Phenol, 4,4'-(1-methylethylidene)bis-, polymer with (chloromethyl)oxirane, reaction products with diethylenetriamine and 4-methyl-2-pentanone in xylene and butanol

Skin sensitisation

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:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

Tested according to GPMT as described by B. Magnussen and A.M. Kligman (allergic contact dermatitits in the guinea pig. Publ. Ch.C. Tomas: Springfiled, Illinois, USA, 1970)

Deviations:

not applicable

GLP compliance:

no

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The LLNA method was not adopted at the time of testing (1987)

Test material

Test material information

Constituent 1

Reference substance name:

Phenol, 4,4'-(1-methylethylidene)bis-, polymer with (chloromethyl)oxirane, reaction products with diethylenetriamine and 4-methyl-2-pentanone in xylene and butanol

EC Number:

947-984-6

Molecular formula:

The substance is a UVCB, of which a major component can be represented by the following: C28H46ClN3O4

IUPAC Name:

Phenol, 4,4'-(1-methylethylidene)bis-, polymer with (chloromethyl)oxirane, reaction products with diethylenetriamine and 4-methyl-2-pentanone in xylene and butanol

Constituent 2

Reference substance name:

Phenol, 4,4'-(1-methylethylidene)bis-, oligomer with (chloromethyl)oxirane, reaction products with diethylenetriamine and 4-methyl-2-pentanone, n=2

IUPAC Name:

Phenol, 4,4'-(1-methylethylidene)bis-, oligomer with (chloromethyl)oxirane, reaction products with diethylenetriamine and 4-methyl-2-pentanone, n=2

Constituent 3

Reference substance name:

1,2-xylene; 1,3-xylene; 1,4-xylene; ethylbenzene

EC Number:

905-588-0

Molecular formula:

C32H40

IUPAC Name:

1,2-xylene; 1,3-xylene; 1,4-xylene; ethylbenzene

Constituent 4

Reference substance name:

Butan-1-ol

EC Number:

200-751-6

EC Name:

Butan-1-ol

Cas Number:

71-36-3

Molecular formula:

C4H10O

IUPAC Name:

butan-1-ol

Test material form:

liquid: viscous

In vivo test system

Test animals

Species:

guinea pig

Strain:

other:

Remarks:

albino (DHPW)

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: F. Winkelmann, Instutute ofr Breeding of LAboratory Animals GmbH and Co. KG, Borchen, Germany
- Females nulliparous and non-pregnant: not specified
- Microbiological status of animals, when known: not specified
- Age at study initiation: not specified
- Weight at study initiation: 330-390 gram (male) and 280-375 gram (female)
- Housing: The animal were kept individually under conventional conditions un suspended, stainless steel cages, fitted with wire mesh floor and front
- Diet: Pelleted, natural ingredient diet, ad libitum
- Water: Unfluorinated tap water, ad libitum
- Acclimation period: 8 days
- Indication of any skin lesions: No visible skin abnormalities observed

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18±2 ºC
- Humidity (%): 40-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12
- IN-LIFE DATES: From: Marc 18, 1987 To: April 11, 1987

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

propylene glycol

Concentration / amount:

10% dilution (v/v)

Day(s)/duration:

7

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Induction 2

Route:

intradermal

Vehicle:

other: FCA and PG (1:1)

Concentration / amount:

10% dilution (v/v)

Day(s)/duration:

7

Adequacy of induction:

not specified

Induction 3

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

25% mixture (w/w)

Day(s)/duration:

2

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

10% mixture (w/w)

Day(s)/duration:

1 and 2

Adequacy of challenge:

not specified

No. of animals per dose:

20 (10 males and 10 females)

Details on study design:

RANGE FINDING TESTS:
The irritation response to intradermal injection of various concentrations was examined in 3 guinea pigs. A sufficiently large area of the scapular region was clipped free from hair with electric clippers. Subsequeantly 0.1 mL portions of 5, 10 and 25% (v/v) dilutions of the test substance in propylene glycol, were applied to each animal by intradermal injection.

