1H-Benz[e]indolium, 2-[2-[2-chloro-3-[2-[1,3-dihydro-1,1-dimethyl-3-(4-sulfobutyl)-2H-benz[e]indol-2-ylidene]ethylidene]-1-cyclohexen-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfobutyl)-, inner salt, ammonium salt (1:1)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

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-[(1E)-2-[(3Z)-2-chloro-3-{2-[(2E)-1,1-dimethyl-3-(4-sulfobutyl)-1H,2H,3H-benzo[e]indol-2-ylidene]ethylidene}cyclohex-1-en-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfonatobutyl)-1H-benzo[e]indol-3-ium amine

EC Number:

700-626-0

Cas Number:

1073607-50-7

Molecular formula:

C46 H50 Cl N2 O6 S2 . H4 N

IUPAC Name:

2-[(1E)-2-[(3Z)-2-chloro-3-{2-[(2E)-1,1-dimethyl-3-(4-sulfobutyl)-1H,2H,3H-benzo[e]indol-2-ylidene]ethylidene}cyclohex-1-en-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfonatobutyl)-1H-benzo[e]indol-3-ium amine

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

Details on test animals: 20 females (nulliparous and non-pregnant), five females per group, young adult animals (approx. 9 weeks ols), body weight variation was within +/- 20% of the sex mean.

Study design: in vivo (non-LLNA)

Challenge controls:

vehicle control: 0% test substance, 100% propylene glycol

Study design: in vivo (LLNA)

Vehicle:

propylene glycol

Concentration:

10%, 25% and 50%

No. of animals per dose:

5

Positive control substance(s):

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

Results and discussion

Positive control results:

The SI values calculated for the substance concentrations 5, 10 and 25% were 1.5, 1.9 and 6.1 respectively. An EC3 value of 13.9% was calculated using linear interpolation. The calculated EC3 value was found to be in the acceptable range of 2 and 20%. The results of the 6 monthly HCA reliability checks of the recent years were 9.5, 13.1, 14.1 and 13.8%.
Based on the results, it was concluded that the LLNA as performed at Notox is an appropriate model for testing for contact hypersensitivity.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: The SI values calculated for the substance concentrations 10, 25 and 50% were 1.1, 2.1 and 2.7 respectively.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Mean DPM/animal values for the experimental groups treated with test substance concentrations 10, 25 and 50% were 311, 590 and 782 respectively. The mean DPM/animal value for the vehicle control group was 287.

Any other information on results incl. tables

Skin reactions/irritation: Green/brown staining by the test substance prevented scoring for erythema. No oedema was observed in any of the animals examined.

Macroscopy of the auricular lymph nodes and surrounding area: All nodes of the experimental and control groups were considered normal in size, except for the lymph nodes of one animal at 50%. No macroscopic abnormalities of the surrounding area were noted.

Body weights: Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

Toxicity and mortality: No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: other: OECD GHS, EC No1272/2008, Directive 67/548/EEC

Conclusions:

Since there was no indication that the test substance elicits an SI>=3 when tested up to 50%, the test substance was considered not to be a skin sensitizer.