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Description of key information

No repeated dose toxicity data on the substance itself is available. Therefore, the sub-chronic toxicity study on the structural analogue with CAS# 151661-88-0 was used. Based on these observations the NOAEL was determined to be 1000 mg/kg bw/day in male and female rat. However, since a developmental toxicity study is available for a close analogue, in which the NOAEL for maternal toxicity was determined to be 200 mg/kg bw/day, this value is adapted as starting point for hazard assessment, as a conservative approach.


To further improve the toxicological data available for this substance and to further improve the read-across hypothesis, an OECD 422 for substance itself is ordered. This study can then be used as bridging study and further support the read-across hypothesis.


Additionally, a subchronic study is ordered for the substance itself according to the final decision (CCH-DL01_DEV-01-2119977075-30-0000).

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-06-18 until 1990-09-21
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
Application was performed on 5 days per week as opposed to 7 days per week with no analytical verification of test concentrations.
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- Name of test material: Stabiol VP 1711
- Substance type: fatty acid ester
- Physical state: white powder
- Lot/batch No.: 041/8/055 (1988-02-04)
- Stability under test conditions: stable under test conditions
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Sulzfeld, Germany
- Age at study initiation: 4 weeks
- Weight at study initiation: 62 - 82 g (females); 58 - 80 g (males)
- Housing: Makrolon Cage type III
- Diet: Altromin 1324, ad libitum, supplied by Altromin GmbH
- Water: tap water (drinking water quality analytically verified), ad libitum
- Acclimation period: 13 days
Route of administration:
oral: gavage
Vehicle:
peanut oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The testing solution was prepared daily just before administration by weighing out an appropriate amount of test substance and dispersing it in the vehicle.

VEHICLE
- The administration volume was 5 mL/kg bw.
- The concentration of test item in the vehicle was: 200, 60, 20 and 0 mg/mL
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
107 to 108 days starting with animal acclimation, with a total treatment of 68 to 69 administrations
Frequency of treatment:
5 days per week
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10 animals/ sex/ dose.
Satellite groups of 5 male and 5 female animals were used for the control and 1000 mg/kg bw day dose groups.
Control animals:
yes, concurrent vehicle
Details on study design:
- On the day of arrival the animals were subjected to an acclimatization period during which they received ground diet and drinking water ad libitum.
- The total number of application was 68 to 69, corresponding to a total administered amount of 6800 to 6900, 20400 to 20700, 68000 to 69000 mg/kg bw for the respective 100, 300, 1000 mg/kg bw/day dose groups. -
- Satellite groups of 5 male and 5 female animals were used for the control and 1000 mg/kg bw day dose groups. Following concurrent treatment these satellite animals were not treated for an additional observation period of 33 days in order to investigate the reversibility of potential toxic responses.
Positive control:
Not required
Observations and examinations performed and frequency:
MORTALITY
A check for moribund and dead animals was made twice daily on working days. If animals were in a moribund state, they were sacrificed and necropsied.

CLINICAL OBSERVATIONS
All animals were checked twice daily for any abnormal clinically signs on working days.

FOOD CONSUMPTION
Group food consumption was determined weekly for each cage.

DRINKING WATER CONSUMPTION
Drinking water consumption was determined weekly for each cage.

BODY WEIGHT
Body weight was determined before the start of the administration period. During the administration period body weight was determined on study day 0 (start of the administration period) and thereafter at weekly intervals.

HAEMATOLOGY
The following parameters were determined in blood: Leukocyte count (WBC), Erythrocyte count (RBC), Hemoglobin (HGB), Hematocrit (HCT), Mean corpuscular volume (MCV), Platelet count (PLT), Differential blood count

CLINICAL CHEMISTRY
Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatase (AP), γ-Glutamyltransferase (GGT), Sodium (NA), Potassium (K), Chloride (CL), Calcium (CA), Urea (UREA), Creatinine (CREA), Glucose (GLUC), Total bilirubin (TBIL), Total protein (TPROT), Cholesterol (CHOL)

OPHTHALMOSCOPY
Prior to the start of necropsy the eyes of all animals of the control and high dose animals were treated with Mydriaticum and examined for any changes using a slit lamp.


Sacrifice and pathology:
NECROPSY
The animals were sacrificed by exsanguination under an overdose of ether. The exsanguinated
animals were necropsied and assessed by gross pathology.

