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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002-07-11 to 2003-03-03

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 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

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:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan UK Ltd., Bicester, Oxon, England
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 188 to 228 g
- Housing: individually in metal cages (RS Biotech Sub-Dividable Rodent Cages - polished stainless steel)
- Diet (e.g. ad libitum): standard laboratory rodent diet (Special Diet Services RM1(E) SQC expanded pellet) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°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

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: One day prior to treatment, hair was removed from the dorso-lumbar region of each rat with electric clippers taking care to avoid damaging the skin, exposing an area equivalent to approximately 10% of the total body surface area.
- % coverage: approximately 50 mm x 50 mm was covered with substance
- Type of wrap if used: covered with porous gauze held in place with a non-irritating dressing and further covered by a waterproof dressing encircled firmly around the trunk of the animal

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated area of skin was washed with warm water (30 - 40°C) to remove any residual test substance. The treated area was blotted dry with absorbent paper.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bodyweight
- Concentration (if solution): 20% test substance
- Constant volume or concentration used: yes

Duration of exposure:

24 hours

Doses:

2000 mg/kg bodyweight

No. of animals per sex per dose:

5 males
5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed soon after dosing and at frequent intervals for the remainder of Day 1. On subsequent days animals were observed once in the morning and again at the end of the experimental day (with the exception of Day 15 - morning only). The nature and severity of the clinical signs and time were recorded at each observation. The bodyweight was recorded on day 1 (prior to dosing), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: mortality, clinical signs, dermal responses, body weight, organ weights
- Scoring System for dermal observations:
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) to 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 mm): 3
Severe oedema (raised more than 1 mm and extending beyond the area of exposure): 4

Results and discussion

Effect levels

Effect levels 1

Sex:

male/female

Dose descriptor:

LD0

Effect level:

>= 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

no deaths

Clinical signs:

other: - no systemic response - Very slight to moderate to severe dermal irritation (Grade 1 to 3 erythema with or without Grade 1 or 2 oedema) was observed in all animals following removal of the dressings, resolving completely by either Day 7 in males and Day

Gross pathology:

Dryness of the dose site was seen at the macroscopic examination of one male and three females with scabbing of the dose site seen in four females. No other abnormalities were revealed at the macroscopic examination at study termination on Day 15.

Any other information on results incl. tables

Table 1 - Dermal reactions

Dose (mg/kg)	Sex	Animal No	E=Erythema O=Oedema	Day
				2	3	4	5	6	7	8	9	10	11	12	13 to 15
2000	Male	CCI	E	1	1a	1a	0a	0a	0a	0a	0	0	0	0	0
			O	1	1	0	0	0	0	0	0	0	0	0	0
		CC2	E	1	1	0	0	0	0	0	0	0	0	0	0
			O	0	0	0	0	0	0	0	0	0	0	0	0
		CC3	E	1	1	1	1a	0a	0	0	0	0	0	0	0
			O	1	1	1	0	0	0	0	0	0	0	0	0
		CC4	E	2	1	1	1a	0a	0	0	0a	0a	0a	0	0
			O	1	2	2	2	1	0	0	0	0	0	0	0
		CC5	E	2	1a	1a	1a	0a	0a	0a	0a	0a	0a	0a	0a
			O	1	2	1	1	1	0b	0b	0	0	0	0	0
	Female	CC6	E	2	2	2a	1a	0a	0a	0a	0a	0a	0a	0a	0a
			O	1	2	1	1	1	1b	1b	0b	0b	0b	0b	0b
		CC7	F	3	2	2a	2a	1a	0a	0a	0a	0a	0a	0a	0a
			O	2	2	2	1	1	1b	0b	0b	0b	0b	0b	0b
		CC8	E	2	2	2a	1a	0a	0a	0a	0	0	0	0	0
			O	1	1	1	1	1	0	0	0	0	0	0	0
		CC9	E	1	1	1a	1a	0a	0a	0a	0a	0a	0a	0a	0b
			O	1	1	1	1	1	0b	0b	0b	0b	0b	0b	0
		CC10	E	1	1	1a	1a	0a	0a	0a	0a	0a	0a	0a	0a
			O	1	1	1	1	1	1	0	0	0	0b	0b	0b

a Desquamation (characterised by dryness/exfoliation)

b Spots and/or scabbing (confined to small area of dose site)

Table 2- Individual and group mean bodyweights (g)

Dose (mg/kg)	Sex	Animal Number	Bodyweight (g) at Day
			1*	8	15
2000	Male	CC1	200	236	275
		CC2	190	224	260
		CC3	192	223	253
		CC4	188	232	289
		CC5	201	238	277
	Mean		194	231	271
	Female	CC6	228	238	251
		CC7	220	231	237
		CC8	209	221	238
		CC9	207	207	207
		CC10	206	212	225
	Mean		214	222	232

 *       Prior to dosing

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute lethal dermal dose to rats of DV6850 was demonstrated to be greater than 2000 mg/kg bodyweight. DV6850 is in accordance with DSD (Directive 67/548/EEC) or CLP Regulation (EC) No 1272/2008 "unclassified" and will not require labelling

Executive summary:

A study was performed to assess the acute dermal toxicity of DV6850 to the rat. The method followed was 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 B3. "Acute toxicity (dermal)" and OECD Guideline for Testing of Chemicals No, 402 "Acute Dermal Toxicity". Adopted: 24 February 1987.

A group of ten rats (five males and five females) received a single topical application of the test substance, in water for irrigation, at a dose level of 2000 mg/kg bodyweight. All animals were killed and examined macroscopically on Day 15, the end of the observation period.

There were no deaths and no systemic response to treatment in any animal throughout the study.

Very slight to moderate to severe dermal irritation (Grade 1 to 3 erythema with or without Grade 1 or 2 oedema) was observed in all animals following removal of the dressings, resolving completely by either Day 7 in males and Day 9 in females. In addition, desquamation (characterised by dryness/exfoliation) was seen in four males and all females from Day 3, resolving in three males by Day 12 and two females by Day 14 and persisting in one male and three females until study termination on Day 15. Spots and/or scabbing (confined to a small area of the dose site) were also observed in one male and four females from Day 7, resolving in the male by Day 9 and persisting in all the females until study termination on Day 15.

No bodyweight gain was recorded for one female on both Day 8 and Day 15. A notably low bodyweight gain was seen in one female on Day 8 and one female on Day 15. All other animals were considered to have achieved satisfactory bodyweight gains throughout the study.

Dryness of the dose site was seen at the macroscopic examination of one male and three females with scabbing of the dose site seen in four females. No other abnormalities were revealed at the macroscopic examination at study termination on Day 15.

The acute lethal dermal dose to rats of DV6850 was demonstrated to be greater than 2000 mg/kg bodyweight.

DV6850 is in accordance with DSD (Directive 67/548/EEC) or CLP Regulation (EC) No 1272/2008 "unclassified" and will not require labelling