Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report Date:
2019

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Specific details on test material used for the study:
Name: Ethanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanamine
Batch/Lot number: 0002293298
Appearance: High-viscous, colourless to yellowish liquid
Purity: Considered as 100%
Expiry date: 22 November 2019
Storage conditions: Controlled room temperature (15-25 ºC, below 70 RH%)
Safety precautions: Routine safety precautions (lab coat, gloves, safety glasses, face mask) for unknown materials were applied to assure personnel health and safety.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Species and strain: Crl:WI Wistar rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7,
D-97633 Sulzfeld, Germany
Hygienic level at arrival: SPF
Hygienic level during
the study: Standard housing conditions
Justification of strain: The Wistar rat is one of the standard rodent species used in acute toxicity studies.
Number of animals: 3 animals
Sex: Female, nulliparous and non-pregnant.
Age of animals at study start: ~10-11 weeks old
Body weight range at dosing: Between 240 g and 250 g
Acclimation time: 26 or 28 days

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Based on the outcome in the range-finding test, two female animals were dosed approximately 48 hours later in the main test to confirm the classification outcome. The dose progression followed the Test Guideline OECD 402.

The amount of test item applied for each one of the animals is given in the table below:

Animal ID /Sex : 6620 Female; Body weight (g): 240; Applied amount (g): 0.48
Animal ID /Sex : 6621 Female; Body weight (g): 250; Applied amount (g): 0.50
Animal ID /Sex : 6622 Female; Body weight (g): 246; Applied amount (g): 0.49



The back of each animal was shaved (approximately 10% area of the total body surface) approximately 24 hours prior to treatment. The test item was applied to the shaved skin as a single dose and remained in contact with the skin for the
24-hour exposure period. Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin using a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.

At the end of the exposure period, the treated area of skin with the test item was washed with water at body temperature.
Duration of exposure:
24H
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
OBSERVATIONS
1/ Clinical Observations
Animals were inspected for signs of morbidity and mortality twice daily. Clinical observations were performed on the day of treatment at 30 minutes, 1, 2 and 5 hours after application of the test item and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

2/ Skin Irritation
Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.

3/ Measurement of Body Weight
The body weights were recorded on Day 0 (before the test item administration) and on Days 7 and 14 (before necropsy).

NECROPSY
Macroscopic examination was performed on all animals. All animals were anaesthetised with sodium pentobarbital (details in 3.3) and exsanguinated. Following confirmation of death, after examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.

Skin tissue from the treatment area was saved in formalin for further investigation if needed for other studies (eg. skin irritation or skin corrosion study).

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
The test item did not cause mortality at a dose level of 2000 mg/kg bw
Clinical signs:
Systemic Clinical Signs

There were no systemic clinical signs noted in any animal throughout the study.

Local Dermal Signs

Animal 1 (animal no. 6620)
The following adverse local dermal signs were observed on the treatment area after treatment with the test item: oedema (slight, Day 1), white skin with brown spots (Day 1), dark brown coloured alteration (Day 2), crust (Day 2-12) and erosion (Day 11-14).

Animal 2 (animal no. 6621)
The following adverse local dermal signs were observed on the treatment area after treatment with the test item: oedema (slight, Day 1), white skin with brown spots (Day 1), erythema (slight, Day 2) brown coloured alteration (Day 2), crust (Day 3-12) and erosion (Day 11-14).

Animal 3 (animal no. 6622)
The following adverse local dermal signs were observed on the treatment area after treatment with the test item: oedema (slight, Day 1), white skin with brown spots (Day 1), brown coloured alteration (Day 2), crust (Day 3-12) and erosion (Day 11-14).
Body weight:
In one animal (animal ID: 6622), a decreased body weight was observed between Days 0 and 7 and considering the clinical signs, a test item effect could not be excluded. However, the body weight gain was normal between Day 7 and 14. Body weights were within the range commonly recorded for this strain and age for the other two animals.
Gross pathology:
Erosion of the skin at the treated area was observed in 3/3 females administered 2000 mg/kg bw.

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Executive summary:

The acute dermal median lethal dose (LD50) of the test itemEthanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanaminewas found to be greater than 2000 mg/kg body weight in female Crl:WI rats.

According to the GHS criteria,Ethanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanaminecan be ranked as "Category 5" for acute dermal exposure.