4-methylmorpholine

Endpoint summary

Description of key information

Skin irritation: Mallory VT (1990) determined in a reliable K2 study, the skin irritation potential of the test substance in New Zealand White rabbits according to OECD Guideline 404. The test substance was observed to be corrosive to the skin. This study was selected as key study.

Eye irritation: BASF (1964) performed an eye irritation test in Angora rabbits according to a method equivalent to OECD Guideline 405. It is possible that the results in this eye irritation test were not as severe as expected as only 0.05 ml of the test substance instead of 0.1 ml as recommended in OECD guideline 405 was applied to the eye. On the basis of the findings of this study, the test substance would have to be classified as eye irritant Cat. 2B according to CLP. The test substance is however corrosive to the skin (Cat. 1B) and therefore is classified for eye damage category 1.

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:

key study

Study period:

1990-08-22 - 1990-09-05

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Well documented GLP study performed according to a method similar to OECD Guideline 404 with the following deviations: an occlusive dressing was used (instead of semi-occlusive). Furthermore, next to application of the test substance on intact shaved skin, also abraded skin was tested.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

occlusive dressing used (instead of semi-occlusive); abraded skin was tested

GLP compliance:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): 6398-24-20, Project #90-005
- Substance type: clear, colorless liquid
- Physical state: liquid
- Lot No.: 90-005
- Purity: responsibility of the sponsor
- Stability under test conditions: There was no apparent change in the physical state of the test article during administration.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hare-Marland, Hewitt, NJ, USA
- Age at study initiation: adult animals
- Weight at study initiation: males 2129-2507 g (mean 2351 g); females 2170-2633 g (mean 2468 g)
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3 °C
- Humidity (%): 30-70%
- Air changes (per hr): not indicated
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Preparation of test site:

other: abraded and intact shaved skin

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL/site

Duration of treatment / exposure:

The test substance was kept in contact with the skin on the upper dorsal site (intact) for four hours and at the lower dorsal sites (intact and abraded) for twenty four hours.

Observation period:

Dermal irritation was scored at 30-60 minutes, 24, 48 and 72 hours and on Days 4 through 14 after patch removal. Dermal irritation of the lower dorsal sites (intact and abraded) were scored at 24, 48 and 72 hours and on Days 4 through 14 after application of the test material.

Number of animals:

6 (3 males and 3 females)

Details on study design:

TEST SITE
- Area of exposure: one intact site on the upper dorsal trunk and at two sites, one intact and one abraded, on the lower dorsal trunk of the animal.
- Type of wrap if used: All application sites were clipped free of hair. The abraded site was prepared using a burred needle; the abrasion penetrated the stratum corneum but not the derma. Following the application of the test material, gauze patches were applied to each of the sites then wrapped with a rubber dam and an elastic bandage to retard evaporation.

REMOVAL OF TEST SUBSTANCE
At the end of each exposure period, the sites were wiped with water and gauze and observed for erythema and edema.

SCORING SYSTEM: Draize evaluation of dermal irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

edema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: application on intact shaved skin (upper dorsal area)

Irritant / corrosive response data:

Moderate to severe erythema and edema were observed at the 30-60 minute observation period on the upper dorsal site. Severe erythema and edema were observed at the 24, 48 and 72 hours and on days 4 through 14 on the upper dorsal site. Severe erythema and edema were observed at the 24, 48 and 72 hours and on days 4 through 14 on the lower dorsal sites. Necrosis of the skin was observed at each application site at the 24, 48 and 72 hours and on days 4 through 14 observation periods at each application site. Sloughing of the skin at the application site was observed at the day 14 observation period on the upper dorsal site and at the day 13 and 14 observation periods at the lower dorsal sites. The study was terminated following the day 14 observation period.

