N,N'-ethylenebis[N-acetylacetamide]

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: according to guideline and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-ethylenebis[N-acetylacetamide]

EC Number:

234-123-8

EC Name:

N,N'-ethylenebis[N-acetylacetamide]

Cas Number:

10543-57-4

Molecular formula:

C10H16N2O4

IUPAC Name:

N,N'-ethylenebis[N-acetylacetamide]

Details on test material:

- Name of test material (as cited in study report): Tetraacetylethylenediamine(TAED)

- Substance type:
- Physical state:
- Analytical purity: The test article, TAED, was received from the Sponsor on March 3, 2000, stored at room temperature, and was reported to
be greater than 99% pure.

PRODUCT: TAED POWDER
DATE: 24th March2000
BATCH NUMBER: 13405002

ANALYSIS

Parameter Test Specification Results
APPEARANCE(METHOD1637) WhitePowder WhitePowder
COLOUR(METHOD1634 0.2 A Max 0.078 A
% TAED(METHOD1631) 99.0% Min 99% Min
% WATER(METHOD1632) 0.2%Max 0.01%
% ACETICACID/ANHYDRIDE
As ACID (METHOD 1633) 0.2% Max 0.02%
PARTICLE SIZE (METHOD 1636)
% retained above 0.125mm 50.0% Max 11.7%
IRON (METHOD1635) 15.0 ppm Max 0.2 ppm

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Three approximately twelve-week-old, male HsdOkd:NZW New Zealand White Rabbits were received on March 7, 2000 from Harlan Sprague Dawley,
Inc., Indianapolis, Indiana. They were assigned temporary animal numbers, acclimated to laboratory conditions for eight days, and released for study
use by a staff veterinarian.
Upon receipt, animals were housed individually in stainless steel cages measuring 20x28x16inches (widthxdepthxheight). Animals were
provided with filtered tap water via the S.E. Lab Group, Inc. (Napa, CA) Recirculating Automatic Watering System. Tap water and TEKLAD (Tm) 7015 Certified Rabbit Diet were provided ad libitum.The water is routinely analyzed for contaminants and specific microbes. The feed is analyzed by the
manufacturer for concentrations of specified heavy metals, aflatoxin, chlorinated hydrocarbons, organophosphates, and specific nutrients. The
results of the feed and water analyses are on file at Therlmmune. No contaminants were known to be present in the diet or water at levels which might interfere with achieving the objectives of the study.
During the study period, the temperature and relative humidity in the animal rooms were monitored continuously using the ReesTM Scientific
Monitoring System and recorded twice daily using a Bacharach@ sling psychrometer. The environmental controls in the animal room were set to
maintain temperatures between 62 and 73°F and relative humidity between 30 and 70%. Ten or greater air changes/hour and a 12-hour light/
12-hour dark cycle were maintained.

Test system

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

other: Carboxymethylcellulose

Controls:

other: second shaved region on each rabbit

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500mg

Dosing formulations were prepared in the animal room immediately prior to dosing of each animal. The test material was assumed to be 100% pure
for formulating purposes.

Duration of treatment / exposure:

4 hours

Observation period:

6 days

Number of animals:

3 males

Details on study design:

At least 24 hours prior to treatment, two areas of approximately 6cm² each, in the dorsal thoracic region, were shaved on each rabbit. One area
was treated with the test article and the other served as the untreated control.
Thea ppropriate amount of test material was moistened with l% CMC in water until a paste was achieved. The paste was then applied uniformly on to
the shaved skin (one of the two shaved areas) of the rabbit, covered with a porous gauze dressing and then occluded. Following approximately
4 hours of exposure, the occlusive wrapping was removed.
The test material was administered dermally because this is the expected route of human exposure.

SCORING SYSTEM:
Observations and Records
All of the rabbits were observed for mortality, moribundity and clinical signs of toxicity by cageside observation twice daily, at least sixhours
apart each day. Observations included skin and fur,eyes and mucous membrane, respiratory system, circulatory system, autonomic and
central nervous system, somatomotor pattern, and behavior pattern.
Special Clinical Observations for dermal irritation, including edema and erythema, were recorded at approximately 0.5, 1, 24, 48, and 72 hours
after the end of exposure. Derm alirritation was scored and recorded for both edema and erythema according to the grades listed below:

Score Grade Edema Erythema
0 None No swelling Normalcolor
1 Minimal Slight swelling; indistinct border Light pink; indistinct
2 Mild Defined swelling; distinct border Bright pink / pale red; distinct
3 Moderate Defined swelling; raised border (ca. 1mm) Bright red; distinct
4 Severe Pronounced swelling; raised border (>1mm) Dark red; pronounced

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

edema score

Basis:

mean

Remarks:

all animals

Time point:

other: 0.5h / 1h/ 24h / 48h / 72h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

erythema score

Basis:

mean

Remarks:

all animals

Time point:

other: 0.5h / 1h/ 24h / 48h / 72h

Score:

0

Max. score:

4

Irritant / corrosive response data:

none

Other effects:

none

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

A single, 4-hour dermal treatment with 500 mg of TAED had no effect on mortality and caused no dermal irritation in New Zealand White Rabbits.
There was no edema or erythema noted in any animal at any time after treatment.

Executive summary:

The study was designed to determine the irritant and / or corrosive effects of a single, 4 -hour dermal application of 500mg

Tetraacetylethylenediamine (TAED) in New Zealand White Rabbits. Three male rabbits were treated with TAED on a shaved area of

the back. An adjacent area of the skin was shaved and served as the control. Clinical observations for dermal irritation were performed at approximately 0.5, 1, 24, 48, and 72 hours after treatment.

Treatment had no effect on mortality and caused no dermal irritation.