3-ETHYL-5,5-DIMETHYL-GAMMA BUTYROLACTONE

Endpoint summary

Description of key information

Skin irritation: not irritating (OECD 404, GLP, K, Rel.1)

Eye irritation: not irritating (OECD 405, GLP, K, Rel.1)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 24 June and 15 July 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 404.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

24 April 2002.

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

Directive n° 2004/73/EC.

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 2008-04-14&15 / Signed on 2008-07-16.

Specific details on test material used for the study:

- The test item was considered at 100% for the study
- Date received: 02 June 2008

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage de Gérome (Quartier Labaste – F40260 Linxe).
- Weight at study initiation: 2.17 - 3.07 kg
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: foodstuff (SDS –C15), ad libitum
- Water: tap-water from public distribution, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-24°C
- Humidity: 40-66%
- Air changes: at least ten changes per hour.
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: From 24 June To 15 July 2008.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): Test item was applied as supplied.

Duration of treatment / exposure:

4 hours

Observation period:

Skin reactions were appreciated 1, 24, 48 and 72 hours after removal of the patch.

Number of animals:

3 females

Details on study design:

PRETEST
Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated.

TEST SITE
- Area of exposure: Undamaged skin area of one flank of each animal
- Type of wrap if used: Patch was secured in position with a strip of surgical adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Washing: After removal of the patch, the treated area was rinsed with distilled water.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
1, 24, 48 and 72 hours after removal of the patch and observations were continued from Days 4 to 14 (in case of persistent reactions were observed).

SCORING SYSTEM:
- Method of calculation: Skin irritant reaction was scored as per Draize scale according to OECD guideline 404

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 3 days

Remarks on result:

probability of weak irritation

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritant / corrosive response data:

- It was noted, 24 hours after the test item application, on the treated area, a slight to well defined erythema associated with a slight oedema in two animals. The oedematous reactions were totally reversible at D2 and the erythematous reactions were totally reversible between D2 and D3.
- On the cutaneous structure, slight dryness was noted on the treated area, from D2 in two animals and from D3 in the last one. Slight dryness was still noted at the end of the observation time (D14) in two animals.

Other effects:

None

Table 7.3.1/1: Individual and mean skin reactions/Erythema - Eschar formation following 4 hour exposure

Skin reaction	Observation time (following patch removal)	Individual Scores – Rabbit Number and Sex
		A9134/Female	A9149/Female	A9150/Female
Erythema/Eschar formation	24 h	1	1	2
	48 h	0	0	1
	72 h	0	0	0
	Total (24, 48 and 72 hours)	1	1	3
	Mean (24, 48 and 72 hours)	0.3	0.3	1

Table 7.3.1/2: Individual and mean skin reactions/Oedema formation following 4 hour exposure

Skin reaction	Observation time (following patch removal)	Individual Scores – Rabbit Number and Sex
		A9134/Female	A9149/Female	A9150/Female
Oedema formation	24 h	0	2	2
	48 h	0	0	0
	72 h	0	0	0
	Total (24, 48 and 72 hours)	0	2	2
	Mean (24, 48 and 72 hours)	0	0.7	0.7

Note

A9134: slight dryness between D2 and D5

A9149: slight dryness between D2 and D3, D6 and D10; D13 and D14

A9150: slight dryness at D3 between D6 and D10; D13 and D14

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the test material is not classified as irritant to the skin according to the Annex I of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

Executive summary:

In a dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 0.5 mL of undiluted test item was applied on an undamaged skin area of the flank of 3 female New Zealand White rabbits. On the other flank an untreated area was served as the control. Test sites were covered with a semi-occlusive dressing for 4 h. Skin irritation was assessed and scored according to the Draize scale at 1, 24, 48 and 72 hours after removal of the patch. Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated during 4 hours.

It was noted, 24 hours after the test item application, on the treated area, a slight to well defined erythema associated with a slight oedema in two animals. The oedematous reactions were totally reversible at D2 and the erythematous reactions were totally reversible between D2 and D3. On the cutaneous structure, slight dryness was noted on the treated area, from D2 in two animals and from D3 in the last one. Slight dryness was still noted at the end of the observation time (D14) in two animals.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 0.3 / 0.3 / 1 for erythema and 0.0/ 0.7/ 0.7 for oedema.

Therefore, the test material is not classified as irritant to the skin according to the Annex I of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for skin irritation endpoint.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 30 June and 04 July 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 404 without any deviation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24 April 2002.

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Directive 2004/73/EC

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 2008-04-14&15 / Signed on 2008-07-16.

Specific details on test material used for the study:

Date received: 02 June 2008
The test item was considered at 100% for the study

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage de Gérome (Quartier Labaste – F40260 Linxe).
- Weight at study initiation: 3.04 - 3.34 kg
- Housing: the animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: foodstuff (SDS-C15), ad libitum
- Water: tap-water from public distribution, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18-23 °C
- Humidity: 41-70 %
- Air changes: at least ten changes
- Photoperiod: 12 hours light/12 hours dark.

IN-LIFE DATES: from 30 June to 04 July 2008.

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

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

- Concentration (if solution): Test item was instilled, as supplied

Duration of treatment / exposure:

No washing was done

Observation period (in vivo):

Ocular examinations were performed on both right and left eyes 1, 24, 48 and 72 h following treatment, according to a numerical evaluation.

