3-amino-2-chlor-6-methylphenol

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24.11. - 08.12.2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2004

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:

EPA OPPTS 870.2400 (Acute Eye Irritation)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-amino-2-chlor-6-methylphenol

EC Number:

283-144-9

EC Name:

3-amino-2-chlor-6-methylphenol

Cas Number:

84540-50-1

Molecular formula:

C7H8ClNO

IUPAC Name:

3-amino-2-chloro-6-methylphenol

Test material form:

other: beige powder

Details on test material:

molecular weight: 157.5

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Males
Age: at least 6 weeks
Body weight: at least 1.0 kg
Conditions:
Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0°C (actual range: 18.1 -21.5°C), a relative humidity of 30-70% (actual range: 38 - 74%) and 12 hours artificial fluorescent light and 12 hours darkness per day. Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70% for relative humidity. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.
Accommodation:
lndividually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm). Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.
Diet:
Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany) approx. 100 g. per day. Certificates of analysis were examined and retained in the NOTOX archives. In addition, pressed hay (BMI, Helmond, the Netherlands) was provided twice a week.
Water:
Free access to tap-water.

Test system

Amount / concentration applied:

38.3 (38.0 - 38.7) mg / approx. 0.1 ml

Duration of treatment / exposure:

24 h

Observation period (in vivo):

24 h

Number of animals or in vitro replicates:

3

Details on study design:

All available data relevant to the potential eye irritation/corrosivity of the substance indicated that no severe effects were to be expected. No skin reactions were noted in the skin irritation study (Notox Project 395786). 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. Since no severe harm for the animals was to be expected, this in-vivo eye irritation study was performed and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 8 days later, after considering the degree of eye irritation observed in the first animal.

A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the eyes, which were free from any abnormality.

Each animal was treated by instillation of 38.3 (38.0 - 38.7) mg of the test substance (a volume of approximately 0.1 ml) in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.

lmmediately after the 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. When considered necessary, this procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

lmmediately after fluorescein examination on day 2, the treated eye was rinsed with approx 50 ml tepid tap-water, using a velocity of flow which did not affect the eye, to remove any residual test substance. For reference control the other eye was also rinsed.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

sentinel

Time point:

other: 1 h

Score:

0

Max. score:

4

Remarks on result:

other: Slight dulling of the normal lustre of the comea.

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

sentinel

Time point:

24 h

Score:

1

Max. score:

4

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

sentinel

Time point:

48 h

Score:

1

Max. score:

4

Results 4

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

sentinel

Time point:

72 h

Score:

1

Max. score:

4

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

sentinel

Time point:

7 d

Score:

0

Max. score:

4

Results 6

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

sentinel

Time point:

14 d

Score:

0

Max. score:

4

Results 7

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

sentinel

Time point:

other: 1 h

Score:

0

Max. score:

2

Results 8

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

sentinel

Time point:

24 h

Score:

0

Max. score:

2

Results 9

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

sentinel

Time point:

48 h

Score:

0

Max. score:

2

Results 10

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

sentinel

Time point:

72 h

Score:

0

Max. score:

2

Results 11

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

sentinel

Time point:

7 d

Score:

0

Max. score:

2

Results 12

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

sentinel

Time point:

14 d

Score:

0

Max. score:

2

Results 13

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

sentinel

Time point:

other: 1 h

Score:

2

Max. score:

3

Results 14

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

sentinel

Time point:

24 h

Score:

3

Max. score:

3

Results 15

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

sentinel

Time point:

48 h

Score:

2

Max. score:

3

Results 16

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

sentinel

Time point:

72 h

Score:

2

Max. score:

3

Results 17

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

sentinel

Time point:

7 d

Score:

1

Max. score:

3

Results 18

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

sentinel

Time point:

14 d

Score:

0

Max. score:

3

Results 19

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

sentinel

Time point:

other: 1 h

Score:

3

Max. score:

4

Results 20

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

sentinel

Time point:

24 h

Score:

2

Max. score:

4

Results 21

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

sentinel

Time point:

48 h

Score:

1

Max. score:

4

Results 22

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

sentinel

Time point:

72 h

Score:

1

Max. score:

4

Results 23

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

sentinel

Time point:

7 d

Score:

0

Max. score:

4

Results 24

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

sentinel

Time point:

14 d

Score:

0

Max. score:

4

Results 25

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #1

Remarks:

sentinel

Time point:

other: 1 h

Score:

2

Max. score:

3

Results 26

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #1

Remarks:

sentinel

Time point:

24 h

Score:

1

Max. score:

3

Results 27

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #1

Remarks:

sentinel

Time point:

48 h

Score:

1

Max. score:

3

Results 28

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #1

Remarks:

sentinel

Time point:

72 h

Score:

1

Max. score:

3

Results 29

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #1

Remarks:

sentinel

Time point:

7 d

Score:

0

Max. score:

3

Results 30

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #1

Remarks:

sentinal

Time point:

14 d

Score:

0

Max. score:

3

Results 31

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 1 h

Score:

0

Max. score:

4

Remarks on result:

other: Slight dulling of the normal lustre of the comea.

