2-isopropyl-N-hydroxyethyl 1,3-oxazolidine;methyl carbonato-N-ethyl-2-isopropyl-1,3-oxazolidine;reaction mass of: carbonato-bis-N-ethyl-2-isopropyl-1,3-oxazolidine

Endpoint summary

Description of key information

The test item is considered to be a non-irritant to the skin of the rabbit based on both, the erythema and edema score. The ocular irritancy of Incozol LV was sufficient for the classification as corrosive for the eye in Cat. 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:

from 1998-02-18 to 1998-04-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Version / remarks:

cited as: Directive 92/69/EEC method B4, from 29 Dec 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted: 17 Jul 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River UK Ltd. Margate
- Age at study initiation: 10 to 14 weeks old
- Weight at study initiation: 2.35 to 2.58 kg
- Housing: floor-pens, minimum floor area of 0.6 sq m
- Diet: ad libitum, SQC TRB Rabbit Diet 9603 (pelleted) from Harlan Teklad. Bicester
- Water: tab water, ad libitum
- Acclimation period: 15 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 to 22 °C.
- Humidity (%): 40 - 80 %
- Air changes (per hr): 10 air changes per hour
- Photoperiod: 12 hours light, from 6.00 a.m to 6.00 p.m.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount applied : 500 mg

Duration of treatment / exposure:

4 h

Observation period:

15 days

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 10x8 cm
- % coverage: not specified

REMOVAL OF TEST SUBSTANCE
- Washing: not performed
- Time after start of exposure: 4 hours

SCORING SYSTEM: Macroscopic Dermal Grading System based on Draize (erythema /edema score).

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

(mean)

Time point:

24/48/72 h

Score:

1.66

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

(mean)

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

(mean)

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

1.8

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Max. duration: 8 d; Max. value at end of observation period: 0 (related to all animals)

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

(mean)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

(mean)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

(mean)

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 14 days

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0.1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Max. duration: 8 d; Max. value at end of observation period: 0 (related to all animals)

Irritant / corrosive response data:

Reversibility of any observed effect: Changes fully reversible within 14 days. Based on the Regulation (EC) No 1272/2008 (CLP) , the test item is considered to be a non-irritant to the skin of the rabbit based on both, the erythema and edema score.

Interpretation of results:

GHS criteria not met

Conclusions:

A single semi-occluded application of Incozol LV to intact rabbit skin for four hours caused marked inflammation and other dermal changes including petechial haemorrhage. However, all overt signs of irritation resolved within fourteen days of treatment. Thus, based on the Regulation (EC) No 1272/2008 (CLP), the test item is considered to be a non-irritant to the skin of the rabbit based on both, the erythema and edema score.

Executive summary:

A study was conducted according to OECD TG 404 and Directive 92/69/EEC method B.4 to assess the irritation or corrosion caused to the intact dorsal skin of rabbit following a single (4 hour) semi-occluded topical application of Incozol LV. Therefore, 3 New Zealand White rabbits were administrated to the test item Incozol LV. A single semi-occluded application of Incozol LV to intact rabbit skin for four hours caused marked inflammation and other dermal changes including petechial haemorrhage. However, all overt signs of irritation resolved within fourteen days of treatment. Thus, based on the Regulation (EC) No 1272/2008 (CLP), the test item is considered to be a non-irritant to the skin of the rabbit based on both, the erythema and edema score.

Reference 2

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:

no adverse effect observed (not irritating)

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:

from 1998-03-04 to 1998-03-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

cited as: Directive 92/69/EEC, from 29 Dec 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted: 24 Feb 1987

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River UK Ltd. Margate
- Age at study initiation: 11 to 16 weeks
- Weight at study initiation: 2.54 to 3.38 kg
- Housing: single pen with minimum floor area of 0.6 sq m
- Diet: ad libitum, SQC TRB Rabbit Diet 9603 (pelleted) from Harlan Teklad. Bicester
- Water: tab water, ad libitum
- Acclimation period: two weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 to 22 °C.
- Humidity (%): 40 - 80 % RH
- Air changes: 10 air changes per hour
- Photoperiod: 12 hours light from 6:00 a.m. to 6:00 p.m.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL

Duration of treatment / exposure:

single treatment

Observation period (in vivo):

1, 24, 48 and 72 hours and up to 4 days after dosing

Number of animals or in vitro replicates:

3 femals

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: the treated eyes were not washed after instillation

SCORING SYSTEM:
Ocular Grading System based on Draize. EEC Ocular Scores (Corneal Opacity, Iris Lesions, Conjunctival Redness, Conjunctival Edema)

TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Remarks:

(mean)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Remarks:

(mean)

Time point:

24/48 h

Score:

1

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no values presented because the rabbit was terminated on Day 2

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Remarks:

(mean)

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no values presented because the rabbit was terminated on Day 2

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

(mean score 1)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

(mean)

Time point:

24/48 h

Score:

1

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no values presented because the rabbit was terminated on Day 2

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

(mean)

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no values presented because the rabbit was terminated on Day 2

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

(mean)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

(mean)

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no values presented because the rabbit was terminated on Day 2

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

(mean)

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no values presented because the rabbit was terminated on Day 2

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

(mean)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

(mean)

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no values presented because the rabbit was terminated on Day 2

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

(mean)

Time point:

24/48 h

Score:

2

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no values presented because the rabbit was terminated on Day 2

Other effects:

With animal 1, there was improvement over the study period and all observations had resolved by 24 hours. Two further rabbits showed marked irritant responses from 4 hours after instillation, including translucent cornea, partial eversion of the eyelids and moderate ocular discharge. At 24 hours these two animals showed no iridial response and they were therefore killed on humane grounds.

Interpretation of results:

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

Conclusions:

Based on the results of this study, the ocular irritancy of Incozol LV was sufficient for the substance to be classified as Eye Irritant in Cat. 1, according to Regulation (EC) 1272/2008, as amended for the tenth time in Regulation (EC) 2017/776.

Executive summary:

This study was conducted to determine the irritation caused to the rabbit eye following a single instillation of Incozol LV into the conjunctival sac. The method followed was in compliance with Directive 92/69/EEC method B5 and OHCD TG 405. The undiluted test article (0.1 mL) was instilled into one conjunctival sac of each of three New Zealand White rabbits on Day 1. Ocular reactions were assessed for up to four days after treatment.

Instillation of undiluted Incozol LV into one ocular sac of the sentinel rabbit caused a moderate initial sting response. Injection of the conjunctival blood vasculature and inflammation of the iris were apparent immediately after treatment. Although iridial inflammation resolved within one-half hour of treatment, conjunctival irritation reactions persisted for up to 24 hours; injection of the conjunctival blood vasculature, slight chemosis and a slight ocular discharge were apparent one-half hour and one hour after instillation of the test article and a moderate ocular discharge was apparent 4 hours and 24 hours after treatment. The cornea remained overtly unaffected by Incozol LV and was impermeable to applied fluorescein on Day 2. All irritation reactions resolved within 48 hours of instillation of Incozol LV into the non-anaesthetised eye.

The initial sting reaction observed in the sentinel rabbit was sufficiently marked that both rabbits subsequently committed to the study were prepared for treatment by administration of a corneal anaesthetic. Diffuse opacity of the greater part of the cornea and inflammation of the iris and conjunctivae were apparent in both rabbits one-half hour after treatment. The irritant effects of Incozol LV intensified such that four hours after treatment the treated cornea appeared translucent and conjunctival reactions included a crimson red appearance, chemosis sufficient to cause partial aversion of the eyelids and a moderate ocular discharge. Conjunctival irritation reactions included a crimson red appearance, chemosis sufficient to cause partial eversion of the eyelids and a marked ocular discharge. The iridial reflex of both rabbits (meiosis in response to bright incident light) was found to be absent. The loss of the iridial reaction necessitated immediate termination of the study. Both rabbits were killed on humane grounds.

Irritant effects of Incozol LV in the anaesthetised eye included translucency of the cornea, disruption of the corneal epithelium, marked conjunctivitis and loss of the iridial reflex. Although it is accepted that use of anaesthetic appears to have exacerbated the irritant effects of the test article by impairing the normal clearance mechanisms provided by the tearing mechanism and the nictitating membrane, it has been concluded that Incozol LV is severely irritating to the eye.

