Fatty acids, C8-10, oxybis(2-hydroxy-3,1-propanediyl) esters

Endpoint summary

Description of key information

Combined repeated dose and reproduction / developmental screening (OECD 422): NOAEL = 1000 mg/kg bw/day

RA from source substance (CAS 63705-03-3)

Subacute oral repeated dose toxicity (OECD 401): NOAEL = 1800 mg/kg bw/day

RA from source substance (CAS 130905-60-1)

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:

experimental study

Adequacy of study:

weight of evidence

Study period:

12 Jul - 9 Aug 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 1982

Deviations:

yes

Remarks:

no analytical purity reported

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Wistar Albino rats of the Crl: Wi/Br strain

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH (Laboratory Animal Breeding), Sulzfeld, Germany
- Age at study initiation: about 6 weeks
- Weight at study initiation: 121-152 g (males); 123-143 g (females)
- Housing: individual in Makrolon type II cages, bedding was produced from pure soft wood, dried, freed from dust and sterilized at 180 °C.
- Diet: “Ssniff R” diet in pellet form (laboratory standard rat diet), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.5 ± 1.5
- Humidity (%): 60 ±10
- Air changes (per hr): 16 times per hour
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The test material was diluted with corn oil to give the final concentrations. For this purpose, the test material was heated to about 70°C to liquefy the test article and appropriate amounts were weighed and mixed with corn oil in volume/volume proportions.

VEHICLE
- Concentration in vehicle: 2.0, 20.0 and 50%
- Amount of vehicle (if gavage): 10 mL

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability of the test material was determined in the vehicle prior to the initiation of the study. In fact a 20% suspension of the test article was analysed at preparation and 2, 4 and 24 hours thereafter.
Analytical results indicated that the dosing suspensions were stable for at least 24 hours.
These analyses provided information on the homogeneity of the test item in suspension as well.
The concentration and identity of the dosing solutions were determined at the start and at the end of the study. All values obtained were in reasonable accordance with the nominal values.

Duration of treatment / exposure:

4 weeks

Frequency of treatment:

once daily, 7 days/week

Dose / conc.:

0.2 other: mL/kg bw/day

Remarks:

actual ingested

Dose / conc.:

2 other: mL/kg bw/day

Remarks:

actual ingested

Dose / conc.:

5 other: mL/kg bw/day

Remarks:

actual ingested

Dose / conc.:

180 mg/kg bw/day (actual dose received)

Dose / conc.:

1 800 mg/kg bw/day (actual dose received)

Dose / conc.:

4 500 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 used in the present study were chosen on the basis of results obtained during a 7-day range finding study. A dosage of 5 mL/kg bw was used and did not cause overt signs of toxicity. On the basis of these results, 5 mL/kg bw was selected as the highest dose in the present study with the purpose to get information on target organ toxicity. The low and mid doses were chosen in order to obtain a clear no adverse effect level.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Clinical observations were conducted daily. Mortality checks were performed twice daily.
- Cage side observations checked in table [animals´ sensory and motor behaviour, coat, body orifices, urine, feces and general health status] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Special clinical examinations were conducted prior to study initiation (week 0) and thereafter on week 2 and 4.

BODY WEIGHT: Yes
- Time schedule for examinations: Measurements of body weight were made at weekly intervals.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Time schedule for examinations: at weekly intervals
- Food conversion ratio was calculated from the change in weight and the food consumption over the same periods of time: Yes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: cornea, episcleral vessels, anterior chamber, pupil, lens and, if possible, retina were examined with a slit lamp and an ophthalmoscope fitted with a selection of lenses prior to study initiation (week 0) and thereafter on week 2 and 4.
- Dose groups that were examined: on male and female rats of each test group.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of the administration period
- Anaesthetic used for blood collection: No
- Animals fasted: No data
- How many animals: on male and female rats of each test group
- Parameters examined: erythrocytes (RBC), leukocytes (WBC), thrombocytes, haemoglobin, hematocrit, MCV, MCH and MCHC, differential blood count, reticulocytes, inclusion bodies, prothrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the administration period
- Animals fasted: No
- How many animals: on male and female rats of each test group
- Parameters examined: Glucose, triglyceride, cholesterol, protein (total), albumin, bilirubin (total), creatinine, urea nitrogen (BUN), uric acid, calcium, chloride, inorg. Phosphorus, iron, potassium, sodium, Na/K ratio by statistical evaluation, alanine aminotransferase (ALT), alkaline phosphatase, aspartate aminotransferase (AST), creatinine kinase (CK), glutamate dehydrogenase (GLDH), AST/ALT ratio by statistical evaluation

