2-[[4-(dimethylamino)phenyl]azo]-1,3-dimethyl-1H-imidazolium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Reported 17Apr2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: OECD (2014a) Draft Proposal for a New Test Guideline (EpiOcular™)

Version / remarks:

(2014a) Draft Proposal

Deviations:

not specified

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-[[4-(dimethylamino)phenyl]azo]-1,3-dimethyl-1H-imidazolium chloride

EC Number:

278-601-4

EC Name:

2-[[4-(dimethylamino)phenyl]azo]-1,3-dimethyl-1H-imidazolium chloride

Cas Number:

77061-58-6

Molecular formula:

C13H18N5.Cl

IUPAC Name:

2-[[4-(dimethylamino)phenyl]azo]-1,3-dimethyl-1H-imidazolium chloride

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Test Substance name: Vibracolor ruby red
Test Substance: 10/0383-3
Batch: 0009051093
CAS: 77061-58-6

Test animals / tissue source

Species:

other: Two test systems utilised: 1) Isolated corneas from the eyes of freshly slaughtered cattle 2) human reconstructed cornea after a short term topical exposure

Details on test animals or tissues and environmental conditions:

Two test systems utilised:
1) Isolated corneas from the eyes of freshly slaughtered cattle
2) human reconstructed cornea after a short term topical exposure

Test system

Vehicle:

other: Both diluted in de-ionized water and undiluted (unchanged; no vehicle)

Amount / concentration applied:

BCOP: Three corneas were treated with 750 μL of 50% and 5% test-substance preparations in deionized water for 10 minutes followed by a 2-hours post-incubation period.
EpiOcular: Two EpiOcular™ tissue samples were incubated with each, 50 μL bulk volume (about 14 mg) of the undiluted test substance or 50 μL of a 5% test-substance preparation for 6 hours followed by an 18-hours post-incubation period.

Duration of treatment / exposure:

BCOP: 10 minutes followed by a 2-hours post-incubation period.
EpiOcular: 6 hours followed by an 18-hours post-incubation period.

Duration of post- treatment incubation (in vitro):

BCOP: 2-hours post-incubation period.
EpiOcular: 18-hours post-incubation period.

Number of animals or in vitro replicates:

BCOP: Three corneas
EpiOcular: Two EpiOcular™ tissue samples

Details on study design:

BCOP: Three corneas were treated with 750 μL of 50% and 5% test-substance preparations in deionized water for 10 minutes followed by a 2-hours post-incubation period.
EpiOcular: Two EpiOcular™ tissue samples were incubated with each, 50 μL bulk volume (about 14 mg) of the undiluted test substance or 50 μL of a 5% test-substance preparation for 6 hours followed by an 18-hours post-incubation period.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Run / experiment:

BCOP TEST with test item at 5% in de-ionised water

Value:

1.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: Potential for irritation - refer to EIPOCULAR test

Results 2

Irritation parameter:

cornea opacity score

Run / experiment:

BCOP TEST with test item at 50% in de-ionised water

Value:

0

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: Potential for irritation - refer to EIPOCULAR test

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

BCOP TEST with test item at 5% in de-ionised water

Value:

1.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: Potential for irritation - refer to EIPOCULAR test

Results 4

Irritation parameter:

in vitro irritation score

Run / experiment:

BCOP TEST with test item at 50% in de-ionised water

Value:

0

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: Potential for irritation - refer to EIPOCULAR test

Results 5

Irritation parameter:

other: viability values

Run / experiment:

EPIOCULAR TEST - Undiluted test substance

Value:

2.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 6

Irritation parameter:

other: viabilty values

Run / experiment:

EPIOCULAR TEST - 5% test-substance de-ionised in urified water

Value:

4.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Any other information on results incl. tables

Applying the criteria of the OECD 437 fortheassessment of the BCOP test results,theIVISscores ofthe testsubstance of 0.0 (50%testsubstance preparation) or 1.9 (5% test substance preparation) would classify it as a substance that does not cause eye irritation or serious eye damage.However,the Guideline also states,that due to limited accuracy of the BCOP test to correctlyidentifysubstances that do not require classification for eyeirritationor serious eye damage,thistestmethodshould not bethe firstchoice toinitiatea bottom-up approach. Based onthisstatement and the experience ofthe testfacility,testsubstances leading to IVIS <3 are generally examinedin the EpiOculartest as well.In thecurrent case, the EpiOculartestwas positive and thetestsubstance was assessedtobe irritating.

 

Based on the results forBCOPandEpiOcular Testand applying the evaluation criteria describedinsection 3,Vibracolor ruby red showsaneye irritation potentialinthein vitroeye irritation test strategy underthe testconditions chosen.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Based on the results for BCOP and EpiOcular Test and applying the evaluation criteria described in section 3, Vibracolor ruby red shows an eye irritation potential in the in vitro eye irritation test strategy under the test conditions chosen .

Executive summary:

Based on the results for BCOP and EpiOcular Test and applying the evaluation criteria described in section 3,Vibracolor ruby red shows an eye irritation potential in the in vitro eye irritation test strategy under the test conditions chosen.