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

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

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 conducted according to OECD TG 425 and U.S. EPA Health Effects TG, OPPTS 870.1100 and in accordance with the Principles of Good Laboratory Practice (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 425 (Acute Oral Toxicity: Up-and-Down Procedure)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes

Test type:

up-and-down procedure

Limit test:

yes

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:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ace Animals, Inc., Boyertown, PA
- Age at study initiation: 11 weeks
- Weight at study initiation: 192-220 grams
- Fasting period before study: Animals were fasted overnight.
- 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 DHEW (NIH).
- Diet (e.g. ad libitum): Purina Rodent Chow #5012
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 20-24 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-22°C
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle

IN-LIFE DATES: From: November 2, 2004 To: November 22, 2004

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

Dose Calculations:
Individual doses were calculated based on the initial body weights, taking into account the specific gravity (determined by PSL) and concentration of the test mixture.

Dosing:
The test substance was administered as a 18% w/w mixture in corn oil using a stainless steel balltipped gavage needle attached to an appropriate syringe. Due to the high volume of test suspension to be administered (11.42 ml/kg ) , each animal's dose was divided into two approximately equal portions, administered approximately two hours apart. During dosing, the test suspension was kept on a magnetic stir plate.

Doses:

2,000 mg/kg bodyweight

No. of animals per sex per dose:

5 female rats/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (or other?) 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

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

2000 mg/kg - 0/5 female

Clinical signs:

other: Following administration, all animals had blue staining at the base of the tail, and one animal exhibited facial staining. However, these symptoms cleared by Day 3, and along with the other animals, appeared active and healthy for the reminder of the 14-d

Gross pathology:

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

Any other information on results incl. tables

INDIVDUAL BODY WEIGHTS AND DOSES

Animal No.	Sex	Dose Level (mg/kg)	Body Weight (g)			Dose
			Initial	Day 7	Day 14	ml
8331	F	2,000	197	219	244	2.2
8356	F		192	216	260	2.2
8478	F		192	228	259	2.2
8523	F		204	221	253	2.3
8580	F		220	233	254	2.5

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 this study, the acute oral LD50 of the test substance is greater than 2,000 mg/kg of body weight in female rats.

Executive summary:

An acute oral toxicity test (Up and Down Procedure) was conducted with rats to determine the potential for Spectrace™MD-810 Marker (Solvent Free) to produce toxicity from a single dose via the oral route. All animals survived exposure to the test substance and gained body weight during the study. Following administration, all animals had blue staining at the base of the tail, and one animal exhibited facial staining. However, these symptoms cleared by Day 3, and along with the other animals, appeared active and healthy for the remainder of the 14-day observation period. No gross abnormalities were noted for any of the animals when necropsied at the conclusion of the 14-day observation period.

Under the conditions of this study, the acute oral LD50 of the test substance is greater than 2,000 mg/kg of body weight in female rats. Based on these results and according to the EEC criteria for classification and labelling requirements for dangerous substances and preparations (EEC Directive 91/325/EEC, Amendment to Annex VI of the EEC Directive 67/548/EEC), Spectrace MD-810 Marker (Solvent Free) would not be classified and has no obligatory labelling requirement.