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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:
1 Mar - 1 Apr 2004
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
/ no information on reliability check, less than 20 test and 10 control guinea pigs, no information on positive control reported

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
17 July 1992
GLP compliance:
not specified
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was conducted in 2004, where no validated protocol for the LLNA was available.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, protected from light

FORM AS APPLIED IN THE TEST: liquid

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Remarks:
Albino
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Japan SLC, Inc., S, Shizuoaka, Japan
- Age at study initiation: 5 weeks
- Weight at study initiation: 336 g
- Housing: 5 animals per cage in aluminium cages (Natsume Seisakusho Co., Ltd), placed on automatic water washing shelves
- Diet: solid food (RC4: Oriental Yeast Co., Ltd.), ad libitum
- Water: filtered tap water (5 µm cartridge filter), ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 2
- Humidity (%): 50 ± 10
- Air changes (per hr): 17
- Photoperiod (hrs dark / hrs light): 12/12
- IN-LIFE DATES: From: 1 Mar To: 1 Apr 2004

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
0.01 % (v/v)
0.1 mL/ site
Day(s)/duration:
single injection
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: ethanol
Concentration / amount:
30 % (v/v) in ethanol (50% in water)
Day(s)/duration:
2
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: ethanol
Concentration / amount:
0.5, 1, 3, 5, 10, 30% in ethanol (50% in water)
Day(s)/duration:
1
No. of animals per dose:
5 females (control), 10 females (test group)
8 females (range finding)
Details on study design:
RANGE FINDING TESTS:
- intradermal injections:
doses: 0.01, 0.03, 0.05, 0.1, 0.5, 1 and 5% (in physiological saline)
Injections (0.1 mL of each) were made into the lateral abdomen of the guinea pigs.
- epidermal applications: 0.3, 0.5, 1, 3, 5 and 10 % (in ethanol (50% in water). 0.05 mL of the test article were applied to lateral abdomen of the guinea pigs via the cloth portion of an adhesive plaster.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal:
Injection 1: 1:1 mixture (v/v) FCA/ physiological saline
Injection 2: 0.01% test substance (in physiological saline)
Injection 3: 0.02% test substance and FCA, in a 1:1 mixture (v/v)
Epicutaneous:
30 % test substance in ethanol (50% in water)

- Control group:
Intradermal: the vehicle (physiological saline) was used instead of the test substance
Epicutaneous: ethanol (50% in water) was used
- Site: lateral abdomen (intradermal and epicutaneous)
- Frequency of applications: every 7 days
- Duration: Days 0 - 10
- Concentrations: intradermal 0.01%, epicutaneous 30%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day of challenge: 21
- Exposure period: 24 h
- Test groups: test substance in ethanol (50% in water)
- Control group: test substance in ethanol (50% in water)
- Site: lateral abdomen
- Concentrations: 0.5, 1, 3, 5, 10 and 30% in ethanol (50% in water)
- Evaluation (hr after challenge): 3, 24 and 48 h
Challenge controls:
the control group is actually a challenge control
Positive control substance(s):
no

Results and discussion

Positive control results:
No positive control was used

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
3
Group:
negative control
Dose level:
intradermal induction: 0%; challenge: 0.5, 1, 3, 5, 10 and 30% in ethanol (50% in H2O)
No. with + reactions:
0
Total no. in group:
5
Key result
Reading:
1st reading
Hours after challenge:
3
Group:
test group
Dose level:
intradermal induction: 0.01% challenge: 0.5, 1, 3, 5, 10 and 30% in ethanol (50% in H2O)
No. with + reactions:
0
Total no. in group:
10
Key result
Reading:
2nd reading
Hours after challenge:
24
Group:
negative control
Dose level:
intradermal induction: 0%; challenge: 0.5, 1, 3, 5, 10 and 30% in ethanol (50% in H2O)
No. with + reactions:
0
Total no. in group:
5
Key result
Reading:
2nd reading
Hours after challenge:
24
Group:
test group
Dose level:
intradermal induction: 0.01%; challenge: 0.5, 1, 3, 5, 10 and 30% in ethanol (50% in H2O)
No. with + reactions:
0
Total no. in group:
10
Key result
Reading:
other: 3rd reading
Hours after challenge:
48
Group:
negative control
Dose level:
intradermal induction: 0%; challenge: 0.5, 1, 3, 5, 10 and 30% in ethanol (50% in H2O)
No. with + reactions:
0
Total no. in group:
5
Key result
Reading:
other: 3rd reading
Hours after challenge:
48
Group:
test group
Dose level:
intradermal induction: 0.01%; challenge: 0.5, 1, 3, 5, 10 and 30% in ethanol (50% in H2O)
No. with + reactions:
0
Total no. in group:
10

Any other information on results incl. tables

Range finding test

 

Intradermal injection

4 female guinea pigs were tested using different concentrations (0.01, 0.03, 0.05, 0.1, 0.5, 1 and 5 % in physiological saline) of the test substance and were checked for erythema and oedema after 24, 48 and 72h after the application.

24 h after the injections with 0.01% of the test substance 2 animals showed slight erythema and 2 animals an apparent erythema. At a concentration of 0.03% in 2 animals an apparent erythema was observed and in 2 animals an apparent erythema with oedema, whereas 1 animal showed an apparent erythema and 3 animals an apparent erythema with oedema after treatment with 0.05% of the test substance. 

All 4 animals showed eschar formation or necrosis starting at a dose of 0.1% of the test substance.

48 h after the injections with 0.01% of the test substance all 4 animals showed slight erythema. At a concentration of 0.03% in 2 animals an apparent erythema was observed and in 2 animals an apparent erythema with oedema, whereas 1 animal showed an apparent erythema and 3 animals an apparent erythema with oedema after treatment with 0.05% of the test substance. 

All 4 animals showed eschar formation or necrosis starting at a dose of 0.1% of the test substance.

72 h after the injections with 0.01% of the test substance all 4 animals showed slight erythema. At a concentration of 0.03% in 1 animal slight erythema was observed, in 1 animal an apparent erythema and the other 2 animals an apparent erythema with oedema, whereas 1 animal showed an apparent erythema and 3 animals an apparent erythema with oedema after treatment with 0.05% of the test substance. 

All 4 animals showed eschar formation or necrosis starting at a dose of 0.1% of the test substance.

 

Epicutaneous application

4 female guinea pigs were treated per dose ( 0.3, 0.5, 1, 3, 5 and 10% test substance in ethanol (50% in water)). They and were checked for erythema and oedema after 3, 24 and 48 h after removal of the patch.

After 3 h, all 4 animals treated with the highest dose (10%) showed a very slight erythema which remained until 72 h after patch removal.

No erythema or oedema was detected in any other animal at any time point and using any other concentration (0.3 – 5%).

Body weight

The body weight gain of the the animals was not affected adversely during the study.

Applicant's summary and conclusion

Interpretation of results:
other: CLP/ EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
CLP: not classified