Sodium long chain alkyl salicylate

Endpoint summary

Description of key information

SKIN IRRITATION/CORROSION
In vitro: not corrosive (human three dimensional epidermal model (EpiDerm (EPI-200)); OECD 431 and EU Method B.40 BIS
In vivo: corrosive (rabbit); equivalent to OECD 404
EYE IRRITATION 
In vivo: causes serious eye damage (rabbit); equivalent to OECD 405

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985-07-22 to 1985-08-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age on receipt from supplier: 3-6 months
- Age at study initiation: 4-9 months
- Weight at study initiation: 4790-4862 g (female); 4008-4106 g (male)
- Housing: Singly housed in hanging, stainless-steel cages with all-mesh floors and tops and half-mesh fronts; each cage measured 67 x 43 x 45 cm. The cages were mounted in three rows of two on a single-sided rack (a total of six cages).
- Diet: ad libitum by means of a top-loading food hopper
- Water: Filtered but untreated from water from the public supply by means of an automatic valve-regulated drinking line
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature: 16-19 °C
- Humidity (%): Monitored but values not stated
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): Light 06:00 to 18:00 GMT; Dark 18:00 to 06:00 GMT via fluorescent tubes

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

21 days

Number of animals:

Three males and three females

Details on study design:

A semi-occlusive patch test was used to assess the primary skin irritation induced by the undiluted test material. Each animal was identified by means of a number metal ear tag and/or a number written on the pinna.

Dorsal hair between the shoulders and hindquarters was closely shorn with fine electric clippers. A test site was selected and a 2 x 2 cm lint patch dosed with 0.5 mL of the test material applied to it. The patch and surrounding skin were covered by a single layer of gauze held in place by means of an elastic adhesive bandage.

After 4 hours the wrapping and patch were removed. The site was examined and scored for erythema and oedema on a graded scale of 0 to 4 using the criteria given in the applicable tables (attached). Observations were made at 30 minutes after removal of the patch (i.e. approximately 4 hours after application) and at 24, 48 and 72 hours plus 7, 14, and 21 days after application.

Irritation parameter:

erythema score

Basis:

animal: 804M

Time point:

24 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 804M

Time point:

48 h

Score:

2.5

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 804M

Time point:

72 h

Score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 800M

Time point:

24 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 800M

Time point:

48 h

Score:

2.5

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 800M

Time point:

72 h

Score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 805M

Time point:

24 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 805M

Time point:

48 h

Score:

2.5

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 805M

Time point:

72 h

Score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 833F

Time point:

24 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 833F

Time point:

48 h

Score:

2.5

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 833F

Time point:

72 h

Score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 801F

Time point:

24 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 801F

Time point:

48 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 801F

Time point:

72 h

Score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 836F

Time point:

24 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 836F

Time point:

48 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal: 836F

Time point:

72 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: 804M

Time point:

24 h

Score:

1

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: 804M

Time point:

48 h

Score:

1

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: 804M

Time point:

72 h

Score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: 800M

Time point:

24 h

Score:

1

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: 800M

Time point:

48 h

Score:

2

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: 800M

Time point:

72 h

Score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: 805M

Time point:

24 h

Score:

1

Irritation parameter:

edema score

Basis:

animal: 805M

Time point:

48 h

Score:

1

Irritation parameter:

edema score

Basis:

animal: 805M

Time point:

72 h

Score:

4

Irritation parameter:

edema score

Basis:

animal: 833F

Time point:

24 h

Score:

1

Irritation parameter:

edema score

Basis:

animal: 833F

Time point:

48 h

Score:

1

Irritation parameter:

edema score

Basis:

animal: 833F

Time point:

72 h

Score:

4

Irritation parameter:

edema score

Basis:

animal: 801F

Time point:

24 h

Score:

1

Irritation parameter:

edema score

Basis:

animal: 801F

Time point:

48 h

Score:

2

Irritation parameter:

edema score

Basis:

animal: 801F

Time point:

72 h

Score:

4

Irritation parameter:

edema score

Basis:

animal: 836F

Time point:

24 h

Score:

1

Irritation parameter:

edema score

Basis:

animal: 836F

Time point:

48 h

Score:

1

Irritation parameter:

edema score

Basis:

animal: 836F

Time point:

72 h

Score:

1

Max. score:

4

Irritant / corrosive response data:

All rabbits had slight inflammation of the treated site by 24 hours post-dosing which progressively increased until, by 72 hours, five rabbits had necrosis; by 7 days all six rabbits were thus affected. By 14 days the treated skins were either lifting or had lifted to reveal slightly discoloured new skin with small, semi-raw areas in three animals (see Table 2, attached). There are additional time points included in the study which have not been included in the Results table above as this is an older study conducted out to 21 days. the maximum results were however obtained after around 72hrs and did not reverse so only timepoints to 72hrs are relevant. All data has been provided in the attachment for reference.