URINALYSIS: Yes
- Time schedule for collection of urine: at the end of the administration period
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- Parameters examined: color, protein, pH value, glucose, bilirubin, urobilinogen, blood, nitrite, ketones, sediment, specific gravity

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: prior to study initiation (week 0) and thereafter on week 2 and 4.
- Dose groups that were examined: on male and female rats of each test group
- Battery of functions tested: hearing activity (tested by simple noise production)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes. All animals were sacrificed at the end of the experimental phase by CO2 asphyxiation. Rapid exsanguination was carried out by cutting the A.carotis. The following organs were examined: cranial, thoracic, abdominal and pelvis cavities.

HISTOPATHOLOGY: Yes. A complete histopathological examination was performed on all male and female animals of control group and high dose group on the following organs: adrenals (2x), heart, kidneys (2x), liver, spleen, prostate.

Other examinations:

The following organ weights were determined: liver, kidneys, adrenals, spleen, prostate gland, uterus, testes (left and right) with epididymides

Statistics:

Statistical analyses were performed separately on data from male and female animals. One or two-factorial analysis of variance was conducted on weight changes and food consumption. Group means were compared by the “Scheffe´” test.
Organ weights were evaluated by analysis of covariance and the mean values were compared by the “Scheffe´” test of covariance.
Values form chemical chemistry and hematology were analysed by analysis of variance with subsequent the “Scheffe´” test of variance.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

significant increase in prostate weight in high dose animals. However, this increase was not attended by any histopathologcal changes.

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

HISTOPATHOLOGY
All histopathological findings noted in the study were considered to be unrelated to the administration of the test article.

CLINICAL SIGNS
There were no deviations from normal in any group. All animals showed a healthy habit and behaviour throughout the study.
Reflex, ophthalmoscopic and auditory examinations revealed no findings related to the administration of the test article.
In individual animals the righting reflex was slightly reduced. This finding was attributed to coincidence, since there was no evidence of a dose-relationship.

PRE-TERMINAL DEATHS
No pre-terminal deaths occurred during the test period.

BODY WEIGHT DEVELOPMENT, FOOD CONSUMPTION AND FOOD CONVERSION RATIO
Body weights, food consumption and food conversion ratio were comparable to the controls and there were no significant intergroup differences.

HEMATOLOGY
All mean values were within the limits of the respective normal range. Hence the test article administration did not influence hematology parameters in any group.

CLINICAL CHEMISTRY
Clinical chemistry parameters did not reveal any changes which were connected with test article administration. The only significant difference was a slightly reduced total bilirubin content in high dose male animals compared to the respective control animals. Since this slight decline was without dose-relation and within the limits of the normal range, it is not considered biologically relevant and is attributed to coincidence.

URINALYSIS
The results of urinalysis gave no indication of treatment-related changes in any group. All findings were within the respective normal range and comparable to the controls.

NECROPSY - MACROSCOPIC FINDINGS
There were no macroscopic changes which were connected with test article administration.

ORGAN WEIGHTS
The only significant finding was an increase in prostate weight in high dose males. In amendment to the original study protocol this organ was therefore subjected to histo­ pathological examination.
Since nothing abnormal was found microscopically in this organ, the weight increase of prostates in high dose males may be considered either a coincidental finding or a result of a test article induced load reaction without histopathological manifestation.

Dose descriptor:

NOAEL

Effect level:

1 800 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Under the experimental conditions of this study a daily oral administration of 1800 mg/kg bw/day had no adverse effects on rats after 4 weeks

Critical effects observed:

no

Reference 2

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3650 “Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test”

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: RccHan: WIST(SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories, B.V., Kreuzelweg 53, 5961 NM Horst / Netherlands
- Age at study initiation: 11 weeks
- Weight at study initiation: 260 - 390 g (males), 175 - 240 g (females)
- Fasting period before study:
- Housing: in groups of three to five in Makrolon type-4 cages with wire mesh tops during acclimatisation and afterwards individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding with paper enrichment
- Diet (ad libitum): pelleted standard Harlan Teklad 2018C rodent maintenance diet (Provimi Kliba SA, 4303 Kaiseraugst / Switzerland)
- Water (ad libitum): community tap-water
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: daily. The test substance was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration.
Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.

