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: Study conducted according to OECD Guideline under GLP conditions.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
Twenty-one female albino Dunkin Hartley guinea pigs supplied by David Hall Limited were used. At the start of the main study the animals weighed 322 to 383 g, and were approx 8 to 12 wks old. After an acclimisation period of at least five days, each animal was selected at random and given a number unique within the study which was written on a small area of clipped rum using a black indelible marker-pen. Animals were housed singly or in pairs in cages. Free access to tap water and food was allowed throughout the study. Animal room was maintained at 19 to 22 degrees C and relative humidity of 42 to 61%. 15 air changes per hour and lighting was controlled by a time switch to give 12 hour continuous light and darkness.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
unchanged (no vehicle)
Remarks:
also with liquid paraffin BP
Concentration / amount:
Intradermal Induction: 1% w/v in liquid paraffin BP; 1% w/v in a mixture of Freund's Complete Adjuvant plus distilled water (1:1)
Topical Induction: undiluted as supplied
Topical Challenge: 75% and 50% v/v in liquid paraffin BP
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Remarks:
also with liquid paraffin BP
Concentration / amount:
Intradermal Induction: 1% w/v in liquid paraffin BP; 1% w/v in a mixture of Freund's Complete Adjuvant plus distilled water (1:1)
Topical Induction: undiluted as supplied
Topical Challenge: 75% and 50% v/v in liquid paraffin BP
No. of animals per dose:
Ten test and five control animals (Total 15)
Details on study design:
Two main phases were involved in the Main Study; (1) an induction of a response and (b) a challenge of that response.

A row of three injections (0.1 ml each ) was made on each side of the mid-line. The injections were: a) Freund's Complete Adjuvant plus distilled water in the ratio 1:1; b) a 1% w/v emulsion of the test material in liquid paraffin BP; c) a 1% w/v emulsion of the test material in a 1:1 preparation of Freund's Complete Adjuvant plus distilled water.
Approximately 24 and 48 hrs after intradermal injection the degree of erythema at the test material injection sites (injection site b) was evaluated according to scale. One week later the shoulder region used previously for intradermal injections was clipped again and treated with a topical application of the undiluted test material. A filter paper patch was applied to the skin and held in place with surgical tape covered with an overlapping length of aluminum foil. The patch and foil were further secured with a strip of elastic bandage. The occlusive dressing was kept in place for 48 hours. Degree of erythema and oedema was quantified one and 24 hrs following removal of the patches. Other reactions were also recorded.
Challenge controls:
Controls: Identical procedure to that used for the test animals except that injections were: a) Freund's Complete Adjuvant plus distilled water in the ratio 1:1; b) liquid paraffin BP; c) a 50% w/v formulation of liquid paraffin BP in Freund's Complete Adjuvant/distilled water. The topical application followed the same procedure except that nothing was applied to the filter paper.

Challenge: Shortly before treatment day 21, both flanks of each animal was clipped free of hair with veterinary clippers. A square filter patch was loaded with the maximum non-irritant concentration (75% v/v in liquid paraffin BP) and applied to the shorn right flank of each animal and was held in place by surgical adhesive tape. To ensure that the maximum non-irritant concentration was used at challenge, the test material at a concentration of 50% v/v in liquid paraffin BP was similarly applied to a skin site on the left shorn flank. The patches were occluded with an overlapping length of aluminum foil and secured with a strip of elastic adhesive bandage. After 24 hours the dressing was carefully removed and discarded. The challenge sites were swabbed with cotton wool soaked in diethyl ether to remove residual material. The position of the treatment sites was identified by using a black indelible marker.
Approximately 24 and 48 hours after challenge dressing removed, the degree of erythema and oedema was quantified according to scale. Other reactions were also recorded.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50% v/v
No. with + reactions:
0
Total no. in group:
15
Clinical observations:
None
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50% v/v. No with. + reactions: 0.0. Total no. in groups: 15.0. Clinical observations: None.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
75% v/v
No. with + reactions:
0
Total no. in group:
15
Clinical observations:
brown/green coloured staining
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 75% v/v. No with. + reactions: 0.0. Total no. in groups: 15.0. Clinical observations: brown/green coloured staining.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
50% v/v
No. with + reactions:
0
Total no. in group:
15
Clinical observations:
None
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50% v/v. No with. + reactions: 0.0. Total no. in groups: 15.0. Clinical observations: None.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
75% v/v
No. with + reactions:
0
Total no. in group:
15
Clinical observations:
brown/green coloured staining
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 75% v/v. No with. + reactions: 0.0. Total no. in groups: 15.0. Clinical observations: brown/green coloured staining.

Any other information on results incl. tables

Intradermal: Well defined erythema was noted at the intradermal induction sites of all test group animals at the 24 hr observation with very slight to well-defined erythema at the 48 hr observation. Very slight erythema was noted at the intradermal induction sites of all control group animals at the 24 and 48 hour observations.

Topical Induction: Brown/green coloured staining was noted at the induction sites of all test group animals at the 1 and 24 hour observations. The staining did not affect evaluation of skin responses. Well defined erythema with or without very slight to slight oedema was noted at the induction sites of all test group animals at the 1 hour observation. Very slight to well-defined erythema with or without very slight to slight oedema was noted at the induction sited of 8 test group animals at the 24 hr observation. Other skin reactions noted in test group animals were bleeding from the intradermal induction sites, hardened dark brown/black coloured scabs and small superficial scattered scabs. Adverse dermal reactions prevented accurate evaluation of erythema and oedema at the induction sites of two test group animals at 24 hr observation.

Bleeding from the intradermal induction sites was noted in two control group animals at the 1 -hr observation. No signs of erythema or oedema were noted at the treatment sites of control group animals at the 1 or 24 hr observation.

Topical Challenge: 75% v/v in Liquid Paraffin BP: Brown/green coloured staining was noted at the challenge sites of all test and control group animals at the 24 and 48 hr observations. The staining did not affect evaluation of skin responses. No skin reactions were noted at the Challenge sites of the test or control group animals at 24 or 48 hr observations.

50% v/v in Liquid Paraffin BP: Brown/green coloured staining was not described in the Table of observations at 24 and 48 hr observations. No skin reactions were noted at the Challenge sites of the test or control group animals at 24 or 48 hr observations.

Bodyweight gains were comparable to those observed in the control group animals over the same period.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an OECD 406 study conducted under GLP conditions Molyvan 855 produced a 0% (0/10) sensitisation rate and was found to be non-sensitiser to guinea pig skin.