[2R-(2α,3Z,5α)]-3-(2-hydroxyethylidene)-7-oxo-4-oxa-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, compound with tert-butylamine (1:1)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2 October 1991 and 5 October 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

[2R-(2α,3Z,5α)]-3-(2-hydroxyethylidene)-7-oxo-4-oxa-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, compound with tert-butylamine (1:1)

EC Number:

266-104-5

EC Name:

[2R-(2α,3Z,5α)]-3-(2-hydroxyethylidene)-7-oxo-4-oxa-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, compound with tert-butylamine (1:1)

Cas Number:

66069-34-9

Molecular formula:

C8H9NO5.C4H11N

IUPAC Name:

(2R,3Z,5R)-3-(2-hydroxyethylidene)-7-oxo-4-oxa-1-azabicyclo[3.2.0]heptane-2-carboxylic acid; 2-methylpropan-2-amine

Details on test material:

- Name of test material (as cited in study report): BRL 14151C
- Substance type: pure substance
- Physical state: cream powder
- Storage condition of test material: room temperature over silica gel
- Other:
Container: clear glass jar
Date recieved: 14 August 1991

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: supplied by David Percival Ltd., Moston, Sandbach, Cheshire, U.K.
- Age at study initiation:12-16 weeks
- Weight at study initiation: 2.31-2.68 kg
- Housing:suspended metal cages.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-22°C
- Humidity (%): 59-69%
- Air changes (per hr): 15 changes/hour
- Photoperiod (hrs dark / hrs light): 12 light / 12 dark

Test system

Type of coverage:

semiocclusive

Preparation of test site:

abraded

Vehicle:

water

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5g of substance moistened with 0.5ml of distilled water.

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5g of substance moistened with 0.5ml of distilled water.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: gauze patches secured with surgical adhesive tape, the trunk of each animal was wrapped in an elasticated corset (Turbigrip)

REMOVAL OF TEST SUBSTANCE
- Washing (if done):residual test material removed by gentle swabbing with cotton wool soaked in distilled water followed by diethyl ether.
- Time after start of exposure: 4 hours

SCORING SYSTEM:
Evaluation of skin Reactions
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 slight 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

Interpretation of results
The scores for erythema and oedema for each intact and abraded skin site at the 24 and 72 hour readings were totalled for the three test rabbits (24 values) and this total was divided by 12 to give the primary irritation index of the test material.
The test material was classified based on the criteria layed out by Regulation (EC) 1272/2008 on classification, labelling and packaging of substances and mixtures (CLP) and 67/548/EEC Dangerous Substances Directive.

Results and discussion

In vivo

Results

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 72 hours

Score:

0.3

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Irritant / corrosive response data:

The individual scores for erythema /eschar and oedema, are given in Table 1. Mean Values Required for EEC labelling regulations are given in Table 2.
Yellow-coloured staining was commonly noted during the study. this did not affect evaluation of the skin responses.
Very slight erythema was noted at one intact treated skin site one hour after patch removal. very slight erythema was noted at two intact and two abbraded treated skin sites at the 24 hour observation and persisted at one abraded skin site at the 48 hour observation.
No signs of skin irritation were noted 72 hours after treatment.

Any other information on results incl. tables

Table 1 Individual Skin Reactions

Skin Reaction	Skin Site	Observation Time Hour(s)	Rabbit Number and Sec (Bodyweight Kg)
			40 Male (2.38)	41 Male (2.38)	42 Female (2.31)	Total
Erythema & Eschar Formation	Intact	1	0 STA	0 STA	0 STA	( 1 )
	Intact	24	0 STA	0 STA	0 STA	2
	Intact	48	0 STA	0 STA	0 STA	( 0 )
	Intact	72	0 STA	0 STA	0 STA	0
	Abraded	1	0 STA	0 STA	0 STA	( 0 )
	Abraded	24	0 STA	0 STA	0 STA	2
	Abraded	48	0 STA	0 STA	0 STA	( 1 )
	Abraded	72	0 STA	0 STA	0 STA	0
Oedema Formation	Intact	1	0	0	0	( 0 )
	Intact	24	0	0	0	0
	Intact	48	0	0	0	( 0 )
	Intact	72	0	0	0	0
	Abraded	1	0	0	0	( 0 )
	Abraded	24	0	0	0	0
	Abraded	48	0	0	0	( 0 )
	Abraded	72	0	0	0	0
Sum of 24 and 72 hour readings (5)  :         4
Primary Irritation Index ( 5/12 )          :         4/12 = 0.3
() = total values not used for calculation of primary irritation index                         STA = yellow-coloured staining

Table 2: Individual Daily and Idividual Mean Scores for Dermal Irritation 4 -hour exposure Required for EEC labelling regulations

Skin Reaction	Reading (hours)	Rabbit Number and Sex (Bodyweight Kg)
		40 Male (2.68)	41 Male (2.38)	42 Female (2.31)
Erythema/Eschar Formation	24	1	1	0
	48	0	0	0
	72	0	0	0
Total		1	1	0
Mean Score		0.3	0.3	0.0
Oedema Formation	24	0	0	0
	48	0	0	0
	72	0	0	0
Total		0	0	0
Mean Score		0.0	0.0	0.0

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test material, BRL 14151C, produced a primary irritation index of 0.3. No corrosive effects were noted during the study.
The test material did not produce positive criteria in any rabbit according to the EEC labelling regulations and was determined to be not irritating to rabbit skin. No symbol and risk phrase are therefore required.

Executive summary:

This study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit.

A single 4 hour, semi-occluded application of tet material to intact and abraded skin of three rabbits produced incidents of very slight erythema. No signs of skin irritation were noted 72 hours after treatment.

No Corrosive effects were noted during the study. The test material was determined to be not irritating to rabbit skin according to criteria layed out by Regulation (EC) 1272/2008 on classification, labelling and packaging of substances and mixtures (CLP) and 67/548/EEC Dangerous Substances Directive.