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Description of key information

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:
Between 11th January and 11th February 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted according to OECD guidelines and in compliance with GLP.
Reference:
Composition 0
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Test material information:
Composition 1
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
Fifteen healthy male albino guinea-pigs of the Dunkin/Hartley strain were obtained from D. Hall, Newchurch, Staffordshire, England.
The animals were in the weight range of 211 to 295 g on arrival and approximately six to seven weeks of age. All the guinea-pigs were acclimatised to the experimental environment for five days prior to allocation to the main study.
An additional eight animals, from the same supplier, were used for the preliminary investigations.
The animals in the main study were allocated without conscious bias to two groups as follows:

Group Number of animals Animal numbers
Control animals 5 15 to 19
Test animals 10 20 to 29

The guinea-pigs were housed in groups of five in suspended metal cages with wire mesh floors in Building R 17 Room 14.
A vitamin C enriched guinea-pig diet FD1 and drinking water were provided ad libitum. Hay was given weekly as a dietary supplement. The batch of diet used for the study was not analysed for nutrients, possible contaminants or microorganisms.
However, other batches of diet are periodically tested, by the Supplier, and were found to be within set limits.
Results of routine physical and chemical examination of drinking water at source, as conducted usually weekly by the supplier, are made available to Huntingdon Research Centre Ltd. as quarterly summaries.
Animal room temperature was maintained within the range 19.0 to 20.0 °C and relative humidity within the range 38 to 56 % . These environmental parameters were recorded daily. Air exchange was maintained at approximately 15 air changes per hour and lighting was controlled by means of a time
switch to give 12 hours of artificial light (0700 - 1900 hours) in each 24 hours period.
Each animal was identified by ear tattoo number. This number was unique within the HRC Industrial Toxicology Department throughout the duration of the study. Each cage was identified by a coloured label displaying the study schedule number, animal numbers and the initials of the Study Director and Home Office licensee.
Route:
intradermal and epicutaneous
Vehicle:
other: Alembicol D
Concentration / amount:
Induction intradermal injection - 40% v/v in Alembicol D
Induction topical application - as supplied
Topical challenge - as supplied and 50% v/v in Alembicol D
Route:
epicutaneous, occlusive
Vehicle:
other: Alembicol D
Concentration / amount:
Induction intradermal injection - 40% v/v in Alembicol D
Induction topical application - as supplied
Topical challenge - as supplied and 50% v/v in Alembicol D
No. of animals per dose:
5 - control
10 - test
Details on study design:
TREATMENT PROCEDURE
Preliminary study
The intradermal and topical irritancy of a range of dilutions of the test substance was investigated to identify where possible (a) concentrations of the test substance that would produce irritation suitable for the induction phase of the main study and (b) a maximum non-irritant concentration by the topical route of administration for the challenge phase.

Selection of concentrations of test substance for the main study
Based on the results of the preliminary investigations, the following concentrations of Salicynalva were selected:
Induction intradermal injection - 40% v/v in Alembicol D
This was the highest concentration dosed intradermally that caused irritation but did not adversely affect the animals.
Induction topical application - as supplied
Topical challenge - as supplied and 50% v/v in Alembicol D
From preliminary investigations administration of the test substance as supplied did not give rise to irritating effects.

Main study
The procedure may be considered in two parts, Induction and Challenge.
Induction
Induction intradermal injections - test animals
A 40 x 60 mm area of dorsal skin on the scapular region of the guinea-pig was clipped free of hair with electric clippers. Three pairs of intradermal injections were made into a 20 x 40 mm area within the clipped area.
Injectables for the test animals were prepared as follows:
1. Freund's complete adjuvant was diluted with an equal volume of water for irrigation (Ph. Eur.).
2. Salicynalva, 40% v/v in Alembicol D.
3. Salicynalva, 40 % v/v in a 50 : 50 mixture of Freund's complete adjuvant and Alembicol D.

