Disodium; 4-Amino-3-{4-[4-(2,4-diamino-phenylazo)-benzenesulfonylamino]-phenylazo}-5-hydroxy-6-phenylazo-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:

2020

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:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

605-104-5

Cas Number:

157577-99-6

Molecular formula:

C34H26N10Na2O9S3

IUPAC Name:

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

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Species: laboratory albino rat (it is the preferred rodent species according to the guideline), females, nulliparous and non-pregnant
Strain: Wistar Han, monitored quality
Source: breeding farm VELAZ s.r.o., Koleč u Kladna, Czech Republic, RČH CZ 21760118
Sex: females
Age: 6-7 weeks at the time application
Acclimatisation: 6 days
Total number: 5 females
Housing: animal room with monitoring conditions, plastic breeding cages Velaz T4, sighting study 1 animal/cage, main study - 4 animals/cage
Diet: Altromin for rats, Manufacturer: Altromin Spezialfutter GmbH & Co. KG, Germany supplied via VELAZ
Water: drinking tap water ad libitum (quality corresponding to Regulation No. 252/2004 Czech Coll. of Law)
Microclimatic conditions: room temperature 22 + 3°C, permanently monitored
relative humidity 30 – 70 %, permanently monitored
light period 12-hour light/12 hour dark
Bedding: sterilized shavings of soft wood
Randomisation: according to the internal rule, at the start of the study the weight variation of animals was minimal
Identification of animals: colour marks on tail of animals, each cage was marked with the number of study, sex and dose of the test item
Health condition: certificate of good health condition – from breeding farm; no signs of diseases were observed at clinical check-in, during the acclimatisation period and before the start of study

About twenty hours before oral administration the animals were not fed, water was given ad libitum. Immediately before application the animals were weighed. The feed was given to animals 3 hours after application of the test item.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Immediately before application the test item was weighed and mixed in vehicle (Aqua pro iniectione) and resulting suspension was administered to the stomach by tube. The single volume of administered suspension was 1 ml/100 g of animal body weight

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 females

Control animals:

no

Details on study design:

Sighting study - the dose level of 2000 mg/kg was used as the starting dose - no death of animal was recorded.
Main study - on the basis of result of sighting study dose level 2000 mg/kg with four animals was used. No death of animals was detected.
Final number of animals for dose level 2000 mg/kg – 5 animals (1 animal – sighting study and 4 animals - main study).

Animals were weighed before application, at the 8th day of study and at the 15th day, before euthanasia of animals. Average body weight in a group was calculated from individual body weights. Body weight increments were calculated from body weight at the start of the study, the first week and at the end of the study.

After application the animals were observed individually:
- the first day: twice (30 minutes and 3 hours after application)
- the second day: twice (in the morning and in the afternoon) and daily thereafter for 14 days.
Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotoric activity, reactions to stimuli, and presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system.
The results of the observations were recorded on special data sheets.

All test animals survived to the end of study were sacrificed on the 15th day and gross necropsy was carried out. Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated. All gross macroscopic changes of organs and tissues were recorded on special data sheets

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No death recorded

Clinical signs:

other: No clinical sings of intoxication were observed during the study

Gross pathology:

No pathologic macroscopic changes were diagnosed during pathological examination.

Applicant's summary and conclusion

Interpretation of results:

other: not classified for acute oral toxicity

Conclusions:

The test item toxicity was evaluated on the basis of mortality, clinical signs of intoxication, body weight increments during the observation period and necropsy findings at the end of study.
The test item administered at the dose of 2000 mg/kg caused no death of animals. No clinical sings of intoxication were observed.
Body weight and weight increments were adequate to species, sex and age of animals in experiment. No pathologic macroscopic changes were diagnosed during pathological examination.
According to the study results the value of LD50 of the test item, Acid Black 234, for female rats is higher than 2000 mg/kg of body weight

Executive summary:

The aim of the study was to investigate acute toxic effects of the test item Acid Black 234, after a single oral administration to Wistar rats.

The testing was performed according to the OECD Test Guideline No. 420: Acute Oral Toxicity – Fixed Dose Procedure, adopted 17th December 2001.

With respect to information available indicating that the test item is likely to be nontoxic, the study was performed as limit test.

First the sighting study was performed, using the starting dose of 2000 mg/kg of body weight with one female, then followed main study with group of four females, dosed by the same dose of 2000 mg/kg of body weight.

The test item was administered in a single dose by gavage using a stomach tube to female Wistar CRL rats. After dosing the animals were observed individually for a total of 14 day. At the end of the test surviving animals were killed. Necropsy of all animals were carried out, and all gross pathological changes were recorded.

The test item administered at the dose level of 2000 mg/kg did not cause the death of animals. No clinical signs of intoxication were recorded. No pathologic macroscopic changes were diagnosed during pathological examination.

According to the study results the value of LD50 of the test item, Acid Black 234, for female rats is higher than 2000 mg/kg of body weight.