VEHICLE
- Amount of vehicle (if gavage): 4 mL/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

On the first dose formulation day samples from the control group as well as three samples (top, middle and bottom) of about 1 g of each concentration were taken prior to dosing for analysis of concentration and homogeneity. Samples of about 1 g of each test item concentration were taken from the middle to confirm the stability (4 hours). Towards the end of the study, samples were taken from the middle to confirm concentration. The aliquots for analysis of dose formulations were frozen (-20 ± 5 °C) and stored at -20 ± 5 °C until analysis.
The samples were analyzed by derivatisation with MSTFA and analysis by GC-FID. The test item was used as the analytical standard. Analyzed samples were not discarded without written consent from the study director.
Duplicates were taken of all samples and stored at Harlan Laboratories Ltd., Füllinsdorf / Switzerland. The samples were not discarded without written consent from the study director.

Duration of treatment / exposure:

Males: Minimum 28 days
Females: Approximately 6 weeks

Frequency of treatment:

once daily (within four hours)

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

12

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on single dose toxicity data of the test compound and repeated dose toxicity data of chemically similar compounds, the results of a 14-day range-finding study the dose levels of 100, 300, and 1000 mg/kg bw per day were selected for the present study.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once prior to the first administration of the test item and weekly thereafter (in the gestation period on day 0, 6, 13 and 20 post coitum).
- detailed clinical observations were performed outside the home cage in a standard arena and the animals were observed for the following changes in: skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported.

BODY WEIGHT: Yes
- Time schedule for examinations: daily from treatment start to day of necropsy.

FOOD CONSUMPTION: Yes
- Males: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 - 14; after pairing period weekly.
- Females: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 - 14; gestation days 0 – 7, 7 14 and 14 – 21 and days 1 - 4 of the lactation
No food consumption was recorded during the pairing period.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood samples will be obtained on the day before or on the day of the scheduled necropsy from 5 males randomly selected from each group. Blood samples from 5 lactating females from each group will be obtained on day 5 postpartum.
- Anaesthetic used for blood collection: Yes (light isoflurane anesthesia)
- Animals fasted: Yes, for approximately 18 hours
- How many animals: 5 males randomly selected from each group and from 5 lactating females from each group
- Parameters checked: erythrocyte count, haemoglobin, haematocrit, mean corpuscular volume, red cell volume distribution width, mean corpuscular haemoglobin, mean corpuscular hemoglobin concentration, haemoglobin concentration distribution width, leukocyte count (total), differential leukocyte count: platelet count, reticulocytes; prothrombin time (= thromboplastin time), activated partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: see haematology
- Animals fasted: Yes, for approximately 18 hours
- How many animals: see haematology
- Parameters checked: alanine aminotransferase, alkaline phosphatase, aspartate aminotransferase, bile acids, bilirubin (total), cholesterol (total), creatinine, gamma-glutamyl-transferase, glucose, triglycerides, and urea

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at one time during the study following the daily dose administration (males shortly before the scheduled sacrifice and females on day 3 or 4 postpartum)
- Dose groups that were examined: five P generation males and five P generation females randomly selected from each group
- Battery of functions tested:
• Cage-side observations: faeces-balls, urine and posture as well as resistance to removal.
• Hand-held observations: muscle tone, constituation, skin, pupile size, palpebral closure, lacrimation, salivation, reaction to handling and general abnormalities.
• Open field observations: level of ambulatory activity including rearing (one minute evaluation), unusual body movements (e.g. spasms, convulsions), gait evaluation, behavior, hair coat, respiration, quantity of faeces-balls and urine.
• Reflexes: blinking, palpebral closure, pinna reflex, extensor thrust response, paw pinch, responsiveness to sharp noise, righting reflex and hearing ability (Preyer’s reflex).
• Measurements / Counts: hind limb / fore limb grip strength, rectal temperature.
Additionally, locomotor activity was measured quantitatively for the same animals. Activity was measured with an Activity Monitor AMS-0151 (FMI, Germany). Activity of the animals (based on beam count) was recorded for 6-minute intervals over a period of 30 minutes.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
The following organs were trimmed from any adherent tissue, as appropriate, and their wet weight was taken: adrenal glands (weighed as pairs), brain, epididymides, heart, kidneys (weighed as pairs), liver, ovaries (weighed as pairs), prostate, seminal vesicles (inclusive coagulating gland), spleen, testes, thymus, thyroid (after fixation), uterus (including cervix)
HISTOPATHOLOGY: Yes
The following tissues from all parental animals were preserved in neutral phosphate buffered 4% formaldehyde solution if not otherwise described: adrenals, brain, bone marrow (femur), epididymides (in Bouin’s fixative), gross lesions, heart, kidneys, liver, lymph nodes (axillary and mesenteric), ovaries (with oviduct), peripheral nerve (sciatic), prostate, seminal vesicles with coagulating gland, small and large intestines (incl. Peyer’spatches; duodenum, jejunum, ileum, colon, caecum, rectum), spinal chord (cervical, thoracic, lumbar), spleen, stomach (forestomach and glandular stomach), testes (in Bouin’s fixative), thymus, thyroids, and parathyroids if possible, trachea and lungs (preserved by inflation with fixative and then immersion), uterus (with vagina), urinary bladder
The following tissues were only examined by histopathology in case of macroscopic findings indicative of potential toxicity: aorta, esophagus, eyes with optic nerve and harderian gland, femur with knee joint, lacrimal gland, larynx, mammary gland (male and female), nasal cavity, pancreas, pharynx, pituitary gland, salivary glands – mandibular, sublingual, skeletal muscle, sternum with bone marrow.

