[[2,2',2''-[29H,31H-phthalocyaninetriyltris(methylene)]tris[1H-isoindole-1,3(2H)-dionato]](2-)-N29,N30,N31,N32]copper

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

[[2,2',2''-[29H,31H-phthalocyaninetriyltris(methylene)]tris[1H-isoindole-1,3(2H)-dionato]](2-)-N29,N30,N31,N32]copper

EC Number:

261-638-5

EC Name:

[[2,2',2''-[29H,31H-phthalocyaninetriyltris(methylene)]tris[1H-isoindole-1,3(2H)-dionato]](2-)-N29,N30,N31,N32]copper

Cas Number:

59160-79-1

Molecular formula:

C59 H31 Cu N11 O6

IUPAC Name:

[[2,2',2''-[(29H,31H-phthalocyanine-C,C,C-triyl-kN29,kN30,kN31,kN32)tris(methylene)]tris[1H-isoindole-1,3(2H)-dionato]](2-)]-copper

Test material form:

solid: nanoform

Details on test material:

BET: 17.5 m2/g
TEM (min. Feret): 46.2 nm (D50)

Specific details on test material used for the study:

- Substance type: blue solid
- Physical state: solid
- Analytical purity: concentration ≥ 97%
- Purity test date: 2014
- Lot/batch No.: 120001P040
- Expiration date of the lot/batch: Unlimited stability
- Stability under test conditions: stable
- Storage condition of test material: At room temperature, protect against humidity, dry storage
- Other: Identifier in report: CAS 59160-79-1

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River UK
- Age at study initiation: 7-8 weeks
- Weight at study initiation: 17.8 - 20.5 g
- Housing: all animals in one group were housed in a cage together as a group
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2°C
- Humidity (%): 45 – 65%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2015-06-05 To 2014-07-01

Study design: in vivo (LLNA)

Vehicle:

propylene glycol

Concentration:

2, 5 and 10% (w/w)

No. of animals per dose:

5

Details on study design:

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: standard H-Methylthymidine incorporation
- Criteria used to consider a positive response: SI > 3

TREATMENT PREPARATION AND ADMINISTRATION: Each test group of mice was treated by topical (epidermal) application to the dorsal surface
of each ear. The application volume, 25 μL/ear/day, was spread over the entire dorsal surface (diameter 8 mm) of each ear once daily for three consecutive days. A further group of mice (control animals) was treated with an equivalent volume of the relevant vehicle alone.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

SI = 13.7%
The periodic positive control experiment was performed in April 2014
Positive control substance: α-Hexylcinnamaldehyde; Vehicle: acetone:olive oil (4+1 v/v)

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks:

(2%)

Value:

0.69

Results 2

Parameter:

SI

Remarks:

(5%)

Value:

0.95

Results 3

Parameter:

SI

Remarks:

(10%)

Value:

1.24

Results 4

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: See table

Any other information on results incl. tables

Table 1: Lymph Node Cell Counts after Sacrifice

Test item concentration % (w/w)	Animal No.	Lymph Node Cell Count x10E06 per animal	Mean Lymph Node Cell Count x10E06 per animal	SD	Index (Value Test Group versus Value Control
Vehicle Control (PG)	1	7.65	8.4	1.4	1.00
	2	7.99
	3	9.45
	4	10.18
	5	6.61
2	6	6.84	6.9	1.3	0.82
	7	6.42
	8	5.01
	9	8.30
	10	7.86
5	11	8.67	7.1	2.4	0.85
	12	7.99
	13	8.56
	14	7.29
	15	2.95
10	16	7.45	9.2	1.6	1.10
	17	10.13
	18	7.65
	19	10.91
	20	10.00

Table 2: Lymph Node Weights after Sacrifice

Test item concentration % (w/w)	Animal No.	Lymph Node weight mg per animal	Mean Lymph Node weight (mg)	SD	Index (Value Test Group versus Value Control
Vehicle Control (PG)	1	4.67	4.7	0.5	1.0
	2	4.47
	3	5.09
	4	5.30
	5	4.09
2	6	4.39	4.3	0.5	0.9
	7	4.41
	8	3.39
	9	4.46
	10	4.75
5	11	4.92	3.9	1.3	0.83
	12	4.72
	13	4.25
	14	4.11
	15	1.63
10	16	4.19	5.4	0.9	1.14
	17	6.41
	18	4.88
	19	6.27
	20	5.22

Table 3: Ear Weights after Sacrifice

Test item concentration % (w/w)	Animal No.	Ear weight mg per animal	Mean Ear weight (mg)	SD	Index (Value Test Group versus Value Control)
Vehicle Control (PG)	1	23.74	24.4	0.9	1.00
	2	24.87
	3	25.81
	4	23.78
	5	23.95
2.0%	6	23.89	23.2	0.6	0.95
	7	22.44
	8	23.60
	9	23.09
	10	22.80
5%	11	23.29	25.6	2.5	1.05
	12	24.89
	13	29.90
	14	25.25
	15	24.81
10%	16	27.50	27.1	0.8	1.11
	17	26.10
	18	28.34
	19	26.87
	20	26.85

No signs of systemic toxicity or local skin irritation were observed during the study period. A possible erythema of the ear skin could not be evaluated due to the inherent colour of the test item.

The body weight of the animals, recordedprior to the first application and prior to treatment with³HTdR, was within the range commonly recorded for animals of this strain and age.

A statistically significant increase in ear weights was observed in the high dose group in comparison to the vehicle control group (p<0.05). Furthermore, for BALB/c mice, a cut-off value of 1.1 for the ear weight index was reported for a positive response regarding ear skin irritation (see Ref. 9). The index determined for the test item treated high dose group slightly exceeded this threshold (index of 1.11). However, this was considered to be not biologically relevant, as the observed increase did not exceedthe threshold value of 25% for excessive local skin irritation mentioned in OECD guideline 429. Nevertheless, the increased ear weights indicated an irritant property of the test item.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met