Hydrogen [tris[[[3-[(2-ethylhexyl)oxy]propyl]amino]sulphonyl]-29H,31H-phthalocyaninesulphonato(3-)-N29,N30,N31,N32]cuprate(1-), compound with 3-[(2-ethylhexyl)oxy]propylamine (1:1)

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

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:

other: GLP and OECD guideline compliant study

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)

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River UK
- Age at study initiation: 8-9 weeks
- Weight at study initiation: 15.9 - 19.3 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: 2014-06-24 to 2014-07-29

Vehicle:

dimethylformamide

Concentration:

5, 10, 25 % (w/w).

No. of animals per dose:

5

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: max 25% in DMF (suspension)
- Irritation: none (blue staining)
- Lymph node proliferation response:

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 with test item in dimethylformamide. 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)

Parameter:

SI

Remarks on result:

other: 1.95 (25%) - 1.74 (10%) - 1.8 (5%)

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 (DMF)	1	6.21	9.7	2.3	1.00
	2	10.16
	3	11.23
	4	12.12
	5	8.70
5	6	9.45	13.1	7.7	1.35
	7	9.61
	8	26.84
	9	9.58
	10	9.92
10	11	10.44	11.9	2.2	1.23
	12	9.71
	13	15.17
	14	13.26
	15	11.02
25	16	10.99	13.9	2.7	1.43
	17	13.37
	18	17.81
	19	15.12
	20	12.14

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 (DMF)	1	24.22	27.0	1.9	1.00
	2	27.56
	3	27.88
	4	26.40
	5	29.15
5.0%	6	29.12	27.8	1.2	1.03
	7	28.20
	8	26.06
	9	28.49
	10	27.05
10%	11	25.74	26.9	1.9	1.00
	12	24.36
	13	29.02
	14	28.01
	15	27.41
25%	16	26.33	27.2	1.6	1.01
	17	29.49
	18	28.08
	19	26.31
	20	25.71

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 (DMF)	6	4.89	5.6	0.7	1.00
	7	5.32
	8	6.29
	9	6.36
	10	5.14
5	16	5.45	5.5	0.3	0.98
	17	5.63
	18	5.03
	19	5.60
	20	5.78
10	21	6.23	6.5	1.1	1.16
	22	5.40
	23	8.16
	24	7.04
	25	5.58
25	26	6.71	6.4	0.8	1.14
	27	6.39
	28	7.27
	29	6.50
	30	5.13

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

The substance was assessed for its skin sensitising potential using the Local Lymph Node Assay (LLNA) in mice. Test item solution at different concentrations was prepared in the vehicle DMF using test item concentrations of 5, 10 and 25% (w/w). The highest concentration tested was the highest concentration that could be achieved whilst avoiding systemic toxicity and excessive local skin irritation (as determined by a pre-experiment). The animals did not show any signs of systemic toxicity during the course of the study and no cases of mortality were observed. A possible erythema of the ears could not be determined in all 3 dose groups due to the inherent blue colour of the test item. Additionally, all three dose groups showed blue discolored faeces from days 2 to day 5. A statistically significant or biologically relevant increase in ear weights was not observed in any treated group in comparison to the vehicle control group.

A test item is regarded as a sensitiser in the LLNA if exposure to one or more test item concentration results in a 3-fold or greater increase in incorporation of 3HTdR compared with concurrent controls, as indicated by the Stimulation Index (S.I.). The estimated test item concentration required to produce a S.I. of 3 is referred to as the EC3 value.

In this study Stimulation Indices (S.I.) of 1.80, 1.74 and 1.95 were determined with the test item at concentrations of 5, 10 and 25% (w/w) in DMF, respectively. A dose response was not observed.

A statistically significant and biologically relevant increase in DPM value and also in lymph node weight and cell count was not observed in any test item treated group in comparison to the vehicle control group. Furthermore, the cut-off value of 1.55 for a positive response regarding the lymph node cell count index reported for BALB/c mice (see Ref. 8) was not exceeded in any dose group.

Migrated from Short description of key information:
The substance was non sensitizing in the LLNA (OECD 429, GLP).

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for skin sensitization under Directive 67/548/EEC, as amended for the 31st time in Directive2009/2/EG.

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. The SI is less than 3. As a result the substance is not considered to be classified for skin sensitization under Regulation (EC) No. 1272/2008, as amended for the fifth time in Directive EC944/2013.