By 21 days the skin reaction had cleared completely in two rabbits but three animals still had a light brown discolouration of the skin; it was anticipated that this would clear in time. However, the skin of 805 M was discoloured (purple) and thickened, which was taken as a permanent alteration to the skin of this animal.

Interpretation of results:

Category 1C (corrosive)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the test material caused irreversible damage to the skin of one rabbit and is therefore considered to be corrosive.

Executive summary:

The potential of the test material to cause skin irritation was assessed in a study conducted using methodology equivalent to that outlined in the standardised guideline OECD 404 under GLP conditions.

Three males and three female New Zealand White rabbits were exposed to the test material in a semi-occlusive fashion for 4 hours. A lint patch dosed with 0.5 mL of the undiluted test material was applied to the shaved skin of the rabbits. Following the exposure, rabbits were observed 30 minutes after removal of the patch (i.e. approximately 4 hours after application) and at 24, 48 and 72 hours plus 7, 14, and 21 days after application. The site was examined and scored for erythema and oedema on a graded scale of 0 to 4.

There was so little inflammation (erythema) half an hour after removal of the patch that a score between 0 and 1 was assigned for each animal. Although not a standard procedure, if the response was considered to be between the scores usually used (i.e., 0 to 4 for both erythema and oedema) an intermediate score was allocated. Inflammation progressed afterwards, reaching a maximal point on day 7 when all animals were assigned 4 for both erythema and oedema and a note was added in the report to state “All treated skins thickened with light brown, necrotic appearance so maximum scores given.“ At 14 days there was some recovery, but 3 out of the 6 rabbits still had erythema values > 2. At 21 days 2 animals appeared to have completely recovered, 3 animals presented “Light brown discolouration of skin; probably not permanent effect” but the remaining one presented “treated skin thickened and discoloured (purple); probably permanent effect”. Although on the basis of the group mean 24, 48 and 72 hour scores for erythema and oedema (2.7 and 1.9, respectively) the test material would be classified as a moderate irritant, this scheme is clearly inappropriate in this case where the skin responses increased with time up to and including 7 days post-dosing and one animal developed irreversible skin effects.

Under the conditions of this study the test material caused irreversible damage to the skin of one rabbit and is therefore considered to be corrosive.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985-07-22 to 1985-08-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

The method of Draize, as described in the Federal Register (1963) was used

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age on receipt from supplier: 3-6 months
- Age at study initiation: 4-9 months
- Weight at study initiation: 4312-4682 g (female); 4220-4360 g (male)
- Housing: Singly housed in hanging, stainless-steel cages with all-mesh floors and tops and half-mesh fronts; each cage measured 67 x 43 x 45 cm. The cages were mounted in three rows of two on a single-sided rack (a total of six cages).
- Diet: ad libitum by means of a top-loading food hopper
- Water: Filtered but untreated from water from the public supply by means of an automatic valve-regulated drinking line
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature: 16-19 °C
- Humidity: Monitored but values not stated
- Air changes: No data
- Photoperiod: Light 06:00 to 18:00 GMT; Dark 18:00 to 06:00 GMT via fluorescent tubes

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

Single application

Observation period (in vivo):

7 days (all animals); 14 days (5 out of 6 animals)

Number of animals or in vitro replicates:

Three males and three females

Details on study design:

Each rabbit was identified by means of a numbered metal ear tag and/or a number written on the pinna.

The day before testing, the eyes of the rabbits were carefully examined for any damage, and any animals showing damage were replaced. A dose of 0.1 mL of the undiluted test material was placed into the lower conjunctival sac of one eye of each rabbit and the lids held together for a few seconds to prevent loss of material. The eyes were not washed.

The reactions of each animal were observed immediately after instillation and the initial pain response graded (see table, attached). A visual assessment of eye irritancy was then made at 1 hour, 24 hours, 48 hours, 72 hours and 7 days after instillation. Five of the six rabbits were retained and observed at 14 days post-dosing. Irritancy was scored for the cornea, iris and conjunctivae using standard scores detailed in the attached listings. In the event of any corneal damage visualisation was aided by the instillation of one drop of 2 % fluorescein solution.

