Reaction mass of 2,4-dimethyl-6-phenyl-3,6-dihydro-2H-pyran and 2-methyl-4-methylene-6-phenyltetrahydro-2H-pyran and 4,6-dimethyl-2-phenyl-3,6-dihydro-2H-pyran

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

The study was conducted between 11 May 1977 and 02 August 1977.

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Documentation insufficient for assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1977

Report date:

1977

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Sensitisation was induced in guinea pigs by intradermal injections of both test substance and Freunds Complete Adjuvant and the induction process supplemented seven days later by test substance applied to the shoulder injection sites under occlusion. Fourteen days later, the animals were challenged by occluded patch. Further challenges were made at weekly intervals as required.

GLP compliance:

no

Remarks:

Study pre-dates GLP

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2,4-dimethyl-6-phenyl-3,6-dihydro-2H-pyran and 2-methyl-4-methylene-6-phenyltetrahydro-2H-pyran and 4,6-dimethyl-2-phenyl-3,6-dihydro-2H-pyran

Molecular formula:

C13H160

IUPAC Name:

Reaction mass of 2,4-dimethyl-6-phenyl-3,6-dihydro-2H-pyran and 2-methyl-4-methylene-6-phenyltetrahydro-2H-pyran and 4,6-dimethyl-2-phenyl-3,6-dihydro-2H-pyran

Test material form:

other: Liquid

Details on test material:

Name: Pelargene
Chemical name: 2,4-dimethyl-6-phenyldihydropyran
Appearance: Colourless liquid

In vivo test system

Test animals

Species:

guinea pig

Strain:

not specified

Sex:

male/female

Details on test animals and environmental conditions:

Weight - Approximately 320 g

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

no data

Induction 2

Concentration / amount:

Induction (intradermal injection): 0.5 %
Induction (covered patch application): 15 %
Challenge (covered patch application): 5 %

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

no data

Challenge 2

Concentration / amount:

Induction (intradermal injection): 0.5 %
Induction (covered patch application): 15 %
Challenge (covered patch application): 5 %

No. of animals per dose:

10 test animals

Details on study design:

Preliminary irritation tests were performed in guinea pigs to determine concentrations of test substance suitable for induction of sensitisation and for sensitisation challenge. Guinea pigs are then treated by intradermal injection in the shoulder region to induce sensitisation and seven days later the sensitisation is boosted by an occluded patch placed over the injection site. 14 days later the animals are challenged on the flank by occluded patch, with further challenges made at weekly or greater intervals as required.

The method is intended to follow as closely as possible that described by Magnusson and Kligman, except that 10 test animals. 4 treated controls and 4 untreated controls were used.
For each of the ten test animals six 0.1 mL intradermal injections were made close together within a 2 x 4 cm area of the shoulder region as follows:
Two injections of test substance in solvent at the chosen intradermal injection concentration,
Two injections of test substance at the chosen intradermal injection concentration in 50 % Complete Freunds Adjuvant in saline.
Two injections of 50 % Complete Freunds Adjuvant in saline.
Seven days later, sensitisation was boosted by placing over the shoulder injections site a 2 x 4 cm filter paper patch saturated with test substance at the selected concentration. The patch was occluded with Blendtherm and held in place by Poroplast, and was left in place for 48 hours.
Fourteen days after application of the shoulder patch, the guinea pigs were challenged on the flank by occluded patch. For each animal an 8 mm diameter filter paper patch in a patch test cup was saturated with test solution and the cup applied to the shaved flank. It was held in place by Poroplast wound around the trunk. 24 hours later, the patch was removed and the reaction site examined 24 and 248 hours after removal of the patch.

Scoring
Reactions were examined under constant artificial daylight 24 hours and 48 hours after removal of the challenge patches and are scored on a scale from 0 to +++. Reactions were considered positive if they were + or greater and there was no irritation reactions in controls.

Challenge controls:

Treated controls
Four guinea pigs of the same sex and weighing approximately 320 g were treated exactly as for the test animals, except that the test substance was omitted from the intradermal injection and covered patch induction procedures. The animals were challenged exactly as for the test animals.

Untreated controls
At each challenge four previously untreated animals of the same sex and weighing the same as the test animals were “challenged” as for the test animals.

Positive control substance(s):

no

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

5 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

One animal died during the study.

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: One animal died during the study..

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

5 %

No. with + reactions:

0

Total no. in group:

9

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: 9.0.

Results 3

Reading:

rechallenge

Hours after challenge:

24

Group:

test chemical

Dose level:

5 %

No. with + reactions:

1

Total no. in group:

9

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 5 %. No with. + reactions: 1.0. Total no. in groups: 9.0.

Results 4

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

5 %

No. with + reactions:

1

Total no. in group:

9

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 5 %. No with. + reactions: 1.0. Total no. in groups: 9.0.

Results 5

Reading:

other: Challenge 3

Hours after challenge:

24

Group:

test chemical

Dose level:

2.5 %

No. with + reactions:

0

Total no. in group:

9

Remarks on result:

other: Reading: other: Challenge 3. . Hours after challenge: 24.0. Group: test group. Dose level: 2.5 %. No with. + reactions: 0.0. Total no. in groups: 9.0.

