Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989-07-18 to 1989-08-11
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 guideline
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
Cited as Directive 84/449/EEC, B.6
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Constituent 1
Chemical structure
Reference substance name:
2-chloroaniline
EC Number:
202-426-4
EC Name:
2-chloroaniline
Cas Number:
95-51-2
Molecular formula:
C6H6ClN
IUPAC Name:
2-chloroaniline
Details on test material:
- Name of test material (as cited in study report): o-Chloraniline
- Physical state: yellow-orange, clear liquid
- Analytical purity: 99.6 % (IR spectroscopy)
- Impurities (identity and concentrations):
<0.01 % cyclohexylamine
<0.01 % N-isopropylaniline
<0.01 % 2-chloro-N-isopropylaniline
0.08% aniline
<0.01% 2-isopropylaniline
<0.01% o-nitrochlorobenzene
0.24% p-chloroaniline
<0.01% m-chloroaniline
<0.01% 2,4-dichloroaniline
<0.01% 2,5-dichloroaniline
<0.01% 2,3-dichloroaniline
<0.01% 2,3-dichloroaniline
<0.01% o-phenylendiamine
0.03 % unknown impurities
- Purity test date: 1 Feb 1988
- Lot/batch No.: Lagerkessel 15/ 594 488
- Stability under test conditions: stable and homogen throughout the test period
- Storage condition of test material: 3-7°C, dark

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Bor:DHPW
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: 5 - 7 weeks
- Weight at study initiation: 310-395 g
- Housing: 5 animals per Macrolon cage type IV on wood chips (Ssniff), cleaned 4 times a week
- Diet: ad libitum, Altromin 3020
- Water: ad libitum, tap water
- Acclimation period: 7 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): ~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: 5 % (w/v)
epicutaneous induction: 100 %
epicutaneous challenge: 100 %
Challengeopen allclose all
Route:
epicutaneous, semiocclusive
Vehicle:
propylene glycol
Concentration / amount:
intradermal induction: 5 % (w/v)
epicutaneous induction: 100 %
epicutaneous challenge: 100 %
No. of animals per dose:
test group: 20 male randomized guinea pigs
control group 1: 10 male randomized guinea pigs
control group 2: 10 male randomized guinea pigs
Details on study design:
RANGE FINDING TESTS:
intradermal: 1-5 % o-chloroaniline (w/v), injection site showed slight irritation after 48 h
epicutaneous: yes, no irritation was observed at any concentration (12, 25, 50 and 100 % (w/v) tested after 24 h


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/v)= 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)
- Test group:
day 0: 1st injection, cranial, 0.1 mL, FCA : 0.9 % saline solution= 1 : 1
2nd injection, medial, 0.1 mL, 5 % o-chloroaniline in propylene glycol (w/v)
3rd injection, caudal, 0.1 mL, FCA : 5 % o-chloroaniline in propylene glycol (w/v)= 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 100 % o-chloroaniline

B. CHALLENGE EXPOSURE
- No. of exposures: 3 ( epicutaneous)
- Day(s) of challenge: 3 weeks after intradermal induction
- Exposure period: 24 h
- Test groups:
challenge: 21st day, 1 semiocclusive gauze patch (2 x 4 cm) at the left flank treated
with 0.5 mL 100 % o-chloroaniline


- Control group:
1st control group: 21st day, 1 occlusive gauze patch (2 x 4 cm) at the left flank treated
with 0.5 mL 100 % o-dichloroaniline


- Site: left flank challenge, right flank control (gauze patch)

- Evaluation (hr after challenge): 48 and 72 h after challenge according to OECD guideline 406
Challenge controls:
yes
Positive control substance(s):
not required

Results and discussion

Positive control results:
no

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100 % . No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
72
Group:
test chemical
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 100 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 100 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 100 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Any other information on results incl. tables

No influence on body weight gain was observed. After challenge skin sensitisation was observed neither in the control nor in the test group.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
Classification: not sensitizing
Executive summary:

Diesing, L, 1990

o-Chloroaniline was tested for skin sensitisation according to OECD guideline 406 in a Magnusson and Kligman test in male guinea pigs. No irritation was evident in the control group and test group after challenge with 100 % o-chloroaniline. Therefore o-chloroaniline was categorised as not sensitising.