Interpretation of results:

Category 1 (corrosive) based on GHS criteria

Conclusions:

Based on the test results and according to the criteria of the CLP Regulation, the substance is classified as category 1 corrosive.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1964

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Acceptable study comparable to guideline (non GLP; occlusive conditions; TS purity not specified, 1 min, 5 min, 15 min or 20 hours treatment duration; TS preparation: 50% in water, up to 8 days observation period)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

occlusive conditions; 1 min, 5 min, 15 min or 20 hours treatment duration; TS preparation: 50% in water, up to 8 days observation period

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): 4-Methyl-morpholine
no further data.

Species:

rabbit

Strain:

not specified

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.37 and 2.13 kg

Type of coverage:

occlusive

Preparation of test site:

other: intact skin

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml
- Concentration (if solution): 50%

Duration of treatment / exposure:

1 min, 5 min, 15 min and 20 hours

Observation period:

up to 8 days

Number of animals:

2 (all males)

Details on study design:

TEST SITE
- Area of exposure: 2.5x2.5 cm

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The skin was washed with Lutrol (conc.) and Lutrol/water (1:1)
- Time after start of exposure: 1, 5 or 15 min / 1 hour

SCORING SYSTEM: comparable to the OECD scoring system

Irritation parameter:

other: necrosis

Time point:

other: after 1 and 5 min exposure

Reversibility:

not specified

Remarks on result:

other: Exposure of rabbit skin to the test substance for 1 to 5 min under occlusive conditions did not lead to necrosis

Irritation parameter:

other: necrosis

Time point:

other: after 15 min and 1 hour exposure

Reversibility:

not fully reversible within: 8 days

Remarks on result:

other: Exposure of rabbit skin to the test substance for 15 min or 20 hours under occlusive conditions led to full thickness necrosis.

Irritant / corrosive response data:

Strong signs of irritation reactions were also seen, but are not reported here because corrosivity was the leading effect.

Exposure of rabbit skin to the test substance for 15 min or 20 hours under occlusive conditions led to full thickness necrosis of the skin. No full thickness necrosis was observed, when the test substance was applied for 1 or 5 minutes.

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Conclusions:

In this study, necrosis was observed after 15 min and 1 hour of exposure. Exposure of rabbit skin to the test substance for 15 min or 20 hours under occlusive conditions led to full thickness necrosis.

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Non GLP study performed according to a method similar to OECD Guideline 404 with the following deviations: an occlusive dressing was used (instead of semi-occlusive), no irritation score readings at 48h and later than 72h, application to abraded skin sites (next to intact skin sites).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

occlusive dressing used, abraded skin sites tested, no 48h score readings and no observations after 72h

Qualifier:

according to guideline

Guideline:

other: Hazardous Materials Regulations of the Department of Transportation (1976)

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): Sample No. 4236-13-34, AUS-0099

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Pel-Freez, Inc. USDA No. 7I-B-16, Rogers, Arkansas 72756
- Age at study initiation: approximately 10-13 weeks of age
- Housing: The animals were housed in suspended, wire-bottomed steel cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 7 days prior to experimental use

Type of coverage:

occlusive

Preparation of test site:

other: one abraded and one intact shaved site

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL/test site

Duration of treatment / exposure:

4 hours

Observation period:

One hour after removal of the test material, each abraded and intact test site was examined and scored separately for erythema and edema on a graded scale of 0 to 4. After 24 and 72 hours, the sites were again examined and scored.

Number of animals:

6 animals

Details on study design:

TEST SITE
- Area of exposure: back and flank from each rabbit
- Two test sites located lateral to the midline of the back approximately 10 centimeters apart were selected. One of the test sites was abraded (with a slighly bent tip of a hypodermic needle) by making 4 epidermal abrasions, 2 perpendicular to the other 2, while the other test site remained intact. Each site was immediately occluded with a 2-inch square gauze patch which was secured with masking tape. The trunk of each animal was then wrapped with impervious plastic sheeting. The wrap held the patches in position but did not retard evaporation of the test material during the 4-hour exposure period.

REMOVAL OF TEST SUBSTANCE
At the end of the 4 hours, the plastic wrappings and patches were removed. In addition, all residual test material was removed by flooding the test sites with lukewarm tap water while gently rubbing the sites.

