4-(4-nitrophenylazo)-2,6-di-sec-butyl-phenol

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to OECD TG 405 and EEC Directive 84/449/EEC Part B.5 and in accordance with the Principles of Good Laboratory Practices (GLP)

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

MORTRACE SB CONC.

IUPAC Name:

MORTRACE SB CONC.

Constituent 2

Reference substance name:

4-(4-Nitrophenylazo)-2,6-di- sec.butylphenol

IUPAC Name:

4-(4-Nitrophenylazo)-2,6-di- sec.butylphenol

Constituent 3

Reference substance name:

4-(4-nitrophenylazo)-2,6-di-sec-butyl-phenol

EC Number:

410-610-2

EC Name:

4-(4-nitrophenylazo)-2,6-di-sec-butyl-phenol

Cas Number:

111850-24-9

Molecular formula:

C20 H25 N3 O3

IUPAC Name:

2,6-bis(butan-2-yl)-4-[2-(4-nitrophenyl)diazen-1-yl]phenol

Test material form:

other: dark red-brown liquid

Details on test material:

- Name of test material (as cited in study report): Mortrace SB Conc.
- Physical state: dark red-brown liquid
- Analytical purity: concentrate
- Lot/batch No.: MR 26592 SBC
- Expiry date: June 03, 1993
- Stability under storage conditions: stable
- Storage condition of test material: at room temperature protected from light

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: approximately 14-18 weeks
- Weight at study initiation: 3003 grams - 3325 grams
- Housing: individually housed
- Diet: standard rabbit pelleted diet provided approximately 100 grams/day
- Water: ad libitum
- Acclimation period: at least 5 days before start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C
- Humidity (%): 55%
- Air changes (per hr): 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/dark cycle

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

100 ± 2 mg/animal

Duration of treatment / exposure:

as per guideline

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 and 14 days after instillation of the test substance.

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

Prior to test substance administration, each of the animals were physically examined and the eyes were found to be normal.
The test substance (100 ± 0.2 mg per animal) was instilled in the conjunctival sac of the left eye of one animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about ten seconds to prevent loss of the test substance. The other eye remained untreated and served as the reference control
Immediately after 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively
determine corneal epithelial damage. This procedure was repeated in on day 4, until recovery was completed. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.
The two other animals were treated in a similar manner one week later, after considering the degree of eye irritation observed in the first animal. Treatment of the eyes with a fluorescein solution was performed immediately after the observations on days 2, 4 and/or day 8 in these animals.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

mean

Basis:

animal: rabbit 3972

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Results 2

Irritation parameter:

cornea opacity score

Remarks:

mean

Basis:

animal: rabbit 3979

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Results 3

Irritation parameter:

cornea opacity score

Remarks:

mean

Basis:

animal: rabbit 4011

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Results 4

Irritation parameter:

iris score

Remarks:

mean

Basis:

animal: rabbit 3972

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Results 5

Irritation parameter:

iris score

Remarks:

mean

Basis:

animal: rabbit 3979

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Results 6

Irritation parameter:

iris score

Remarks:

mean

Basis:

animal: rabbit 4011

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Results 7

Irritation parameter:

conjunctivae score

Remarks:

mean

Basis:

animal: rabbit 3972

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 8

Irritation parameter:

conjunctivae score

Remarks:

mean

Basis:

animal: rabbit 3979

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 9

Irritation parameter:

conjunctivae score

Remarks:

mean

Basis:

animal: rabbit 4011

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 10

Irritation parameter:

chemosis score

Remarks:

mean

Basis:

animal: rabbit 3972

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Results 11

Irritation parameter:

chemosis score

Remarks:

mean

Basis:

animal: rabbit 3979

Time point:

24/48/72 h

Score:

1.3

Max. score:

2

Reversibility:

fully reversible within: 7 days

Results 12

Irritation parameter:

chemosis score

Remarks:

mean

Basis:

animal: rabbit 4011

Time point:

24/48/72 h

Score:

1.3

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

One hour after instillation of MORTRACE SB CONC. into one eye of each of the three male albino rabbits resulted in slight redness and moderate to severe swelling in the conjunctivae. Increase of the conjunctival redness was noted at the 24-hour observation, whereas iridic irritation and slight dulling of normal lustre in the cornea (in two animals only) were also observed. Corneal epithelial damage, determined by fluorescein, was seen on day 2 in all three animals and persisted on day 4 in animal 4011. The corneal damage and iridic irritation had resolved within 48 hours after instillation in animals 3972 and 3979 and within 7 days in animal 4011. The irritation of the conjunctivae had resolved within 14 days in all three animals.
There was no evidence of ocular corrosion.

Other effects:

The outside of the eyelids were covered with a brown, sticky layer of test substance during the study period. The layer had disappeared at the 14-day observation and a yellow staining of the eyelids remained. No toxic symptoms were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Individual Scores

Cornea opacity
Animal	Observation time			Mean value	Mean value (over all animals)
	24 hours	48 hours	72 hours
3972	0	0	0	0.0	0.0
3979	0	0	0	0.0
4011	0	0	0	0.0
Iris lesion
Animal	Observation time			Mean value	Mean value (over all animals)
	24 hours	48 hours	72 hours
3972	1	0	0	0.3	0.6
3979	1	0	0	0.3
4011	1	1	1	1.0
Conjunctivae (redness)
Animal	Observation time			Mean value	Mean value (over all animals)
	24 hours	48 hours	72 hours
3972	3	3	2	2.7	2.8
3979	3	3	3	3.0
4011	2	3	3	2.7
Conjunctivae (chemosis)
Animal	Observation time			Mean value	Mean value (over all animals)
	24 hours	48 hours	72 hours
3972	1	1	1	1.0	1.2
3979	2	1	1	1.3
4011	2	1	1	1.3

Applicant's summary and conclusion

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

MORTRACE SB CONC. was considered mildly irritating (M2) to the rabbit eye (Kay and Calandra interpretation of the calculated maximum Draize score: 12.3). According to the EEC criteria for general classification and labelling requirements for dangerous substances and preparations (EEC Directive 91/325/EEC, Amendment to Annex VI of the Directive 67/548/EEC), MORTRACE SB CONC. should be labelled as an eye irritant.

Executive summary:

The purpose of this study was to assess the possible irritation or corrosion potential when a single dose of MORTRACE SB CONC. was placed in the conjunctival sac of a rabbit eye.

This study was carried out in accordance with OECD Guideline No. 405, “Acute Eye Irritation / Corrosion” and EEC Directive 84/449/EEC, Part 8.5, “Acute Toxicity — Eye Irritation”.

MORTRACE SB CONC. was instilled into one eye of each of three male albino rabbits, followed by six observations at approximately 1, 24, 48 and 72 hours and 7 and 14 days after test substance administration.

Under the conditions of this study, MORTRACE SB CONC. resulted in adverse effects on the cornea, iris and the conjunctivae. Corrieal epithelial damage was observed in all three animals, which resulted in slight dulling of normal lustre in two animals. The corneal damage had resolved within 2 or 7 days. The iridic irritation had resolved within 2-7 days and the irritation of the conjunctivae had resolved within 14 days after instillation in the animals. Ocular corrosion was not observed in any of the rabbits.

Brown, sticky remnants of and yellow staining by the test substance were noted on the outside of the eyelids during the study period.

No signs of systemic intoxication were observed during the study period.

MORTRACE SB CONC. was considered mildly irritating (N2) to the rabbit eye (Kay and Calancira interpretation of the calculated maximum Draize score: 12.3). According to the EEC criteria for classification and labelling requirements for dangerous substances and preparations (EEC Directive 91/325/EEC, Amendment to Annex VI of the EEC Council Directive 67/548/EEC), NORTRACE SB CONC. should be labelled as an eye irritant.