3-(methylthio)propionaldehyde

Endpoint summary

Description of key information

Skin irritation (in vivo, similar to OECD 404): irritating

Eye irritation (in vivo, OECD 405): irreversible damage

Respiratory irritation: not irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

No information on analytical purity, no experimental details, no details given on animals and environmental conditions, observation period restricted to 5 days in which the effects were not fully reversible, type of coverage and amount of test substance not specified, no separate scoring for erythema and edema for individual animals, only numerical evaluation reported

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Primary skin irritation in rabbits
- Short description of test conditions: Undiluted test substance was applied to the skin of 3 rabbits, followed by an observation period of 5 days.
- Parameters analysed / observed: Skin reactions were assessed according to Draize criteria 2, 24, 48 and 72 h, and 5 days after test substance administration.

GLP compliance:

no

Species:

rabbit

Strain:

not specified

Type of coverage:

not specified

Preparation of test site:

not specified

Vehicle:

not specified

Controls:

no

Observation period:

5 days
Reading time points: 2, 24, 48, 72 h and 5 days

Number of animals:

3 (1 male and 2 female)

Details on study design:

OBSERVATION TIME POINTS: 2, 24, 48 and 72 h, and 5 days

SCORING SYSTEM:
- Method of calculation: Draize, Woodard and Calvery (Journal of Pharm. and Exp. Therapeutics, 82:4, 1944)

Irritation parameter:

overall irritation score

Remarks:

Numerical evaluation of skin reactions

Basis:

other: average score over all animals and all skin reactions

Time point:

24 h

Score:

4

Reversibility:

not fully reversible within: 5 days

Irritation parameter:

overall irritation score

Remarks:

Numerical evaluation of skin reactions

Basis:

other: average score over all animals and skin reactions

Time point:

48 h

Score:

3

Reversibility:

not fully reversible within: 5 days

Irritation parameter:

overall irritation score

Remarks:

Numerical evaluation of skin reactions

Basis:

animal: average score over all animals and all skin reactions

Time point:

72 h

Score:

2

Reversibility:

not fully reversible within: 5 days

Skin reactions were assessed based on numerical evaluation. After 2 h of exposure, the skin developed a blush tint and an average score of 2.6 out of 8 possible was observed. The animals showed well defined erythema and two instances of slight edema were observed. After 24 h inflammation in form of blistering increased to an average score of 4.0. Afterwards, there was a gradual decrease and the average score dropped to 0.6 by observation Day 5.

Interpretation of results:

study cannot be used for classification

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

other: Draize test

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 28 Jul 2015

Deviations:

yes

Remarks:

24 h instead of 4 h application, occlusive instead of semi-occlusive application, no details on animals and environmental conditions, reading time points 24 and 72 h only, test item administration application onto abraded and to intact skin area

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.5 - 3.5 kg
- Housing: Individually in cages measuring 600 x 540 x 315 cm
- Diet: granulated rabbit food (SANDERS), 200 g per rabbit per day
- Water: ad libitum

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g

Duration of treatment / exposure:

24 h

Observation period:

3 days
Reading time points: 24 and 72 h

Number of animals:

6

Irritation parameter:

erythema score

Basis:

animal: #1, #3, #4, #5 and #6

Time point:

other: 24/72 h

Score:

0.5

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 24/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility not applicable

Irritation parameter:

edema score

Basis:

animal: #1 and #3

Time point:

other: 24/72 h

Score:

1.5

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: #2, #5 and #6

Time point:

other: 24/72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal: #4

Time point:

other: 24/72 h

Score:

0.5

Max. score:

4

Reversibility:

not reversible

Irritant / corrosive response data:

Barely perceptible erythema (score 0.5) was observed in 5/6 rabbits, which was fully reversible in all animals within 72 hours after test item instillation. Barely perceptible edema (score 0.5) was observed in 1/6 rabbits, slight edema (score 1) in 3/6 rabbits and slight to well defined edema (score 1.5) was observed in 2/6 rabbits and remained unreversible until study termination 72 hours after test item instillation.

Interpretation of results:

study cannot be used for classification

Conclusions:

The test conditions (24 h exposure under occlusive conditions) may have led to more detrimental skin reactions than those specified in current guidelines. In the 3-days observation period the reversibility of the local skin reactions could not fully be assessed.

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

no information on analytical purity, only a short summary is available, 24 h exposure, type of coverage not specified, no experimental details, no information on animals and environmental conditions

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

- Principle of test: Primary skin irritation in rabbits
- Short description of test conditions: 0.5 mL of undiluted test substance was applied to abraded and intact skin of 6 rabbits. Afterwards, the animals were observed for 7 days.
- Parameters analysed / observed: Skin reactions were assessed according to Draize criteria after 4, 24, 48 and 72 h and 7 days.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

24 h

Observation period:

7 days
Reading time points: 4, 24, 48 and 72 h and 7 days

Number of animals:

6

Details on study design:

TEST SITE: The test material was applied to intact and abraded skin.

OBSERVATION TIME POINTS : 4, 24, 48 and 72 h and 7 days

SCORING SYSTEM:
- Method of calculation: Draize scoring system

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: intact skin

Irritation parameter:

erythema score

Basis:

animal: #2, #3 and #5

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: intact skin

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: intact skin

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: intact skin

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: intact skin

Irritation parameter:

edema score

Basis:

animal: #2 and #4

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: intact skin

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: intact skin

Irritation parameter:

edema score

Basis:

animal: #5 and #6

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: intact skin

Other effects:

Defatting effect: The skin sloughed off in 10-14 days. There was no injury in depth.

