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Diss Factsheets

Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 FEB 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
please see attached justification statement
Cross-reference
Reason / purpose for cross-reference:
read-across source
Reference

Key, guinea pig maximisation test, read across to 3,4 dichloroaniline, positive


Supp, QSAR, Danish QSAR database, battery model, positive

Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

Guinea pig maximisation Test - Read across to 3,4 -dichloroaniline


3,4 -dichloroaniline was tested for skin sensitisation according to OECD Guideline 406 in a Magnusson and Kligman test in male guinea pigs. Although irritation was also evident in the control group after challenge or rechallenge with either 50, 25 or 5 % 3,4 -dichloroaniline in propylene glycol (w/w) the incidence and severity of irritation was always higher in the induced test group. Therefore 3,4 -dichloroaniline was concluded to be sensitising in this guinea pig maximisation test. Based on the read across approach, this result is also fully applicable to the registered substance 2,4 -dichloroaniline (for more information please refer to the Read Across Statement attached in Section 13).


QSAR prediction


Skin sensitisation was predicted using the models Sci QSAR and CASE ULtra implemented in the Danish data base and the OECD QSAR toolbox v4.5. Positive indication of skin sensitisation was given by these QSAR predictions. The final result was drawn from the overall prediction of the battery model that contains 3 different models, for skin sensitisation implemented in the Danish QSAR database. For the Sci QSAR model and the CASE Ultra QSAR model the substance was predicted positive and the predictions fall in the applicability domain. The third model of the battery (Leadscope) could not make a prediction, as the substance was out of the applicability domain of the model.

Endpoint conclusion:
no study available

Based on the available information, the registered substance is classified as skin sens 1 (H317) in accordance with Regulation (EC) No 1272/2008.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
12 May 1981
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The study was performed before the LLNA was established as the standard method for skin sensitisation.

Test material

Constituent 1
Reference substance name:
3,4-dichloroaniline
Cas Number:
95-76-1
Molecular formula:
C6H5Cl2N
IUPAC Name:
3,4-dichloroaniline

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: DHPW, SPF
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: 5 to 8 weeks
- Weight at study initiation: 313 to 428 g
- Housing: 5 animals per Macrolon cage type IV
- Diet: ad libitum, Altromin 3022
- Water: ad libitum, tap water
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): approx. 50
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
propylene glycol
Concentration / amount:
intradermal induction 2.5 % (w/w) 3,4-dichloroaniline in propylene glycol
epicutaneous induction 50 % (w/w) 3,4-dichloroaniline in propylene glycol
Day(s)/duration:
day 0 intradermal induction
day 7 epicutaneous induction for 48 h
Challenge
Route:
epicutaneous, occlusive
Vehicle:
propylene glycol
Concentration / amount:
epicutaneous challenge 50 % (w/w) 3,4-dichloroaniline in propylene glycol
epicutaneous rechallenge 25 % (w/w) 3,4-dichloroaniline in propylene glycol
epicutaneous rechallenge 5 % (w/w) 3,4-dichloroaniline in propylene glycol
Day(s)/duration:
3 and 4 weeks after intradermal induction
No. of animals per dose:
two individual control groups: 10 animals each
one test group: 20 animals
Details on study design:

RANGE FINDING TESTS: yes, one female guinea pig received intra dermal injections of 0, 1, 2.5 and 5 % 3,4-dichloroaniline in propylene glycol (w/w), all injection sites displayed redness after 24 and 48 h, the 5 % (w/w) injection site also became encrusted after 48 h. For the range finding for epicutaneous induction 4 female guineapigs received 3 occlusive gauze patchs with either 6, 12 or 25 % 3,4-dichloroaniline in propylene glycol for 24 h. No redness was observed after 48 and 72 h. 4 additional female guinea pigs then received occlusive gauze patchs with 50 % 3,4-dichloroaniline in propylene glycol for 24 h. Again no local irritation was observed after 48 and 72 h. Therefore the animals were pretreated with 10 % SDS in paraffine oil one day prior to epicutaneous induction to create a local irritation.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous)
- Exposure period: day 0 intradermal induction
day 7 epicutaneous induction for 48 h
- Control groups:
day 0: 1st injection, cranial, 0.1 mL, FCA : 0.9 % saline solution= 1 : 1
2nd injection, medial, 0.1 mL, propylene glycol
3rd injection, caudal, 0.1 mL, FCA : propylene glycol (w/w)= 1 : 1
day 6: epicutaneous treatment of the same areas with 0.2 mL 10 % SDS in paraffine oil
day 7: 3 occlusive gauze patches (2 x 4 cm) treated with 0.5 mL propylene glycol
- Test group:
day 0: 1st injection, cranial, 0.1 mL, FCA : 0.9 % saline solution= 1 : 1
2nd injection, medial, 0.1 mL, 2.5 % 3,4-dichloroaniline in propylene glycol (w/w)
3rd injection, caudal, 0.1 mL, FCA : 2.5 % 3,4-dichloroaniline in propylene glycol (w/w)= 1 : 1
day 6: epicutaneous treatment of the same areas with 0.2 mL 10 % SDS in paraffine oil
day 7: 3 occlusive gauze patches (2 x 4 cm) treated with 0.5 mL 50 % 3,4-dichloroaniline in propylene glycol (w/w)

