N-(carboxymethyl)-3-[(2Z)-docos-2-enoylamino]-N,N-dimethylpropan-1-aminium

Endpoint summary

Description of key information

The test item DV6850 was determined to be irritating to the skin. It is not irritating to the eyes. 

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:

from 2 October 2002 to 7 November 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

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: Highgate Farm, Market Rasen, Lincolnshire, England
- Age at study initiation: >= 8 weeks
- Weight at study initiation: 2.84 - 3.07 kg
- Housing: individually in stainless steel cages with perforated floors
- Diet (e.g. ad libitum): 125 g/day standard laboratory rabbit diet (Special Diet Services STANRAB (P) SQC pellets ), dietary supplement of hay
- Water (e.g. ad libitum): ad libitum
- Acclimation period: >= 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-20°C. The study protocol indicated that the target temperature range was 15-23°C, however, the range was changed to 16-20°C on 4th November 2002. The change was to accord with the UK Home Office Animals (Scientific Procedures) Act 1986 Code of Practice for the Housing and Care of Animals used in Scientific Procedures (1989).
- Humidity (%): 40-70%
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12 hours continuous artificial light in each 24 hour period

Type of coverage:

semiocclusive

Preparation of test site:

other: Intact skin, clipped hair

Vehicle:

unchanged (no vehicle)

Remarks:

The treatement site was wetted with 0.5 mL of reverse osmosis water.

Controls:

other: An additional site of the test animals was similarly treated with the exception of test substance and acted as a control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g on wetted skin

Duration of treatment / exposure:

4 hours (a single animal received three exposures (of three minutes, one or four hours duration) in a step-wise manner and acted as a preliminary screen)

Observation period:

15 days

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: On the day before application of the test substance, hair was removed with clippers from the dorso-lumbar region of each rabbit exposing an appropriate sized area of skin.
- % Coverage: approximately 25 mm x 25 mm was covered with substance
- Type of wrap if used: Test substance was applied under a 2-ply porous gauze pad. For exposures of one hour or more each treatment site was covered with cotton wool and "Tubigrip" elasticated bandage dressing for the duration of the exposure period.
- Preparation of test site: The skin treatment site was wetted with 0.5 mL of water before application of the powder.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period the semi-occlusive dressing and gauze pad were removed and the treatment site was washed with lukewarm water (30-40°C) to remove any residual test substance. The treated area was blotted dry with absorbent paper.
- Time after start of exposure: 4 hours

SCORING SYSTEM:
Erythema and eschar formation:
No erythema: 0
Very slight erythema (barely perceptible): 1
Well-defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) or eschar formation (injuries in depth): 4

Oedema formation:
No oedema: 0
Very slight oedema (barely perceptible): 1
Slight oedema (edges of area well-defined by definite raising): 2
Moderate oedema (raised approximately 1 millimetre): 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure): 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.7

Max. score:

4

Reversibility:

fully reversible within: 15 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

not fully reversible within: 15 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 15 days

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 15 days

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

not fully reversible within: 15 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritant / corrosive response data:

Very-slight to moderate to severe erythema with or without very-slight or, in one case, slight oedema was apparent during the first week after bandage removal, persisting in two cases until termination on Day 15. Loss of flexibility was evident in two animals 72 hours after bandage removal and in one case on Day 8; eschar formation or exfoliation was apparent in two animals on Day 8 or 15.

Other effects:

There was no sign of toxicity or ill health in any rabbit during the observation period.

Table 1 - Scoring of irritance responses

Test site: 0.5 g DV6850, semi occluded for four hours, dorso lumbar

Control site: No treatment

Animal number and sex	Type of response	1 hour		24 hours		48 hours		72 hours		Day 8		Day 15
		Test site	Control site	Test site	Control site	Test site	Control site	Test site	Control site	Test site	Control site	Test site	Control site
4617 M *	Erythema	0	0	2	0	3	0	3	0	1 Lf	0	0	0
	Oedema	0	0	1	0	1	0	1	0	1	0	0	0
4660 M	Erythema	1	0	1	0	1	0	2 Lf	0	2 E	0	1 D	0
	Oedema	0	0	1	0	1	0	2	0	1	0	1	0
4662 M	Erythema	1	0	1	0	1	0	1 Lf	0	1 D	0	1 D	0
	Oedema	1	0	1	0	1	0	1	0	0	0	0	0

M: Male

*: Sentinel animal

Lf: Loss of flexibility

D: Exfoliation

E: Eschar

Classification Criteria

The study protocol indicated that the criteria regarding classification, packaging and labelling of dangerous substances of the European Communities as outlined in Directive 93/21/EEC would be employed. However, the updated criteria as documented in 2001/59/EC was employed.

