Benzoic acid, 2-ethoxy-5-[(4-methyl-1-piperazinyl)sulfonyl]-, ethyl ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2017-03-12 to 2017-04-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2012

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

ethyl 2-ethoxy-5-[(4-methylpiperazin-1-yl)sulfonyl]benzoate

EC Number:

814-360-9

Cas Number:

304435-84-5

Molecular formula:

C16H24N2O5S

IUPAC Name:

ethyl 2-ethoxy-5-[(4-methylpiperazin-1-yl)sulfonyl]benzoate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Batch No.: APM0005851606003
Purity/Composition: 99.8% w/w
Test item storage: At room temperature

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approximately 29-35 weeks old
- Weight at study initiation: 3926 to 4316 g
- Housing: housed individually in labeled cages with perforated floors (dimensions 67 x 62 x 55 cm) equipped with water bottles
- Diet: Pelleted diet for rabbits was provided once daily throughout the study. In addition, hay was available during the study period.
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20°C
- Humidity: 58 to 63%
- Air changes: Ten or greater per hour
- Photoperiod: 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 2017-03-12 To: 2017-04-11

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 99.7 mg (range 99.1 – 100.0 mg)

Duration of treatment / exposure:

14 days

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

3 rabbits

Details on study design:

In-life Procedures, Observations, and Measurements
Mortality
Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day.
Clinical Observations
Postdose Observations
Postdose observations were performed once daily throughout the study, the observation period was 14 days.
Body Weights
Animals were weighed individually on Day 1 (predose) and on the day of the final observation.
Irritation
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 and 14 days after instillation of the test item. The irritation scores and a description of all other (local) effects were recorded.

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0.56

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0.44

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

1.44

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Instillation of test substance resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity and epithelial damage. The corneal injury resolved within 72 hours in one animal and within 7 days in the other two animals.
Iridial irritation was observed and resolved within 48 hours in two animals and within 72 hours in the third animal.
The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 14 days.
There was no evidence of ocular corrosion.

Other effects:

No staining of (peri) ocular tissues by the test item was observed and no test item remnants were seen.
No signs of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Individual Eye Irritation Scores:

Eye reaction	Time after treatment	Individual scores-rabbit number and sex
		236 Male	219 Male	221 Male
Cornea opacity	1 hour 24 hours 48 hours 72 hours	0 1 1 0	0 1 1 0	0 1 0 0
Iris	1 hour 24 hours 48 hours 72 hours	0 1 0 0	0 1 1 0	0 1 0 0
Conjunctivae redness	1 hour 24 hours 48 hours 72 hours	2 2 2 2	2 2 2 2	2 2 2 2
Chemosis	1 hour 24 hours 48 hours 72 hours	2 2 1 1	2 2 1 1	2 3 1 1

Applicant's summary and conclusion

Interpretation of results:

Category 2A (irritating to eyes) based on GHS criteria

Conclusions:

Based on the test results:
• according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), test item should be classified as : irritating to eyes (Category 2A).
• according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), test item should be classified as Irritating to eyes (Category 2) and labeled as H319: Causes serious eye irritation.
Based on the maximum group mean of 27.3, test item should be classified as moderate irritant to the rabbit eye according to the Kay and Calandra classification system.

Executive summary:

The objective of this acute eye irritation study was to assess the possible irritation or corrosion potential when a single dose of test item was placed in the conjunctival sac of the rabbit eye according to OECD No.405 (2012) and EC No 440/2008, part B.

Single samples of approximately 99.7 mg of test item (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 and 14 days after instillation.

Instillation of the test item resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity and epithelial damage. The corneal injury resolved within 72 hours in one animal and within 7 days in the other two animals. Iridial irritation was observed and resolved within 48 hours in two animals and within 72 hours in the third animal.

The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 14 days.

Based on the test results:

• according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), test item should be classified as : irritating to eyes (Category 2A).

• according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), test item should be classified as Irritating to eyes (Category 2) and labeled as H319: Causes serious eye irritation.

Based on the maximum group mean of 27.3, test item should be classified as moderate irritant to the rabbit eye according to the Kay and Calandra classification system.