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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:
1988-02-23 to 1988-03-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report Date:
1988

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted 12 May 1981
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
25. April 1984
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The guinea pig test method described in OECD TG 406 provides suitable information for hazard identification.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Test material form:
liquid

In vivo test system

Test animals

Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: HOECHST AG, Kastengrund, SPF-breed, Germany
- Weight at study initiation: 204g - 286 g
- Housing: groups of five animals each
- Diet: ad libitum, ERKA feed Number 8300 for guinea pigs and rabbits
- Water: tap water, ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +/- 3°C
- Humidity: 50 +/- 20 %
- Photoperiod: 12 hrs dark / 12 hrs light

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
5%
Day(s)/duration:
Day 1 / 8 days
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
10%
Day(s)/duration:
Day 9 / 2 days
Adequacy of induction:
other: selected concentration induced no mild-to-moderate irritation
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
10%
Day(s)/duration:
Day 22 / 1 day
No. of animals per dose:
Pretest:
6 animals for the determination of the non-irritating concentration
3 animals for the determination of the intradermal tolerance
Main test:
5 animals satellite group
5 animals control group
10 animals treated goup
Details on study design:
RANGE FINDING TESTS:

A. Determination of the non-irritating concentration
0.5 mL on the right and left flank, occlusive, exposure time: 24h

- undiluted solution (30 % Sodium ethylensulphonate)
- 10% test solution in 0.9 % NaCl (3 % Sodium ethylensulphonate)
- 1% test item in 0.9 % NaCl (0.3 % Sodium ethylensulphonate)

At the end of the exposure period the skin of the animals was observed for eythema and oedema formation. Signs of skin irritation were scored according to the Draize scoring system.

B. Determination of the intradermal tolerance
Intradermal injection on the back of the animals (3 areas per animal, 2 injections per area)

1. area: 2x0.1 mL 5% test solution in 0.9 % NaCl (1.5 % Sodium ethylensulphonate)
2. area: 2x0.1 mL 1% test solution in 0.9 % NaCl (0.3 % Sodium ethylensulphonate)
3. area: 2x0.1 mL 0.2% test solution in 0.9 % NaCl (0.06 % Sodium ethylensulphonate)

MAIN STUDY

A. Intradermal induction (day 1)
Intradermal injection on the back of the animals (3 areas, 2 injections per area)

1. area: 2x0.1 mL 50 % Freund´s Adjuvant
2. aerea: 2x0.1 mL 5 % test solution in 0.9 % NaCl (1.5 % Sodium ethylensulphonate)
3. aerea: 2x0.1 mL 5 % test item in Freund´s Adjuvant (1.5 % Sodium ethylensulphonate)

B. Epicutaneous induction (day 9)
0.5 mL on the intradermal injection area (back), occlusive, exposure time: 48 h

Treated group: 10 % test solution in 0. 9% NaCl (3 % Sodium ethylensulphonate)
Control/satellite group: 0.9 % NaCl

C. CHALLENGE EXPOSURE (day 22, epicutaneous)
0.5 mL on the left flank, occlusive, exposure time: 24 h

Treated group and control group: 10 % test solution in 0.9 % NaCl (3 % Sodium ethylensulphonate)
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10 % test solution in 0.9 % NaCl (3 % Sodium ethylenesulphonate)
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no clinical signs were observed
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10 % test item in 0.9 % NaCl (3 % Sodium ethylenesulphonate)
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no clinical signs were observed
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10 % test item in 0.9 % NaCl (3 % Sodium ethylenesulphonate)
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no clinical signs were observed
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10 % test item in 0.9 % NaCl (3 % Sodium ethylenesulphonate)
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no clincal signs were observed
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no clincal signs were observed
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no clincal signs were observed
Group:
positive control
Remarks on result:
not measured/tested

Any other information on results incl. tables

Evaluation:

A response to challenge of at least 30 % of the animals is considered as positive.