ORGAN WEIGHTS
The following weights were determined in all animals sacrificed on schedule: Anesthetized animals, Adrenal glands, Brain, Heart, Kidneys, Liver, Spleen, Testes, Thymus

HISTOPATHOLOGY
Fixation was followed by histotechnical processing, examination by light microscopy and assessment of findings on following organs: Adrenal glands, Aorta, Skeletal muscle, Brain, Cecum, Uterus, Coagulating glands, Colon, Duodenum, Epididymis (left), Esophagus, Heart, Ileum, Jejunum, Kidneys, Liver, Lung, Lymph nodes (mesenteric and axillary lymph nodes), Pancreas, Tounge, Pituitary gland, Prostate, Salivary gland, Seminal vesicles, Skin, Semen vesicles, Spleen, Forestomach, Testis, Thymus, Thyroid glands, Trachea, Uterus
Statistics:
- A comparison of inter-group differences (clinical chemistry, hematology and body weight) was performed using the t-test.
- DUNNETT's test was used to test the hypothesis of equal means.
- Steel-test was used to evaluate the inter-group differences relating to organ weights of each dose group.
Clinical signs:
no effects observed
Description (incidence and severity):
No test substance-related findings were observed
Mortality:
no mortality observed
Description (incidence):
In the present study no animal died ahead of schedule.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No test substance-related findings were observed in all parameters under investigation.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No test substance-related findings were observed.
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No test substance-related findings were observed.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No test substance-related findings were observed in all parameters under investigation.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
A decrease of the haematocrit (HCT) value in the male groups 2 - 4 was observed. The decrease of the HCT seems to be compound-and partially dose-related. Additionally the intermediate analysis showed slightly reduced Red Blood Cell (RBC)-value for the male group 4. The observed deviation of the HCT is considered to be incidental/ because the diagnosed values are within the ranges of the historical control. In addition to this, there are no corresponding deviations of the RBC- or MCV-values to prove the biological relevance of this findings.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
No test substance-related findings were observed in all parameters under investigation.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
- Absolute organ weights: No test substance-related findings were observed in all parameters under investigation.
- Relative organ weights: No test substance-related findings were observed in all parameters under investigation.
Gross pathological findings:
no effects observed
Description (incidence and severity):
The macroscopical examination of the organs displayed some observations like discolouration of the thymus deformation of the spleen, cyst of the kidney, hydronephrosis, atrophy of the testes, hydrometra and necrosis of the fatty tissue which were considered to be spontaneous. Compound-related macroscopical effects were not observed. The male and female animals of the recovery group 4 (recovery period 33 days) showed no macroscopical compound-related alterations.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
No test substance-related findings were observed in all parameters under investigation. However, in the male and female animals of all groups (including the recovery group 1 and 4) the livers, the heart and the mandibulary lymph node showed effects which were due to a bacteriosis of unknown etiology. The observed germinal hyperplasia of the mandibulary lymph node can be interpreted as a consequence of this bacteriosis.
Histopathological findings: neoplastic:
no effects observed
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed at the highest dose tested.
Critical effects observed:
no
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat

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

No data on repeated dose toxicity of the substance itself are available. Therefore, investigations on repeated dose toxicity of the structural analogous substances with CAS# 151661-88-0 and Oleic methyl ester, epoxidized, reaction products with glycerol (OEG) were used. Since a developmental toxicity study is available the close analogue with CAS# 211450 -54 -3, in which the NOAEL for maternal toxicity was determined to be 200 mg/kg bw/day, this value is adapted as starting point for hazard assessment, as a conservative approach.


 


90-day repeated dose toxicity (CAS 151661-88-0)