SCORING SYSTEM: In evaluating the average irritation present, the mean scores for erythema and edema of the intact test sites after 4, 24 and 72 hours were added. Similarly, the mean scores for erythema and edema of the abraded test sites after 4, 24 and 72 hours were added. These 2 values were totaled and divided by 6 to obtain the mean primary irritation score.
The test material was classified corrosive if corrosion occurs when the material is tested on intact rabbit skin. Corrosion will be considered to have resulted if the substance in contact with the rabbit skin has caused destruction or irreversible alteration of the tissue after an exposure period of 4 hours or less. Tissue destruction is considered to have occurred if, at any of the readings, there is ulceration or necrosis. Tissue destruction does not include merely sloughing of the epidermis, or erythema, edema or fissuring.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: 24 and 72h

Score:

4

Max. score:

4

Reversibility:

no data

Irritation parameter:

edema score

Basis:

mean

Remarks:

of 6 animals

Time point:

other: 24 and 72h

Score:

4

Max. score:

4

Reversibility:

no data

Irritant / corrosive response data:

Burns were severe and diffuse. Second degree chemical burns were observed after 4 hours (not resulting in fibrosis). Third degree chemical burns were observed after 24 and 72h (resulting in fibrosis).
Primary irritation score: 8 (max possible)

Interpretation of results:

Category 1 (corrosive) based on GHS criteria

Conclusions:

A mean primary irritation score of 8.0 was observed after 4 hours application of the substance to New Zealand White rabbit skin. The substance shows chemical burns and is considered corrosive.

Reference 4

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1979-01-05 - 1979-04-05

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Non GLP study performed according to a method similar to OECD Guideline 404 with the following deviations: an occlusive dressing was used (instead of semi-occlusive), abraded skin sites were tested, no scoring at 48h or later than 72h, 24h exposure (instead of 4h).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

occlusive dressing used, abraded skin sites tested, no readings at 48h and later than 72h, 24h exposure time.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): sample label 4236-24-30
- Substance type: clear liquid
- Physical state: liquid

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ace Animals
- Age at study initiation: at least 8 weeks old
- Housing: The animals were housed 2/cage in suspended wire mesh cages (30" x 18" x 18").
- Diet (e.g. ad libitum) and water (e.g. ad libitum): Fresh Purina rabbit chow and water were freely available.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-21°C

Type of coverage:

occlusive

Preparation of test site:

other: intact and abraded

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of treatment / exposure:

The test material was kept in contact with the skin for 24 hours.

Observation period:

Dermal reactions were scored at 24 and 72 hours

Number of animals:

6 rabbits (2 males, 4 females)

Details on study design:

TEST SITE
The fur was clipped from the backs and sides of the animals. The back from the left of the spinal column of all animals was abraded. The abrasions, extending the length of the exposure site, scratched the stratum corneum but did not reach the derma or produce bleeding. The back to the right of the spinal cord column remained intact.

REMOVAL OF TEST SUBSTANCE
The test material was kept in contact with the skin for 24 hours, at which time the wrappings were removed.

SCORING SYSTEM: Draize scoring system

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

2.29

Max. score:

8

Reversibility:

no data

Remarks:

72 hours

Irritant / corrosive response data:

Average erythema scores were very slight to well defined.
Average edema scores were very slight to slight.

Interpretation of results:

GHS criteria not met

Conclusions:

The primary dermal index of the substance is determined to be 2.29 after 24 hours application to New Zealand White rabbit skin.