Irritation parameter:

cornea opacity score

Basis:

animal: 838M

Time point:

other: 1h

Score:

0

Max. score:

2.5

Remarks on result:

other:

Remarks:

additional timepoints used in study (1h-14d)

Irritation parameter:

cornea opacity score

Basis:

animal: 838M

Time point:

24 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 838M

Time point:

48 h

Score:

0.5

Irritation parameter:

cornea opacity score

Basis:

animal: 838M

Time point:

72 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 838M

Time point:

7 d

Score:

2.5

Remarks on result:

other:

Remarks:

additional timepoints in older study

Irritation parameter:

cornea opacity score

Basis:

animal: 838M

Time point:

14 d

Score:

2.5

Remarks on result:

other:

Remarks:

additional timepoints in older study

Irritation parameter:

iris score

Basis:

animal: 838M

Time point:

24/48/72 h

Score:

0

Max. score:

0.5

Irritation parameter:

conjunctivae score

Basis:

animal: 838M

Time point:

other: 1h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 838M

Time point:

24 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 838M

Time point:

48 h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 838M

Time point:

72 h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 838M

Time point:

7 d

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 838M

Time point:

14 d

Score:

1

Irritation parameter:

chemosis score

Basis:

animal: 838M

Time point:

other: 1h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 838M

Time point:

24 h

Score:

1.5

Irritation parameter:

chemosis score

Basis:

animal: 838M

Time point:

48 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 838M

Time point:

72 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 838M

Time point:

7 d

Score:

0

Irritation parameter:

chemosis score

Basis:

animal: 838M

Time point:

14 d

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal: 839M

Time point:

other: 1h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 839M

Time point:

24 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 839M

Time point:

48 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 839M

Time point:

72 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 839M

Time point:

7 d

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 839M

Time point:

14 d

Score:

1

Irritation parameter:

chemosis score

Basis:

animal: 839M

Time point:

other: 1h

Score:

1

Irritation parameter:

chemosis score

Basis:

animal: 839M

Time point:

24 h

Score:

1

Irritation parameter:

chemosis score

Basis:

animal: 839M

Time point:

48 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 839M

Time point:

72 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 839M

Time point:

7 d

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 839M

Time point:

14 d

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 839M

Time point:

other: 1h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 839M

Time point:

24 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 839M

Time point:

48 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 839M

Time point:

72 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 839M

Time point:

7 d

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 839M

Time point:

14 d

Score:

3

Irritation parameter:

iris score

Basis:

animal: 839M

Time point:

other: 1h

Score:

0

Irritation parameter:

iris score

Basis:

animal: 839M

Time point:

24 h

Score:

0.5

Irritation parameter:

iris score

Basis:

animal: 839M

Time point:

48 h

Score:

0.5

Irritation parameter:

iris score

Basis:

animal: 839M

Time point:

72 h

Score:

0.5

Irritation parameter:

iris score

Basis:

animal: 839M

Time point:

7 d

Score:

1

Irritation parameter:

iris score

Basis:

animal: 839M

Time point:

14 d

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal: 840M

Time point:

other: 1h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 840M

Time point:

24 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 840M

Time point:

48 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 840M

Time point:

72 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 840M

Time point:

7 d

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 840M

Time point:

14 d

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 840M

Time point:

other: 1h

Score:

1

Irritation parameter:

chemosis score

Basis:

animal: 840M

Time point:

24 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 840M

Time point:

48 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 840M

Time point:

72 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 840M

Time point:

7 d

Score:

1

Irritation parameter:

chemosis score

Basis:

animal: 840M

Time point:

14 d

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 840M

Time point:

other: 1h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 840M

Time point:

24 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 840M

Time point:

48 h

Score:

0.5

Irritation parameter:

cornea opacity score

Basis:

animal: 840M

Time point:

72 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 840M

Time point:

7 d

Score:

2

Irritation parameter:

cornea opacity score

Basis:

animal: 840M

Time point:

14 d

Score:

3

Irritation parameter:

iris score

Basis:

animal: 840M

Time point:

other: 1h

Score:

0

Irritation parameter:

iris score

Basis:

animal: 840M

Time point:

24 h

Score:

0

Irritation parameter:

iris score

Basis:

animal: 840M

Time point:

48 h

Score:

0

Irritation parameter:

iris score

Basis:

animal: 840M

Time point:

72 h

Score:

0.5

Irritation parameter:

iris score

Basis:

animal: 840M

Time point:

7 d

Score:

0.5

Irritation parameter:

iris score

Basis:

animal: 840M

Time point:

14 d

Score:

0.5

Irritation parameter:

conjunctivae score

Basis:

animal: 843F

Time point:

other: 1h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 843F

Time point:

24 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 843F

Time point:

48 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 843F

Time point:

72 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 843F

Time point:

7 d

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 843F

Time point:

14 d

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 843F

Time point:

other: 1h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 843F

Time point:

24 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 843F

Time point:

48 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 843F

Time point:

72 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 843F

Time point:

7 d

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 843F

Time point:

14 d

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 843F

Time point:

other: 1h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 843F

Time point:

24 h

Score:

0.5

Irritation parameter:

cornea opacity score

Basis:

animal: 843F

Time point:

48 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 843F

Time point:

72 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 843F

Time point:

7 d

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 843F

Time point:

14 d

Score:

1.5

Irritation parameter:

iris score

Basis:

animal: 843F

Time point:

24/48/72 h

Score:

0

Max. score:

0.5

Irritation parameter:

conjunctivae score

Basis:

animal: 844F

Time point:

other: 1h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 844F

Time point:

24/48/72 h

Score:

2.5

Max. score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 844F

Time point:

7 d

Score:

2.5

Irritation parameter:

chemosis score

Basis:

animal: 844F

Time point:

other: 1h

Score:

1.5

Irritation parameter:

chemosis score

Basis:

animal: 844F

Time point:

24 h

Score:

1.5

Irritation parameter:

chemosis score

Basis:

animal: 844F

Time point:

48 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 844F

Time point:

72 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 844F

Time point:

7 d

Score:

2

Irritation parameter:

cornea opacity score

Basis:

animal: 844F

Time point:

other: 1h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 844F

Time point:

24 h

Score:

0.5

Irritation parameter:

cornea opacity score

Basis:

animal: 844F

Time point:

48 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 844F

Time point:

72 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 844F

Time point:

7 d

Score:

2.5

Remarks on result:

other: vascularisation

Remarks:

vascularisation considered permanent and test terminated at 7d

Irritation parameter:

iris score

Basis:

animal: 844F

Time point:

24/48/72 h

Score:

0

Max. score:

0.5

Irritation parameter:

conjunctivae score

Basis:

animal: 845F

Time point:

other: 1h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 845F

Time point:

24 h

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 845F

Time point:

48 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 845F

Time point:

72 h

Score:

2.5

Irritation parameter:

conjunctivae score

Basis:

animal: 845F

Time point:

7 d

Score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: 845F

Time point:

14 d

Score:

0.5

Irritation parameter:

chemosis score

Basis:

animal: 845F

Time point:

other: 1h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 845F

Time point:

24 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 845F

Time point:

48 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 845F

Time point:

72 h

Score:

2

Irritation parameter:

chemosis score

Basis:

animal: 845F

Time point:

7 d

Score:

1

Irritation parameter:

chemosis score

Basis:

animal: 845F

Time point:

14 d

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 845F

Time point:

other: 1h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 845F

Time point:

24 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: 845F

Time point:

48 h

Score:

0.5

Irritation parameter:

cornea opacity score

Basis:

animal: 845F

Time point:

72 h

Score:

1

Irritation parameter:

cornea opacity score

Basis:

animal: 845F

Time point:

7 d

Score:

1.5

Irritation parameter:

cornea opacity score

Basis:

animal: 845F

Time point:

14 d

Score:

2

Irritation parameter:

iris score

Basis:

animal: 845F

Time point:

other: 1h

Score:

0

Irritation parameter:

iris score

Basis:

animal: 845F

Time point:

24/48/72 h

Score:

0

Max. score:

0.5

Irritant / corrosive response data:

The instillation of the undiluted test material into the conjunctival sac of one eye of each of six rabbits resulted in severe initial pain. The conjunctival redness, chemosis and discharge, corneal opacity and damage to the iris were assessed, and the mean total scores calculated. The results are shown in Table 3 (attached).

All rabbits had moderate inflammation of the conjunctivae within one hour of dosing, which persisted up to 7 days. During this period all animals developed corneal opacity and had transient iritis. At 7 days 844 F had extensive vascularisation of the cornea, which is considered a permanent effect; this animal was therefore killed. Of the remaining five rabbits, four also developed extensive corneal vascularisation by day 14 so the test was terminated.

