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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1993-12-23 to 1994-02-14
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report date:
1994

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was performed before LLNA method was described as OECD test guideline and before it could be established in the EU-REACH regulation.

Test material

Constituent 1
Reference substance name:
Acetic acid, oxo-, sodium salt, reaction products with ethylenediamine and phenol, iron sodium salts
EC Number:
283-044-5
EC Name:
Acetic acid, oxo-, sodium salt, reaction products with ethylenediamine and phenol, iron sodium salts
Cas Number:
84539-55-9
Molecular formula:
non specified (UVCB substance)
IUPAC Name:
Acetic acid, oxo-, sodium salt, reaction products with ethylenediamine and phenol, iron sodium salts
Details on test material:
- Test article: CGA65047 SG 100, (A-5787 A); identical to FeNaEDDHA
- Additional specification: Sesquestrene 138 Fe 100 SG
- Physical state: granules; miscible in water
- Analytical purity: 100% (UVCB)
- Lot/batch No.: P.201845
- Storage condition of test material: room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pirbright White Strain
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: CIBA-GEIGY Limited, Animal Production, 4332 Stein, Switzerland
- Weight at study initiation: 326 to 424 g
- Housing: individually in Macrolon cages (Type 3)
- Diet (e.g. ad libitum): ad libitum, standard guinea pig pellets - NAFAG No. 845
- Water (e.g. ad libitum): ad libitum, fresh water

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 degree C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12 hrs light cycle

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
physiological saline
Day(s)/duration:
Day 0 and Day 8 (48 h)
Adequacy of induction:
other: control
Route:
intradermal and epicutaneous
Vehicle:
other: physiological saline and vaseline
Concentration / amount:
intradermal: 5% in physiological saline; epicutaneous: 50% in vaseline
Day(s)/duration:
Day 0 and Day 8 (48 h)
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: vaseline
Concentration / amount:
10%
Day(s)/duration:
Day 21 / 24 h
Adequacy of challenge:
highest non-irritant concentration
Route:
epicutaneous, occlusive
Vehicle:
other: vasline
Concentration / amount:
10%
Day(s)/duration:
Day 21 / 24 h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
10 male/10 female (test)
5 male/5 female (control)
Details on study design:
RANGE FINDING TESTS:
The concentration for the intradermal injection was selected on account of the solubility of the test article in standard vehicles and its local and systemic tolerability in a pretest. Since 5% test substance in physiological saline could be injected and was well tolerated, this concentration was used for the intradermal induction.
The concentration for the epidermal application was selected on account of the primary irritation potential of the test article. The following concentrations of the test item have been examined on separate animals for the determination of the maximum sub-irritant concentration: 10, 20, 30 and 50% in vaseline. 50% in vaseline was the highest applicable concentration of the test article. Erythema was observed at 20, 30 and 50% test article in vaseline.

MAIN STUDY
A. INDUCTION EXPOSURE
Day 0: Induction, intradermal injection:
3 pairs of intradermal injections (0.1 mL per injection) were made simultaneously into the left and right side of the shaved neck of the test and control group animals.
Test group:
- adjuvant/saline mixture 1:1 (v/v)
- 5% test substance in phys. saline (w/v)
- 5% test substance in the adjuvant/saline mixture (w/v)
Control group:
- adjuvant/saline mixture 1:1 (v/v)
- adjuvant/saline mixture 1:1 (v/v)
- physiol. saline

Day 8: Induction, epidermal application
In the test group test substance was incorporated in vaseline (w/v) and applied on a filter paper patch to the neck of the animals (patch 2x4 cm; approx. 0.4 g per patch; occluded administration for 48 hrs). The control group was treated with the vehicle only.
Test group:
- 50% test substance in vaseline
Control group:
- vaseline only

Day 21: Challenge
The test and control animals were tested on one flank with the test substance in vaseline (w/w) and on the other flank with the vehicle alone (patch 2x2 cm; approx. 0.2 g per patch; occluded for 24 hrs).
Test and control group:
- 10% test substance in vaseline
- vaseline only

The test was performed on 10 male and 10 female guinea pigs in the test group and 5 males and 5 females in the control group, respectively.
Challenge controls:
The test and control animals were tested on one flank with 10% test substance in vaseline (w/w) and on the other flank with vaseline alone (patch 2x2 cm; approx. 0.2 g per patch; occluded for 24 hrs).
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10% test test substance in vaseline
No. with + reactions:
4
Total no. in group:
20
Clinical observations:
erythema, edema
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10% test test substance in vaseline
No. with + reactions:
11
Total no. in group:
20
Clinical observations:
erythema, edema
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10% test test substance in vaseline
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10% test test substance in vaseline
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Remarks:
see read across documentation in section 13; positive result most probably due to presence of a sensitising impurity.
Conclusions:
Under the experimental conditions employed, 20% and 55% of the animals of the test group showed skin reactions 24 and 48 hours after removing the dressings, respectively. However, the positive reactions observed for erythema and oedema were ''very slight/barely perceptible". With only slight/barely perceptible reactions needing 48 h to develop for a major part of the animals, and based on a challenge concentration that was only half of what was needed to induce irritation in the animals (pre-test), the skin sensitizing potential, if any, is only very minimal at most.
Executive summary:

A dermal sensitisation test was performed in albino guinea pigs to determine the contact allergenic potency of the test article. This test was based on the OECD Guideline No. 406 (Skin Sensitisation) and the EU method B.5 (Skin Sensitisation).


On day 0 three pairs of intradermal injections (0.1 mL per injection) were made simultaneously into the left and right side of the shaved neck of the test and control group animals.


On day 8 test substance was incorporated in vaseline (w/v) and applied on a filter paper patch to the neck of the animals (patch 2x4 cm; approx. 0.4 g per patch; occluded administration for 48 hrs) in the test group. The control group was treated with the vehicle only.


On day 21 (Challenge) the test and control animals were tested on one flank with the test substance in vaseline (w/w) and on the other flank with the vehicle alone (patch 2x2 cm; approx. 0.2 g per patch; occluded for 24 hrs).


Under the experimental conditions employed, 20% and 55% of the animals of the test group showed skin reactions 24 and 48 hours after removing the dressings, respectively. However, the following remarks can be made to this study:


- Unfortunately there are no observations reported of the skin reactions following the intra- and epidermal applications during induction.


- All positive reactions observed for erythema and oedema were ''very slight/barely perceptible". This offers a possibility for subjective readings. It is not indicated whether evaluations of the individual animals were performed in a blinded way.


- With only slight/barely perceptible reactions needing 48 h to develop for a major part of the animals, and based on a challenge concentration that was only half of what was needed to induce irritation in the animals in this study, the skin sensitizing potential, if any, is only very minimal at most.