Number of animals or in vitro replicates:

3 males

Details on study design:

TREATMENT
- 0.1 mL of the test item was instilled, as supplied, into the conjunctival sac of one eye. The other eye remained untreated serving as control.
- Initially, a single animal was treated. After consideration of the responses produced in the first treated animal at D1, two additional animals were treated.

REMOVAL OF TEST SUBSTANCE
- Washing: No

SCORING SYSTEM: According to OECD guideline 405

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 3 days.

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable.

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 3 days

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

mean individual score

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 2 days

Remarks on result:

probability of weak irritation

Irritant / corrosive response data:

- The ocular conjunctivae reactions observed during the study have been slight to moderate and totally reversible in the three animals.
- A slight redness, noted 1 hour after the test item instillation and totally reversible between D2 and D3.
- A slight to moderate chemosis noted 1 hour after the test item instillation and totally reversible between D1 and D2.

Other effects:

None

Table 7.3.2/1: Individual and Mean Scores for Cornea, Iris and Conjunctivae

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A9115 Male	1 h	0	0	1	1
	24 h	0	0	1	0
	48 h	0	0	0	0
	72 h	0	0	0	0
	Total (24, 48 and 72 h)	0	0	1	0
	Mean (24, 48 and 72 h)	0	0	0.3	0

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A9118 Male	1 h	0	0	1	1
	24 h	0	0	1	0
	48 h	0	0	1	0
	72 h	0	0	0	0
	Total (24, 48 and 72 h)	0	0	2	0
	Mean (24, 48 and 72 h)	0	0	0.7	0

Rabbit Number and Sex	Time After Treatment	Corneal Opacity	Iris lesion	Conjunctival Redness	Conjunctival Chemosis
A9119 Male	1 h	0	0	1	2
	24 h	0	0	1	1
	48 h	0	0	1	0
	72 h	0	0	0	0
	Total (24, 48 and 72 h)	0	0	2	1
	Mean (24, 48 and 72 h)	0.0	0.0	0.7	0.3

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the test substance is not classified as irritating to eyes according to the criteria of annex I of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

Executive summary:

In an eye irritation study conducted according to OECD 405 guideline and in compliance with GLP, 3 New Zealand White male rabbits were exposed to 0.1 mL of test item in one eye while the contralateral eye remained untreated and served as control. The eyes were examined and the changes were observed at 1, 24, 48 and 72 h following treatment and graded according to the Draize method. Initially, a single animal was treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

 

The ocular conjunctivae reactions observed during the study have been slight to moderate and totally reversible in the three animals. A slight redness, noted 1 hour after the test item instillation and totally reversible between D2 and D3. A slight to moderate chemosis noted 1 hour after the test item instillation and totally reversible between D1 and D2.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.0, 0.0, 0.0 for cornea score; 0.0, 0.0, 0.0 for iris score; 0.3, 0.7, 0.7 for conjunctivae score and 0.0, 0.0, 0.3 for chemosis score.

 

Under the test conditions, the test substance is not classified as irritating to eyes according to the criteria of annex I of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

A key study was identified (Phycher, 2008). In this dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 3 female New Zealand White rabbits were exposed for 4 hours to a semi-occluded application of 0.5 mL of undiluted test material to the intact skin. Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated.

It was noted, 24 hours after the test item application, on the treated area, a slight to well defined erythema associated with a slight oedema in two animals. The oedematous reactions were totally reversible at D2 and the erythematous reactions were totally reversible between D2 and D3. On the cutaneous structure, slight dryness was noted on the treated area, from D2 in two animals and from D3 in the last one. Slight dryness was still noted at the end of the observation time (D14) in two animals.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 0.3 / 0.3 / 1 for erythema and 0.0/ 0.7/ 0.7 for oedema.

Eye irritation:

A key study was identified (Phycher, 2008). In this eye irritation study performed according to the OECD guideline 405, and in compliance with GLP, 0.1 mL of undiluted test material was instilled into one eye of 3 male New Zealand White rabbits. The other eye remained untreated and served as control. The eyes were examined and the changes were observed at 1, 24, 48 and 72 h following treatment and graded according to the Draize method. Initially, a single animal was treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

The ocular conjunctivae reactions observed during the study have been slight to moderate and totally reversible in the three animals. A slight redness, noted 1 hour after the test item instillation and totally reversible between D2 and D3. A slight to moderate chemosis noted 1 hour after the test item instillation and totally reversible between D1 and D2.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.0, 0.0, 0.0 for cornea score; 0.0, 0.0, 0.0 for iris score; 0.3, 0.7, 0.7 for conjunctivae score and 0.0, 0.0, 0.3 for chemosis score.

 

Justification for classification or non-classification

 

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008.

 

 

Self classification:

Skin irritation:

Based on the available data, no additional self-classification is proposed regarding skin irritation according to the CLP and to the GHS.

However, the supplemental hazard statement, EUH066, should be applied based on the extrapolation of the results of the OECD 404 study, i.e. slight dryness noted until the end of the observation time (D14) in two out of three animals.

 

Eye irritation:

Based on the available data,no additional self-classification is proposed regarding eye irritation according to the CLP and to the GHS.

 

Respiratory irritation:

No data was available regarding respiratory irritation.