hexasodium 5-[(4-{3-[(8-benzamido-1-hydroxy-3,6-disulfonato-2-naphthyl)diazenyl]-4-sulfonatoanilino}-6-[(2-sulfonatoethyl)amino]-1,3,5-triazin-2-yl)amino]isophthalate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was conducted between 05 February 2007 and 28 February 2007.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Magenta T-43

IUPAC Name:

Magenta T-43

Test material form:

solid: particulate/powder

Details on test material:

Sponsor's identification: JPD Magenta T-43
Description: dark red solid
Storage conditions: room temperature in the dark
Batch number: H186-5

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

Animals and Animal Husbandry
Female Sprague-Dawley CD (Crl : CD® (SD) IGS BR) strain rats were supplied by Charles River (UK) Ltd, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non-pregnant. After an acclimatisation period of at least five days the animals were selected at random and given a number unique within the study by indelible ink-marking on the tail and a number written on a cage card. At the start of the study the animals were eight to twelve weeks of age. The bodyweight variation did not exceed ±20% of the mean initial bodyweight of any previously dosed animals.

The animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing, free access to mains drinking water and food (Certified Rat and Mouse Diet) was allowed throughout the study. The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.

The temperature and relative humidity were set to achieve limits of 19 to 25°C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.

Doses:

2000 mg/kg

No. of animals per sex per dose:

Preliminary sighting study: 1 female rat
Main study: 4 female rats

Control animals:

no

Details on study design:

Preliminary sighting study:
In the absence of data suggesting the test material was toxic, 2000 mg/kg was chosen as the starting dose. A single female animal was treated at a dose level of 2000 mg/kg.

Main study:
In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of 4 animals was treated at a dose level of 2000 mg/kg.

All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.

Clinical observations were made 0.5, 1, 2, and 4 hours after dosing and subsequently once daily for fourteen days. Morbidity and mortality checks were made twice daily.

Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.

At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.


Evaluation of Data:
Evaluation of data included identification of the number of animals that died during the study (or that were killed for humane reasons), and determination of the nature, severity, onset and duration of the toxic effects. If possible, the signs of evident toxicity were described. Evident toxicity
refers to the toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probable mortality. Effects on bodyweights and abnormalities noted at necropsy were also identified.

Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test material was made

Results and discussion

Preliminary study:

Following a sighting test in which there was no mortality at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a solution in distilled water at a dose level of 2000 mg/kg bodyweight.

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths.

Clinical signs:

other: No signs of systemic toxicity were noted. Red stained urine was noted in all animals during the day of dosing and one day after dosing.

Gross pathology:

Individual necropsy findings are provided.
No abnormalities were noted at necropsy.

Other findings:

None reported.

Any other information on results incl. tables

Individual Clinical Observations and Mortality Data

 

Dose mg/kg	Animal No. & sex	Effects After Dosing (hours)    Effects During period After Dosing (Days)
		0.5	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	1-0 Female	0	OU	OU	OU	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-0 Female	0	0	0	OU	OU	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-1 Female	0	0	0	OU	OU	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-2 Female	0	0	0	OU	OU	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-3 Female	0	0	0	OU	OU	0	0	0	0	0	0	0	0	0	0	0	0	0

 

0 = No signs of systemic toxicity

U = Red stained urine

 

 

Individual Bodyweights and Bodyweight Changes

 

Dose Level Mg/kg	Animal No. And Sex	Bodyweight (g) at Day			Bodyweight Gain (g) During Week
		0	7	14	1	2
2000	1-0 Female	229	256	272	27	16
	2-0 Female	212	241	253	29	12
	2-1 Female	220	256	278	36	22
	2-2 Female	195	222	252	27	30
	2-3 Female	207	235	270	28	35

 

 

Individual Necropsy Findings

 

Dose Level (mg/kg)	Animal No. and Sex	Time of Death	Macroscopic Observations
2000	1-0 Female	Killed Day 14	No abnormalities detected
	2-0 Female	Killed Day 14	No abnormalities detected
	2-1 Female	Killed Day 14	No abnormalities detected
	2-2 Female	Killed Day 14	No abnormalities detected
	2-3 Female	Killed Day 14	No abnormalities detected

                                                                                                                                        

Applicant's summary and conclusion

Interpretation of results:

other: Not classified according to EU criteria.

Conclusions:

The acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System Category - Unclassified).

Executive summary:

Introduction

The study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley CD strain rat. The method was designed to meet the requirements of the following:

• OECD Guidelines for Testing of Chemicals No 420 "Acute Oral Toxicity - Fixed Dose Method" (adopted 17 December 2001)

• Method B1 his Acute Toxicity (Oral) of Commission Directive 2004/73/EC

Method

Following a sighting test in which there was no mortality at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a solution in distilled water at a dose level of 2000 mg/kg bodyweight. Clinical signs and

bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity. Red stained urine was noted in all animals during the day of dosing and one day after dosing.

Bodyweight. All animals showed expected gains in bodyweight.

Necropsy. No abnormalities were noted at necropsy.

Conclusion.

The acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System Category - Unclassified).