p-phenylenebis(methylamine)

Endpoint summary

Description of key information

Skin sensitisation:

One in vivo study (GPMT) is available which was similar to OECD 406 study and resulted the substance is skin sensitizer.

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:

1981

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Qualifier:

no guideline followed

Principles of method if other than guideline:

In vivo skin sensitisation test according to the maximisation test after Magnusson and Kligman.
On the basis of Magnusson, B . and Kligman A .M.: The Identification of Contact Allergens by Animal Assay. The Guinea Pig Maximization Test. J. Invest. Dermatol. 52, 268 - 276 (1969).

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

This study was initiated before the REACH regulation effectived.

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Species: Pirbright White, Dunkin Hartley HOE DHPK (SPF-LAC)
- Source: Lippische Versuchstierzucht, Hagemann GmbH 8c Co . KG, 4923 Extertal 1
- Age at study initiation: not specified
- Weight at study initiation: 307 - 422 g (1. experiment); 343 - 423 g (2. experiment)
- Fasting period before study: not reported
- Housing: Animals were housed 6 to a tage (same sex) in Makrolon-cages, type IV
- Diet: Ssniff GK 4 mm ; standard diet for rabbits and guinea pigs (Firma Ssniff-Versuchstierdiäten GmbH, 4770 Soest)
- Water (e.g. ad libitum): Tap water, ad libitum (twice a week 2 g ascorbic acid in 10 l water)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 30-70%
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12 hours artificial light / 12 hours dark

- Acclimation period: At least 7 days.

RANDOMISATION :
After Salfi, R . : A Long-Period Random Number Generator with Application to Permutation . Compstat 1974, (28 - 35)

Route:

intradermal

Vehicle:

water

Concentration / amount:

10% of test substance

Day(s)/duration:

7

Adequacy of induction:

other: highest concentration used in pretest and minimal concentration to trigger an irritation

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

7 days after intradermal induction application of approx. 0.3 g of a 20% solution of the test substance

Day(s)/duration:

2

Adequacy of induction:

not specified

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

1% and 3% test substance

Day(s)/duration:

1

Adequacy of challenge:

other: non irritant concentration

No.:

#2

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

1% and 3% test substance

Day(s)/duration:

1

Adequacy of challenge:

other: non irritant concentration

No.:

#3

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

1% and 3% test substance

Day(s)/duration:

1

Adequacy of challenge:

other: non irritant concentration

No. of animals per dose:

6 per control group
12 per test group

Details on study design:

Body weight:
- before the application of the test substance and berfore the end of the test

Clipping of the animals:
- If deemed necessary approx. 3 to 5 hours before the readings and application of test substance at the respective testing sites.

Clinical observations:
- no detaiiled investigatins, daily control for illness and general health conditins

Type of application:
- intradermal and percutaneous occlusive

Preparation of the test solution:
- shortly before teh applicatio of the test substance with Ultraturrax and magnetic stirrer

PRETEST:
Applied quantity:
- filterpaper stripes of 2 x 2 cm with approx. 0.15 g test substance solution were applied under occlusion in the region of the flanks
Application time :
- 24 h
Site of application:
- region of the flanks
Number of animals :
- min. 4 per test concentration
Readings :
- after 24, 48, 72 h after start of application

MAIN EXPERIMENT:
INDUCTION:
Intradermal induction :
- 6 intradermal injections in groups of two per animal
- Injektions for the experimental group:
a) first row: 2 injections at 0.1 ml Freund'sches Adjuvans without test substance in water (1 : 1)
b) middle row: 2 injections at 0.1 ml test solution preparation
c) posterior row: 2 injections at 0.1 ml Freund'sches Adjuvans /water (1 : 1) with test substance
Injections for the control groups 1 and 2:
- the animals received the same injections (a, b, c) but without test substance.
- no treatment of control group 3
Site of application:
- region of the shoulder
Readings:
- 24 h after start of application