Results 6

Reading:

other: Challenge 3

Hours after challenge:

48

Group:

test chemical

Dose level:

2.5 %

No. with + reactions:

0

Total no. in group:

9

Remarks on result:

other: Reading: other: Challenge 3. . Hours after challenge: 48.0. Group: test group. Dose level: 2.5 %. No with. + reactions: 0.0. Total no. in groups: 9.0.

Results 7

Reading:

other: Challenge 3

Hours after challenge:

24

Group:

test chemical

Dose level:

5 %

No. with + reactions:

1

Total no. in group:

9

Remarks on result:

other: Reading: other: Challenge 3. . Hours after challenge: 24.0. Group: test group. Dose level: 5 %. No with. + reactions: 1.0. Total no. in groups: 9.0.

Results 8

Reading:

other: Challenge 3

Hours after challenge:

48

Group:

test chemical

Dose level:

5 %

No. with + reactions:

1

Total no. in group:

9

Remarks on result:

other: Reading: other: Challenge 3. . Hours after challenge: 48.0. Group: test group. Dose level: 5 %. No with. + reactions: 1.0. Total no. in groups: 9.0.

Results 9

Reading:

other: Challenge 4

Hours after challenge:

24

Group:

test chemical

Dose level:

2.5 %

No. with + reactions:

0

Total no. in group:

9

Remarks on result:

other: Reading: other: Challenge 4. . Hours after challenge: 24.0. Group: test group. Dose level: 2.5 %. No with. + reactions: 0.0. Total no. in groups: 9.0.

Results 10

Reading:

other: Challenge 4

Hours after challenge:

48

Group:

test chemical

Dose level:

2.5 %

No. with + reactions:

0

Total no. in group:

9

Remarks on result:

other: Reading: other: Challenge 4. . Hours after challenge: 48.0. Group: test group. Dose level: 2.5 %. No with. + reactions: 0.0. Total no. in groups: 9.0.

Results 11

Reading:

other: Challenge 4

Hours after challenge:

24

Group:

test chemical

Dose level:

5 %

No. with + reactions:

0

Total no. in group:

9

Remarks on result:

other: Reading: other: Challenge 4. . Hours after challenge: 24.0. Group: test group. Dose level: 5 %. No with. + reactions: 0.0. Total no. in groups: 9.0.

Results 12

Reading:

other: Challenge 4

Hours after challenge:

48

Group:

test chemical

Dose level:

5 %

No. with + reactions:

0

Total no. in group:

9

Remarks on result:

other: Reading: other: Challenge 4. . Hours after challenge: 48.0. Group: test group. Dose level: 5 %. No with. + reactions: 0.0. Total no. in groups: 9.0.

Any other information on results incl. tables

None out of 10 guinea pigs tested were sensitised after four challenges.

Pelargene is probably not a sensitiser, although two challenges at 5 % suggest sensitisation, this response was not consistent.

Guinea pigs			Challenge 1		Challenge 2		Challenge 3				Challenge 4
No.	Ear No.	Sex	5 %		5 %		2.5 %		5 %		2.5 %		5 %
			24 h	48 h	24 h	48 h	24 h	48 h	24 h	48 h	24 h	48 h	24 h	48 h
Test Substance
1	4459	F	±	±	±	+	0	0	0	±	?0	0	0	0
2	4268	F	0	0	±	0	0	±	0	±	?0	0	0	0
3	R4167	F	0	0	±	±	0	0	0	0	?0	0	?0	0
4	4622	F	0	0	0	0	0	0	±	±	0	0	0	0
5	4106	F	?0	±	?0	0	0	0	0	0	±	0	0	0
6	4421	F	0	0	0	0	0	0	0	0	0	0	0	0
7	5695	M	0	0	±	0	0	0	?0	0	?0	0	0	0
8	4625	M	±	±	+	+ / ++	0	0	+	+ / ++	?0	0	0	0
9	4690	M
10	4224	M	?0	0	±	±	0	?0	0	0	±	?0	0	0
Treated Controls
1	R599	F	0	±
2	4280	F	?0	0
3	4402	F	0	0
4	4255	F	0	±
Untreated Controls
1	4353	M	0	0
2	4436	M	0	0
3	R4187	M	?±	?±
4	4094	M	±	±
1	4513	F			0	?0
2	4475	F			±	±
3	3282	F			0	0
4	4726	F			?±	?±
1	5551	M					0	0	0	0
2	5274	M					0	0	0	0
3	5164	M					0	0	0	?0
4	5051	M					0	0	0	0
1	4956	M									0	0	0	0
2	5425	M									0	0	0	0
3	5344	M									0	0	?0	0
4	5566	M									0	0	0	0

0 = No reaction

±= barely perceptible erythema

+= Scattered, mild erythema (faint pink)

++ = Moderate and diffuse erythema (pale pink)

+++ = Intense erythema (deep pink) and oedema

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

None of the 10 guinea pigs tested were sensitised after four challenges.

Executive summary:

The skin sensitisation potential of the test substance was assessed. None of the 10 guinea pigs tested were sensitised after four challenges.