Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: According to the 67/548/EEC directive, the test preparation Montanov 82 - batch 1261MP can be considered as not sensitizing by contact with skin. No deviation occured.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Batch: 1261MP
Appearence: white flakes
Storage: at room temperature, away from the light
Condition of use: the material is used diluted in distilled water

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
female
Details on test animals and environmental conditions:
Species: strain, supplier: Guinea pigs, Charles-River
Number and sex: 15, females
Weight between 336 and 391g at the beginning of the study
Acclimatization: at least 5 days
Housing:, diet: in individual polypropylene cage (310*465*190). Complete pelleted diet UAR 106 (91360-Epinay sur orge, France.)

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Remarks:
distilled water
Concentration / amount:
Determination of MNIC
injection of 0.1mL of the following preparations:
Freund's complete adjuvant (FCA) diluted to 50% in distilled water
vehicle: distilled water
50/50 mixture of the 2 solutions
Seven days later, occlusive patch were applied for 24 hours:
0.2 mL of the test material diluted to 40%, 20%, 10% and 5% in distilled water.

sensitizing potential
-Induction by intradermal route with 0.1 mL of the test maerial diluted to 10% in distilled water
epicutaneous application with 0.5mL of the product diluted to 50% in distilled water
-chllenge test by dermal route with 0.2mL at MNIC (10% in distilled water)and at 5%

Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Remarks:
distilled water
Concentration / amount:
Determination of MNIC
injection of 0.1mL of the following preparations:
Freund's complete adjuvant (FCA) diluted to 50% in distilled water
vehicle: distilled water
50/50 mixture of the 2 solutions
Seven days later, occlusive patch were applied for 24 hours:
0.2 mL of the test material diluted to 40%, 20%, 10% and 5% in distilled water.

sensitizing potential
-Induction by intradermal route with 0.1 mL of the test maerial diluted to 10% in distilled water
epicutaneous application with 0.5mL of the product diluted to 50% in distilled water
-chllenge test by dermal route with 0.2mL at MNIC (10% in distilled water)and at 5%

No. of animals per dose:
10
Details on study design:
Determination of the MNIC
0.1mL in intradermal injection of the following preparation:
- Freund's complete adjuvant. diluted to 50% in distilled water,
- vehicle: distilled water
- a 50/50 mixture of the 2 solutions
7 days later, the animals are clipped again and we applied for 24 hours under an occlusive patch 0.2 mL of the test material diluted to 40%, 20%, 10% and 5% in distilled water.
The cutaneous irritation reactions were assessed 24 and 48 hours after removal of patches.
The concentration retained for the challenge test is the maximal concentration that produces no primary cutaneous irritation in the animals.

Determination of sensitizing potential
this determination is carried out on a treated group of 10 guinea pigs compared with a control group of 5 animals.
Induction:
- intrademal route
The guinea pig is clipped at the dorasl level. It receives, on both sides of the spinal column, at the scapular level, 3 symmetrical injections of 0.1 mL of the following preparations:
for group 1
injection 1 mixture 1:1 (v/v) of Freund's complete adjuvant and distilled water,
injection 2: vehicle: distilled water (heated at 40°C)
injection 3: 50% pf distilled water (heated at 40°C) in a mixture 1:1 (v/v) of FCA and distilled water
for group 2
injection1: mixture 1:1 (v/v) of FCA and distilled water
injection2: test material diluted to 10% in distilled water (heated at 40°C)
injection 3: test material diluted to 10% in a mixture 1:1 (v/v) of FCA and distilled water (heated at 40°C)
- Epicutaneous route
After 6 days of rest period, the 15 guinea pigs (group 1 and 2) are reclipped at the injection sites. As the material produces a very light local irritation, the test area of control and treated animals are coated with 0.5 mL of sodium lauryl sulfate at 10 % in mineral oil to produce local irritation.
24 hours later, we applied to the test area level on each of the guinea pig from group 2, a piece of filter paper Whatman completely moistened with 0.5 mL of the product diluted to 50% in distilled water heated at 40°C. It is kept in contact with the skin for 48 hours with an occlusive patch.
the guinea pigs from group1 receive in the same way 0.5 mL of distilled water.

