Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08 April 2015 - 08 September 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was performed at a GLP laboratory in accordance with OECD, EU and US testing guidelines for the assessment of skin sensitization.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
A deviations in the daily mean relative humidity and a procedural deviation during the pre-screening test occured. The study integrity was not adversely affected by the deviations.
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
A deviations in the daily mean relative humidity and a procedural deviation during the pre-screening test occured. The study integrity was not adversely affected by the deviations.
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
yes
Remarks:
A deviations in the daily mean relative humidity and a procedural deviation during the pre-screening test occured. The study integrity was not adversely affected by the deviations.
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): K1600 black dye and 206474/A
- Physical state: Powder

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: Approx. 10 weeks old
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean.
- Housing: Group housed in labeled Makrolon cages containing sterilised sawdust as bedding material
- Diet: Free access to pelleted rodent diet
- Water: Free access to tap water.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%
- Air changes (per hr): 10
- Photoperiod: 12-hour light/12-hour dark cycle

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, open
Vehicle:
propylene glycol
Concentration / amount:
Pre-screening & main study: 0 (control, vehicle only) 10, 25 or 50% w/w
Challengeopen allclose all
Route:
epicutaneous, open
Vehicle:
propylene glycol
Concentration / amount:
Pre-screening & main study: 0 (control, vehicle only) 10, 25 or 50% w/w
No. of animals per dose:
Pre-screening study: 2 animals per concentration
Main study: 5 animals per concentration

Study design: in vivo (LLNA)

Vehicle:
propylene glycol
Concentration:
0 (Vehicle only), 10, 25 and 50 % (w/w)
No. of animals per dose:
Pre-screening study: 2 animals per concentration
Main study: 5 animals per concentration
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: There was no information available regarding the solubility or stability in vehicle.
- Irritation: No irritation and no signs of systemic toxicity were observed in any of the pre-screen animals.
- Lymph node proliferation response: Not performed.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response:
Erythema and eschar formation:
No erythema: 0
Very slight erythema (barely perceptible): 1
Well-defined erythema: 2
Moderate to severe erythema (beet redness) to slight eschar formation (injuries in depth): 3
Severe erythema (beet redness) to eschar formation preventing grading of erythema: 4


TREATMENT PREPARATION AND ADMINISTRATION:
Pre-screening: Each animal was treated with one concentration on three consecutive days.
Main study (induction): Each animal was treated with one concentration on three consecutive days. The dorsal surface of both ears was topically treated (25 μL/ear) with the test substance, at approximately the same time on each day.

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
The SI values calculated for the substance concentrations 10, 25 and 50% were 3.7, 4.9 and 17.9, respectively. These results show that the test substance elicits a SI ≥ 3. The EC3 value (the estimated test substance concentration that will give a SI =3) was established to be between 0 and 10%.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Mean DPM/animal values for the experimental groups treated with test substance concentrations 10, 25 and 50% were 2293, 3072 and 11192 DPM, respectively. The mean DPM/animal value for the vehicle control group was 627 DPM.

Any other information on results incl. tables

DEVIATIONS

1. Deviations from the maximum level of daily mean relative humidity occurred. Evaluation: Laboratory historical data do not indicate an effect of the deviations.

2. Following cleaning of the ears on Day 2 of the pre-screentest, the animals were dosed before scoring for erythema, instead of after. Evaluation: The test substance remnants after dosing did not hamper scoring of the ears therefore the ears could be scored according to the standard procedure (without cleaning of the ears). The purpose of the pre-screen test is to select the highest concentration for the main study.

The main study results showed that the right concentration was selected. The study integrity was not adversely affected by the deviations.

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information Skin Sens. 1 Criteria used for interpretation of results: EU
Conclusions:
The test substance was determined to be classified as Skin Sens. 1 in accordance with Regulation (EC) No 1272/2008.
Executive summary:

The sensitising potential of the test substance was examined in accordance with the test guidance provided in EC, No 440/2008; B42, OECD 429 and EPA, OPPTS 870.2600. The study was performed in a GLP compliant laboratory, minor deviations were observed but were determined not to affect the validity of the study. The test substance was determined to be classified as Skin Sens. 1 in accordance with Regulation (EC) No 1272/2008.