Statistics:

The following statistical methods were used to analyze food consumption, body weights and reproduction data:
• The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables were assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test will be applied if the variables could be dichotomized without loss of information

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

No clinical signs were observed in males. Incidentally, one control female showed a slightly decreased activity, ruffled fur and vaginal bleeding on day 25 post coitum. Furthermore, one female treated at 100 mg/kg bw/day that did not give birth around day 21 post coitum either showed a wound on the anterior dorsum during the last three days prior to necropsy on day 25 post coitum. These isolated findings were not related to treatment with the test item.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

At 1000 mg/kg bw/day, one male died spontaneously on day 1 of the after pairing period. No clinical signs were observed and a normal body weight gain was recorded. The death was considered not treatment-related.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Mean body weight gain in males was lower at 1000 mg/kg bw/day during the pre-pairing period. This was a result of a minor body weight loss (1%) between days 1 to 2. During the pairing period, body weight gain at the high dose level was similar to the control value. Due to this fact, the reduction in mean body weight gain and body weights were considered to be not adverse. No effect of the test item on mean body weights and mean body weight gain was noted for females at any dose level.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

At 1000 mg/kg bw/day, mean food consumption was reduced during the first 11 days of treatment in males. Thereafter, the food consumption remained slightly lower than in controls. Since this reduction in food consumption occurred only transiently, the reduction was considered to be not adverse. There were no effects on mean food consumption at any dose level and in any study phase in females.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Treatment at 1000 mg/kg bw/day resulted in higher absolute and relative liver and kidney weights in animals of both genders. Since there was no evidence for an impairment of organ function by clinical pathology and histopathology, these findings were not considered to be adverse. Additionally, in females, absolute and relative heart weights were also increased. This finding was without histopathological correlate, and was therefore considered to be incidental.

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

VIABILITY/MORTALITIY
At a dose level of 1000 mg/kg bw/day, one male (no. 171, third animal delivery) died spontaneously on day 1 of the after pairing period. No clinical signs were observed and a normal body weight gain was recorded until day 13 of the paring period. One day before he died he showed a body weight loss of 3%. During necropsy, a dark red discolored thymus was recorded.

DAILY AND DETAILED WEEKLY CLINICAL OBSERVATIONS
No clinical signs were noted in males during the course of this study. Incidentally, one control female (no. 45) showed a slightly decreased activity, ruffled fur and vaginal bleeding on day 25 post coitum. Necropsy of this female revealed masses in the uterus and vagina. Furthermore, one female (no. 66) treated at 100 mg/kg bw/day that did not give birth around day 21 post coitum either showed a wound on the anterior dorsum during the last three days prior to necropsy on day 25 post coitum. During necropsy reddish discolored ovaries and enlarged axillary lymph nodes were noted. Both females were pregnant.

These isolated findings were not related to treatment with the test item.

No findings were noted at detailed weekly clinical observation during the whole course of the study.

FUNCTIONAL OBSERVATIONAL BATTERY
There were no treatment-related changes noted at functional observational battery.