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Although the mean scores were below the labelling threshold of Commission Directive 2001 /59/EC the persistent nature of the treatment related findings in two cases indicates significant irritation. Accordingly, DV6850 required labelling with the risk phrase R38, "Irritating to skin" (DSD) or Skin irrit. 2, H315 (CLP).

Executive summary:

A study was performed to assess the skin irritation potential of DV6850 to the rabbit. The methods followed were that described in EEC Methods for the determination of toxicity, Annex to Directive 92/69/EEC (Official Journal No. L383A, 29.12.92), Part B, Method B.4. Acute toxicity (Skin Irritation), OECD Guideline for Testing of Chemicals No. 404 "Acute Dermal Irritation/Corrosion", adopted 17 July 1992 and EPA Health Effects Test Guidelines OPPTS 870.2500 Acute Dermal Irritation EPA 712 -C-98-196, August 1998.

Three rabbits received a single four hour, semi-occlusive, dermal administration of approximately 0.5 g of the test substance as supplied on wetted skin and were observed for fifteen days.

Very-slight to moderate to severe erythema with or without very-slight or, in one case, slight oedema was apparent during the first week after bandage removal, persisting in two cases until termination on Day 15. Loss of flexibility was evident in two animals 72 hours after bandage removal and in one case on Day 8; eschar formation or exfoliation was apparent in two animals on Day 8 or 15. Mean individual scores at 24, 48 and 72 h after application were 2.7, 1.3, 1.0 for erythema and 1.0, 1.3, 1.0 for edema, for rabbits No. 1, 2, 3, respectively. Although the mean scores were below the labelling threshold of Comission Directive 2001 /59/EC the persistent nature of the treatment related findings in two cases indicates significant irritation. Accordingly, DV6850 required labelling with the risk phrase R38, "Irritating to skin" (DSD) or Skin irrit. 2, H315 (CLP).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (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:

from 30 October 2002 to 19 November 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Highgate Farm, Market Rasen, Lincolnshire, England
- Age at study initiation: >= 8 weeks
- Weight at study initiation: 3.12 - 3.60 kg
- Housing: individually in stainless steel cages with perforated floors
- Diet (e.g. ad libitum): 125 g/d of a standard laboratory diet (Special Diet Services STANRAB (P) SQC pellet), dietary supplement of hay until at least two days prior to dose instillation
- Water (e.g. ad libitum): ad libitum
- Acclimation period: >= 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15-23°C until 4 November 2002, thereafter 16-20°C
- Humidity (%): 40-70%
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12 hours of continuous artificial light in each 24 hour period

IN-LIFE DATES: From: To:

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL weighing approximately 100 mg

Duration of treatment / exposure:

single instillation

Observation period (in vivo):

4 days

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
The substance was not removed.

SCORING SYSTEM:
See Table 1 and Table 2 in "Any other information on materials and methods incl. tables"

TOOL USED TO ASSESS SCORE: An ophthalmoscope and a pencil beam torch were used to facilitate inspection of the eyes.

Irritation parameter:

cornea opacity score

Remarks:

corneal opacity

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

iris score

Remarks:

Iridal lesions

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Time point:

24/48/72 h

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: 3 days

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

A crimson-red conjunctival appearance, very-slight discharge and in two cases very-slight chemosis were apparent one hour after instillation. Injection of the conjunctival blood vessels was evident in all animals 24 and 48 hours later; all animals were overtly normal 72 hours after instillation.

Other effects:

There was no sign of toxicity or ill health in any rabbit during the observation period.

Table 1 - Grades for ocular irritation responses following instillation of DV6850

