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Description of key information

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
other: read across from analogue substance
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
performed on analogue substance
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
LLNA method was not available at the time of testing
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
EST ANIMALS
- Source: Kleintierfarm Madörin, 4414 Füllinsdorf, CH
- Females nulliparous and non-pregnant: yes
- Microbiological status of animals: not specified
- Age at study initiation: 7 to 8 weeks
- Weight at study initiation: males: 383 to 447 g; females: 425 to 486 g
- Housing: individually in Makrolon type-3 cages with standard softwood bedding ("Lignocell", Schill AG, 4132 Muttenz, CH)
- Diet: ad libitum; Pelleted standard Kliba 342, batch 38/87 guinea pig breeding/maintenance diet ("Kliba", Klingentalmühle AG, 4303 Kaiseraugst, CH); subjected to analysis for contaminants (results included in report)
- Water: ad libitum; community tap water from Itingen, subjected to analyses for contaminants (results included in report)
- Acclimation period: one week under test conditions after veterinary health examination
- Indication of any skin lesions: not specified
- Identification: by unique cage number and corresponding ear tags
- Randomisation: randomly selected at time of delivery

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 40 to 70 %
- Air changes: 10 to 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12
Route:
intradermal
Vehicle:
other: 50:50 Freunds' complete adjuvant and physiological saline
Concentration / amount:
0%
Day(s)/duration:
single injection
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
1%
Day(s)/duration:
single injection
Route:
intradermal
Vehicle:
other: 50:50 Freunds' complete adjuvant and physiological saline
Concentration / amount:
1%
Day(s)/duration:
single injection
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
25%
Day(s)/duration:
48 hours
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
left flank: 25%
Right flank: 0%
Day(s)/duration:
24 hours
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
Left flank: 0%
Right flank: 25%
Day(s)/duration:
24h
No. of animals per dose:
Preliminary study: 6 animals (intradermal testing: 1 male and 1 female; epidermal testing: 2 males and 2 females)
Control: 10 animals (5 males, 5 females)
Test: 20 animals (10 males, 10 females)
Total: 36 animals (18 males, 18 females)
Details on study design:
RANGE FINDING TESTS: PRELIMINARY STUDY
- The objective of this investigation was to identify irritant test item concentrations for the induction phase of the main study. In addition, a suitable non-irritant concentration of test item, by topical administration, was identified for the challenge application.
- INTRADERMAL INJECTIONS: 0.1 ml was administrated into the clipped flank of two guinea pigs (1 male and 1 female) at concentrations of 0.1, 0.3, 0.5, 1.0, 3.0 and 5.0 % test item in physiological saline and assessed for dermal reactions (erythema, oedema and diameter in mm) after 24 hours.
- EPIDERMAL APPLICATIONS: 2 x 2 cm patches of filter paper were saturated with concentrations of 3, 5, 10 and 25 % test item in physiological saline and applied to the clipped, shaved flanks of four guinea pigs. The patches were covered by a strip of aluminium foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. The dressings were removed after 24 hours and the reaction sites were assessed for erythema and oedema. Further examinations of the sites were performed 24 and 48 hours after the removal of the dressing.
- RESULTS AND CONCLUSIONS: following intradermal injections, no erythema was recorded at any concentration at either 0 or 24 hours, and oedema was observed in 1 animal at 1.0 % test item, and both animals at concentrations of 3.0 and 5.0 % 24 hours after injections. 1.0 % was the lowest concentration to produce positive reaction. Following epidermal applications, no erythema and no oedema was observed among any animals or any test item concentrations 24 hours after application of the test item. 25 % was the highest concentration of test item tested, and it produced no positive reactions.

