Reaction mass of 29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper and Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, (dimethylamino)methyl derivs.

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 August 2016 - 16 August 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Due to technical reasons, the actual maximum relative humidity was 88 % instead of 70 %
and the actual maximum temperature was 25.8 °C instead of 22 ± 3 °C as it was indicated in
the Study Plan. Due to a new Certificate of Analysis an incorrect purity data of the test item
showed in the Study Plan. The Draft Report was issued later than indicated in the Study Plan.

Deviations:

yes

Remarks:

These deviations are considered not to adversely affect the results or integrity of the study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

Due to technical reasons, the actual maximum relative humidity was 88 % instead of 70 %
and the actual maximum temperature was 25.8 °C instead of 22 ± 3 °C as it was indicated in
the Study Plan. Due to a new Certificate of Analysis an incorrect purity data of the test item
showed in the Study Plan. The Draft Report was issued later than indicated in the Study Plan.

Deviations:

yes

Remarks:

These deviations are considered not to adversely affect the results or integrity of the study.

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:

2λ⁵,11,20,29,38,39λ⁵,40,41-octaaza-1-cupradodecacyclo[19.17.1.1¹,¹².1³,³⁷.0²,¹⁰.0⁴,⁹.0¹³,¹⁸.0²²,²⁷.0²⁸,³⁹.0³⁰,³⁸.0³¹,³⁶.0¹⁹,⁴¹]hentetraconta-2,4,6,8,10,12,14,16,18,20,22,24,26,28(39),29,31,33,35,37(40)-nonadecaene-2,39-bis(ylium)-1,1-diuide

EC Number:

908-081-2

IUPAC Name:

2λ⁵,11,20,29,38,39λ⁵,40,41-octaaza-1-cupradodecacyclo[19.17.1.1¹,¹².1³,³⁷.0²,¹⁰.0⁴,⁹.0¹³,¹⁸.0²²,²⁷.0²⁸,³⁹.0³⁰,³⁸.0³¹,³⁶.0¹⁹,⁴¹]hentetraconta-2,4,6,8,10,12,14,16,18,20,22,24,26,28(39),29,31,33,35,37(40)-nonadecaene-2,39-bis(ylium)-1,1-diuide

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Batch number: 150205
Description: Blue powder
Expiry date: 05 February 2019
Purity1: As a UVCB, purity is considered as 100%
Storage conditions: Controlled room temperature (15-25°C, below 70% RH%)

In vivo test system

Test animals

Species:

mouse

Strain:

CBA/Ca

Remarks:

CBA/CaOlaHsd mice

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation:10 weeks old (age-matched, within one week)
- Weight at study initiation: 20.6 – 21.9 grams
- Housing: polypropylene / polycarbonate cages and bedding was available to animals during the study
- Enrichment: mice were provided with glass tunnel tubes
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 21 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.0 - 25.8°C
- Humidity (%): 30 - 88 %
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

Test item concentration: 2.5, 5 and 10 %(w/v)

No. of animals per dose:

4

Details on study design:

PRE-SCREEN TESTS:
- Compound solubility: formulations at 100 % (w/v) concentrations were not achievable
- Irritation: no
- Systemic toxicity: no
- Ear thickness measurements: ear thickness values and ear punch weights were within the acceptable range

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response:
That exposure to at least one concentration of the test item resulted in an incorporation of 3 HTdR at least 3-fold or greater than recorded in control mice, as indicated by the stimulation index.
The data are compatible with a conventional dose response, although allowance must be made (especially at high topical concentrations) for either local toxicity or immunological suppression.

TREATMENT PREPARATION AND ADMINISTRATION:
Topical application: on the dorsal surface of each ear. Each animal was dosed once a day for three consecutive days

Positive control substance(s):

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

Results and discussion

Positive control results:

No mortality, cutaneous reactions or signs of toxicity were observed for the positive control substance in the study. A lymphoproliferative response in line with historic positive control data (stimulation index value of 15.6*) was noted for HCA in the main experiment. This value was considered to confirm the appropriate performance of the assay.
*The stimulation index (SI) value was slightly above the upper limit of the historic positive control data, however it has not affect to the results.

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

1.3

Test group / Remarks:

2.5 % (w/v) in acetone:olive oil 4:1 (v:v) mixture

Results 2

Key result

Parameter:

SI

Value:

1.3

Test group / Remarks:

5 %(w/v) in acetone:olive oil 4:1 (v:v) mixture

Results 3

Key result

Parameter:

SI

Value:

1.2

Test group / Remarks:

10 %(w/v) in acetone:olive oil 4:1 (v:v) mixture

Cellular proliferation data / Observations:

DETAILS ON STIMULATION INDEX CALCULATION
SI = DPN value of a treated group divided by the DPN value of the negative control group) for each treatment group was also calculated. A stimulation index of 3 or greater is an indication of a positive result.

CLINICAL OBSERVATIONS:
No mortality or signs of systemic toxicity were observed during the study. Test item precipitate or minimal amount of test item precipitate was observed for all animals of the 10% (w/v), for all animals of the 5% (w/v) and for all animals of the 2.5% (w/v) dose groups on Days 1-6. There were no indications of any irritancy at the site of application.

BODY WEIGHTS
Marked body weight loss (>5% reduction of body weight) was observed for one animal of the negative control group, positive control group and 2.5% (w/v) dose group, for two animals of the 10 % (w/v) dose group; for three animals of the 5% (w/v) dose group, thus the mean body weight loss of the 5% (w/v) dose group was more than 5 %. However there was no clear dose relationship and all changes were considered as individual variability.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, under the conditions of the present assay, the test item, tested in a suitable vehicle, was shown to have no sensitisation potential (non-sensitizer) in the Local Lymph Node Assay.

The following classification/labelling is triggered:
Regulation (EC) No 1272/2008 (CLP) / GHS (rev. 6) 2015: none.