[8-[(4-amino-3-nitrophenyl)azo]-7-hydroxy-2-naphthyl]trimethylammonium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06 July 2004 - 19 July 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from study report

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EC Commission Directive 92/69/EEC, B.5, "Acute Eye Irritation/Corrosion" (1992)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF guidelines (2000).

Principles of method if other than guideline:

The purpose of this acute eye irritation/corrosion study was to assess the possible irritation or corrosion potential when a single dose of the test substance was placed in the conjunctival sac of the rabbit eye.
The absence of eye pigmentation in the albino rabbit facilitates the evaluation of induced eye reactions. The ocular route was selected because the test substance may accidentally come into contact with the eyes during manufacture, handling and/or use.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

[8-[(4-amino-3-nitrophenyl)azo]-7-hydroxy-2-naphthyl]trimethylammonium chloride

EC Number:

269-944-0

EC Name:

[8-[(4-amino-3-nitrophenyl)azo]-7-hydroxy-2-naphthyl]trimethylammonium chloride

Cas Number:

68391-32-2

Molecular formula:

ClN5O3

IUPAC Name:

8-[(4-amino-3-nitrophenyl)diazenyl]-7-hydroxy-N,N,N-trimethylnaphthalen-2-aminium chloride

Constituent 2

Reference substance name:

(8-((4-Amino-3-nitrophenyl)azo)-7-hydroxy-2-naphthyl)trimethylammonium chloride

IUPAC Name:

(8-((4-Amino-3-nitrophenyl)azo)-7-hydroxy-2-naphthyl)trimethylammonium chloride

Test material form:

other: Dark brown powder

Details on test material:

- Name of test material (as cited in study report): Basic brown 17
- Molecular formula (if other than submission substance): C19H20N5O3.Cl
- Molecular weight (if other than submission substance): 401.852 g/mol
- Smiles notation (if other than submission substance):[N+](c1cc(\N=N\c2c3cc([N+](C)(C)C)ccc3ccc2O)ccc1N)([O])=O.[ClH]
- InChl (if other than submission substance): 1S/C19H19N5O3.ClH/c1-24(2,3)14-7-4-12-5-9-18(25)19(15(12)11-14)22-21-13-6-8-16(20)17(10-13)23(26)27;/h4-11H,1-3H3,(H2-,20,21,22,25);1H
- Structural formula attached as image file (if other than submission substance): No data available
- Substance type: Organic
- Physical state: Solid
- Analytical purity: No data available
- Impurities (identity and concentrations): No data available
- Composition of test material, percentage of components: No data available
- Isomers composition: No data available
- Purity test date: No data available
- Lot/batch No.: No data available
- Expiration date of the lot/batch: No data available
- Radiochemical purity (if radiolabelling): No data available
- Specific activity (if radiolabelling): No data available
- Locations of the label (if radiolabelling): No data available
- Expiration date of radiochemical substance (if radiolabelling): No data available
- Stability under test conditions: No data available
- Storage condition of test material: No data available
- Other: No data available

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Test Item Identification: B 007 ([8-[(4-amino-3-nitrophenyl)azo]-7-hydroxy-2-naphthyl]trimethylammonium chloride)
- Batch No.of test material: NDKS 1944
- Expiration date of the lot/batch: 31 December 2013
- Purity: 96.3% (HPLC)

RADIOLABELLING INFORMATION (Not applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In refrigerator in the dark.
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: No data
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The powdery test substance was instilled as delivered by the sponsor.
- Preliminary purification step (if any): No data
- Final dilution of a dissolved solid, stock liquid or gel: No data
- Final preparation of a solid: No data

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

(SPF-Quality)

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany
- Age at study initiation: Animals used within the study were at least 6 weeks old
- Weight at study initiation: Body weights were at least 1.0 kg.
- Identification: Earmark.
- Housing: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm).
- Diet (e.g. ad libitum):Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits,Altromin, Lage, Germany) approx. 100 g. per day. In addition, hay was provided at least three times a week.
- Water (e.g. ad libitum): Free access to tap-water.
- Acclimation period: 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0°C (actual range: 19.0-20.7°C)
- Humidity (%): 30-70% (actual range: 38 - 80%)
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkne ss per day.

IN-LIFE DATES: From:06 July 2004 To:19 July 2004

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 45.4 (43.8 - 47.1) mg of the test substance (a volume of appraximately 0.1 ml).

VEHICLE (No vehicle)
- Amount(s) applied (volume or weight with unit): None

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 days after instillation of the test substance.

Duration of post- treatment incubation (in vitro):

No data

Number of animals or in vitro replicates:

3 Males.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data
- Time after start of exposure: No data

SCORING SYSTEM:The irritation was assessed according to the numerical scoring system.

TOOL USED TO ASSESS SCORE: No data

Results and discussion

In vitro

Other effects / acceptance of results:

All available data relevant to the potential eye irritation/corrosivity of the substance indicated that no severe effects were to be expected. No severe reactions were noted in the skin irritation study. An in-vitro test was considered, but a negative test result was anticipated that still would have to be confirmed in an in-vivo study.

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 39,64,65

Time point:

7 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

animal: 39,64,65

Time point:

7 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 39,64,65

Time point:

7 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 39,64,65

Time point:

7 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal: 39

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

iris score

Basis:

animal: 39

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal: 39

Time point:

24/48/72 h

Score:

1.7

Reversibility:

not specified

Results 8

Irritation parameter:

chemosis score

Basis:

animal: 39

Time point:

24/48/72 h

Score:

0.3

Reversibility:

not specified

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal: 64

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 10

Irritation parameter:

iris score

Basis:

animal: 64

Time point:

24/48/72 h

Score:

0.3

Reversibility:

not specified

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal: 64

Time point:

24/48/72 h

Score:

2

Reversibility:

not specified

Results 12

Irritation parameter:

chemosis score

Basis:

animal: 64

Time point:

24/48/72 h

Score:

1

Reversibility:

not specified

Results 13

Irritation parameter:

cornea opacity score

Basis:

animal: 65

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 14

Irritation parameter:

iris score

Basis:

animal: 65

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 15

Irritation parameter:

conjunctivae score

Basis:

animal: 65

Time point:

24/48/72 h

Score:

1.7

Reversibility:

not specified

Results 16

Irritation parameter:

chemosis score

Basis:

animal: 65

Time point:

24/48/72 h

Score:

0.3

Reversibility:

not specified

Irritant / corrosive response data:

Irritation: Instillation of 45 mg of B 007 (a volume of approximately 0.1 ml) into one eye of each of three rabbits resulted in effects on the iris in two animals and on the conjunc tivae in all animals. Iridial irritation grade 1 was observed and had resolved within 24 or 72 hours.
The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 7 days in all animals.
No corneaI opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneaI epithelial damage.

Corrosion
There was no evidence of ocular corrosion.

Other effects:

- clinical observations:Colouration / Remnants
Remnants of the test substance were present in the eye 1 hour after instillation. And remnants were present on the outside of the eyelids 24 and 48 hours after instillation in one animal.Yellow-brown staining of the fur on the head and paws, caused by the test substance, was noted during the observation period.

- Other observations:Toxicity / Mortality
No symptoms of systemic toxicity were observed in the animals during the test period and no
mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

other: Not irritating

Remarks:

based on CLP criteria

Conclusions:

Based on the above study the test item B 007 ([8-[(4-amino-3-nitrophenyl)azo]-7-hydroxy-2-naphthyl]trimethylammonium chloride) does not have to be classified for eye irritation.

Executive summary:

Acute eye irritation/corrosion study with B 007 ([8-[(4-amino-3-nitrophenyl)azo]-7-hydroxy-2-naphthyl]trimethylammonium chloride) in the rabbit. The study was carried out based on the guidelines described in: "Acute Toxicity - Eye irritation",OECD NoA05 (2002); EC Commission Directive 92169/EEC, B.5, "Acute Eye Irritation/Corrosion" (1992); US EPA, OPPTS 870.2400, Acute Eye Irritation, (1998) and JMAFF guidelines (2000). Single samples of approximately 45 mg of B 007 (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours and 7 days after instillation. Instillation of the test substance resulted in effects on the iris in two animals and on the conjunctivae in all animals. Iridial irritation grade 1 was observed and had resolved within 24 or 72 hours. The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 7 days in all animals. Remnants of the test substance were present in the eye 1 hour after instillation. And remnants were present on the outside of the eyelids 24 and 48 hours after instillation in one animal.Yellow-brown staining of the fur on the head and paws, caused by the test substance, was noted during the observation period. Based on these results and according to the CLP classification criteria: B 007 ([8-[(4-amino-3-nitrophenyl)azo]-7-hydroxy-2-naphthyl]trimethylammonium chloride) does not have to be classified for eye irritation.