures the test of equality of means was performed using the Kruskal-Wallis Test. If significant differences among the means were indicated, Dunn's Summed Rank Test was used to determine which treatment groups differed significantly from the control. In addition to the Kruskal-Wallis Test, Jonckheere's Test for monotonic trend in the dose response was performed.
Bartlett's Test for equal variance was conducted at the 1% level of significance. All other tests were conducted at the 5% and 1% level of significance.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
but no treatment-related effects
Mortality:
mortality observed, treatment-related
Description (incidence):
but no treatment-related effects
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
but no dose-response pattern
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
but no dose-response pattern
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
but no dose-response pattern
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
but no dose-response pattern
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
but no treatment-related effects
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
but no treatment-related effects
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
One male animal of the 500 mg/kg bw group died after the first application due to a gavage error. This animal was replaced on Day 1 with another animal, that received one less dose than the other animals. All animals survived the study period to the scheduled termination.
No clinical signs occurred that were judged to be related to the treatment with the test substance. In one or more groups including the controls, there were single or very low incidences of scabs (1/5 males in the control and in the mid dose group), sores (1/5 males in the control and in the mid dose group), dental abnormalities (1/5 males in the mid and high dose group), red material penis (1/5 males in the mid dose group), dry rales (1/5 females in the low and high dose group) and alopecia (1/5 males in the control group) observed. Soft stool was observed in 1/5 males and 2/5 females of the control group, 3/5 females of the low dose group, 3/5 males and 1/5 females of the mid dose group and 4/5 females of the high dose group.

BODY WEIGHT AND WEIGHT GAIN
No effects on the body weight were observed after treatment of the animals with the test substance.

FOOD CONSUMPTION AND COMPOUND INTAKE
No effects on the food consumption of the test substance treated animals were observed.

HAEMATOLOGY
There were no significant differences in the hematology parameters between the negative control and the animals received the test substance. The hemoglobin of the females of the low dose group was slightly decreased compared to the negative control. This effect was not considered to be clinically significant due to the absence of similar findings in other red blood cell parameters and a dose-response pattern.

CLINICAL CHEMISTRY
There were no significant differences in the clinical chemistry parameters between test substance treated and negative control animals which were judged to be test substance related. In male animals, statistically significant decreases in mean calcium, total protein and albumin in the mid dose group and an increase in alanine aminotransferase in the low dose group were observed. These effects were not considered to be clinically significant due to the absence of a clear dose-response pattern.

NEUROBEHAVIOUR
The neurobehavioural observations made on Day 8 and Day 27 were unremarkable for the animals treated with the test substance.
On Day 8, the following anomalities in neurobehaviour were observed: increased respiration in 1/5 males at 100 mg/kg bw/d and 1/5 females at 500 mg/kg bw/d, decreased toe pinch response in 1/5 females at 100 mg/kg bw/d and 2/5 males at 1000 mg/kg bw/d, increased toe pinch response in 1/5 females of the negative control and abnormal air righting in 1/5 males at 1000 mg/kg bw/d.
On Day 27, there was an increased incidence of decreased arousal in 3/5 males and 4/5 females of the negative control group, 5/5 males and 3/5 females at 100 mg/kg bw/d, 2/5 males and 1/5 females at 500 mg/kg bw/d and 4/5 males and 1/5 females at 1000 mg/kg bw/d. Irregular breathing was observed in 4/5 males and 2/5 females of the negative control group, 3/5 males and 1/5 females at 100 mg/kg bw/d, 2/5 males and 1/5 females at 500 mg/kg bw/d and 1/5 males and 1/5 females at 1000 mg/kg bw/d. Additionally, there occurred increased muscle tone in one male of the negative control group, slightly impaired gait in 1/5 males of the negative control group, decreased startle response in 1/5 males of the negative control group, increased arousal in 1/5 males at 1000 mg/kg bw/d and decreased toe pinch response in 1/5 males of the negative control group and 1/5 females at 1000 mg/kg bw/d.
Due to the absence of a dose-response pattern, these effects were judged to be incidental and not related to treatment.

ORGAN WEIGHTS
There were no significant differences in mean absolute organ weight between the test substance treated and negative control animals. In the mid dose group, there was a statistically significant increase in the mean relative testes weight. Due to the absence of a dose-response pattern, this effect was judged to be incidental.

GROSS PATHOLOGY
The necropsy revealed no treatment-related findings. Single incidences of dilated renal pelvis (1/5 males at 100 mg/kg bw/d), scabs (1/5 males at 500 mg/kg bw/d), thickened/roughened stomach (1/5 females at 500 mg/kg bw/d) and distended uterus (1/5 females at 500 mg/kg bw/d) were observed. These findings were considered to be incidental and not treatment-related.

HISTOPATHOLOGY: NON-NEOPLASTIC
No treatment-related histopathologic changes were observed in any of the organs or tissues examined. The most common spontaneously occurring incidental findings were focal mononuclear or mixed inflammatory cell infiltrations in the liver (5/5 males and 5/5 females of the negative control, 5/5 males and 1/5 females at 100 mg/kg bw/d, 3/5 males and 5/5 females at 500 mg/kg bw/d, 2/5 males and 4/5 females at 1000 mg/kg bw/d) and the heart (2/5 males and 0/5 females of the negative control and 2/5 males and 0/5 females at 1000 mg/kg bw/d). Furthermore, focal tubular degeneration in the kidneys of male rats of all groups (4/5 of the negative control, 5/5 at 100 mg/kg bw/d, 2/5 at 500 mg/kg bw/d, 4/5 males at 1000 mg/kg bw/d) was observed. This effect was not observed in any dose group in the kidneys of the female rats.
The changes observed were considered to have been spontaneous in origin and were typical of incidental findings commonly encountered in laboratory rats of this age and strain. Because these changes occurred at similar incidence and intensity among all groups, they were not considered to be related to treatment.

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: NOAEL corresponds to the highest dose tested
Critical effects observed:
no
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
19 Feb 1998 - 30 March 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Version / remarks:
adopted in 1995
Qualifier:
according to
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
GLP compliance:
yes (incl. certificate)
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 and environmental conditions:
TEST ANIMALS
- Source: Charles River Limited, Margate, Kent, UK
- Age at study initiation: 5 to 6 weeks
- Weight at study initiation: 127 – 161 g (males) and 117 – 152 g (females)
- Housing: animals were housed in groups of 5 of the same sex per cage in polypropylene grid-floor cages
- Diet: Rat and Mouse SQC Expanded Diet No.1, pelleted, ad libitum
- Water: ad libitum
- Acclimation period: 7 days

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

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

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

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

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

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

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

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

BEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: prior to the start of treatment and on days 2, 8, 15 and 23.
- Dose groups that were examined: all animals
- Parameters observed: gait, tremors, twiches, convulsions, bizarre/abnormal/stereotypic behaviour, pilo-erection, exophthalmia, lachrymation, hyper/hypothermia, skin colour, respiration, palpebral closure, urination, defecation, transfer arousal, tail elevation.

Sacrifice and pathology:
GROSS PATHOLOGY: Yes. All animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.
HISTOPATHOLOGY: Yes. Adrenals, aorta (thoracic), bone and bone marrow (femur including stifle joint), bone and bone marrow (sternum), muscle (skeletal), oesophagus, ovaries, pancreas, pituitary, brain (including cerebrum, cerebellum and pons), caecum, colon duodenum, epididymides, eyes, gross lesions, heart, ileum, jejunum, kidneys, liver, lungs (with bronchi), lymph nodes (cervical and mesenteric), prostate, rectum, salivary glands (submaxillary), sciatic nerve, seminal vesicles, skin (hind limb), spinal cord (cervical), spleen, stomach, testes, thymus, thyroid/parathyroid, trachea, urinary bladder, uterus.
Other examinations:
Organ weights: adrenals, kidneys, testes, brain, liver, thymus, epididymes, ovaries, heart, spleen
Statistics:
Haematological, blood chemical, organ weight (absolute and relative to terminal body weight), weekly body weight gain and quantitative functional performance and sensory reactivity data were considered for dose response relationships by linear regression analysis followed by one way analysis of variance (ANOVA) including Levene’s test for homogeneity of variance. Where variances were shown to be homogeneous pair wise comparisons were conducted by means of Dunnett’s test. Where Levene’s test showed unequal variances the data were analysed using non-parametric methods: Kruskal-Wallis ANOVA and Mann Whitney “U” test.

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

Probability values (p) are presented as follows:

p< 0.001***
p < 0.01**
p< 0.05*
p ≥0.05 (not significant)


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

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

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

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

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

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

HISTOPATHOLOGY: NON-NEOPLASTIC
Three males treated with 1000 mg/kg/day demonstrated globular accumulations of eosinophilic material in the proximal tubular epithelium which should be regarded as a possible effect of treatment. The authors considered this finding consistent with the appearance of hydrocarbon nephropathy, which results from the excessive accumulation of α2-microglobulin in renal proximal tubular epithelium of adult male rats, which does not represent a hazard to human health.
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: overall effects, NOAEL corresponding to the highest dose tested
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 (109/L)

Group 1

0 mg/kg bw

34.6 ± 0.4

792 ± 67

Group 2

150 mg/kg bw

34.3 ± 0.5

928 ± 115

Group 3

500 mg/kg bw

34.2 ± 0.3

944 ± 68*

Group 4

1000 mg/kg bw

33.7 ± 0.7*

920 ± 76

*: 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

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

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

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

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

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked: changes in clinical condition and behaviour and significant changes were recorded.

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

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

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

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

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

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

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

MORTALITY
There were no mortalities.

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

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

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

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

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

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

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

Dose descriptor:
NOAEL
Effect level:
1 613 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: no adverse effects observed
Dose descriptor:
NOAEL
Effect level:
1 450 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: no adverse effects observed
Critical effects observed:
no
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The available information comprise adequate and reliable (Klimisch score 1 and 2) studies from various source substances with similar structures and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected studies are thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Repeated dose toxicity: inhalation - systemic effects

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

Justification for grouping of substances and read-across

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

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

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

Oral repeated dose toxicity

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

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

In a short-term (28 day) repeated dose toxicity study Fatty acids, C5-10, esters with pentaerythritol (CAS No. 68424-31-7) was investigated according to OECD Guideline 407 and under GLP conditions (WoE, RA-A, 68424-31-7, 1993). The test substance was administered in the diet in concentrations of 1000 ppm, 5000 ppm and 12500 ppm (corresponding to 112, 562 and 1450 mg/kg bw/day for male and 119, 586 and 1613 mg/kg bw/day for female rats) to 5 Alpk:APfSD rats per sex and dose for 28 consecutive days. Control animals (5 per sex and dose) received the plain diet. There were no toxicologically significant effects on body weight, food consumption and clinical condition and mortality up to and including the highest dose level. Changes in clinical chemistry and red cell-related parameters were observed in male rats at 12500 ppm, but these were minor and considered not to be of toxicological significance. A minimal hepatocyte hypertrophy present in males of the 12500 ppm group was observed and considered to be evidence of an adaptive response. Microscopic examination of the kidneys from male animals from all dose groups revealed an increase in hyaline droplet formation (the main constituent of which is alpha-2µ-globulin) and tubular basophilia. This phenomenon is widely accepted to be specific to the male rat and as such is considered to have no relevance to man. A NOAEL of 1450 and 1613 mg/kg bw/day could be identified for male and female rats, respectively.

In another 28-day subacute repeated dose toxicity study performed with Fatty acids, C8-10 mixed esters with dipenaterythritol, isooctanoic acid, pentaerythritol and tripentaerythritol (CAS No. 189200-42-8) equivalent or similar to OECD Guideline 407 under GLP conditions, male and female Crl:CD BR rats were dosed with 100, 500 and 1000 mg/kg bw/day by oral gavage once daily, 7 day/week for a total of 28 days (Exxon, 1995d). 5 animals per dose and sex were used. A control group receiving the vehicle polyethylene glycol (PEG 400) only was also included in the study design. One male animal of the 500 mg/kg bw group died after the first application due to a gavage error. This animal was replaced on day 1 with another animal that received one dose less than the other animals. All animals survived the study period until scheduled termination. No clinical signs occurred that were judged to be treatment-related, no effects on body weights and on food consumption of the treated animals were observed. Moreover, there were no significant differences in the hematology parameters between the negative control and the animals treated. There were also no significant differences in the clinical chemistry parameters which were judged to be test substance related. Neuro-behavioural observations made on day 8 and day 27 were unremarkable. The necropsy revealed no treatment-related findings. Single effects were observed but were considered to be incidental and not treatment-related. No significant differences in mean absolute organ weight and no histopathologic changes were observed in any of the organs or tissues examined between the test substance treated and negative control animals. In the mid dose group, there was a statistically significant increase in the mean relative testes weight. Due to the absence of a dose-response pattern, this effect was judged to be incidental. The oral NOAEL was therefore determined to be ≥ 1000 mg/kg bw/day.

Conclusion on repeated dose toxicity

In conclusion, the available studies clearly indicate a very low level of repeated dose toxicity after oral exposure. Since no toxicologically relevant findings were detected in any of the studies and hence all NOAELs determined correspond to the highest doses tested, no hazard with respect to oral repeated dose toxicity is identified for the target substance Octadecanoic acid, 1,1'-[2-[[3-[(1-oxooctadecyl)oxy]-2,2-bis[[(1-oxooctadecyl)oxy]methyl]propoxy]methyl]-2-[[(1-oxooctadecyl)oxy]methyl]-1,3-propanediyl] ester (CAS No. 70967-57-2) and a NOAEL of ≥ 1000 mg/kg bw/day is used for risk assessment and C&L purposes of the target substance.

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 information on intrinsic properties of substances may be generated by means other than tests, e.g. using information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI states that “substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or ‘category’ of substances. This avoids the need to test every substance for every endpoint". Since the read-across concept is applied to the target substance Octadecanoic acid, 1,1'-[2-[[3-[(1-oxooctadecyl)oxy]-2,2-bis[[(1-oxooctadecyl)oxy]methyl]propoxy]methyl]-2-[[(1-oxooctadecyl)oxy]methyl]-1,3-propanediyl] ester (CAS No. 70967-57-2), data gaps can be filled by interpolation from representative structural analogue source substances to avoid unnecessary animal testing.

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