MAIN STUDY

A. INDUCTION EXPOSURE

I) INTRADERMAL INJECTIONS: 6 x 8 cm area was clipped free of hair and administered 3 pairs of 0.1 ml intradermal injections per site at the border of a 4 x 4 cm area within the clipped area.
- No. of exposures: 3 pairs of injections
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: clipped dorsal skin from the scapular region
- Frequency of injections: once
- Concentrations: 1st pair: Freunds' complete adjunct 50:50 with physiological saline; 2nd pair: 1 % test item in physiological saline; 3rd pair: 1 % test item in Freunds' complete adjunct 50:50 with physiological saline (control group: 0 % test item in 2nd and 3rd pair of injections).
- Duration: day 1

II) EPIDERMAL APPLICATIONS: one week after the intradermal injections, the same region was re-clipped and a a patch of 4 x 4 cm filter paper was saturated with 25 % test item in physiological saline (control group: 0% test item) and placed over the injection sites on the test animals, covered with aluminium foil and firmly secured by elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape for 48 hours.
- No. of exposures: 1
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: clipped dorsal skin from the scapular region, as for intradermal injections
- Frequency of applications: once
- Exposure period: 48 hours
- Concentrations: 25 %
- Duration: days 8 to 10

B. CHALLENGE EXPOSURES: EPIDERMAL APPLICATIONS
- No. of exposures: 2
- Timing of challenge: two and four weeks after the epidermal applications
- Exposure periods: 24 hours
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: left and right flanks
- Concentration: 25 % test item
- Evaluation: 0, 24 and 48 hours after removal of dressing
- Duration: days 22 to 38

I) FIRST CHALLENGE: hair from a 5 x 5 cm area on the left and right flanks of all animals was clipped and a 2 x 2 cm patch of filter paper was saturated with 25 % test item in physiological saline (non-irritant concentration) and applied to the left flank of all animals (test and control) and secured with a dressing for 24 hours. A second 2 x 2 cm patch of filter paper, saturated in vehicle only (physiological saline), was simultaneously applied to the right flanks of all animals. Test sites were assessed for erythema and oedema locally at 0, 24 and 48 hours after the removal of the dressings.

II) SECOND CHALLENGE: hair was re-clipped and a 2 x 2 cm patch of filter paper, saturated with 25 % test item in physiological saline, was applied to the right flank of each test animal with a secured dressing for 24 hours. A second patch of filter paper, saturated with vehicle (physiological saline), was simultaneously applied to the left flank of each test animal. A patch of filter paper saturated in vehicle only (physiological saline) was applied to the left flank of each control animal for 24 hours, and the right flank received no treatment. Test sites were assessed for erythema and oedema locally at 0, 24 and 48 hours after the removal of the dressings.
Challenge controls:
1st challenge: controls received same treatment as test animals (application of 25 % test item to the left flank; vehicle to the right flank)
2nd challenge: controls received vehicle to left flank
Positive control substance(s):
yes
Remarks:
dinitrochlorobenzenol (concurrent)
Positive control results:
positive
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
intradermal induction 1% in physiological saline
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25%. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25%
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25%. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
25%
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 25%. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
25%
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 25%. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Interpretation of results:
other: not classified under Regulation 1272/2008
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

The analogue substance was tested for skin sensitisation following EU Method B6 -Guine Pig Maximisation Test (1989). The substance did not show any potential to be a skin sensitizer.

Based on the read across considerations, the same applies to Acid Black 233:1

Migrated from Short description of key information:

Skin sensitisation, GPMT, not sensitising

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

According to the CLP Regulation (EC n. 1272/2008) the following classifications apply:

skin sensitizer Cat 1A, with GPMT:

≥ 30 % is responsive to ≤ 0,1 % of the intradermal induction dose or

> 60 % is responsive to > 0,1 %- ≤ 1 % of the intradermal induction dose

 

skin sensitizer Cat 1B, with GPMT

≥ 30 % to < 60 % is responsive to > 0,1 % - ≤ 1 % of the intradermal induction dose or,

≥ 30 % is responsive to > 1 % of the intradermal induction dose

 

Based on the results of the skin sensitization test (GPMT) the substance is not classified as skin sensitizer