Results 32

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24 h

Score:

0

Max. score:

4

Results 33

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

48 h

Score:

0

Max. score:

4

Results 34

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

4

Results 35

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

7 d

Score:

0

Max. score:

4

Results 36

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

14 d

Score:

0

Max. score:

4

Results 37

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 1 h

Score:

0

Max. score:

2

Results 38

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24 h

Score:

0

Max. score:

2

Results 39

Irritation parameter:

iris score

Basis:

animal #2

Time point:

48 h

Score:

0

Max. score:

2

Results 40

Irritation parameter:

iris score

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

2

Results 41

Irritation parameter:

iris score

Basis:

animal #2

Time point:

7 d

Score:

0

Max. score:

2

Results 42

Irritation parameter:

iris score

Basis:

animal #2

Time point:

14 d

Score:

0

Max. score:

2

Results 43

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

other: 1 h

Score:

2

Max. score:

3

Results 44

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24 h

Score:

2

Max. score:

3

Results 45

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

48 h

Score:

2

Max. score:

3

Results 46

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

72 h

Score:

1

Max. score:

3

Results 47

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

7 d

Score:

0

Max. score:

3

Results 48

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 1 h

Score:

3

Max. score:

4

Results 49

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24 h

Score:

2

Max. score:

4

Results 50

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

48 h

Score:

1

Max. score:

4

Results 51

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

4

Results 52

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

7 d

Score:

0

Max. score:

4

Results 53

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #2

Time point:

other: 1 h

Score:

2

Max. score:

3

Results 54

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #2

Time point:

24 h

Score:

1

Max. score:

3

Results 55

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #2

Time point:

48 h

Score:

0

Max. score:

3

Results 56

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

3

Results 57

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: 1 h

Score:

1

Max. score:

4

Results 58

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24 h

Score:

1

Max. score:

4

Results 59

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

48 h

Score:

1

Max. score:

4

Results 60

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

other: Slight dulling of the normal lustre of the comea.

Results 61

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

7 d

Score:

0

Max. score:

4

Results 62

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: 1 h

Score:

0

Max. score:

2

Results 63

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24 h

Score:

1

Max. score:

2

Results 64

Irritation parameter:

iris score

Basis:

animal #3

Time point:

48 h

Score:

0

Max. score:

2

Results 65

Irritation parameter:

iris score

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

2

Results 66

Irritation parameter:

iris score

Basis:

animal #3

Time point:

7 d

Score:

0

Max. score:

2

Results 67

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

other: 1 h

Score:

2

Max. score:

3

Results 68

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24 h

Score:

3

Max. score:

3

Results 69

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

48 h

Score:

3

Max. score:

3

Results 70

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

72 h

Score:

2

Max. score:

3

Results 71

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

7 d

Score:

0

Max. score:

3

Results 72

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: 1 h

Score:

3

Max. score:

4

Results 73

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24 h

Score:

2

Max. score:

4

Results 74

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

48 h

Score:

1

Max. score:

4

Results 75

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

4

Results 76

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

7 d

Score:

0

Max. score:

4

Results 77

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #3

Time point:

other: 1 h

Score:

1

Max. score:

3

Results 78

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #3

Time point:

24 h

Score:

1

Max. score:

3

Results 79

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #3

Time point:

48 h

Score:

1

Max. score:

3

Results 80

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

3

Results 81

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

animal #3

Time point:

7 d

Score:

0

Max. score:

3

Irritant / corrosive response data:

Irritation
Instillation of approximately 38 mg of A 094 (a volume of approximately 0.1 ml) into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity (maximum grade 1) and epithelial damage (maximum 55% of the corneal area). The corneal injury had resolved within 24 hours in one animal, and within 7 days in the other animals. lridial irritation grade 1 was observed in one animal only 24 hours after instillation. The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 7 days in two animals and within 14 days in the other animal.

Corrosion
There was no evidence of ocular corrosion.

Other effects:

Colouration / Remnants
No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen.

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

Conclusions:

Based on these results and according to the:
- EC criteria for classification and labelling requirements for dangerous substances and preparations (Council Directive 67/548/EEC), A 094 / SAT 030633 does not have to be classified and has no obligatory labelling requirement for eye irritation.

Executive summary:

Acute eye irritation/corrosion study with A 094 in the rabbit.

The study was carried out based on the guidelines described in: "Acute Taxicity- Eye irritation", OECD No.405 (2002); EC Commission Directive 92/69/EEC, 8.5, "Acute Eye lrritation/Corrosion" (1992); US EPA, OPPTS 870.2400, Acute Eye Irritation, (1998) and JMAFF guidelines (2000).

Single samples of approximately 38 mg of A 094 (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/or 14 days after instillation.

Instillation of the test substance resulted in effects on the cornea, iris and conjunctivae.

The corneal injury consisted of opacity (maximum grade 1) and epithelial damage (maximum 55% of the corneal area). The corneal injury had resolved within 24 hours in one animal, and within 7 days in the other animals. lridial irritation grade 1 was observed in one animal only 24 hours after instillation. The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 7 days in two animals and within 14 days in the other animal.

Based on these results and according to the:

EC criteria for classification and labelling requirements for dangerous substances and preparations (Council Directive 67/548/EEC),

A 094 does not have to be classified and has no obligatory labelling requirement for eye irritation.