Induction topical application - test animals
The preliminary investigations indicated that the maximum practical concentration of the test substance for topical application (as supplied) did not produce skin irritation. Therefore, six days after the injections, the same 40 x 60 mm interscapular area was clipped and shaved free of hair and the site was pre-treated by gentle rubbing with 0.2 ml per site of 10% w/w sodium lauryl sulphate in petrolatum. Twenty-four hours later a 20 x 40 mm patch of Whatman No. 3 paper was saturated with approximately 0.4 ml of Salicynalva, as supplied. The patch was placed on the skin of the test animals and covered by a length of impermeable plastic adhesive tape (50 mm width "Blenderm"). This in turn was firmly secured by elastic adhesive bandage (50 mm width "Elastoplast") wound round the torso of the animal and fixed with "Sleek" impervious plastic adhesive tape. The dressing was left in place for 48 hours.
Induction - control animals
During the induction phase, the control animals were treated similarly to the test animals with the exception that the test substance was omitted from the intradermal injections and topical application.

Challenge
Challenge - control and test animals
The control and test animals were challenged topically two weeks after the topical induction application using Salicynalva, as supplied and 50% v/v in Alembicol D.
Hair was removed by clipping and then shaving from an area on the left flank of each guinea-pig.
A 20 x 20 mm patch of Whatman No. 3 paper was saturated with approximately 0.2 ml of Salicynalva, as supplied and applied to an anterior site on the flank. Salicynalva, 50% v/v in Alembicol D was applied in a similar manner to a posterior site. The patches were sealed to the flank for 24 hours under strips of "Blenderm" covered by "Elastoplast" wound round the trunk and secured with "Sleek"impervious plastic adhesive tape. The dressing was left in place for 48 hours.
Challenge controls:
The control and test animals were challenged topically two weeks after the topical induction application using Salicynalva, as supplied and 50% v/v in Alembicol D.
Positive control substance(s):
yes
Remarks:
hexyl cinnamic aldehyde
Positive control results:
The sensitivity of the guinea-pig strain used is checked periodically at HRC with hexyl cinnamic aldehyde, a known sensitiser.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
Salicynalva, as supplied
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Salicynalva, as supplied. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
Salicynalva, as supplied
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Salicynalva, as supplied. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test group
Dose level:
Salicynalva, as supplied
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
Salicynalva, 50% in Alembicol D
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Salicynalva, 50% in Alembicol D. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
Salicynalva, 50% in Alembicol D
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Salicynalva, 50% in Alembicol D. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test group
Dose level:
Salicynalva, 50% in Alembicol D
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
Salicynalva, unchanged
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: Salicynalva, unchanged. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
Salicynalva, unchanged
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: Salicynalva, unchanged. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
negative control
Dose level:
Salicynalva, unchanged
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading:
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
Salicynalva, 50% in Alembicol D
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: Salicynalva, 50% in Alembicol D. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
Salicynalva, 50% in Alembicol D
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: Salicynalva, 50% in Alembicol D. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
other:
Hours after challenge:
3
Group:
negative control
Dose level:
Salicynalva, 50% in Alembicol D
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: other:. . Hours after challenge: 3.0. Group: negative control. Dose level: Salicynalva, 50% in Alembicol D. No with. + reactions: 0.0. Total no. in groups: 5.0.

INTERPRETATION OF RESULTS:

Dermal reactions in the test animals elicited by the challenge application were compared with the findings simultaneously obtained in the control animals. A test animal was considered to show positive evidence of delayed contact hypersensitivity if the observed dermal reaction at challenge was definitely more marked and/or persistent than the maximum reaction seen in animals of the control group. If the dermal reaction seen in a test animal at challenge was slightly more marked and/or persistent than (but not clearly distinguishable from) the maximum reaction seen in control animals, the result for that test animal was classified as inconclusive. A test animal was considered to show no evidence of delayed contact hypersensitivity if the dermal reaction resulting from the challenge application was the same as, or less marked and/or persistent than the maximum reaction seen in animals of the control group.

CLINICAL SIGNS:

No signs of ill health or toxicity were recorded.

 

BODYWEIGHT:

Anticipated bodyweight increases were recorded for all guinea-pigs over the period of the study.

