disodium 4-amino-6-((4-((4-(2,4-diaminophenyl)azo)phenylsulfamoyl)phenyl)azo)-5-hydroxy-3-((4-nitrophenyl)azo)naphthalene-2,7-disulfonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 august 1995 - 14 september 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

421-880-6

EC Name:

-

Cas Number:

201792-73-6

Molecular formula:

C34H25N11Na2O11S3

IUPAC Name:

disodium 4-amino-6-{2-[4-({4-[2-(2,4-diaminophenyl)diazen-1-yl]phenyl}sulfamoyl)phenyl]diazen-1-yl}-5-hydroxy-3-[2-(4-nitrophenyl)diazen-1-yl]naphthalene-2,7-disulfonate

Test animals

Species:

rat

Strain:

other: Alpk: APfSD (Wistar)

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animals:
species: rat
strain: Alpk: APfSD (Wistar)
source: Barriered animal Breeding unit, Zeneca Pharmaceuticals, Alderley Park
Sex: 5 male and 5 female
Specification: Young Adults, Males weighed 279-294g and the females weighed 200-256g at the beginning of the study.

Accomodation and husbandry:
A maximum of 5 rats was housed per cage, sexes separately, in multiple rat sacks suitable for animals of this strain and the weight range expected during the course of the study.
The rats were fasted overnight immediately prior to dosing to ensure that the stomachs were void of food (the presence of food could affect the rate of absorption of the test substance)
The rats were transferred to clean cages and racks,as necessary, during the study.
The animal room was designed to give the environmental conditions shows as follows:
temperature: 21±2°C
Relative humidity: 40-70%
Air: approximately 25-30 changes/hour
Light cycle: artifical giving 12 hours light, 12 hours dark

Both temperature and relative humidity were monitored continuously using an automated system which triggers an alarm if values are outside specified ranges. In general, the recorded values were within the specified ranges and any deviations that were observed are considered not to have affected the integrity of the study.

Diet (PCD) supplied by Special Diet Services Limited, Witham, Essex, UK and mains water, supplied by an automatic system were available ad libitum.

Each batch of diet is routinely analysed for composition and for the presence of contaminants. Water is also periodically analysed for the presence of contaminants. No contaminants were found to be present in the diet or water at levels considered to be capable of interering with the puropose or outcome of the study.


Acclimatisation:
The animals were housed under the experimental conditions for at least 6 days, prior the start of the study.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

A standard volume of 10 ml/kg of the dosing preparation was administered by oral gavage using a plastic catheter attached to a sterile disposable plastic syringe of suitable size.

Doses:

The dose levels selected for this study were based on the results of a preliminary study (speculative oral dosing study) in the rat.
The animals were given a single dose.
As no deaths were noted in the speculaive study, a single, limit dose level of 2000 mg/kg was administered.
The volume of the dose was calculated for each animal according to its weight at the time of dosing.

No. of animals per sex per dose:

5 males and 5 females

Control animals:

not specified

Details on study design:

Five male and five female rats were allocated to the study. animals were individually identified with a number, unique within the study, by ear punching.
On the front of each cage of animals was a card identifying the contained animals by procedure code, test substance,date of administration, dose level, individual number, sex and study.
A card was also displayed on the cage to indicate the date of the pre-dose fasting period.

Prior to the start of the sudy, all rats were examined to ensure that they were physically normal and exhibited normal activity. The animlas were observed for signs of systemic toxicity once within 2 hours of dosing and again between 4 and 7 hours after dosing. Subsequent observations were made once daily up to day 15.

The animals were weighed on the day before dosing (day -1), immediately before dosing (day 1) and at days 1, 3, 6, 8, 15.

All rats were killed by exsanguination under terminal anaesthesia induced by halothane vapour.

All animals were subjected to an examination post mortem. This involved an external observation and a careful examination of all thoracic and abdominal viscera. All abnormalities were recorded but tissues were not submitted for histopathological examination.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

no deaths

Clinical signs:

other: no significant signs of toxicity. Upward curvature of the spine and piloerection were noted in all animals for up to 2 days. All animals showed yellow staining of the fur throughout the study and two males and one female had coloured faeces on day 1.

Gross pathology:

Post mortem examination revealed mottled areas in the lung of one female and red areas in the thymus of another female. These are common spontaneus findings in rats of this age and strain and are considered not to be treatment-related. There were no macroscopic finding in the males.

Applicant's summary and conclusion

Interpretation of results:

other: not classified

Conclusions:

The acute oral medina lethal dose of the tested substance is in excess of 2000 mg/kg to male and female rats.

Executive summary:

Groups of five male and five female Alpk:APfSD(Wistar-derived) rats received a single dose of 2000 mg/ks of the test sample.

The animals were assessed daily for the following 15 days for any signs of systemic toxicity and their bodyweights were recorded at intervals throughout the study. At the end of the study all the animals were killed and subjected to a macroscopic examination post mortem. Following a single oral dose of 2000mg/kg, none of the animals died and there were no significant signs of toxicity.

The acute oral median lethal dose of the tested substance is in excess of 2000mg/kg to male and female rats.