B. CHALLENGE EXPOSURE
- No. of exposures: 3 ( epicutaneous)
- Day(s) of challenge: 3 and 4 weeks after intradermal induction
- Exposure period: 24 h
- Test groups:
challenge: 21st day, 1 occlusive gauze patch (2 x 4 cm) at the left flank treated
with 0.5 mL 50 % 3,4-dichloroaniline in propylene glycol (w/w)
rechallenge 28th day, 2 occlusive gauze patches (2 x 4 cm) at the left flank treated
with either 0.5 mL 25 % or 5 % 3,4-dichloroaniline in propylene glycol (w/w)

- Control group:
1st control group: 21st day, 1 occlusive gauze patch (2 x 4 cm) at the left flank treated
with 0.5 mL 50 % 3,4-dichloroaniline in propylene glycol (w/w)
2nd control group 28th day, 2 occlusive gauze patches (2 x 4 cm) at the left flank treated
with either 0.5 mL 25 % or 5 % 3,4-dichloroaniline in propylene glycol (w/w)

- Site: left flank challenge, right flank control (gauze patch treated with propylene glycol alone)

- Evaluation (hr after challenge): 48 and 72 h after challenge according to OECD Guideline 406
Challenge controls:

see details on study design
Positive control substance(s):
yes
Remarks:
routinely checked by the testing laboratory, no further data

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
rechallenge
Hours after challenge:
72
Group:
negative control
Dose level:

5 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
0
Total no. in group:
10
Clinical observations:

none
Reading:
rechallenge
Hours after challenge:
72
Group:
test chemical
Dose level:

5 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
4
Total no. in group:
20
Clinical observations:

4 animals with discrete or patchy erythema
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:

5 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
1
Total no. in group:
10
Clinical observations:

1 animal with dicrete or patchy erythema
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:

5 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
14
Total no. in group:
20
Clinical observations:

14 animals with discrete or patchy erythema
Reading:
rechallenge
Hours after challenge:
72
Group:
negative control
Dose level:

25 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
5
Total no. in group:
10
Clinical observations:

5 animals with discrete or patchy erythema
Reading:
rechallenge
Hours after challenge:
72
Group:
test chemical
Dose level:

25 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
17
Total no. in group:
20
Clinical observations:
17 animals with discrete or patchy erythema
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:

25 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
8
Total no. in group:
10
Clinical observations:

7 animals with discrete or patchy erythema, 1 animal with moderate and confluent erythema
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:

25 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
19
Total no. in group:
20
Clinical observations:

13 animals with discrete or patchy erythema, 6 animals with moderate and confluent erythema
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:

50 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
7
Total no. in group:
10
Clinical observations:

7 animals with discrete or patchy erythema
Reading:
2nd reading
Hours after challenge:
72
Group:
test chemical
Dose level:

50 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
18
Total no. in group:
20
Clinical observations:

11 animals with discrete or patchy erythema, 4 animals with moderate and confluent erythema and 3 animals with intense erythema and swelling
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:

50 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
9
Total no. in group:
10
Clinical observations:

9 animals with discrete or patchy erythema
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:

50 % 3,4-dichloroaniline in propylene glycol
No. with + reactions:
20
Total no. in group:
20
Clinical observations:

15 animals with moderate to confluent erythema; 5 animals with discrete or patchy erythema

Any other information on results incl. tables


During the range finding test for the challenge concentration acute dermal toxicity was observed after simultaneous exposure of each of five guinea pigs to 4 occlusive gauze patches treated with either 0.5 mL 6, 12, 25 or 50 % 3,4 -dichloroaniline in propylene glycol. Two animals were found dead after 24 h and the remaining three were euthanized for animal welfare reasons due to severe signs of toxicity.



 

Applicant's summary and conclusion

Conclusions:
3,4 -dichloroaniline was found to be sensitising in this guinea pig maximisation test.
Executive summary:

3,4 -dichloroaniline was tested for skin sensitisation according to OECD Guideline 406 in a Magnusson and Kligman test in male guinea pigs. Although irritation was also evident in the control group after challenge or rechallenge with either 50, 25 or 5 % 3,4 -dichloroaniline in propylene glycol (w/w) the incidence and severity of irritation was always higher in the induced test group. Therefore 3,4 -dichloroaniline was concluded to be sensitising in this guinea pig maximisation test.