 

INDUCTION:

Intradermal injections:

Necrosis was recorded at sites receiving Freund's Complete Adjuvant in test and control animals. Slight irritation was seen in test animals at sites receiving Salicynalva, 40% v/v in Alembicol D and slight irritation was observed in control animals receiving Alembicol D alone.

 

Topical application:

Slight erythema was observed in test animals following topical application with Salicynalva, as supplied and slight erythema was seen in the control guinea-pigs.

 

CHALLENGE:

At the challenge there were no dermal reactions seen in any of the test or control animals.

 

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In this study, Salicynalva did not produce evidence of skin sensitisation (delayed contact hypersensitivity) in any of the ten test animals.
Executive summary:

This study was performed to assess the skin sensitisation potential of Salicynalva using the guinea-pig. The method followed was that described in: EEC Methods for the determination of toxicity, Annex of Directive 92/69/EEC (OJ No. L383A, 29. 12. 92), Part B, Method B. 6. Skin sensitisation. OECD Guideline for Testing of Chemicals No. 406 "Skin Sensitisation". Adopted 17 July 1992. Based on the results of a preliminary study and in compliance with the guidelines, the following dose levels were selected: Intradermal injection: 40% v/v in Alembicol D. Topical application: as supplied, Challenge application: as supplied and 50% v/v in Alembicol D. Ten test and five control guinea-pigs were used in this study. In the preliminary study irritation and necrosis was seen whith increasing doses using intradermal injection using 0.1 -100% of the test substance. Therefore in the main study a 40% induction concentration for intradermal application was considered justified showing a score 2 reaction in the preliminary study. In the preliminary study no skin irritation effects were seen up to 100% of the test substance. Therefore in the main study a 50 and 100% an induction concentration for topical application was considered justified. In this study Salicynalva did not produce evidence of skin sensitisation (delayed contact hypersensitivity) in any of the ten test animals.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

An available HRIPT test using 4% Salicynalva and 56 subjects did not show any effects. This information was not summarised because it limitedly adds to the negative result of the GPMT test.


Migrated from Short description of key information:
A GPMT was performed to assess the skin sensitisation potential of Salicynalva. The method followed was that described in: EEC Methods for the determination of toxicity, Annex of Directive 92/69/EEC (OJ No. L383A, 29. 12. 92), Part B, Method B. 6. Skin sensitisation. OECD Guideline for Testing of Chemicals No. 406 "Skin Sensitisation". Adopted 17 July 1992. Based on the results of a preliminary study and in compliance with the guidelines, the following dose levels were selected: Intradermal injection: 40% v/v in Alembicol D. Topical application: as supplied, Challenge application: as supplied and 50% v/v in Alembicol D. Ten test and five control guinea-pigs were used in this study. In the preliminary study irritation and necrosis was seen with increasing doses using intradermal injection using 0.1 -100% of the test substance. Therefore in the main study a 40% induction concentration for intradermal application was considered justified showing a score 2 reaction in the preliminary study. In the preliminary study no skin irritation effects were seen up to 100% of the test substance. Therefore in the main study a 50 and 100% an induction concentration for topical application was considered justified. In this study Salicynalva did not produce evidence of skin sensitisation (delayed contact hypersensitivity) in any of the ten test animals.

Justification for selection of skin sensitisation endpoint:
The GPMT skin sensitisation study is adequate for covering this endpoint.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:
Migrated from Short description of key information:
The substance is not a respiratory sensitiser because it is not a skin sensitiser following the ITS in the ECHA guidance (R7A, Fig. 7.3-2).

Justification for selection of respiratory sensitisation endpoint:
For respiratory sensitisation the results of the integrated evaluation strategy for respiratory sensitization data in the ECHA guidance (R7A, Fig. 7.3-2)is adequate for assessing this endpoint.

Justification for classification or non-classification

Based on the available information, the substance does not need to be classified as sensitising to the skin based on the criteria outlined in Annex VI of 67/548/EEC and Annex I of 1272/2002/EC. Therefore the substance is not a respiratory sensitiser either. This means that the substance does not have to be classified for respiratory sensitisation based on the criteria outlined in Annex I of 1272/2008/EC and Annex VI of 67/548/EEC.