Based on the results of this study, the ocular irritancy of Incozol LV was sufficient for the substance to be classified as Eye Irritant in Cat. 1, according to Regulation (EC) 1272/2008, as amended for the tenth time in Regulation (EC) 2017/776.

Reference 2

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 (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation / corrosion

A study was conducted according to OECD TG 404 and Directive 92/69/EEC method B.4 to assess the irritation or corrosion caused to the intact dorsal skin of rabbit following a single (4 hour) semi-occluded topical application of Incozol LV. Therefore, 3 New Zealand White rabbits were administrated to the test item Incozol LV. A single semi-occluded application of Incozol LV to intact rabbit skin for four hours caused marked inflammation and other dermal changes including petechial haemorrhage. However, all overt signs of irritation resolved within fourteen days of treatment. Thus, based on the Regulation (EC) No 1272/2008 (CLP), the test item is considered to be a non-irritant to the skin of the rabbit based on both, the erythema and edema score.

Eye irritation

This study was conducted to determine the irritation caused to the rabbit eye following a single instillation of Incozol LV into the conjunctival sac. The method followed was in compliance with Directive 92/69/EEC method B5 and OHCD TG 405. The undiluted test article (0.1 mL) was instilled into one conjunctival sac of each of three New Zealand White rabbits on Day 1. Ocular reactions were assessed for up to four days after treatment.

Instillation of undiluted Incozol LV into one ocular sac of the sentinel rabbit caused a moderate initial sting response. Injection of the conjunctival blood vasculature and inflammation of the iris were apparent immediately after treatment. Although iridial inflammation resolved within one-half hour of treatment, conjunctival irritation reactions persisted for up to 24 hours; injection of the conjunctival blood vasculature, slight chemosis and a slight ocular discharge were apparent one-half hour and one hour after instillation of the test article and a moderate ocular discharge was apparent 4 hours and 24 hours after treatment. The cornea remained overtly unaffected by Incozol LV and was impermeable to applied fluorescein on Day 2. All irritation reactions resolved within 48 hours of instillation of Incozol LV into the non-anaesthetised eye.

The initial sting reaction observed in the sentinel rabbit was sufficiently marked that both rabbits subsequently committed to the study were prepared for treatment by administration of a corneal anaesthetic. Diffuse opacity of the greater part of the cornea and inflammation of the iris and conjunctivae were apparent in both rabbits one-half hour after treatment. The irritant effects of Incozol LV intensified such that four hours after treatment the treated cornea appeared translucent and conjunctival reactions included a crimson red appearance, chemosis sufficient to cause partial aversion of the eyelids and a moderate ocular discharge. Conjunctival irritation reactions included a crimson red appearance, chemosis sufficient to cause partial eversion of the eyelids and a marked ocular discharge. The iridial reflex of both rabbits (meiosis in response to bright incident light) was found to be absent. The loss of the iridial reaction necessitated immediate termination of the study. Both rabbits were killed on humane grounds.

Irritant effects of Incozol LV in the anaesthetised eye included translucency of the cornea, disruption of the corneal epithelium, marked conjunctivitis and loss of the iridial reflex. Although it is accepted that use of anaesthetic appears to have exacerbated the irritant effects of the test article by impairing the normal clearance mechanisms provided by the tearing mechanism and the nictitating membrane, it has been concluded that Incozol LV is severely irritating to the eye.

Based on the results of this study, the ocular irritancy of Incozol LV was sufficient for the substance to be classified as Eye Irritant in Cat. 1, according to Regulation (EC) 1272/2008, as amended for the tenth time in Regulation (EC) 2017/776.

Justification for classification or non-classification

Based on the findings of the skin and eye irritation studies Incozol LV is not irritating to skin but is classified as eye dam cat 1 with H318 ( Causes serious eye damage) according to Regulation (EC) No 1272/2008 (CLP), as amended for the tenth time in Regulation (EC) 2017/776.