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

On the basis of the modified Kay and Callandra rating system, the test material can be classified as severely irritating to rabbit eyes; on the basis of the permanent effects seen (extensive vascularisation of the cornea) the test material would be classified as Category 1 (1272/2008).

Executive summary:

The potential of the test material to cause eye irritation was assessed in a study conducted using methodology equivalent to that outlined in the standardised guideline OECD 405 under GLP conditions.

0.1 mL of the test material was instilled into the eye of three male and three female New Zealand White rabbits. The dose of the undiluted test material was placed into the lower conjunctival sac of one eye of each rabbit and the lids held together for a few seconds to prevent loss of material. The eyes were not washed.

The reactions of each animal were observed immediately after instillation and the initial pain response graded (see table, attached). A visual assessment of eye irritancy was then made at 1 hour, 24 hours, 48 hours, 72 hours and 7 days after instillation. Five of the six rabbits were retained and observed at 14 days post-dosing. Irritancy was scored for the cornea, iris and conjunctivae using standard scores detailed in the attached listings. In the event of any corneal damage visualisation was aided by the instillation of one drop of 2 % fluorescein solution.

The instillation of the undiluted test material into the conjunctival sac of one eye of each of six rabbits resulted in severe initial pain. The conjunctival redness, chemosis and discharge, corneal opacity and damage to the iris were assessed, and the mean total scores calculated.

All rabbits had moderate inflammation of the conjunctivae within one hour of dosing, which persisted up to 7 days. During this period all animals developed corneal opacity and had transient iritis. At 7 days 844 F had extensive vascularisation of the cornea, which is considered a permanent effect; this animal was therefore killed. Of the remaining five rabbits, four also developed extensive corneal vascularisation by day 14 so the test was terminated.

On the basis of the modified Kay and Callandra rating system, the test material can be classified as severely irritating to rabbit eyes; on the basis of the permanent effects seen (extensive vascularisation of the cornea) the test material would be classified as Category 1 (1272/2008).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion

Two key studies are provided to address this endpoint; one in vitro and one in vivo.

In vitro

The corrosion potential of the test material was investigated in an in vitro skin corrosion test using a human skin model in accordance with the standardised guidelines OECD 431 and EU Method B.40 BIS under GLP conditions. The study was awarded a reliability score of 1 in accordance with the principles for assessing data quality set forth by Klimisch et al. (1997).

The study determined the ability of the test material to induce skin corrosion on a human three dimensional epidermal model (EpiDerm (EPI-200)). The possible corrosive potential was tested through topical application for 3 minutes and 1 hour. The skin was moistened with 25 µL of Milli-Q and the test material was applied (25 mg) directly on top of the skin tissue. Tissues were also exposed to both positive (8 N potassium hydroxide) and negative (Milli-Q water) concurrent controls.

The positive control had a mean relative tissue viability of 8 % after 3 minutes of exposure. The absolute mean OD540 (optical density at 540 nm) of the negative control tissues was within the laboratory historical control data range. The maximum inter-tissue variability in viability between two tissues treated identically was less or equal to 4 % and the maximum difference in percentage between the mean viability of two tissues and one of the two tissues was less than 2 % for the reference materials, indicating that the test system functioned properly.

Although the inter-tissue variability in viability between two tissues and the maximum difference in percentage between the mean viability of two tissues and one of the two tissues treated with the test material were above the acceptability criteria (up to 58 and 29 %, respectively, at the 1 hour treatment period), all individual results obtained were clearly negative.

A repeat experiment was performed to verify the obtained results.

In the second experiment, the absolute mean OD540 of the negative control tissues was within the laboratory historical control data range. The positive control had a mean relative tissue viability of 9 % after 3 minutes of exposure. The maximum inter-tissue variability in viability between two tissues treated identically was less than 30 % and the maximum difference in percentage between the mean viability of two tissues and one of the two tissues was less than or equal to 5 % for the test material and less than 18 % for the positive control.

Although at the 1 hour exposure time the difference in percentage between the mean viability of two tissues and one of the two tissues treated with the positive control was above the acceptability criteria for one of the two tissues (17.6 %), both individual results obtained were clearly positive. Consequently, this deviation has no impact on the outcome of the study.

Skin corrosion is expressed as the remaining cell viability after exposure to the test material. The relative mean tissue viability obtained after 3-minute and 1-hour treatments with the test material compared to the negative control tissues was 88 and 53 %, respectively in the first experiment. In the second experiment, the relative mean tissue viability obtained after the 3-minute and 1-hour treatments with the test material compared to the negative control tissues was 97 and 87 %, respectively. As the mean relative tissue viability was not below 50 % after the3-minute treatment and not below 15 % after the 1-hour treatment, the test material is considered to be not corrosive.