Reference 5

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1978-08 - 1978-10

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

occlusive dressing used, abraded skin sites tested, no readings at 48h and later than 72h, 24h exposure time, limited data on test material, animals and environmental conditions.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): RC-106 or RC-107
- Substance type: clear liquid
- Physical state: liquid
- Analytical purity: no data
- Lot/batch No.: no data
- Stability under test conditions: no data
- Storage condition of test material: no data

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Plummer Rabbitry
- Age at study initiation: no data
- Weight at study initiation: no data
- Housing: no data
- Diet (e.g. ad libitum): no data
- Water (e.g. ad libitum): no data
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

Type of coverage:

occlusive

Preparation of test site:

other: abraded and intact shaved skin

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml

Duration of treatment / exposure:

24h

Observation period:

72h

Number of animals:

6

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Remarks:

of 6 animals, intact and abraded skin

Time point:

other: 24 and 72h

Score:

6.25

Max. score:

8

Reversibility:

no data

Remarks on result:

other: Test substance RC-106

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Remarks:

of 6 animals, intact and abraded skin

Time point:

other: 24 and 72h

Score:

6

Max. score:

8

Reversibility:

no data

Remarks on result:

other: Test substance RC-107

Irritant / corrosive response data:

RC-106: necrosis, peripheral erythema and blanching of the skin was observed after 24h; coriaceous (entire site) and peripheral erythema was observed after 72h
RC-107: necrosis and blanching of the skin was observed after 24h; leathery and thickened texture (still flexible) was observed after 72h

Interpretation of results:

study cannot be used for classification

Conclusions:

Corrosive effects were observed after application of the test substance to the rabbit skin during 24h.

Reference 6

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1964

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Acceptable study comparable to guideline (no GLP, TS purity not specified, 50 µl TS, 8-day observation period).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

50 µl TS, 8-day observation period

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): 4-Methylmorpholin

Species:

rabbit

Strain:

Angora

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.54 and 2.61 kg

ENVIRONMENTAL CONDITIONS
no data

Vehicle:

unchanged (no vehicle)

Controls:

other: left eye of the same rabbits treated with NaCl (0.9%)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): ca. 0.05 ml

Duration of treatment / exposure:

single application

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

2 (all females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

SCORING SYSTEM: comparable to the OECD scoring system

TOOL USED TO ASSESS SCORE: hand-slit lamp

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 8 days

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 8 days

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritant / corrosive response data:

The mean scores (24/48/72 hours) of the individual animals were:
- cornea score: 1 in 2/2 animals
- iris score: 1 in 2/2 animals
- conjunctivae score: 1 in 2/2 animals
- chemosis score: 0 in 2/2 animals

Other effects:

Mucous bleeding, pupil retraction, ciliary injection and misty opacity were generally present, but also reversible within 5-8 days.

In a skin irritation test, the test substance was corrosive. It might be possible that the results in this eye irritation test were not as severe as expected as only 0.05 ml of the test substance instead of 0.1 ml as recommended in OECD guideline 405 were applied to the eye. Due to the findings of this study, the test substance would have to be classified as eye irritant Cat. 2B according to GHS-criteria. The test substance is however corrosive to the skin (Cat. 1B).

Interpretation of results:

Category 2B (mildly irritating to eyes) based on GHS criteria

Conclusions:

In a skin irritation test, the test substance was corrosive.
It might be possible that the results in this eye irritation test were not as severe as expected as only 0.05 ml of the test substance instead of 0.1 ml as recommended in OECD 405 were applied to the eye. Due to the findings of this study, the test substance would have to be classified as eye irritant Cat. 2B according to CLP criteria, although the test substance is corrosive to the skin (Cat. 1B).

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1978-08 - 1978-10

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

limited data on test material, animals and environmental conditions, no 21 day observation period

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): RC-106 or RC-107
- Substance type: clear liquid
- Physical state: liquid
- Analytical purity: no data
- Lot/batch No.: no data
- Stability under test conditions: no data
- Storage condition of test material: no data

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Plummer Rabbitry or Sweetwater Farm, Inc.
- Age at study initiation: no data
- Weight at study initiation: no data
- Housing: no data
- Diet (e.g. ad libitum): no data
- Water (e.g. ad libitum): no data
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

Vehicle:

unchanged (no vehicle)

Controls:

other: left eye were untreated and served as controls

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml

Duration of treatment / exposure:

6 animals: no rinsing
3 animals: rinsed after 30 sec

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

9

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 6 animals (unrinsed)

Time point:

24/48/72 h

Score:

1.78

Max. score:

4

Reversibility:

other: not reversible within 7 days

Remarks on result:

other: Test substance RC-106

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 6 animals (unrinsed)

Time point:

24/48/72 h

Score:

0.5

Max. score:

2

Reversibility:

other: not reversible within 7 days

Remarks on result:

other: test substance RC-106

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

mean

Remarks:

of 6 animals (unrinsed)

Time point:

24/48/72 h

Score:

2.44

Max. score:

3

Reversibility:

other: not reversible within 7 days

Remarks on result:

other: test substance RC-106

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 6 animals (unrinsed)

Time point:

24/48/72 h

Score:

2.22

Max. score:

4

Reversibility:

other: not reversible within 7 days

Remarks on result:

other: test substance RC-106

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 6 animals (unrinsed)

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

other: not reversible within 7 days

Remarks on result:

other: test substance RC-107

Irritation parameter:

conjunctivae score

Remarks:

erythema

Basis:

mean

Remarks:

of 6 animals (unrinsed)

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

other: not reversible within 7 days

Remarks on result:

other: test substance RC-107

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 6 animals (unrinsed)

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

other: not reversible within 7 days

Remarks on result:

other: test substance RC-107

Irritant / corrosive response data:

Corneal opacity in eyes of RC-107 treated animals prevented scoring of iris.

Interpretation of results:

study cannot be used for classification

Conclusions:

According to the criteria of CLP, RC-106 would be classified as irritating to eyes, and RC-107 would be classified as causing serious damage to eyes. However, the study is considered not reliable as the documentation was not sufficient.

Reference 3

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation / corrosion:

Mallory VT (1990c) studied skin irritation/corrosion by applying the test substance on one intact site on the upper dorsal trunk and at two sites, one intact and one abraded, on the lower dorsal trunk of the animal. At the end of each exposure period, the sites were wiped with water and gauze and observed for erythema and edema. Moderate to severe erythema and edema were observed at the 30-60 minute observation period on the upper dorsal site. Severe erythema and edema were observed at the 24, 48 and 72 hours and on days 4 through 14 on the upper dorsal site. Severe erythema and edema were observed at the 24, 48 and 72 hours and on days 4 through 14 on the lower dorsal sites. Necrosis of the skin was observed at each application site at the 24, 48 and 72 hours and on days 4 through 14 observation periods at each application site. Sloughing of the skin at the application site was observed at the day 14 observation period on the upper dorsal site and at the day 13 and 14 observation periods at the lower dorsal sites. The study was terminated following the day 14 observation period.

In addition, the test substance was observed to be corrosive in a non GLP primary skin irritation test (Scibon G, 1977) after 4 hours application of the substance to New Zealand White rabbit skin. Also, BASF (1964) observed necrosis in rabbits after 15 min and 1 hour of exposure, but not after 1 or 5 min of exposure. Exposure of rabbit skin to the test substance for 15 min or 20 hours under occlusive conditions led to full thickness necrosis.These studies were used as supporting evidence.

Based on the test results and according to the criteria of the CLP Regulation, the substance is classified as category 1B corrosive.

Eye irritation:

BASF (1964) performed an eye irritation test in Angora rabbits according to a method equivalent to OECD Guideline 405 (single application in 2 female rabbits). The left eye of the same rabbits treated with NaCl (0.9%) served as control. In a skin irritation test, the test substance was corrosive. It is possible that the results in this eye irritation test were not as severe as expected as only 0.05 ml of the test substance instead of 0.1 ml as recommended in OECD guideline 405 was applied to the eye. Due to the findings of this study, the test substance would have to be classified as eye irritant Cat. 2B according to CLP. The test substance is however corrosive to the skin (Cat. 1B) and therefore is classified for eye damage category 1.

Justification for classification or non-classification

Based on all available data and according to the criteria of the CLP Regulation and as burns were excessive at 24 hours onward the substance is classified as corrosive category 1B (H314, causes severe skin burns and eye damage). In addition, the substance is classified for eye damage category 1 (H318).