Interpretation of results:

study cannot be used for classification

Reference 4

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Remarks:

pre-GLP study, occlusive instead of semi-occlusive application, 24 h instead of 4 h application, pre-GLP study, short documentation, 2 reading times only, observation period of 3 days only

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Primary skin irritation in rabbits with abrarded and intact skin
- Short description of test conditions: 0.5 mL of the test item was administered to the clipped test site of 12 rabbits (6 rabbits with abraded skin, 6 rabbits with intact skin). After 24 hours of exposure under occlusive conditions, the patches and test material were removed.
- Parameters analysed / observed: Skin reactions were assessed according to Draize criteria 24 h (immediately following patch removal) and 72 h after test substance administration (48 h after patch removal).

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

24 h

Observation period:

3 days
Reading time points: 24 h (immediately following patch removal) and 72 h (48 h after patch removal)

Number of animals:

6

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3, #4, #5 and #6

Time point:

other: 24/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks:

within 3 days

Remarks on result:

other: intact skin

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24/72 h

Score:

2.5

Max. score:

4

Reversibility:

not reversible

Remarks:

within 3 days

Remarks on result:

other:

Remarks:

intact skin

Irritation parameter:

edema score

Basis:

animal: #2, #3, #4 and #6

Time point:

other: 24/72 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 3 days

Remarks on result:

other: intact skin

Irritation parameter:

edema score

Basis:

animal #5

Time point:

other: 24/72 h

Score:

1.5

Max. score:

4

Reversibility:

not reversible

Remarks:

within 3 days

Remarks on result:

other: intact skin

Irritant / corrosive response data:

There were no significant difference between the reactions of intact skin and those of the abraded skin. Well-defined erythema and slight to moderate edema were observed at 24 and 72 h after removal of the patch.

Interpretation of results:

study cannot be used for classification

Conclusions:

The test conditions (24 h exposure under occlusive conditions) may have led to more detrimental skin reactions than those specified in current guidelines. In the 3-days observation period the reversibility of the local skin reactions could not fully be assessed.

Reference 5

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

limited documentation, analytical purity of test material not reported, postexposure observation period of 7 days only, no 72 h reading, occlusive application.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 28 Jul 2015

Deviations:

yes

Remarks:

short documentation, analytical purity of test material not reported, postexposure observation period too short, no 72 h reading, occlusive instead of semiocclusive application

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

6 days
Reading time points: 4, 24 and 48 h after patch removal

Number of animals:

6

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: The test substance was brought on the intact or abraded skin under a surgical patch. The patches were fixed to the application sites by means of adhesive tape and the entire trunk of the rabibts was wrapped with an impervious material to maintain the test patches in position and to retard evaporation of volatile substances.

REMOVAL OF TEST SUBSTANCE
- Washing: After exposure the patches and the material applied were removed.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS: 4, 24 and 48 h after patch removal

SCORING SYSTEM:
- Method of calculation: Draize scoring system

Irritation parameter:

erythema score

Basis:

animal: #1, #3, #4, #5 and #6

Time point:

24/48 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks:

within 48 h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 48 h

Irritation parameter:

edema score

Basis:

animal: #1 and #2

Time point:

24/48 h

Score:

2.5

Max. score:

4

Reversibility:

not reversible

Remarks:

within 48 h

Irritation parameter:

edema score

Basis:

animal: #3, #4, #5 and #6

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 48 h

Irritant / corrosive response data:

After 4 hours of exposure all rabbits showed well-defined (score 2, 2/6 rabbits) or moderate to severe erythema (score 3, 4/6 rabbits) and slight (score 2, 1/6 rabbits) or moderate edema (score 3, 5/6 rabbits). At the 24 and 48 h reading time point 5/4 rabbits showed severe erythema (score 4), 1/6 rabbits showed well defined erythema (score 2). At 24 h, 4/6 rabbits showed slight edema (score 2) and 2/6 rabbits showed moderate edema (score 3). At the 48 h reading time point all 6/6 rabbits showed slight edema (score 2). After 6 days the intact, treated skin areas showed distinct scaliness and very slight to distinct necrosis.

Interpretation of results:

study cannot be used for classification

Conclusions:

The test conditions (exposure under occlusive conditions, no 72-h reading time point) may have led to more detrimental skin reactions than those specified in current guidelines. In the 6-days observation period the reversibility of the local skin reactions could not fully be assessed.

Reference 6

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

(limited documentation, analytical purity of test material not reported, postexposure observation period only 48 h, no 72 h reading, occlusive instead of semiocclusive application)

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Primary corrosion to the skin was measured by a patch test technique on the abraded and intact skin of albino rabbits
- Short description of test conditions: 0.5 mL of the test substance were applied to the intact or abraded skin under a surgical patch measuring 1 inch x 1 inch, which were fixed with an occlusive dressing. After 4 h of exposure the patches and test material was removed.
- Parameters analysed / observed: Skin reactions were assessed according to Draize criteria 4, 24 and 48 h after patch removal.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

48 h
reading time points: 4, 24 and 48 h

Number of animals:

6

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: The test substance was brought on the intact or abreaded skin under a surgical patch. The patches were fixed to the application sites by means of adhesive tape and the entire trunk of the rabbibts was wrapped with an impervious material to maintain the test patches in position and to retard evaporation of volatile substances.

