9,10-Anthracenedione, 1,4-bis[(2-ethylhexyl)amino]-5,8-dihydroxy-

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was carried out according to OECD Guideline No. 402 and EEC Directive 84/449/EEC, Part B.3 and in accordance with the Principles of Good Laboratory Practices (GLP).

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

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:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes

Test type:

standard acute method

Test material

Test material information

Constituent 1

Reference substance name:

1,4-bis[(2-ethylhexyl)amino]-5,8-dihydroxy-9,10-dihydroanthracene-9,10-dione

EC Number:

807-717-5

Cas Number:

295800-70-3

Molecular formula:

C30H42N2O4

IUPAC Name:

1,4-bis[(2-ethylhexyl)amino]-5,8-dihydroxy-9,10-dihydroanthracene-9,10-dione

Test material form:

other: blue solid

Details on test material:

- Name of test material (as cited in study report): Spectrace MD-810 Marker (Solvent Free)
- Physical state: Blue solid
- Lot/batch No.: Lot #RWS-2-43, TD#04-049
- Expiration date of the lot/batch: September 23, 2007
- Stability under test conditions: Test substance was expected to be stable for the duration of testing.
- Storage condition of test material: Stored at room tempertaure.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Inc., Boyertown, PA
- Age at study initiation: Young adult (9-10 weeks).
- Fasting period before study:
- Weight at study initiation: males 297-328 grams and females 190-203 grams.
- Housing:The animals were singly housed in suspended stainless steel caging with mesh floors which conform to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals DHE W (hlH). Litter paper was placed beneath the cage and was changed at least three times per week.
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
- Humidity (%):
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):

IN-LIFE DATES: From: To:

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
Prior to application, the test substance was moistened with distilled water to achieve a dry paste by preparing a 40% w/w mixture. Two thousand mag of body weight of the test substance was then applied to a 2-inch x 3-inch, 4-ply gauze pad and placed on a dose area of approximately 2 inches x 3 inches (approximately 10% of the body surface). The gauze pad and entire trunk of each animal were then wrapped with 3-inch Durapore tape to avoid dislocation of the pad and to minimize loss of the test substance. The rats were then returned to their designated cages. The day of application was considered Day 0 of the study.

REMOVAL OF TEST SUBSTANCE
After 24 hours of exposure to the test substance, the pads were removed and the test sites were gently cleansed of any residual test substance.

TEST MATERIAL
The test substance was a blue solid and was stored at room temperature. In order to insure adequate contact with the skin, the sample was applied as a dry paste (40% w/w mixture in distilled water). Preliminary sample preparation conducted by PSL indicated mixtures in excess of 40% (i.e., 45%-90%) were too dry to assure adequate skin contact.

VEHICLE
Prior to application, the test substance was moistened with distilled water to achieve a dry paste by preparing a 40% w/w mixture.

Duration of exposure:

24 Hours

Doses:

2000 mg/kg body weight

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: The animals were observed for mortality, signs of gross toxicity, and behavioral changes during the first several hours post-dosing and at least once daily thereafter. Individual body weights of the animals were recorded prior to test substance administration (initial) and again on Days 7 and 14 (termination) after dosing.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: Observations included gross evaluation of skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavior pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhea, and coma. Gross necropsies were performed on all animals. Tissues and organs of the thoracic and abdominal cavities were examined.

Statistics:

not applicable

Results and discussion

Preliminary study:

not applicable

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000

Based on:

test mat.

Remarks on result:

other: n/a

Mortality:

No mortality occurred during the study period

Clinical signs:

other: There were no signs of gross toxicity, dermal irritation, adverse pharmacologic effects or abnormal behavior.

Gross pathology:

No gross abnormalities were noted for any of the animals when necropsied at the conclusion of the 14-day observation period.

Other findings:

Not applicable

Any other information on results incl. tables

Acute dermal Dose

Animal No.	Sex	Dose Level (mg/kg)	Body Weight (g)			Dose
			Initial	Day 7	Day 14	g
8508	M	2,000	328	351	436	1.6
8509	M		306	332	397	1.5
8510	M		297	325	401	1.5
8511	M		313	342	416	1.6
8512	M		300	330	390	1.5
8513	F		197	236	253	0.99
8514	F		200	235	255	1.0
8515	F		203	241	261	1.0
8516	F		190	217	247	0.95
8517	F		201	226	250	1.0

 

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the study, the dermal LD50 value of Spectrace® MD-810 Marker (Solvent Free) in rats of either sex was established as exceeding 2000 mg/kg body weight.

Executive summary:

An acute dermal toxicity test was conducted with rats to determine the potential for Spectrace® MD-810

Marker (Solvent Free) to produce toxicity from a single topical application. Under the conditions of this

study, the single dose acute dermal of the test substance is greater than 2,000 mg/kg of body weight in

male and female rats. Classification according to HMIS, EPA, EEC, and WHMIS are presented in Table

4.

Two thousand milligrams per kilogram of body weight of the test substance was moistened with distilled

water and applied to the skin of ten healthy rats for 24 hours. The animals were observed for mortality,

signs of gross toxicity, and behavioral changes at least once daily for 14 days. Body weights were

recorded prior to application and again on Days 7 and 14 (termination). Necropsies were performed on all

animals at terminal sacrifice.

All animals survived, gained body weight, and appeared active and healthy. There were no signs of gross

toxicity, dermal irritation, adverse pharmacologic effects or abnormal behavior. No gross abnormalities

were noted for any of the animals when necropsied at the conclusion of the 14-day observation period.