RANGE FINDING TESTS:

A. Determination of the non-irritating concentration

undiluted test solution (30 % Sodium ethylenesulphonate): Erythema and pustule formation, slight edema

10 % test solution in 0.9 % NaCl (3 % Sodium ethylenesulphonate): no skin irritation

1 % test solution in 0.9 % NaCl (0.3 % Sodium ethylenesulphonate): no skin irritation

 

B. Determination of the intradermal tolerance

5 % test solution (1.5 % Sodium ethylenesulphonate): slight redness and edema on the injection site.

1 % test solution (0.3 % Sodium ethylenesulphonate): no skin irritation

0.2 % test solution (0.06 % Sodium ethylenesulphonate): no skin irritation

MAIN STUDY

A. Intradermal induction

50 % Freund´s Adjuvant: Redness and edema on the injection site which turned into indurations

5 % test solution in Freund´s Adjuvant (1.5 % Sodium ethylenesulphonate): Redness and edema on the injection site which turned into indurations

B. Epicutaneous induction

The control and satellite group showed injection sites with open wound, scabbing and necrosis when the occlusive gauze was removed (day 11).

C. CHALLENGE EXPOSURE

24h and 48h after the occlusive gauze was removed the animals of the treated and the control group revealed no signs of skin irritation.

Body weight and clinical signs:

No clinical signs were observed during the study period. The body weight gains of the treated animals were mostly normal. Two animals if the control group showed only a slight increase in body weight development.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the condition of this study a test solution containing 30 % Sodium ethylenesulphonate did not induce skin sensitisation. Even if the concentrations at epicutaneous induction were too low according to the guideline requirements (i.e. the selected concentration induced no mild-to-moderate irritation), in combination with intradermal application irritation was observed and therefore, the study is acceptable and revealed that the substance has no sensitisation potential.
Executive summary:

In a guinea pig maximisation test according to OECD 406 and GLP guideline (Hoechst AG (a), 1988) a test solution containing 30 % Sodium ethylenesulphonate was used. For the determination of the non-irritating concentration and of the intradermal tolerance a range finding test was conducted. Thereby, 0.5 mL undiluted test solution (30 % Sodium ethylenesulphonate) induced erythema, slight oedema and pustles whereas 10 % test solution (3 % Sodium ethylenesulphonate) induced no skin irritations after epicutaneous occlusive exposure (24 hours) on the right and left flank of 6 animals. In the main study ten female Pribright-Hartley guinea pigs were treated on day 1 by two intradermal injections on the back in three areas with 50 % Freund´s Adjuvant (1. area, 2 x 0.1 mL), 5 % test solution in 0.9 % NaCl (2. area 2 x 0.1 mL, 1.5 % Sodium ethylenesulphonate) and 5 % test solution in Freund´s Adjuvant (3. area, 2 x 0.1 mL, 1.5 % Sodium ethylenesulphonate). On day 9, ten animals received 0.5 mL of the test solution by dermal application on the intradermal injection area (back), occlusive, for an exposure time of 48 hours. Thereby, animals were treated with 10 % test solution in 0.9% NaCl (3 % Sodium ethylenesulphonate). Five control animals and five animals of a satellite group were treated with a 0.9 % NaCl solution. On challenge exposure (day 22) animals (ten for the treated group and five for the control group) were treated with 0.5 mL of the test solution on the left flank by an occlusive dressing for an exposure time of 24 hours. All animals received 10 % test solution in 0.9 % NaCl (3 % Sodium ethylenesulphonate). 24h and 48h after the occlusive gauze was removed the animals of the treated and the control group revealed no signs of skin irritation.No clinical signs were observed during the study period. The body weight gains of the treated animals were mostly normal. Two animals of the control group showed only a slight increase in body weight development. After intradermal induction in animals treated with 50 % Freund´s Adjuvant and with 5 % test solution in Freund´s Adjuvant redness and oedema on the injection site were observed which turned into indurations. After the dermal induction, the control and satellite group showed injection sites with open wound, scabbing and necrosis when the occlusive gauze was removed (day 11). In conclusion, even if the concentrations at epicutaneous induction were too low according to the guideline requirements (i.e. the selected concentration induced no mild-to-moderate irritation), in combination with intradermal application skin irritation was observed and therefore, the study is acceptable and revealed that the substance has no sensitisation potential. Furthermore, the numberof 10 animals chosen is considered sufficient since a clear negative result has been observed.