REMOVAL OF TEST SUBSTANCE
- Washing: After exposure the patches and the material applied were removed.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS : 4, 24 and 48 h

SCORING SYSTEM:
- Method of calculation: Draize scoring system

Irritation parameter:

erythema score

Basis:

animal: #1,#3, #4, #5 and #6

Time point:

24/48 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks:

within 48 h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 48 h

Irritation parameter:

edema score

Basis:

animal: #1 and #2

Time point:

24/48 h

Score:

2.5

Max. score:

4

Reversibility:

not reversible

Remarks:

within 48 h

Irritation parameter:

edema score

Basis:

animal: #3, #4, #5 and #6

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 48 h

Irritant / corrosive response data:

After 4 h of exposure all rabbits showed well-defined (score 2, 4/6 rabbits) or moderate to severe erythema (score 3, 2/6 rabbits) and slight (score 2, 1/6) or moderate edema (score 3, 5/6 rabbits). At the 24- and 48- h reading time point 5/4 rabbits showed severe erythema (score 4), 1/6 rabbits showed well defined erythema (score 2). At the 24- h readinig timepoint, 4/6 rabbits showed slight edema (score 2) and 2/6 rabbits showed moderate edema (score 3). At the 48- h reading time point all 6/6 rabbits showed slight edema (score 2).

Other effects:

At the 24- and 48- h reading time point, slight to distinct ischemia and haemorrages were observed.

Interpretation of results:

study cannot be used for classification

Conclusions:

The test conditions (exposure under occlusive conditions, no 72-h reading time point) may have led to more detrimental skin reactions than those specified in current guidelines. In the 48-h observation period the reversibility of the local skin reactions could not fully be assessed.

Reference 7

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985-07-01 to 1985-08-22

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 28 Jul 2015

Deviations:

yes

Remarks:

reading time point 30 min instead of 1 h, 24 h occlusive exposure tested on the same animals at a different test site, no information on analytical purity

Qualifier:

according to guideline

Guideline:

other: FIFRA Pesticide Assessment Guidelines, Subdivision F, Hazard Evaluation: Human and Domestic Animals; Office of Pesticide Programs, U.S. Environmental Protection Agency, Office of Pesticides and Toxic Substances, November 1982; Section 81-5, Primary Dermal

Version / remarks:

adopted Aug 1978

Qualifier:

according to guideline

Guideline:

other: TSCA: Health Effects Test Guidelines, Office of Toxic Substances, Office of Pesticides and Toxic Substances, United States Environmental Protection Agency, August 1982 Acute Exposure, Primary Dermal Irritation

Version / remarks:

adopted 26 Jul 1979

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Source: Hazleton Dutchland, Inc., Denver, Pennsylvania, USA
- Age at study initiation: At least 8 weeks
- Equilibration period: 38 days
- Housing: Individually housed
- Diet: Lab Rabbit Chow HF (Purina #5326), ad libitum
- Water: Tap water supplied by an automatic watering system (Municipal water), ad libitum

Environmental conditions:
- Temperature: 60-70°F (16-21 °C)
- Humidity: 30-70%
- Light cycle: 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

4 h (semi-occlusive dressing) and 24 h (occlusive dressing)

Observation period:

14 days
Reading time points:
4 h exposure: 30 min, 24, 48 and 72 h after patch removal
24 h exposure: 24, 48 and 72 h after patch removal

Number of animals:

6 (The same 6 animals were used for both sets of application sites, 4 and 24 h exposure.)

Details on study design:

TEST SITE
- Area of exposure: closely clipped dorsal area of the trunk
- % coverage: 10
- Type of wrap if used:
4h exposure, semi-occlusive conditions: The test material was applied beneath a gauze square 1x1 cm, placed on each of two test sites nearest the head and held in place with a non-irritating tape. Gauze was then wrapped around the animal and covered with porous tape.
24 h exposure, occlusive conditions: The test material was applied beneath a gauze square 1x1 cm, placed on each of two test sites posterior to the 4 h sites and held in place with a non-irritating tape. Plastic sheeting was then wrapped around the animal and secured with tape to retard evaporation and keep the test material in contact with the skin without pressure.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Not performed.

OBSERVATION TIME POINTS:
4 h exposure, semio-occlusive conditions: 30 minutes, 24, 48 and 72 h after patch removal
24 h exposure, occlusive conditions: 24, 48 and 72 h after patch removal

SCORING SYSTEM:
- Method of calculation: Draize scoring system

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Desquamation on day 7;

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: Desquamation on day 10 & 14;

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: Desquamation on day 10 & 14;

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Desquamation on day 7, 10 & 14;

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Desquamation on day 14;

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Desquamation on day 7, 10 & 14;

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

fully reversible within: 10 days

Remarks on result:

other:

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

fully reversible within: 10 days

Remarks on result:

other:

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

fully reversible within: 10 days

Remarks on result:

other:

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

fully reversible within: 10 days

Remarks on result:

other:

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

edema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other:

Remarks:

4 h semi-occlusive exposure

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other:

Remarks:

4 h semi-occlusive exposure

Irritant / corrosive response data:

4 h exposure: Irritation was generally moderate but transient and consisted of well-defined to moderate erythema (score 2 and 3) and slight to severe edema (score 2 – 4) in all animals at the 4 h and 24 h reading. 48 and 72 h after patch removal, very slight to moderate to severe erythema (score 1 – 3) and very slight to moderate edema (score 1 – 3) was still present in all animals. About 7 days after patch removal, 5/6 rabbits had very slight erythema (score 1) and 1/6 rabbits had well defined erythema (score 2). Very slight erythema (score 1) persisted in 3/6 rabbits until study termination on Day 14. Very slight edema (score 1) was observed up to Day 7 in 4/6 rabbits and fully reversible in all animals until study Day 10.
24 h exposure: All animals showed moderate to severe erythema (score 3-4) during the whole study period and moderate to severe erythema (score 3-4), which remained slight to moderate (score 2-3) until study termination.

Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test - 4 hours semi-occluded exposure (median scores of the two exposure sites)

Time point	Erythema (max score 4)	Edema (max score 4)
4.5 hrs	2/3/2/2/2/2	3/3,5/3/3/2/3
24 hrs	2/3/3/2/3/3	3/2,5/3,5/3/2/2
48 hrs	2/3/3/1/2/2	2/2/2,5/2/2/2
72 hrs	2/2/3/1/2/2	2/2/2/2/1/1
Day 7	1^#/1/1/1,5^#/1/1^#	1/1/1/1,5/0/0
Day 10	1/0^#/0^#/1^#/0/1^#	0/0/0/0/0/0
Day 14	1/0^#/0^#/1^#0,5^#/1^#	0/0/0/0/0/0
Average 24hr/48hr/72hr	2/2.67/3/1.33/2.33/2.33	2.33/2.17/2.67/2.33/1.67/1.67
Reversibility*), **)	nc./c/c/nc/nc/nc	c/c/c/c/c/c
Time for Reversion (day)	-/10/10/-/-/-	10/10/10/10/7/7

*) Reversibility: c = completely reversible; nc = not completely reversible; n = not reversible

# Desquamation

**) The observation time did not extend beyond 14 days, therefore, full reversibility of the skin effects can not be proved. However, skin lesions showed a clear tendency of reversibility. At study termination, reversibility was complete in 2/6 animals and almost complete in the remaining 4/6 animals. Therefore, a full reversibility in all animals seems to be very likely.

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

CLP: Skin Irrit. 2, H315 according to Regulation (EC) No. 1272/2008
 

Reference 8

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 28 Jul 2015

Deviations:

yes

Remarks:

no erythema and edema values for individual animals reported, only average scores given, occlusive application

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.0 - 3.0 kg

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

17 days
Reading time points: 1, 24, 48 and 72 h, and 7, 10, 14 and 17 days

Number of animals:

6 (3 male and 3 female)

Details on study design:

TEST SITE
- Area of exposure: clipped dorsal trunk skin.
- Type of wrap if used: The area was covered with a gauze patch and loosely covered with impervious sheeting.

REMOVAL OF TEST SUBSTANCE
- Occlusive materials were removed and any excess sample wiped away with a moistened gauze swab.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS : 1, 24, 48 and 72 h, and 7, 10, 14 and 17 days

SCORING SYSTEM:
- Method of calculation: Draize scoring system

Irritation parameter:

erythema score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

2.3

Max. score:

4

Reversibility:

fully reversible within: 17 days

Irritation parameter:

edema score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

2.9

Max. score:

4

Reversibility:

fully reversible within: 17 days

Irritant / corrosive response data:

An hour after removal of the occlusive dressing there was mild to moderate erythema (score 1-2) and well defined to severe edema (score 3-4). On study Days 1-3 mild to severe erythema (score 1-4) was observed, which began to resolve between 3 and 7 days and had disappeared in half of the animals by Day 14 and in all animals by Day 17. Mild to severe edema (score 1-4) was observed in the animals up to study Day 3. One week after patch removal up to study Day 10 slight to moderate edema was observed (score 0-2). 14 days after patch removal, only slight edema (score 0-1) was observed, which was fully reversible until study termination. In addition, desquamation and scab formation were seen in all animals from Day 7-17.

Interpretation of results:

study cannot be used for classification

Conclusions:

The test conditions (occlusive conditions) may have led to more detrimental skin reactions than those specified in current guidelines. Individual animal data for erythema and edema were not reported.

Reference 9

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1996

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 28 Jul 2015

Deviations:

yes

Remarks:

Occlusive instead of semi-occlusive application, skin evaluation was performed immediately following patch removal and after 48 h, 7 and 14 days but not after 1, 24 and 72 h

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

14 days
Reading time points: 4 and 48 h, 7 and 14 days

Number of animals:

12 (6 (3 male and 3 female) for uninhibited test material and 6 (3 male and 3 female) for inhibited test material.

Details on study design:

TEST SITE
The hair of 12 (3/sex/test material) New Zealand White rabbits was closely clipped to expose the back. 0.5 ml of each test material (inhibited, uninhibited, and the control, for details please refer to "Any other information on materials and methods incl. tables") was applied, as received, beneath a 6 cm2 gauze square placed directly on the one intact site on each animal. The site was occluded during the 4-h exposure period. At the end of the exposure period, the patch was removed and the site evaluated for skin irritation.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No washing. The site was then wiped free of test material.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS: 4 and 48 h, and 7 and 14 days after test material administration

SCORING SYSTEM:
- Method of calculation: Not specified

Irritation parameter:

erythema score

Remarks on result:

other: please refer to "Irritant / corrosive response data"

Irritation parameter:

edema score

Remarks on result:

other: please refer to "Irritant / corrosive response data"

Irritant / corrosive response data:

The only irritation seen in the animals treated with inhibited test substance was very slight (barely perceptible) erythema in four of the six animals. All animals were free of skin irritation within 7 days. Five of the six animals treated with uninhibited test material exhibited very slight or slight edema. This is unusual since erythema, which usually accompanies edema, was very slight and was only seen in three of the five animals. Four of the five animals were free of all skin irritation within 7 days post-dose; the remaining animal was free of skin irritation by 14 days post-dose. The sixth animal was free of skin irritation  throughout the study.  In conclusion, inhibited test material, under the conditions of the study, was slightly less irritating to the skin than uninhibited test material. Inhibited test material produced very mild, transient skin irritation; uninhibited test material produced mild, transient skin irritation. 

Interpretation of results:

study cannot be used for classification

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

no details on analytical purity, no experimental details, only short summary, no erythema and edema scores for individual animals given, amount of test substance installed not specified

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Acute eye irritation / corrosion in rabbits
- Short description of test conditions: Two drops of undiluted test item samples were placed into the conjunctival sac of the right eyse of each of three albino rabbits.
- Parameters analysed / observed: The animals were observed for 5 days and inflammation (eryhtema, edema, iritis and dullness of the corneal area) was scored according to Draize criteria.

