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Description of key information

A LD50 > 2000 mg/kg bw was observed in the acute oral toxicity study in male and female rats. A LD50 > 5000 mg/kg bw was observed in the acute dermal toxicity study male and female rats. 

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non GLP, non-guideline, animal experimental study. Restrictions in reporting but otherwise adequate for assessment.
Qualifier:
no guideline followed
Principles of method if other than guideline:
The acute oral toxicity of Cetiol R was tested in an approximate manner on 5 male and 5 female rats. A quantity of 2000 mg of the test substance/kg bodyweight in a 2 % aqueous carboxymethyl cellulose preparation (w/w) was administered to the test animals on one single occasion by gavage. After
treatment the animals were observed over a period of 14 days.
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
- Average male body weight: 219 g
- Average female body weight: 165 g
- Animals were fasted for 16 hours prior to dosing
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
Concentration in vehicle: 2%


VEHICLE
- Concentration in vehicle:
- Amount of vehicle (if gavage):
- Justification for choice of vehicle:
- Lot/batch no. (if required):
- Purity:

MAXIMUM DOSE VOLUME APPLIED:

DOSAGE PREPARATION (if unusual):

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of weighing: 48 hours, 7 and 14 days
- Necropsy of survivors performed: yes, by means of an overdose of aranaesthetic and dissected.
- Other examinations performed: clinical signs, body weight, pathology.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
none
Clinical signs:
No intoxication symptoms were observed.
Body weight:
The treatment did not have any unfavourable influence on their bodyweight development.
Gross pathology:
No anatomicopathological changes in the inner organs or in the body cavities were found.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
other: Japan MAFF Testing Guideline of 12 Nosan No. 8147 as this in line with OECD 402.
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
GLP compliance:
yes (incl. certificate)
Remarks:
Bioassay Labor für biologische Analytik GmbH, 69120 Heidelberg
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: male animals approx. 9 weeks, female animals approx. 13 weeks
- Weight at study initiation: Male: 256-268 gram, Female: 212-231 gram. Animals of comparable weight (± 20% of the mean weight)
- Housing: Single housing in Makrolon cage, type III. As bedding material H 15005-29; Ssniff, Spezialitäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany) was used and for enrichment NGM E-022; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria.
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany
- Water: Tap water ad libitum
- Acclimation period: Acclimatization period of at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12h/12h
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Clipping of the fur: About 24 hours before administration
- Route of application: Single application to the clipped epidermis (dorsal and dorsolateral parts of the trunk); covering of the application site with a semi-occlusive dressing for 24 hours. The test item was covered with an air-permeable dressing (4 layers of absorbent gauze (Ph. Eur. supplied by Lohmann GmbH & Co., KG) and stretch bandage (Fixomull® Stretch (adhesive fleece) supplied by Beiersdorf AG). Afterwards removal of the semi-occlusive dressing, rinsing of the application site with warm water.
- Area of exposure: About 40 cm² (corresponds to at least 10% of the body surface)
- Time of day of application: In the morning

TEST MATERIAL
- Amount(s) applied: 4.97 mL/kg bw

Duration of exposure:
14 days
Doses:
5000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. A check for any dead or moribund animals was made at least once each workday, these records are archived by Bioassay. Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation.
- Necropsy of survivors performed: yes. Necropsy with gross-pathology examination on the last day of the observation period after sacrifice with CO2 in a chamber with increasing concentrations over time.
Sex:
male
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality observed
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality observed
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality observed
Mortality:
No mortality occurred.
Clinical signs:
No systemic clinical signs were observed during clinical examination. Furthermore, no local effects were observed.
Body weight:
The mean body weight of the animals increased throughout the study period within the normal range, with the exception of one male and one female, which showed stagnation of body weight during the second post-exposure week.
Gross pathology:
No macroscopic pathologic abnormalities were noted in the animals (5 males and 5 females) examined on the last day of observation.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw

Additional information

In a non-GLP, non guideline study (Henkel AG, 1983) ,5 male and 5 female Wistar rats received a single dose of 2000 mg/kg bw Cetiol R by gavage. The vehicle was carboxymethyl cellulose (CMC), the concentration of the test substance 2% in vehicle. During the post treatment observation period of 14 days no deaths occured. No clinical symptoms were observed. No adverse effects on bodyweight were seen. Based on the study result the LD50 for acute oral exposure is 2000 mg/kg for both male and female rats.

A GLP compliant guideline study (OECD 402) was performed (Bioassay, 2013). In this acute dermal toxicity study, young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 5000 mg/kg bw of the undiluted test item Soybean oil, epoxidized, reaction products with methanol to the clipped skin (dorsal and dorso-lateral parts of the trunk) and covered by semi-occlusive dressing for 24 hours. The application area comprised at least 10% of the total body surface area. The animals were observed for 14 days. No mortality occurred. No signs of systemic toxicity or skin effects were observed. The mean body weight of the animals increased within the normal range throughout the study period, with the exception of one male and one female, which showed stagnation of body weight during the second post-exposure week. There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period. Accordingly, the acute dermal median lethal dose (LD50) was determined to be > 5000 mg/kg bw for both male and female rats.

Justification for classification or non-classification

Based on the observed LD50 of >2000 mg/kg bw in the acute oral and a LD 50 > 5000 mg/kg bw in the dermal toxicity study, the test substance does not need to be classified according to Directive 67/548/EEC and according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.