Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19 March 2018 - 23 April 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
2017
Deviations:
no
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Colourless solidified melt
Details on test material:
Expiry date: 25 October 2018
Specific details on test material used for the study:
Stability in vehicle: The test item is stable in a solution of Propylene glycol for at least 5 hours at room temperature under normal laboratory conditions, for at least 6 days in the refrigerator and, for at least 3 weeks in the freezer (≤ -15°C) over the concentration range 1 to 200 mg/mL.

The test item was heated to approximately 80°C for 15 minutes approximately 2 hours before weighing.

Adjustment was made for specific gravity of the vehicle (1.036).

Test animals

Species:
rat
Strain:
other: Crl: WI(Han)
Remarks:
Outbred, SPF-Quality
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant:yes
- Age at study initiation: approx. 9 - 12 weeks old
- Weight at study initiation: 166-212 g.
- Housing: Group housing (up to 5 animals of the same sex together) on arrival and individual housing during the study. Polycarbonate cages (Makrolon MIII type; height 18 cm.) containing sterilized sawdust were used during the study. For psychological/environmental enrichment, animals were provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom), except when interrupted by study procedures/activities.
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum throughout the study, except during designated procedures.
- Water: Municipal tap-water, ad libitum
- Acclimation period: 5 days

The feed was analyzed by the supplier for nutritional components and environmental contaminants. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
Periodic analysis of the water was performed, and it is considered that there were no known contaminants in the water that would interfere with the objectives of the study.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-21
- Humidity (%): 30-51
- Air changes (per hr): >= 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 20 March 2018 To: 23 April 2018

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
propylene glycol
Remarks:
Specific gravity: 1.036
Details on dermal exposure:
RANGEFINDING STUDY
A range finding study was performed in order to select the dose causing no mortality or significant toxicity to be used in the main study. Initially, one animal was dosed at 1000 mg/kg. A period of at least 48 hours was allowed between the dosing of each animal. Based on the results one additional animal was dosed at 2000 mg/kg. Based on the results of the range finding study, in the main study two animals were dosed at 2000 mg/kg.

TEST SITE
- Area of exposure: 18 cm^2
- % coverage: approx. 10% of the total body surface
- Type of wrap if used: The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages/

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 1000 & 2000 mg/kg bodyweight
- Constant volume or concentration used: yes, 10 mL/kg bodyweight
Duration of exposure:
24 hours
Doses:
1000 mg/kg bodyweight
2000 mg/kg bodyweight
No. of animals per sex per dose:
1 female: 1000 mg/kg bodyweight
3 females: 2000 mg/kg bodyweight
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: Mortality: twice daily; Clinical observations: at periodic intervals on the day of dosing (at least three times) and once daily thereafter; Body weights: on day 1 (pre-administration), day 8 and day 15.
- Necropsy of survivors performed: yes

The skin reactions were assessed approximately 24, 48 and 72 hours after the removal of the dressing and test item. Adjacent areas of untreated skin of each animal served as controls.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Red discoloration of the nose and ptosis were noted for one animal on Day 1.
Body weight:
Body weight loss was noted for one animal at Day 8 and body weight loss or reduced body weight gain was noted for the other two animals at Day 15.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
Irritation and local effects:
No erythema or edema were scored for any of the animals at any time point. Fissures, scales, scabs and/or red discoloration were seen in the treated skin-area of the animals between Days 2 and 15. Scabs were noted on the back of one animal between Days 12 and 15.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Not classified according to Regulation (EC) No. 1272/2008.
Conclusions:
The dermal LD50 value of Ethanol, 2,2’-[(3-methylphenyl)imino]bis- in Wistar rats was determined to exceed 2000 mg/kg body weight. Based on this result, Ethanol, 2,2’-[(3-methylphenyl)imino]bis- is not classified according to GHS and CLP criteria.