The substance was tested for systemic toxicity at repeated doses of 0 (group 1), 100 (group 2), 300 (group 3) and 1000 (group 4) mg/kg body weight/ day. The compound was administered daily by gavage over a period of 90 days. 10 male and 10 female rats were used for each dose. In addition to the groups 1 and 4, 5 male and 5 female animals were used to determine the reversibility of passible compound- related findings (recovery groups). All doses applied were tolerated without lethality. No compound- related symptoms were observed. The mean food consumption in all treated groups was comparable to the control. The increase in food consumption in the female group 4 at the beginning of the week 4 up to the end resulted from food wasting by the animals. The mean water intake of all groups was comparable to the control. The total body weight gain of all groups showed no deviation and was comparable to the control. The intermediate and the final haematological examinations revealed a decrease of the Haematocrit value (HCT) in the male groups 2 - 4. The decrease of the HCT seems to be compound- and partially dose-related. Additionally the intermediate analysis showed slightly reduced (Red Blood Cell count) RBC- value for the male group 4. The observed deviation of the is considered to be incidental/ because the diagnosed values are within the limits of the historical control. In addition to this, there are no corresponding deviations of the RBC- or MCV- values to prove the biological relevance of this findings. All other findings were considered to be spontaneous and therefore not related to the treatment. The biochemical examinations revealed some compound and dose independent findings. The examination of the eyes by slit lamp microscope showed no compound- related effects. The absolute and relative organ weights in all groups showed no deviations and were comparable to the control. The macroscopical examination of the organs displayed some observations like discolouration of the thymus, deformation of the spleen, cyst of the kidney, hydronephrosis, atrophy of the testes, hydrometra and necrosis of the fatty tissue which were considered to be spontaneous. Compound-related macroscopical effects were not observed. The male and female animals of the recovery group 4 (recovery period 33 days) showed no macroscopical compound- related alterations. The microscopical examination revealed no compound- related effects. In the male and female animals of all groups (including the recovery group 1 and 4 ) the livers, the heart and the mandibulary lymph node showed effects which were due to a bacteriosis of unknown etiology. The observed germinal hyperplasia of the mandibulary lymph node can be interpreted as a consequence of this bacteriosis. Possible target organs have not been diagnosed. The liver and the heart of the recovery groups 1 and 4 were examined in order to prove the persistence of the bacteriosis. According to the described study, a daily administration of the test item up to 1000 mg/kg bw/day is not cumulative- systemic toxic to rats. Based on these observations the NOAEL was determined to be 1000 mg/kg bw/day for males and females(Henkel 1991).


 


28-day repeated dose toxicity (Oleic methyl ester, epoxidized, reaction products with glycerol (OEG))


The substance was tested for systemic toxicity at dosages of 100, 500 and 1000 mg/kg body weight/day, administered by gavage for 28 days. 10 male and 10 female rats were used for each dose. The low, middle and high dose was tolerated in all animals without administration related lethality. The only symptom was salivation in few animals of the high dose group. There were some variations in food, water consumption and body weight gain in dosage groups to be without compound and dose related relevance. Haematologically, compound related, decreased haematocrit and erythrocyte values were observed in male animals in the high dose group. In female animals the same findings occurred in the middle dose group. A slight significant increase in GPT-levels in males of the high dose and a significant increase of GPT-values in female animals of the high dose were measured. Eye examination revealed no compound related findings. The relative liver weights were significantly increased in male animals of the high dose. A slight increase of the relative liver weights was seen in female animals of the high dose. These findings were considered to be compound related. Pathologically-anatomically examination revealed in all animals no evidence of compound related damage of inner organs. Microscopic examination in animals of high dose level revealed degenerative alterations in kidneys which were reversible in the recovery group. It was concluded by the study director that a daily administration of 1000 mg/ kg bw/ day is for rats a cumulative toxic level with liver and kidney as target organs. A NOAEL of 100 mg/kg bw/day was suggested as non-cumulative toxic level for rats (BASF 1987).


However, it is arguable whether the effects observed in the study are sufficient to conclude on a NOAEL of 100 mg/kg bw/day. According to a recent publication by Hall et al., the variation of liver weights without histological evidence or distinct alterations in clinical chemistry is not considered to be an adverse effect. Hall et al. reach the conclusion that only a 2-3-fold increase in ALT (GPT) or a biologically significant change in other biomarkers is considered to be an adverse effect. In the study at hand, the only remarkable alteration in liver parameters was an ALT increase in the highest dose group of males and females, but it was less than 2-fold. Regarding the observed degradations in the kidney, they were not accompanied with relevant biochemical changes (urea, creatinine, sodium, potassium) and were reversible in the recovery group. The absence of adverse effects in the 90-day study further supports the argumentation that the alterations found in the 28-day study were temporary and rather a non-adverse adaptation than an evidence for a toxicological action (Henkel 1987).


 


Reference


Hall et al. (2012), Liver hypertrophy: a review of adaptive (adverse and non-adverse) changes – conclusions from the third international ESTP workshop, Toxicol Pathol, 40(7), 971-94


 

Justification for classification or non-classification

Based on the available data, classification for repeated dose toxicity is not warranted in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.