GLP compliance:

no

Species:

rabbit

Strain:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: two drops

Observation period (in vivo):

5 days
reading time points 1, 24, 48 and 72 h and 5 days

Number of animals or in vitro replicates:

3 (1 male and 2 female)

Details on study design:

SCORING SYSTEM: Draize scoring system

Irritation parameter:

overall irritation score

Remarks:

Numerical evaluation of eye irritation

Basis:

other: average score over all animals and all eye irriation parameters

Time point:

24 h

Score:

37.3

Max. score:

110

Reversibility:

not fully reversible within: 5 days

Irritation parameter:

overall irritation score

Remarks:

Numerical evaluation of eye irritation

Basis:

other: average score over all animals and all eye irriation parameters

Time point:

48 h

Score:

22

Max. score:

110

Reversibility:

not fully reversible within: 5 days

Irritation parameter:

overall irritation score

Remarks:

Numerical evaluation of eye irritation

Basis:

other: average score over all animals and all eye irriation parameters

Time point:

72 h

Score:

10.6

Max. score:

110

Reversibility:

not fully reversible within: 5 days

Other effects:

The animals squealed immediately after instillation of the test item.

Eye reactions were assessed based on numerical evaluation. After 1 h of exposure, the average score was 44.0 out of a possible 110. The animals showed moderate to severe erythema, edema leading to nearly closing of the lids, lacrimation and corneal opacity with details of the iris slightly obscured. Cornea and iris began to clear after 48 hours. The score after 72 hours was 10.6. A slight amount of redness remained after 5 days.

Average Draize scores were 37.3 (24 h), 22.0 (48 h), 10.6 (72 h) and 3.3 (5 days).

Interpretation of results:

study cannot be used for classification

Conclusions:

Severe eye reactions evident as moderate to severe erythema, edema, lacrimation and corneal opacity with details of the iris slightly obscured were observed. The effects were not fully reversible during the 5 days observation period. Although strong evidence for severe eye damage was shown, the amount of test item installed into the eyes was not specified. In addition, limited documentation was given on individual eye irritation parameters for the respective reading time points. Therefore, the test result is considered to be inconclusive.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 09 Oct 2017

Deviations:

yes

Remarks:

no details on animals and environmental conditions, post observation period only 7 days, purity not indicated

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.5 kg
- Housing: Individually in cages measuring 600 x 540 x 315 cm
- Diet: granulated rabbit food (SANDERS), 200 g per rabbit per day
- Water: ad libitum

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL

Duration of treatment / exposure:

6 rabbits: single application of undiluted test item without washing
6 rabbits: single application of undiluted test item with rinsing after 30 seconds following test item instillation

Observation period (in vivo):

7 days
Reading time points: 1 h, 1, 2, 4 and 7 days

Number of animals or in vitro replicates:

12 (6 rabbits with eyes rinsed, 6 rabbits without eyes rinsed)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: with rinsing solution DACRYOSERUM (Laboratories CHIBRET, France) in 6/12 animals
- Time after start of exposure: 4 seconds after test item instillation

SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: ophthalmoscope

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3, #4 and #6

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

chemosis score

Basis:

animal #5

Time point:

24/48/72 h

Score:

3.67

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

chemosis score

Basis:

animal: #7

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

fully reversible within: 4 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

chemosis score

Basis:

animal: #8

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

chemosis score

Basis:

animal: #9

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

chemosis score

Basis:

animal: #10

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

chemosis score

Basis:

animal: #11

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

chemosis score

Basis:

animal: #12

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 4 days

Remarks on result:

other: single application without washout

Irritation parameter:

conjunctivae score

Basis:

animal: #4, #5 and #6

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

conjunctivae score

Basis:

animal: #7

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: reversibility not applicable

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

conjunctivae score

Basis:

animal: #8 and #9

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

conjunctivae score

Basis:

animal: #10

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

conjunctivae score

Basis:

animal: #11

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

conjunctivae score

Basis:

animal: #12

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

iris score

Basis:

animal: #1 and #5

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

iris score

Basis:

animal: #2, #3, #4 and #6

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

iris score

Basis:

animal: #7

Time point:

24/48/72 h

Score:

0.67

Max. score:

2

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

iris score

Basis:

animal: #8

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

iris score

Basis:

animal: #9 and #12

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

iris score

Basis:

animal: #10

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 4 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

iris score

Basis:

animal: #11

Time point:

24/48/72 h

Score:

0.67

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

cornea opacity score

Basis:

animal: #4 and #6

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

cornea opacity score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Remarks on result:

other: single application without washout

Irritation parameter:

cornea opacity score

Basis:

animal: #7, #10 and #12

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 4 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

cornea opacity score

Basis:

animal: #8

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

cornea opacity score

Basis:

animal: #9

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritation parameter:

cornea opacity score

Basis:

animal: #11

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eye rinsed 4 sec after instillation

Irritant / corrosive response data:

In rinsed eyes, erythema (conjunctivae redness) was observed in all animals, which fully reversed at the end of the 7 days observation period. In addition, moderate edema (chemosis was observed at the beginning of the observation period, which fully reversed by Day 7. Slight iris lesions and corneal opacity were observed most animals, but the adverse effects were fully reversible in all animals latest by study Day 7.
In non-rinsed eyes, moderate erythema and well defined edema were observed in all animals throughout the whole study period. The adverse effects persistent in all animals until study termination. In addition, slight iris lesions and corneal opacity were observed. Adverse effects on the iris was fully reversible in 4/6 rabbits, but persisted in 2/6 rabbits by study Day 7. Similarly, opacity was reversible in 2/6 rabbits but remained in 4/6 rabbits until study termination.