Under the conditions of this study, the test material is not corrosive in the in vitro skin corrosion test.

In vivo

The potential of the test material to cause skin irritation was assessed in vivo in a study conducted using methodology equivalent to that outlined in the standardised guideline OECD 404 under GLP conditions. The study was awarded a reliability score of 1 in accordance with the principles for assessing data quality set forth by Klimisch et al. (1997).

Three males and three female New Zealand White rabbits were exposed to the test material in a semi-occlusive fashion for 4 hours. A lint patch dosed with 0.5 mL of the undiluted test material was applied to the shaved skin of the rabbits. Following the exposure, rabbits were observed 30 minutes after removal of the patch (i.e. approximately 4 hours after application) and at 24, 48 and 72 hours plus 7, 14, and 21 days after application. The site was examined and scored for erythema and oedema on a graded scale of 0 to 4.

There was so little inflammation (erythema) half an hour after removal of the patch that a score between 0 and 1 was assigned for each animal. Although not a standard procedure, if the response was considered to be between the scores usually used (i.e., 0 to 4 for both erythema and oedema) an intermediate score was allocated. Inflammation progressed afterwards, reaching a maximal point on day 7 when all animals were assigned 4 for both erythema and oedema and a note was added in the report to state “All treated skins thickened with light brown, necrotic appearance so maximum scores given.“ At 14 days there was some recovery, but 3 out of the 6 rabbits still had erythema values > 2. At 21 days 2 animals appeared to have completely recovered, 3 animals presented “Light brown discolouration of skin; probably not permanent effect” but the remaining one presented “treated skin thickened and discoloured (purple); probably permanent effect”. Although on the basis of the group mean 24, 48 and 72 hour scores for erythema and oedema (2.7 and 1.9, respectively) the test material would be classified as a moderate irritant, this scheme is clearly inappropriate in this case where the skin responses increased with time up to and including 7 days post-dosing and one animal developed irreversible skin effects.

Under the conditions of this study the test material caused irreversible damage to the skin of one rabbit and is therefore considered to be corrosive.

Eye Irritation

The potential of the test material to cause eye irritation was assessed in a study conducted using methodology equivalent to that outlined in the standardised guideline OECD 405 under GLP conditions. The study was awarded a reliability score of 1 in accordance with the principles for assessing data quality set forth by Klimisch et al. (1997).

0.1 mL of the test material was instilled into the eye of three male and three female New Zealand White rabbits. The dose of the undiluted test material was placed into the lower conjunctival sac of one eye of each rabbit and the lids held together for a few seconds to prevent loss of material. The eyes were not washed.

The reactions of each animal were observed immediately after instillation and the initial pain response graded. A visual assessment of eye irritancy was then made at 1 hour, 24 hours, 48 hours, 72 hours and 7 days after instillation. Five of the six rabbits were retained and observed at 14 days post-dosing. Irritancy was scored for the cornea, iris and conjunctivae using standard scores detailed in the attached listings. In the event of any corneal damage visualisation was aided by the instillation of one drop of 2 % fluorescein solution.

The instillation of the undiluted test material into the conjunctival sac of one eye of each of six rabbits resulted in severe initial pain. The conjunctival redness, chemosis and discharge, corneal opacity and damage to the iris were assessed, and the mean total scores calculated.

All rabbits had moderate inflammation of the conjunctivae within one hour of dosing, which persisted up to 7 days. During this period all animals developed corneal opacity and had transient iritis. At 7 days 844 F had extensive vascularisation of the cornea, which is considered a permanent effect; this animal was therefore killed. Of the remaining five rabbits, four also developed extensive corneal vascularisation by day 14 so the test was terminated.

Under the conditions of this study, the test material caused serious eye damage following application to the anterior surface of the eye, which is not fully reversible within 21 days of application.

Justification for selection of skin irritation / corrosion endpoint:
The in vivo study was selected on the basis that the positive result in vivo outweighs the negative result obtained in vitro.

Justification for selection of eye irritation endpoint:
Only one study available.

Effects on skin irritation/corrosion: corrosive

Effects on eye irritation: corrosive

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance requires classification with respect to both skin and eye irritation/corrosion.

The substance requires classification for skin corrosion as Category 1C (H314: Causes severe skin burns and eye damage) and for serious eye damage Category 1 (H318: Causes serious eye damage).