Animal number and sex: 4674 M			Pain evaluation response: 2
Region of the eye	Response		Grade of response at time after instillation Hours
			1	24		48	72
Cornea	Opacity		0	0		0	0
	Area		0	0		0	0
	Ulceration		-	-		-	-
	Stippling		-	-		-	-
Iris	Value		0	0		0	0
Conjunctiva	Redness		2	1		1	0
	Chernosis		0	0		0	0
	Discharge		1	0		0	0
	Necrosis		-	-		-	-
	Ulceration		-	-		-	-
Animal number and sex: 4733 M			Pain evaluation response: 2
Region of the eye		Response	Grade of response at time after instillation Hours
			1	24		48	72
Cornea		Opacity	0	0		0	0
		Area	0	0		0	0
		Ulceration	-	-		‑	-
		Stippling	-	-		-	-
Iris		Value	0	0		0	0
Conjunctiva		Redness	2	1		1	0
		Chemosis	0	0		0	0
		Discharge	1	0		0	0
		Necrosis	-	-		-	-
		Ulceration	-	-		-	-
Animal number and sex: 4734 M			Pain evaluation response: 2
Region of the eye	Response		Grade of response at time after instillation Hours
			1	24	48			72
Cornea	Opacity		0	0	0			0
	Area		0	0	0			0
	Ulceration		-	-	-			-
	Stippling		-	-	-			-
Iris	Value		0	0	0			0
Conjunctiva	Redness		2	1	1			0
	Chemosis		1	0	0			0
	Discharge		1	0	0			0
	Necrosis		-	-	-			-
	Ulceration		-	-	-			-

Classification Criteria

The study protocol indicated that the criteria regarding classification, packaging and labelling of dangerous substances of the European Communities as outlined in Directive 93/21/EEC would be employed. However, the updated criteria as documented in 2001/59/EC was employed.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

DV6850 does not require classification as an eye irritant.

Executive summary:

A study was performed to assess the eye irritation potential of DV6850 to the rabbit. The methods followed were that described in EEC Methods for the determination of toxicity, Annex to Directive 92/69/EEC (Official Journal No. L383A, 29.12.92), Part B. Method B.5, acute toxicity (eye irritation), OECD Guideline for the Testing of Chemicals No. 403, "Acute Eye Irritation/Corrosion", adopted 24 February 1987 and EPA Health Effects Test Guidelines OPPTS 870.2400 Acute Eye Irritation EPA 712-C-98-195, August 1998.

Three rabbits were each administered a single ocular dose of a volume of 0.1 mL of the test substance weighing approximately 100 mg and observed for four days after instillation.

A crimson-red conjunctival appearance, very-slight discharge and in two cases very-slight chemosis were apparent one hour after instillation. Injection of the conjunctival blood vessels was evident in all animals 24 and 48 hours later; all animals were overtly normal 72 hours after instillation. Instillation of the test substance gave rise to slight initial pain response. Mean individual scores at 24, 48 and 72 h after application were 0.7, 0.7, 0.7 for conjunctival redness, for rabbits No. 1, 2, 3, respectively, and 0.0 for all other ocular parameters.

DV6850 does not require classification as an eye irritant, according to the criteria of DSD (Directive 67/548/EEC) or CLP (Regulation (EC) 1272/2008).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

The skin irritation and eye irritation potential of the test item DV6850 was assessed in two valid GLP studies according to OECD TG 404 and 405, respectively. To assess the skin irritation potential of the substance, three rabbits received a single four hour, semi-occlusive, dermal administration of approximately 0.5 g of the test substance as supplied and were observed for fifteen days. 24, 48, 72 hours after removal of dressings, erythema and oedema were observed in all animals. The mean scores for erythema were 2.7, 1.3 and 1.0 in animal 1, 2 and 3, respectively, and the mean scores for oedema were 1.0, 1.3 and 1.0. Erythema was still observed in 2 animals and oedema in 1 animal 15 days after exposure.

To assess the eye irritation potential of the substance, three rabbits were each administered a single ocular dose of a volume of 0.1 mL of the test substance weighing approximately 100 mg and observed for four days after instillation. 24 and 48 hours after instillation, no effects except conjunctival redness of the eyes were observed. All animals were overtly normal 72 hours after instillation. The mean scoring of conjunctival redness at 24, 48 and 72 hours was 0.7 for all animals.

Effects on skin irritation/corrosion: irritating

Justification for classification or non-classification

The substance is classified as a skin irritant (Xi; R38 or Skin irrit. 2, H315 according to Directive 67/548/EEC or Regulation (EC) 1272/2008, respectively) because erythema was still observed in 2/3 animals 15 days after exposure.

The substance does not fulfill the criteria for classification as "irritating to eyes" according to Regulation (EC) No 1272/2008 because no effects except conjunctival redness were observed 24, 48 and 72 hours after instillation and the mean scoring of conjunctival redness at these time points was 0.7 for all animals which is below the threshold value for classification (at least 2 of 3 animals with a scoring of >=2 (CLP) or 2.5 (DSD)).