Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

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:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report date:
2001

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
When this study was performed, the OECD guideline for the LLNA did not yet exist.

Test material

Constituent 1
Chemical structure
Reference substance name:
29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper
EC Number:
205-685-1
EC Name:
29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper
Cas Number:
147-14-8
Molecular formula:
C32H16CuN8
IUPAC Name:
[29H,31H-phthalocyaninato(2-)-kappa~2~N~29~,N~31~]copper
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
- Physical state: blue powder
- Analytical purity: > 98 %
- Impurities (identity and concentrations): no data given
- Lot/batch No.: 29122
- Storage condition of test material: room temperature in the dark

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: D. Hall, Newchurch, Staffordshire, UK
- Age at study initiation: four to seven weeks
- Weight at study initiation: 399 - 498 g
- Housing: 5 animals per suspended plastic cage with solid floor and sawdust bedding
- Diet: vitamin C enriched guinea pig diet (Harlan Teklad 9600 FD2 SQC), ad libitum
- Water: ad libitum
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21 +- 3 °C
- Humidity: 30 - 70 %
- Photoperiod: 12 hrs dark / 12 hrs light

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Alembicol D (coconut oil product)
Concentration / amount:
50 % (w/v)
Day(s)/duration:
48h
Adequacy of induction:
highest technically applicable concentration used
Route:
intradermal
Vehicle:
other: Alembicol D (coconut product)
Concentration / amount:
7.5%
Day(s)/duration:
Single injection
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challengeopen allclose all
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: Alembicol D (coconut oil product)
Concentration / amount:
25 % (w/v)
Day(s)/duration:
24h and 48h
Adequacy of challenge:
other: 50% of the highest technically achievable concentration
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
other: Alembicol D (coconut oil product)
Concentration / amount:
50%
Day(s)/duration:
24h and 48h
Adequacy of challenge:
other: highest technically achievable concentration
No. of animals per dose:
5 control animals and 10 test animals
Details on study design:
RANGE FINDING TESTS:
The intradermal and topical irritancy of a range of dilutions of the test substance was investigated to identify the minimum irritant test substance concentration suitable for the induction phase and the maximum non-irritant concentration by the topical route of administration and a dilution of this for the challenge phase.

MAIN STUDY
A. INDUCTION EXPOSURE
- Site: 4 x 6 cm interscapular area
- Intradermal induction:
A clipped, 4 x 6 cm area of dorsal skin on the scapular region was prepared. 3 pairs of intradermal injections (0.1 ml/site) were conducted on day 1 into a 2 x 4 cm area within the clipped area. Injectables for the test animals were 1) Freund´s Complete Adjuvant, diluted with an equal volume of water for irrigation, 2) the test material 7.5 % (w/v) in Alembicol D and 3) the test material 7.5 % (w/v) in a 50 : 50 mixture of Freund´s Complete Adjuvant and Alembicol D.
- Epicutaneous induction:
On day 7 the same 4 x 6 cm area was clipped and the site was prepared by gentle rubbing with 0.5 ml per site of 7.5 % (w/w) sodium lauryl sulphate in petrolatum. On day 8, a 2 x 4 cm patch of Whatman No. 3 paper was saturated with ca. 0.4 ml of the test material (50 % in Alembicol D); the patch was placed over the injection sites and covered by an impermeable plastic adhesive tape which was secured by an elastic adhesive bandage, wound round the torso of the animal and fixed with an impervious plastic adhesive tape. The dressing was left in place for ca. 48 hours.
- Test groups: During the induction phase, the control animals were treated similarly to the test animals with the exception that the test substance was omitted from intradermal injections and topical applications.

B. CHALLENGE EXPOSURE
The control and test animals were challenged topically two weeks after the topical induction application (day 22), using the test material (25 and 50 % w/v in Alembicol D.
Hair were clipped on the left flank of each animal and a 2 x 2 cm patch of Whatman No. 3 paper was saturated with ca. 0.2 ml of the test material. TheWhatman papers containing 50 % w/v in Alembicol D were applied on the anterior site of the flank, the Whatman papers containing 20 % w/v in Alembicol D were applied in a similar manner on the posterior site. The patches were sealed on the flank for 24 hours under impermeable adhesive tape, secured by elastic adhesive bandage, wound round the torso of the animal and fixed with an impervious plastic adhesive tape. The challenged sites were evaluated ca. 24 and 48 h after removal of the patches.

All animals were observed daily for signs of ill health or toxicity. The body weight of each animal was recorded on day 1 of the study and following completion of the study prior to termination.
The dermal reactions resulting from challenge reaction were assessed using a numerical scoring system (Draize scores).
On completion of the study the animals were sacrificed by cervical dislocation.
The sensitivity of the guinea pig strain is checked periodically by the testing laboratory with hexyl cinnamic aldehyde (HCA), a known moderate sensitizer.

Positive control substance(s):
yes
Remarks:
hexyl cinnamic aldehyde (HCA), periodically checkings

Results and discussion

Positive control results:
The following concentrations of HCA were administered: Intradermal injection 7.5 % v/v in Alembicol D; topical application as supplied (neat); challenge application as supplied (neat) and 50 % v/v in Alembicol D.
Slight to well defined dermal reactions were observed for all test animals after challenge reaction compared to no dermal reactions in the controls. The reactions in the test animals represented hypersensitivity and therefore all test animals gave positive sensitization responses.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25 % (w/v) in Alembicol D
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50 % (w/v) in Alembicol D
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
25 % (w/v) in Alembicol D
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
50 % (w/v) in Alembicol D
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
25 % (w/v) in Alembicol D
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50 % (w/v) in Alembicol D
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25 % (w/v) in Alembicol D
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
50 % (w/v) in Alembicol D
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation

Any other information on results incl. tables

No deaths and no signs of ill health or toxicity were observed. The body weight of the animals increased within the normal range over the period of the study.

During the induction period, necrosis was recorded at sites receiving FCA in test animals and also in control animals. No irritation was seen in test animals at sites receiving the test material in vehicle and in control animals receiving vehicle alone after intradermal injection as well as after topical application.

After challenge, no dermal reactions were seen in either animals, all tested animals gave negative responses.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met