Percutaneous induction :
- the percutaneous induction took place one week after the intradermal induction.
Applied quantity:
- application of approx. 0.3 g test substance preparation on a 2 x 4 cm filterpaper stripe (method analogue to the pretest)
- no treatment of control group
Application:
- perkutaneous, occlusive
Application time:
- 48 h
Site of application:
- region of the shoulder, same region like for the intradermal application

CHALLENGE :
Test concentration:
- non irritational dose
- first challenge approx. 14 days after the percutaneous application, second challenge after one week and the third challenge 7 days after the second
Applied quantity:
- filterpaper stripes of 2 x 2 cm with approx. 0.15 g test substance solution were applied
Application:
- first challenge: test substance solution was applied to the control group 1 and experimental group. No treatment for control group 2 and 3
- second challenge: test substance solution was applied to the control group 1, 2 and experimental group. No treatment for control group 3
- third challenge: treatment of all controll groups and of the experimental group
Application:
- perkutaneous, occlusive
Application time:
- 24 h
Site of application:
- region of the flanks
Readings:
- ca . 24, 48, 72 h after the start of the application

EVALUATION:
Only the number of sensitised animals was evaluated. The intensity of the found irritation was not evaluated.

To determine the rate of sensitisation the readings after 48 h after application were considered.

Challenge controls:

without any application

Positive control substance(s):

not specified

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

1%

No. with + reactions:

4

Total no. in group:

12

Clinical observations:

skin reddening and some oedemas

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

3%

No. with + reactions:

5

Total no. in group:

12

Clinical observations:

skin reddening and some oedemas

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

1%

No. with + reactions:

0

Total no. in group:

6

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

3%

No. with + reactions:

0

Total no. in group:

6

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Remarks on result:

not measured/tested

Results after the three challenges in the second experiment

	1. Challenge		2. Challenge		3. Challenge
	1% in b	3% in b	1% in b	3% in b	1% in b	3% in b
Control group 1	0/6	0/6	0/6	0/6	0/6	0/6
Control group 2	a	a	0/6	0/6	0/6	0/6
Control group 3	a	a	a	a	0/6	0/6
Experimental group	0/12	0/12	6/12	12/12	4/12	5/12

a: no application of test substance

b: aqua dest.

x/y: number of positive reactions/number of tested animals, 48 h after application

Results of the first experiment not shown, they were judged as non interpretable by the author.

Interpretation of results:

Category 1 (skin sensitising) based on GHS criteria

Conclusions:

Under the conditions of the test it can be concluded that the test substance is a sensitiser to the skin of guinea pigs.

Executive summary:

To test the skin sensitising potential of the test substance an in vivo skin sensitisation test according to the maximisation test after Magnusson and Kligman was performed on guinea pigs. The test was not performed according to a specific OECD guideline, nevertheless it was performed according to a widely used test protocoll and the results can be regarded as valid and reliable. The test was performed under GLP.

Two experiments were carried out. Because the results of the first experiment were not interpretable a second experiment was performed.

After intradermal induction in the experimental groups reddening, oedemas and some necrotic changes of the skin could be observed. After percutaneous induction reddening and bleeding crusts at the application site were reported.

After the first challenge no reactions could be observed.
After the second challenge in the 3% test substance concentration all animals showed skin reddening and some oedemas. In the 1% test substance group 6/12 animals reacted positively. The control animals remained free of any reactions.

Because of the discrepancy of the test results between the first and the second experiment a third challenge was performed. All negativ control animals remained free of any reaction. In the 3% test substance group 5/12 and in the 1% test substance group 4/12 were tested poitive. The results of the second challenge were confirmed with the third challenge results.

The test substance was tested as skin sensitiser in guinea pigs under the condition of the test.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Justification for classification or non-classification

Skin sensitisation:

Positive result in animal test (GPMT).

According to Regulation (EC) No 1272/2008, table 3.4.2, this substance is classified as skin sensitiser Category 1.