One male at 100 mg/kg bw/day had increased faeces-balls and one female at 300 mg/kg bw/day showed decreased rearings. These isolated findings showing no dose relationship were deemed unrelated to treatment with the test item. A statistically significantly lower mean body temperature was noted in males of the first and second delivery at 1000 mg/kg bw/day (37.8 °C compared to 38.5 °C in the control group). No similar effect was noted for males treated at 100 and 300 mg/kg bw/day or in females. Since the value of 37.8°C was borderline to the range of the historical control data (38.0°C-38.6°C), and no change was noted in females, this finding was considered to be incidental.

LOCOMOTOR ACTIVITY
Locomotor activity was assessed quantitatively in terms of low beam counts in activity monitor.

Locomotor activity was not affected by the treatment with the test item in animals of both genders at any dose level.

FOOD CONSUMPTION
At 1000 mg/kg bw/day, mean food consumption was statistically significantly reduced in males during the first 11 days of treatment in males of the first and second delivery (-9.3% on day 11 of prepairing when compared with controls). Thereafter, the food consumption remained slightly lower than in controls, but without reaching statistical significance. Food consumption was also lower in males of the third delivery, but not to a statistically significant degree.

Since this reduction in food consumption occurred only transiently, the reduction was considered to be not adverse.

There were no effects on mean food consumption of females at any dose level and in any study phase.

BODY WEIGHTS
Mean body weight gain of males was statistically significantly lower at 1000 mg/kg bw/day during the pre-pairing period, resulting in slightly reduced mean body weights (-3% at the end of this period compared to the controls). This was a result of the minor body weight loss (1%) between days 1 to 2. During the pairing period, body weight gain at the high dose level was similar to the control value. Due to this fact, the reduction in mean body weight gain and body weights were considered to be not adverse.

No effect of the test item on mean body weights and mean body weight gain was noted for females at any dose level.

Towards the end of the gestation period, body weights and body weight gain were reduced in females at 1000 mg/kg bw/day (statistically significantly different body weight gains from day 17 post coitum onwards). These differences were considered to be a result of the increased post implantation loss and low number of pups, which were noted for dams at this dose level.

HEMATOLOGY
The assessment of the hematology data did not reveal any test item-related effects in males and females at any dose level.

CLINICAL BIOCHEMISTRY
The assessment of the clinical biochemistry data did not reveal any test item-related effects in males and females at any dose level.

Slightly higher urea levels were noted in males at 300 mg/kg bw/day and in females at 1000 mg/kg bw/day. Additionally, lower alkaline phosphatase levels were noted in females at 300 mg/kg bw/day, higher creatine levels in males and lower glucose levels in females at 1000 mg/kg bw/day, respectively. However, in the absence of dose-dependency and/or since values were additionally within the historical control range, a test item-related effect can be excluded.

ORGAN WEIGHTS
Treatment with the test item at 1000 mg/kg bw/day resulted in statistically significantly higher absolute and relative liver and kidney weights in animals of both genders, which was confirmed in animals of the third delivery except for kidney weights in females. Since there was no evidence for an impairment of organ function by clinical pathology and histopathology, these findings were not considered to be adverse.

Additionally, in females, absolute and relative heart weights were also increased. This finding was without histopathological correlate, and was therefore considered to be incidental.

There were no effects of treatment at 100 and 300 mg/kg bw/day.

MACROSCOPICAL FINDINGS
There were no treatment-related necropsy findings.

All findings were typical of this strain and age of rat and were considered to be incidental.

HISTOPATHOLOGICAL FINDINGS
There were no treatment-related findings. In particular, qualitative examination of the stages of spermatogenesis in the testis did not reveal any treatment-related abnormalities in the integrity of the various cell types present within the different stages of the sperm cycle. No treatment-related microscopic abnormalities were observed in the evaluation of the ovarian follicles and corpora lutea of the ovaries or the evaluation of the uterus.

All findings were typical of this strain and age of rat and were considered to be incidental.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: NOAEL is highest dose tested

Critical effects observed:

no

Please refer to the attached pdf document for detailed summary tables of relevant parameters.

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 comprises two adequate and reliable studies (RL1 and RL2) from reference substances with similar structure. Read-across is justified based on structural similarities and similar chemical behaviour. The selected studies are thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No. 1907/2006.

Repeated dose toxicity: inhalation - systemic effects

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 read-across

No data on repeated dose toxicity are available for the target substance Fatty acids, C8-10, oxybis(2-hydroxy-3,1-propanediyl) esters (CAS 92044-91-2). Therefore, read across from the relevant source substances 1,2,3-Propanetriol, homopolymer, diisooctadecanoate (CAS 63705-03-3) and Hexanedioic acid, mixed esters with decanoic acid, 12-hydroxyoctadecanoic acid, isostearic acid, octanoic acid, 3,3'-oxybis[1,2-propanediol] and stearic acid (CAS 130905-60-1) was applied to obtain information regarding repeated dose toxicity.