The irritation response to topical treatment was tested in 2 guinea pigs were clipped free from hair with electric clippers. Subsequantly several 2x2 pacthes of filter paper (Whatman N0. 3 MM) were loaded with different concentrations of the test substance in vasilne, viz. 10 or 25% (w/w)/ The patched were placed on the clipped skin and covered with a piece of PVC foil and hypo-allergenic paper bandage (Leukopor) which was then secured by elastic adhesive bandage (Tensoplast), 7.5 cm in width, wound aroun the torso of the animal. two concentrations were tested per animal. The dressing was left in place for 24 h. Then the animal were examined for signs of irritation.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: two, one intradermal and one topical
- Exposure period: the topical application was performed 7 days after the intradermal injections, the dressing was removed after 2 days, the resting period after the topical applicatin was 14 days
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: scapular region
- Frequency of applications: one intradermal and one topical
- Duration: the topical application was performed 7 days after the intradermal injections, the dressing was removed after 2 days, the resting period after the topical applicatin was 14 days
- Concentrations: 10% (v/v) for intradermal injections and 25% (w/w) for topical application

B. CHALLENGE EXPOSURE
- No. of exposures: one
- Day(s) of challenge: one
- Exposure period: 24 h
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: scapular region
- Concentrations: 10% (w/w)
- Evaluation (hr after challenge): 24 and 48

Positive control substance(s):

no

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

10% (w/w)

No. with + reactions:

6

Total no. in group:

20

Clinical observations:

All animal remained in good health during the experimental period

Remarks on result:

positive indication of skin sensitisation

Results 2

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

10 % (w/w)

No. with + reactions:

2

Total no. in group:

20

Clinical observations:

All animal remained in good health during the experimental period

Remarks on result:

positive indication of skin sensitisation

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

10% (w/w)

No. with + reactions:

2

Total no. in group:

10

Clinical observations:

All animal remained in good health during the experimental period

Remarks on result:

other:

Remarks:

Since, only very slight skin reactions were observed in controles these observations are considered to be due to iritation rather than to sensitization.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

10 % (w/w)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

All animal remained in good health during the experimental period

Results 5

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

NA

No. with + reactions:

0

Total no. in group:

0

Clinical observations:

NA

Remarks on result:

other: no positive control was added in the study design.

Results 6

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

NA

No. with + reactions:

0

Total no. in group:

0

Clinical observations:

NA

Remarks on result:

other: no positive control was added in the study design.

Any other information on results incl. tables

Dermal reactions elicited by the challenge application of 10% test substance in vaseline

Animal number	Scores at 24 and 48 h after removal of dressing
	Males				Animal number	Females
	24 h		48 h			24 h		48 h
	ER	ED	ER	ED		ER	ED	ER	ED
Control group
7400	0	0	0	0	7401	0	0	0	0
7402	0	0	0	0	7403	0	0	0	0
7404	1	0	0	0	7405	1	0	0	0
7406	0	0	0	0	7407	0	0	0	0
7408	0	0	0	0	7409	0	0	0	0
Test group
7380	2	2	2	2	7381	1	0	1	1
7382	2	1	2	1	7383	1	1	1	1
7384	1	1	0	0	7385	1	1	0	0
7386	2	1	1	1	7387	1	1	0	0
7388	1	1	1	1	7389	0	0	0	0
7390	0	0	0	0	7391	2	2	0	0
7392	0	0	0	0	7393	0	0	0	0
7394	1	1	0	0	7395	2	2	1	1
7396	1	1	1	1	7397	2	2	1	1
7398	1	1	0	0	7399	1	0	1	0