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

CLP: Eye Dam. 1 H318 according to Regulation (EC) No. 1272/2008.

Reference 3

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

no information on analytical purity, only a short summary is available, no experimental details, no information on animals and environmental conditions,

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Acute eye irritation in rabbits
- Short description of test conditions: 0.1 mL of the test material was installed into the eyes of 6 rabbits, followed by a 7 days observation period.
- Parameters analysed / observed: Eye reactions including erythema, edema, corneal opacity and irits were assessed based on numerical evaluation 1, 24, 48 and 72h, and 7 and 5 days after administration.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL

Observation period (in vivo):

7 days
Reading time points: 1, 24, 48 and 72 h, and 5 and 7 days

Number of animals or in vitro replicates:

6

Irritation parameter:

overall irritation score

Basis:

mean

Remarks:

over all animals

Time point:

24 h

Score:

57.3

Irritation parameter:

overall irritation score

Basis:

mean

Remarks:

over all animals

Time point:

48 h

Score:

47.6

Irritation parameter:

overall irritation score

Basis:

mean

Remarks:

over all animals

Time point:

72 h

Score:

43.3

Irritant / corrosive response data:

Immediately after test item instillation the animals showed discomfort by severe pawing, squealing, trashing about the stocks and eyes tightly closed. One hour after test item instillation, severe erythema, severe edema and copious discharge were observed. At the 24 hour reading, areas of slight corneal cloudiness, severe erythema (necrosis), very slight edema and copious discharge containing whitish exudate were noted and iris showed no reaction to light. From Day 2-10 there was a gradual improvement of eye reactions. After 14 days, corneal ulceration was observed. At the 21 day readinig time point slight ulceration persisted in 2/6 animals while 4/6 animals fully recovered.

Eye reactions were assessed based on numerical evaluation. Mean scores over all animals were 20.0 at 1 h, 57.3 at 24 h, 47.6 at 48 h, 43.3 at 72 h, 35.1 at 5 days and 32.5 at 7 days of observation.

Interpretation of results:

study cannot be used for classification

Reference 4

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

pre-GLP study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 09 Oct 2017

Deviations:

yes

Remarks:

limited documentation, observation period of 7 days without full reversibility, no anaesthetic used, no 1 hour reading after test item instillation

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL

Duration of treatment / exposure:

single application without washing

Observation period (in vivo):

7 days
Reading time points: 24, 48 and 72 h and 7 days

Number of animals or in vitro replicates:

1

Details on study design:

SCORING SYSTEM: Draize scoring system

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritant / corrosive response data:

Instillation of the test substance into the eye of one rabbit caused a reaction which suggested pain. The ocular reactions consisted of moderate corneal opacity (score 2), ulcus corneae, slight irritis (score 1), moderate conjunctival redness (score 2) and a severe swelling of the conjunctiva (chemosis score 4), which persisted unchanged for the first three days post instillation. There was no reversibility during the 7-day observation period. After 7 days of observation there was still moderate to severe corneal opacity (score 2), slight irritis (score 1), moderate conjunctial redness (score 2) and moderate conjunctival chemosis (score 2) observed.

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

CLP: Eye Damage 1, H318 according to Regulation (EC) No. 1272/2008

Reference 5

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985-07-22 to 1985-08-26

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 09 Oct 2017

Deviations:

yes

Remarks:

no anaesthetic used

Qualifier:

according to guideline

Guideline:

other: FIFRA Pesticide Assessment Guidelines, Subdivision F, Hazard Evaluation: Human and Domestic Animals; Office of Pesticide Programs, U.S. Environmental Protection Agency, Office of Pesticides and Toxic Substances, November 1982; Section 81-4, Primary Eye Ir

Version / remarks:

adopted Nov 1982

Qualifier:

according to guideline

Guideline:

other: TSCA: Health Effects Test Guidelines, Office of Toxic Substances, Office of Pesticides and Toxic Substances, United States Environmental Protection Agency, August 1982 Acute Exposure, Primary Eye Irritation

Version / remarks:

adopted Aug 1982

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Source: Hazleton Dutchland, Inc., Denver, Pennsylvania, USA
- Age at study initiation: at least 8 weeks
- Acclimatisation period: 14 days
- Housing: Individually housed
- Diet: Lab Rabbit Chow HF (Purina #5326), ad libitum
- Water: automatic watering system (municipal water), ad libitum

Environmental conditions:
- Temperature: 60 - 70°F (16-21 °C)
- Humidity: 30-70%
- Light cycle: 12/12

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL

Duration of treatment / exposure:

24 h

Observation period (in vivo):

21 days
Reading time points: 1, 24, 48 and 72 h, and 7, 14 and 21 days

Number of animals or in vitro replicates:

1

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Approximately 24 h after test item instillation the treated eye was rinsed to remove any residual test material.
- Time after start of exposure: 24 h

SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: Fluorescein dye was used to confirm presence or absence of corneal ulceration in the treated eye starting with the 24 hour observation and at each subsequent observation until there was no stain retention for two observations.

