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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-Guideline study

Data source

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

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)
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Dichloranilin, rein
- Physical state: solid, liquid after melting at 80°C
- Analytical purity: 99.2 %
- Purity test date: 1989-05-17
- Lot/batch No.: 243
- Expiration date of the lot/batch: 1990-03-23
- Stability under test conditions: tested and approved
- Storage condition of test material: dark, 21- 26°C

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 VI
- 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)

Inductionopen allclose all
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
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

Challengeopen allclose all
Route:
epicutaneous, occlusive
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
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

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

Study design: in vivo (LLNA)

Statistics:
no data

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
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
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 9.0. Total no. in groups: 10.0. Clinical observations: 9 animals with discrete or patchy erythema.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
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
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 20.0. Total no. in groups: 20.0. Clinical observations: 15 animals with moderate to confluent erythema; 5 animals with discrete or patchy 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
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 50 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 7.0. Total no. in groups: 10.0. Clinical observations: 7 animals with discrete or patchy erythema.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
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
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 50 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 18.0. Total no. in groups: 20.0. 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:
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
Remarks on result:
other: see Remark
Remarks:
Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 25 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 8.0. Total no. in groups: 10.0. Clinical observations: 7 animals with discrete or patchy erythema, 1 animal with moderate and confluent erythema.
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
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
Remarks on result:
other: see Remark
Remarks:
Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 25 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 19.0. Total no. in groups: 20.0. Clinical observations: 13 animals with discrete or patchy erythema, 6 animals with moderate and confluent 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
Remarks on result:
other: see Remark
Remarks:
Reading: rechallenge. . Hours after challenge: 72.0. Group: negative control. Dose level: 25 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 5.0. Total no. in groups: 10.0. Clinical observations: 5 animals with discrete or patchy erythema.
Reading:
rechallenge
Hours after challenge:
72
Group:
test group
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
Remarks on result:
other: see Remark
Remarks:
Reading: rechallenge. . Hours after challenge: 72.0. Group: test group. Dose level: 25 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 17.0. Total no. in groups: 20.0. Clinical observations: 17 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
Remarks on result:
other: see Remark
Remarks:
Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 5 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: 1 animal with dicrete or patchy erythema.
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
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
Remarks on result:
other: see Remark
Remarks:
Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 5 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 14.0. Total no. in groups: 20.0. Clinical observations: 14 animals with discrete or patchy erythema.
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
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 72.0. Group: negative control. Dose level: 5 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.
Reading:
rechallenge
Hours after challenge:
72
Group:
test group
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
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 72.0. Group: test group. Dose level: 5 % 3,4-dichloroaniline in propylene glycol. No with. + reactions: 4.0. Total no. in groups: 20.0. Clinical observations: 4 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

Interpretation of results:
sensitising
Remarks:
Migrated information
Conclusions:
Classification: sensitising
Executive summary:

Diesing, L, 1990

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 categorised as sensitising.