ER = erythema ED = oedema

Individual skin reactions observed during the induction phase of the study

Animal number	Weight (g)	Induction period
		Initial score at the sites of intradermal injections																		Topical application
		al			ar			bl			br			cl			cr
		D	C	H	D	C	H	D	C	H	D	C	H	D	C	H	D	C	H	ER	ED
Control group - males
7400	355	7	P	2	7	P	1	4	PA	0	5	PA	0	7	PA	2	8	PA	2	0	0
7402	355	7	P	2	8	P	2	4	PA	0	5	PA	0	8	PA	2	7	PA	1	0	0
7404	356	6	P	2	6	P	1	5	PA	0	5	PA	0	7	PA	2	6	PA	1	0	0
7406	360	7	P	2	5	P	1	5	PA	0	6	PA	0	4	PA	1	4	PA	1	0	0
7408	385	8	P	2	7	P	2	5	PA	0	7	PA	0	4	PA	1	-			0	0
Control group - females
7401	330	8	P	2	7	P	2	8	PA	0	5	PA	0	9	PA	2	6	PA	1	0	0
7403	355	8	P	2	8	P	2	5	PA	0	5	PA	0	8	PA	2	8	PA	2	0	0
7405	345	-			7	P	2	5	PA	0	4	PA	0	8	PA	2	5	PA	2	0	0
7407	295	7	P	2	7	P	2	6	PA	0	-			5	PA	2	7	PA	2	0	0
7409	350	8	P	2	9	P	2	5	PA	0	-			9	PA	2	8	PA	2	0	0
Test group – males
7380	363	8	P	2	8	P	2	7	RA	0	6	RA	0	9	RA	1	9	RA	1	0	0
7382	330	8	P	2	8	P	2	5	PA	0	7	RA	0	9	RA	1	10	RA	1	0	0
7384	375	7	P	2	8	P	2	8	RA	0	7	RA	0	9	RA	1	10	RA	1	0	0
7386	345	7	P	2	-			7	PA	0	8	RA	0	9	RA	1	9	RA	1	0	0
7388	380	9	P	2	7	P	2	6	PA	0	7	RA	0	8	RA	1	6	RA	1	0	0
7390	380	8	P	2	7	P	2	7	RA	0	7	RA	0	9	RA	1	9	RA	1	0	0
7392	390	8	P	2	8	P	2	7	RA	0	6	RA	0	9	RA	1	9	RA	1	0	0
7394	380	8	P	2	8	P	2	7	RA	0	7	RA	0	10	RA	1	0	RA	1	0	0
7396	355	8	P	2	7	P	2	6	RA	0	7	RA	0	9	RA	1	10	RA	1	0	0
7398	385	7	P	2	8	P	2	6	RA	0	7	RA	0	7	RA	1	8	RA	1	0	0
Test group – females
7381	310	5	P	2	7	P	2	7	RA	0	8	RA	0	-			11	RA	1	0	0
7383	305	8	P	2	8	P	2	7	RA	0	9	RA	0	9	RA	0	8	RA	0	0	0
7385	280	8	P	2	-			6	RA	0	8	RA	0	9	RA	1	5	RA	0	0	0
7387	320	7	P	2	8	P	2	7	RA	0	7	RA	0	6	RA	1	6	RA	1	0	0
7389	365	8	P	2	8	P	2	7	RA	0	7	RA	0	8	RA	1	9	RA	1	0	0
7391	320	7	P	2	7	P	2	8	RA	0	7	RA	0	10	RA	1	9	RA	1	0	0
7393	350	8	P	2	8	P	2	7	RA	0	7	RA	0	9	RA	1	9	RA	1	0	0
7395	375	7	P	2	6	P	2	7	RA	0	7	RA	0	9	RA	1	10	RA	1	0	0
7397	310	8	P	2	8	P	2	7	RA	0	7	RA	0	9	RA	1	9	RA	1	0	0
7399	365	7	P	2	6	P	2	8	RA	0	7	RA	0	7	RA	1	10	RA	1	0	0

 

al and ar: FCA

bl and br: PG

cl and cr: PC and FCA (1:1)

-              = not scored

D = diameter of skin reaction

H = height of skin reaction

C = color of skin reaction (PP = pale pink; P = pink; R = red; A = abscess; N = necrosi

Applicant's summary and conclusion

Interpretation of results:

Category 1B (indication of skin sensitising potential) based on GHS criteria

Conclusions:

From the results of the challenge treatment with the 10% dilution it appeared that the test substance induced skin reactions in 16 out of 20 test animals, while also in 2 out of 10 controls showed dermal reactions. Since, only very slight skin reactions were observed in controls the very slight skin reactions seen in the test animals are considered due to irritation rather than the senisitzation. However, the more severe reactions observed in 6 out of 20 animals are possible due to sensitization. Therefore, the test substance is considered to be a moderate sensitizer.

Executive summary:

The test substance was examined for possible sensitizing properties by a maximization test in guinea pigs using 20 test animals and 10 controls.

The test comprised of:

a. induction treatment by intradermal injection of a 10% dilution of the test substance in propylene glycol, followed one week later by a topical treatment with 25% test substance in vaseline;

b. challenge treatment, 14 days after the last induction, by topical application of 10% test substance in vaseline.

The challengde treatment with the 10% dilution showed evidence of delayed contact hypersensitivity. It was concluded that the test substance is a moderate sensitizer.