Challenge test: dermal route
After a 11-day rest period, the guinea pigs from both groups are clipped one last time at the dorso-lumbar region, the skin surface chosen corresponding to a zone which had not previously been in contact with the product tested.
We applied under an occlusive patch, to the skin of each of the guinea pigs from group 1 and 2, two (2*2 cm) pieces of filter paper moistened with 0.2 mL of the product tested to its maximal non-irritant concentration (MNIC) and to a lower concentration.
The occlusive patches are held in place for 24 hours then removed.
Approximately 21 hours after patch removal, the chaallenge test area is cleaned and clipped if necessary.
3 hours later, that is 24 hours after patch removal we observe and note the cutaneous reaction which corresponds to an appaisal of erythemae.
Challenge controls:
control group: group 1, 5 animals

Intradermal induction
injection 1 mixture 1:1 (v/v) of Freund's complete adjuvant and distilled water,
injection 2: vehicle: distilled water (heated at 40°C)
injection 3: 50% pf distilled water (heated at 40°C) in a mixture 1:1 (v/v) of FCA and distilled water
Epicutaneous route
After 6 days of rest period, the 15 guinea pigs (group 1 and 2) are reclipped at the injection sites. As the material produces a very light local irritation, the test area of control and treated animals are coated with 0.5 mL of sodium lauryl sulfate at 10 % in mineral oil to produce local irritation.
24 hours later, we applied to the test area level on each of the guinea pig from group 2, a piece of filter paper Whatman completely moistened with 0.5 mL of the product diluted to 50% in distilled water heated at 40°C. It is kept in contact with the skin for 48 hours with an occlusive patch.
the guinea pigs from group1 receive in the same way 0.5 mL of distilled water.

Challenge test: dermal route
After a 11-day rest period, the guinea pigs from both groups are clipped one last time at the dorso-lumbar region, the skin surface chosen corresponding to a zone which had not previously been in contact with the product tested.
We applied under an occlusive patch, to the skin of each of the guinea pigs from group 1 and 2, two (2*2 cm) pieces of filter paper moistened with 0.2 mL of the product tested to its maximal non-irritant concentration (MNIC) and to a lower concentration.
The occlusive patches are held in place for 24 hours then removed.
Approximately 21 hours after patch removal, the chaallenge test area is cleaned and clipped if necessary.
3 hours later, that is 24 hours after patch removal we observe and note the cutaneous reaction which corresponds to an appaisal of erythemae.
Positive control substance(s):
yes
Remarks:
2-mercapto benzothiazole

Results and discussion

Positive control results:
The product 2-mercapto benzothiazole appears moderatly sensitizing for the guinea pigs (60% of reactive animals at the reading 48 hours-class 3) when it is used diluted to 5 % and 2% in mineral oil.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 10%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
According to the 67/548/EEC directive, the test preparation Montanov 82 - batch 1261MP can be considered as not sensitizing by contact with skin.
Executive summary:

Study objective

To determine the skin sensitizing potential of a test material. the induction of the possible sensitizing effect is performed by epicutaneous and intradermal routes; Freund's complete adjuvant is used to increase sensitivity.

Regulatory compliance

This study is conduced according to the 406 OECD guideline (07.17.92) in compliance with Good Laboratory Practices.

This determination is carried out on a treated group of 10 guinea pigs compared with a control group of 5 animals.

Induction: - intrademal route

3 symmetrical injections of 0.1 mL of the following preparations for group the treated group:

injection1: mixture 1:1 (v/v) of FCA and distilled water

injection2: test material diluted to 10% in distilled water (heated at 40°C)

injection 3: test material diluted to 10% in a mixture 1:1 (v/v) of FCA and distilled water (heated at 40°C)

- Epicutaneous route After 6 days of rest period, the guinea pigs are reclipped at the injection sites. As the material produces a very light local irritation, the test area of control and treated animals are coated with 0.5 mL of sodium lauryl sulfate at 10 % in mineral oil to produce local irritation.

24 hours later, we applied to the test area level on each of the guinea pig from treated group, 0.5 mL of the product diluted to 50% in distilled water heated at 40°C. It is kept in contact with the skin for 48 hours with an occlusive patch.

Challenge test: dermal route After a 11-day rest period, the guinea pigs from both groups are clipped one last time at the dorso-lumbar region, the skin surface chosen corresponding to a zone which had not previously been in contact with the product tested. We applied under an occlusive patch, to the skin of each of the guinea pigs from group control and treated, 0.2 mL of the product tested to its maximal non-irritant concentration (MNIC) and to a lower concentration. The occlusive patches are held in place for 24 hours then removed. Approximately 21 hours after patch removal, the chaallenge test area is cleaned and clipped if necessary. 3 hours later, that is 24 hours after patch removal we observe and note the cutaneous reaction which corresponds to an appaisal of erythemae.

Results

the percentage of reactive animals is equal to 0%.

Conclusions

According to the 67/548/EEC directive, the test preparation Montanov 82 - batch 1261MP can be considered as not sensitizing by contact with skin.