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.67

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 21 days

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

not reversible

Irritant / corrosive response data:

Moderate to severe conjunctival redness (score 1-3) was observed within the first 72 hours post test item instillation and remained well-defined (score 2) on study Day 7. The effect was fully reversible until study Day 14. In addition, chemosis (score 1-2) was observed until study Day 14, but was reversible by study Day 21. Iritis (score 1) was observed within the first 72 hours after test item instillation, but the animal recovered from it thereafter. Extensive corneal opacification was seen one hour after dosing and persisted through Day 21. Pannus (neovascularisation of the cornea) occurred from Day 7 through 21, and a red mass (probability vascular in origin) on the cornea was seen on Day 14. Protrusion of the cornea considered indicative of intraocular pressure, was also seen on Day 7 and 21. The study authors mention that also necrosis was observed 48 and 72 hours and 7 days after test item instillation, but not at the the 14- and 21-day reading time point. In general necrosis is irreversibel, therefore it is unclear if the table in the study report is correct.

Irritation/corrosive response data at each reading time point up to removal of animal from the test

	1h	24h	48h	72h	7d	14d	21d	Mean (24h, 48h, 72h)	Max. score:
Conjunctivae   redness	1	2	3	3	2	0	0	2,67	3
Conjunctivae  chemosis	2	2	2	1	1	1	0	1,67	4
Iris	+	1	1	1	0	0	0	1,00	2
Cornea opacity *)	3	2^d,f	3^f	3^f	3^b,c,f	4^b,c,e,f	3^b	2,67	4

*)At all scorings whole cornea (area score 4) affected

b: Pannus; c: Protruding cornea; d: Discharge tinted yellow; e: red mass (2x3 mm) on cornea; f: Observation confirmed with fluorescein

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

CLP: Eye Damage 1, H318 according to Regulation (EC) No. 1272/2008.

Reference 6

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

no details on analytical purity, no details on experimental procedure, no details on animals and environmental conditions, no irritation parameters for individual animals reported, only averaged numbers available

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Eye irritation in rabbits
- Short description of test conditions: 0.1 mL of the test material were installed into the conjunctival sac of one eye of 6 rabbits (3 male and 3 female, first group) or 0.01 mL were placed on the surface of the cornea of 6 rabbits (3 male and 3 female, second group). The test item was installed as single application without washing. Afterwards, the animals were observed for 21 days.
- Parameters analysed / observed: Eye irritation parameters (corneal opacity, iris, conjunctival redness and chemosis) were scored according to Draize criteria. Average scores over all animals were calculated.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.0 - 3.0 kg

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied:
first group: 0.1 mL were instilled into the inferior conjunctival sac of one eye
second group: 0.01 mL were placed on the surface of the cornea

Duration of treatment / exposure:

single application without washing

Observation period (in vivo):

21 days
Reading time points: 1, 24, 48 and 72 h, and 7, 14 and 21 days

Number of animals or in vitro replicates:

0.1 mL test substance instillation into the conjunctival sac of the eye: 6 (3 male and 3 female)
0.01 mL test substance application on the surface of the cornea: 6 (3 male and 3 female)

Details on study design:

SCORING SYSTEM: Draize scoring system

Irritation parameter:

cornea opacity score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

1.5

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: 0.1 mL test substance instillation

Irritation parameter:

iris score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: 0.1 mL test substance instillation,

Remarks:

iris could not be scored in some eyes because of corneal opacity

Irritation parameter:

conjunctivae score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

1.4

Max. score:

3

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: 0.1 mL test substance instillation

Irritation parameter:

chemosis score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

1.83

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: 0.1 mL test substance instillation

Irritation parameter:

cornea opacity score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: 0.01 mL test substance instillation

Irritation parameter:

iris score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

0.37

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: 0.01 mL test substance instillation

Irritation parameter:

conjunctivae score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

1.03

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: 0.01 mL test substance instillation

Irritation parameter:

chemosis score

Basis:

other: average score over all animals

Time point:

24/48/72 h

Score:

1.1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: 0.01 mL test substance instillation

Irritant / corrosive response data:

Instillation of 0.1 mL caused marked hyperemia and obvious to severe chemosis, which persisted as such for about 2 days. Chemosis began to resolve by Day 3 and was still present in 2 animals at Day 7 and in mild form in 1 animal at Day 21. Hyperemia, mild to moderate, was still present in all treated eyes by Day 3 and in 5 animals by Day 6. Three animals had residual conjunctival hyperemia at Day 21. Discharge was moderate to marked up to Day 3, resolving in most by Day 14 but still present in 2 animals by Day 21. There was mild corneal injury affecting 25-50% of the cornea at the 1-h reading time point, spreading to the whole of the cornea by the 24-h reading time point, with 1 animal showing opalescent areas at this time. At the 72-h reading time point, 1 animal had opalescent areas, 2 animals had translucent areas, and 3 had just discernible corneal opacity. Most corneal effects cleared in 3 animals by Day 14 and in 4 animals by Day 21. At final inspection on Day 21 one animal had diffuse opalescent changes and another had diffuse opaque cornea with corneal irregularities and neovascularisation.

Application of 0.01 mL test substance to the surface of the cornea caused minor to moderate corneal injury at 4 h post-instillation, which resolved by Day 3. Mild to moderate conjunctival hyperemia, chemosis and discharge was seen 1 h post instillation, which slowly resolved. At the 72-h reading time point, 1 eye had a normal appearance and the remaining 5 animals showed only minor conjunctival irritation.

At 0.1 mL, 2/6 animals developed haemorrhages of the nicitating membrane within 24 hours. 7 days after test item instillation, the cornea one eye had an irregular shape. In 2/6 rabbit eyes there was corneal vascularisation 14 days after test item instillation.

Interpretation of results:

study cannot be used for classification

Conclusions:

Severe and irreversible eye irritating effects were observed in all animals. However, due to limited documentation of effects in individual animals no conclusion on classification is possible.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation:

Several in vivo studies are available to assess the potential of the test material to cause skin irritation/corrosion.