Read-across from appropriate reference substances is conducted in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. in order to fulfil the standard information requirements defined in Regulation (EC) No. 1907/2006, Annex VIII, 8.6. Structural similarities and similarities in properties and/or activities of the source and target substances are the basis of read-across. A detailed justification for the analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

CAS 63705-03-3

A combined repeated dose and reproduction / developmental screening study according to OECD TG 422 and observing GLP criteria has been performed with 1,2,3-Propanetriol, homopolymer, diisooctadecanoate (CAS 63705-03-3; BASF, 2013). 12 rats per sex and dose have been administered the test item in concentrations of 100, 300 and 1000 mg/kg bw/day by oral gavage. Animals of the control group were treated with the vehicle corn oil only. While males were treated for 28 days, females received the test item for approx. 6 weeks. At 1000 mg/kg bw/day, one male died spontaneously on day 1 of the after pairing period. No clinical signs were observed in that animal and a normal body weight gain was recorded. The death was therefore considered not treatment-related. No other mortalities occurred during the entire study period. Clinical signs observed in several females were considered not related to treatment with the test item. No findings were noted at detailed weekly clinical observation during the whole course of the study. There were no treatment-related changes noted at functional observational battery and locomotor activity. At 1000 mg/kg bw/day, mean food consumption was statistically significantly reduced in males during the first 11 days of treatment. Since the reduction in food consumption occurred only transiently, it was considered to be not adverse. There were no effects on mean food consumption of females at any dose level and in any study phase. Mean body weight gain of males was statistically significantly lower in the high dose group during the pre-pairing period, resulting in slightly reduced mean body weights. The reduction in mean body weight gain and body weights were considered to be not adverse. No effect of the test item on mean body weights and mean body weight gain was noted for females at any dose level. The assessment of the hematology as well as the clinical biochemistry data did not reveal any test item-related effects in males and females at any dose level. Treatment with the test item at 1000 mg/kg bw/day resulted in statistically significantly higher absolute and relative liver and kidney weights in animals of both sexes. Since there was no evidence for an impairment of organ function by clinical pathology and histopathology, these findings were not considered to be adverse. Additionally, in females, absolute and relative heart weights were also increased. This finding was without histopathological correlation and was therefore considered to be incidental. There were no treatment-related necropsy and histopathological findings. Based on the overall results, the NOAEL for repeated dose toxicity after oral exposure in male and female rats was established at 1000 mg/kg bw/day, corresponding to the highest dose tested.

CAS 130905-60-1

Hexanedioic acid, mixed esters with decanoic acid, 12-hydroxyoctadecanoic acid, isostearic acid, octanoic acid, 3,3'-oxybis[1,2-propanediol] and stearic acid (CAS 130905-60-1) has been tested for its short-term (28-day) repeated dose toxicity in a study performed according to OECD TG 407 and under GLP conditions (Sasol, 1990). The test substance was administered daily to groups of 5 rats per sex and dose by oral gavage for 4 weeks (7 days/week) in doses of 180, 1800 and 4500 mg/kg bw/day. A test group received the vehicle corn oil. No unplanned mortalities occurred during the course of the study. All histopathological findings noted in the study were considered to be unrelated to the administration of the test article. There were no abnormal clinical signs in any dose group. All hematology and clinical chemistry parameters were within the limits of the respective normal range and urinalysis gave no indication of treatment-related changes in any group. Macroscopic examination during necropsy revealed no changes which were attributed to the treatment of the test substance. The only significant finding was an increase in prostate weight in high dose males. Since nothing abnormal was found microscopically in this organ, the effect was considered either a coincidental finding or a result of a test article induced load reaction without histopathological  manifestation. The NOAEL for repeated dose toxicity after oral exposure in male and female rats was determined to be 1800 mg/kg bw/day.

Conclusion on repeated dose toxicity

Since no significant adverse effects have been observed in any of the studies performed with adequate source substance, no hazard with respect to repeated dose toxicity after oral exposure is identified for the target substance.

Justification for classification or non-classification

Based on the analogue read-across approach, the available data on oral repeated dose toxicity do not meet the classification criteria according to Regulation (EC) No. 1272/2008, and are therefore conclusive but not sufficient for classification.