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Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1998-10-19 and 1998-11-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study
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:
1996
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The Guinea Pig Test (1999) met the previous requirements before the entry into force of REACH. The test is suitable and reliable to cover this endpoint. For this reason and for animal welfare reasons, no further in vivo study (LLNA test) needs to be performed.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Germany
- Age at study initiation: approx. 4 weeks
- Weight at study initiation: individual bw did not exceed 500 g
- Housing: 5 animals per labelled metal cage with wire-mesh floors
- Diet: standard guinea pig diet, including ascorbic acid (1600 mg/kg), LC-23 B, pellet diameter 4 mm (Hope farms, Woerden, The Netherlands), ad libitum; hay once a week
- Water: tap-water, diluted with decalcified water; ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route:
intradermal and epicutaneous
Vehicle:
corn oil
Concentration / amount:
intradermal injection:
a) 10%
b) 1:1 mixture of FCA and 20% test substance concentration
epidermal application:
50% test substance concentration
epidermal, challenge:
10% test substance concentration
Route:
epicutaneous, semiocclusive
Vehicle:
corn oil
Concentration / amount:
intradermal injection:
a) 10%
b) 1:1 mixture of FCA and 20% test substance concentration
epidermal application:
50% test substance concentration
epidermal, challenge:
10% test substance concentration
No. of animals per dose:
experimental group: 10 females
control group: 5 females
Details on study design:
RANGE FINDING TESTS:
A. INTRADERMAL INJECTIONS:
- Concentrations: 100%, 50%, 20% and 10% test substance
- each of two animals received two different concentrations in duplicate (0.1 mL/site) in the clipped scapular region
- the injection sites were assessed for irritation 24 and 48 hours after treatment

B. EPIDERMAL APPLICATION:
- Concentrations: 100%, 50%, 20% and 10% test substance
- 2 different concentrations were applied (0.5 mL each) per animal to the clipped flank
- animals receiving intradermal injections were treated with the lowest concentrations and two further animals with the highest concentrations
- Time of exposure: 24 hours
- Observation point after exposure: 24 and 48 hours

MAIN STUDY
A.1. INDUCTION EXPOSURE (intradermal), on day 1
- Exposure period: three pairs of intradermal injections (0.1 mL/site)
- Test groups: (A) 1:1 mixture (w/w) of FCA and water, (B) 10% test substance concentration, (C) 1:1 mixture (w/w) of FCA and a 20% test substance concentration
- Control group: were treated as decribed for the experimental animals, except that, instead of the test substance, the vehicle was administered
- Site: scalpular region; one of each pair was on each side of the midline and from cranial (A) to caudal (C)
- Duration: single injection

A.2 INDUCTION EXPOSURE (epidermal), on day 8
- Test group: 50% test substance concentration
- Control group: see comment in section A.1.
- Site: scalpular area between the injection sites
- Duration 48 hours

B. CHALLENGE EXPOSURE; on day 22
- No. of exposures: 1
- Exposure period: 24 hours
- Test group and control group: 10% test substance concentration and the vehicle
- Site: flank
- Evaluation (hr after challenge): 24 and 48 hours
Challenge controls:
yes
Positive control substance(s):
yes
Remarks:
Alpha-Hexylcinnamicaldehyde, Tech. 85%
Positive control results:
The results lead to a sensitisation rate of 100 per cent of the 10% concentration. From these results, it was concluded that the female guinea pig of the albino Dunkin Hartley strain is an appropriate animal model for the performance of studies designed to evaluate the sensitising potential of a substance in a Maximisation type of test.
Reading:
1st reading
Hours after challenge:
24
Group:
other: control group (test substance)
Dose level:
10%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: control group (test substance). Dose level: 10%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
other: control group (vehicle)
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: control group (vehicle). Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
other: control group (test substance)
Dose level:
10%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: control group (test substance). Dose level: 10%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
other: control group (vehicle)
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: control group (vehicle). Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10%
No. with + reactions:
7
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: test group (test substance). Dose level: 10%. No with. + reactions: 7.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
other: test group (vehicle)
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: test group (vehicle). Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10%
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
one animal showed scaliness
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: test group (test substance). Dose level: 10%. No with. + reactions: 7.0. Total no. in groups: 10.0. Clinical observations: one animal showed scaliness.
Reading:
2nd reading
Hours after challenge:
48
Group:
other: test group (vehicle)
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: test group (vehicle). Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Remarks on result:
not measured/tested
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Remarks on result:
other: the vehicle controls were sued as negative controls

No mortality occured and no symptoms of systemic toxicity were observed in the animals of the main study.

Body weights and body weight gain of the experimental animals remained in the same range as controls over the study period.

Seven animals of the 24 readings and seven animals of the 48 readings in the challenge phase were considered indicative of sensitisation. In sum eight different animals were considered indicative of sensitisation.

The skin effects caused by the intradermal injections and epidermal exposure during the induction phase are given in the following table.

Animal Number

Intradermal Injection (Day 3)

A                         B                           C

Epidermal Exposure (Day 10)
                         D

Control

1

2

3

4

5

 

E2

E3

E3

E3

E4

 

E2

E1

NA

NA

NA

 

E3

E3

E2

E3

E3

Erythema

0

0

0

0

0

Oedema

0

0

0

0

0

Experimental

6

7

8

9

10

11

12

13

14

15

 

E2

E3

E2

E2

E3

E3

E3

E3

E2

E2

 

E1

E3

NA

E1

E2

E1

E1

E1

E1

E1

 

E2

E3

E3

E4

E4

E4

E3

E3

E3

E2

 

2

2

0

2

2

2

2

3

1

2

 

0

0

0

0

0

0

0

0

0

0

A. 1:1 Mix of FCA and water

B. 10% test substance concentration (experimental); vehicle (control)

C. 1:1 Mix of FCA and a 20% concentration (experimental) or vehicle (control)

D. 50% test substance concentration (experimental); vehicle (control)

Skin effects intradermal injections: NA no abnormalities; E( ) Erythema (grade)

Interpretation of results:
sensitising
Conclusions:
The skin reactions observed in response to a 10% test substance concentration in 8 (of the 10) experimental animals in the challenge phase were considered indicative of sensitisation, based on the absence of any response in the control animals. The results indicate a sensitisation rate of 80 per cent.
Executive summary:

The test substance (50% in aromatic free mineral spirit) were tested for contact hypersensitivity in the albino guinea pig according to the EU method B6, the OECD guideline 406 and based on the method described by Magnusson and Kligman. Test substance concentrations selected for the Main study were based on the results of a preliminary study. In the main study, 10 experimental animals were intradermally injected with 10% concentration and epidermally exposed to a 50% concentration. 5 control animals were similarly treated, but with the vehicle (corn oil) only. Two weeks after the epidermal application all animals were challenged with a 10% test substance concentration and the vehicle.

No mortality occured and no symptoms of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

Skin reactions of grade 1 were observed in eight experimental animals in response to the 10% test substance concentration. No skin reactions were evident in the control animals. Scaliness was seen in the treated skin site of one experimental animal. The skin reactions observed in response to a 10% test substance concentration in 8 (of the 10) experimental animals in the challenge phase were considered indicative of sensitisation, based on the absence of any response in the control animals. The results indicate a sensitisation rate of 80 per cent.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

Skin sensitisation

The test substance was tested for contact hypersensitivity in the albino guinea pig according to the EU method B6, the OECD guideline 406 and based on the method described by Magnusson and Kligman. Test substance concentrations selected for the main study were based on the results of a preliminary study. In the main study, 10 experimental animals were intradermally injected with 10% concentration and epidermally exposed to a 50% concentration. 5 control animals were similarly treated, but with the vehicle (corn oil) only. Two weeks after the epidermal application all animals were challenged with a 10% test substance concentration and the vehicle.

No mortality occured and no symptoms of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

Skin reactions of grade 1 were observed in eight experimental animals in response to the 10% test substance concentration. No skin reactions were evident in the control animals. Scaliness was seen in the treated skin site of one experimental animal. The skin reactions observed in response to a 10% test substance concentration in 8 (of the 10) experimental animals in the challenge phase were considered indicative of sensitisation, based on the absence of any response in the control animals. The results indicate a sensitisation rate of 80 per cent.


Migrated from Short description of key information:
The test substance (50% in aromatic free mineral spirit) was assessed for skin sensitization in a Guinea pig maximization test according to EU method B.6 and OECD guideline 406. Based on the results obtained the test substance was considered to cause skin sensitisation.

Justification for selection of skin sensitisation endpoint:
Only one study available.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the results of the presented study, the test substance was classified and labelled with R43 (may cause sensitisation by skin contact) and H317 (may cause an allergic skin reaction, cat. 1) according to Regulation (EC) No 1272/2008 (CLP).

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