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

Skin irritation / corrosion

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report):DV6850
- Substance type: powder
- Physical state: solid
- Analytical purity: 83.7%
- Lot/batch No.: R0332-52C
- Expiration date of the lot/batch: 2003/09/30
- Storage condition of test material: ca 4°C in the dark in dry conditions

Test animals

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

Test system

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

Results and discussion

In vivo

Results

Results 1

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

Results 2

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

Results 3

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

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 15 days

Results 5

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

Results 6

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.

Any other information on results incl. tables

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.

Applicant's summary and conclusion

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).