The key study was performed in compliance with GLP and according to US EPA FIFRA Pesticide Assessment Guidelines, US EPA TSCA Heath Effect Test guidelines and equivalent to OECD guideline 404 (86-0245-FKT). An amount of 0.5 mL test substance was applied to the closely clipped area of the trunk of six New Zealand White rabbits for 4 h under semi-occlusive conditions. After exposure, the treated areas were not washed. Dermal reactions were evaluated based on Draize scoring criteria 30 minutes, and 24, 48 and 72 h post administration.

30 minutes after patch removal, well-defined erythema (score 2) was noted in 5/6 animals and moderate to severe erythema (score 3) was present in 1/6 animals. Slight edema (score 2) was noted in 2/6 rabbits, moderate edema in 4/6 rabbits (score 3) and severe edema was noted in 1/6 rabbits (score 4). At the 24-h reading time point, well defined erythema (score 2) was observed in 2/6 rabbits and moderate to severe erythema (score 3) in 4/6 rabbits. 2/6 rabbits showed slight edema (score 2), 3/6 rabbits showed moderate edema (score 3) and severe edema was noted in 1/6 rabbits. 48 and 72 h after patch removal, very slight to moderate to severe erythema (score 1 – 3) and very slight to moderate edema (score 1 – 3) was still present in all animals. About 7 days after patch removal, 5/6 rabbits had very slight erythema (score 1) and 1/6 rabbits had well defined erythema (score 2). Very slight erythema (score 1) persisted in 3/6 rabbits until study termination on Day 14. Very slight edema (score 1) was observed up to Day 7 in 4/6 rabbits and fully reversible in all animals by Day 10. The mean scores over 24, 48 and 72 h for the individual animals were 2.00, 2.67, 3.00, 1.33, 2.33 and 2.33, respectively for erythema. For edema, the mean scores over 24, 48 and 72 h for the individual animals were 2.33, 2.17, 2.67, 2.33, 1.67 and 1.67, respectively.

In the same study, the test item was further applied to 6 separate rabbits, which were treated for 24 h under occlusive conditions. All animals showed moderate to severe erythema (score 3-4) during the whole study period and moderate to severe erythema (score 3-4), which remained slight to moderate (score 2-3) until study termination. This information was not used for classification and labelling purposes.

Based on the results of the study and under the experimental conditions mentioned, the test item is considered to be irritating to the skin. Although the local effects observed were not fully reversible by study termination, a clear tendency of reversibility was observed during the 14-day observation period. Reversibility was complete in 2/6 animals and almost complete in the remaining 4/6 animals. A full reversibility in all animals cannot be proved but seems to be likely if the observation time would have been extended beyond 14 days.

The results obtained in the key study were confirmed by a number of supporting studies, in which even corrosive / irreversible effects of the test item were observed. The studies were considered as supporting information, as the test item was applied under occlusive instead of semi-occlusive conditions and/or for 24 h instead of 4 h exposure. Furthermore, some test results were obtained with abraded instead of intact skin. Therefore, the results obtained with the guideline-conform key study (86-0245-FKT) were used for classification and labelling purposes.

 

Eye irritation:

The eye irritative/corrosive properties of the test material were assessed in several in vivo eye irritation/corrosion studies.

The key study was performed in compliance with GLP and according to US EPA FIFRA Pesticide Assessment Guidelines, US EPA TSCA Heath Effect Test guidelines and equivalent to OECD guideline 405 (86-0246-FKT). 

A single New Zealand White rabbit received a single instillation of 0.1 mL test item into the lower conjunctival sac of the right eye. The left eye remained untreated and served as control. 24 h after treatment, the treated eye was rinsed to remove residual test material. The local irritation effects on the eyes were scored according to Draize criteria 1, 24, 48 and 72 h and 7, 14 and 21 days post instillation. 

Extensive corneal opacity of score was observed 1 h after dosing and persisted through day 21. Conjunctival irritation (moderate to severe redness, chemosis and necrosis), iritis and corneal ulceration were seen for 3 to 7 days after instillation of the test material but subsided by Day 7 or 14. Pannus (neovascularisation of the cornea) occurred from Day 7 through 21, and a red mass, probability of vascular in origin on the cornea was seen on Day 14. Protrusion of the cornea considered indicative of intraocular pressure, was also seen on day 7 and 21.

The mean scores over 24, 48 and 72 h for the animal were 2.67 for conjunctival redness, 1.67 for conjunctival chemosis, 1.0 for iritis and 2.67 for corneal opacity, respectively. The experimental findings clearly show that the test item causes irreversible damage to the eye.

All supporting studies showed similar results, supporting the information obtained in the key study.

 

Respiratory irritation

Local irritation effects were observed in the acute inhalation studies, including clinical signs of toxicity and gross macroscopical changes in the lungs. According to the OECD SIDS Initial Assessment Report (SIAM 17; Arona Nov. 2003; 3-(Methylthio) propionaldehyde) the mode of action is in particular characterized by the local irritation potential of the test item to skin and mucous membranes. With regard to respiratory irritation, it is unclear from the existing studies whether the observed local effects in inhalation studies are attributable to the test item or to acrolein, an impurity which is enriched in the vapor phase under the test conditions. However, at up to 50 ppm (216 mg/m3), the highest concentration tested, no respiratory irritation was detected in a 9 day study with repeated exposure with acrolein-free 3-(methylthio) propionaldehyde.

Justification for classification or non-classification

The available data on skin irritation/corrosion meet the criteria for classification as Skin Irr. 2 (H315) according to Regulation (EC) No. 1272/2008 (CLP) and according to the criteria of the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations.

The available data on eye irritation/corrosion meet the criteria for classification as Eye Damage Cat. 1 (H318) according to Regulation (EC) No. 1272/2008 (CLP) and according to the criteria of the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations.