pentasodium 4-amino-3-[{4-[(2,4-disulfonatophenyl)diazenyl]-3-methylphenyl}diazenyl]-5-hydroxy-6-[(4-nitro-2-sulfonatophenyl)diazenyl]naphthalene-1,7-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:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

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)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

impurity 1

Reference substance name:

Unknown impurities

IUPAC Name:

Unknown impurities

impurity 2

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

Dihydrogen oxide

Constituent 1

Reference substance name:

pentasodium 4-amino-3-(2-{4-[2-(2,4-disulfonatophenyl)diazen-1-yl]-3-methylphenyl}diazen-1-yl)-5-hydroxy-6-[2-(4-nitro-2-sulfonatophenyl)diazen-1-yl]naphthalene-1,7-disulfonate

Cas Number:

2259360-19-3

Molecular formula:

C29H17N8Na5O18S5

IUPAC Name:

pentasodium 4-amino-3-(2-{4-[2-(2,4-disulfonatophenyl)diazen-1-yl]-3-methylphenyl}diazen-1-yl)-5-hydroxy-6-[2-(4-nitro-2-sulfonatophenyl)diazen-1-yl]naphthalene-1,7-disulfonate

Test material form:

solid: particulate/powder

Details on test material:

Batch XMEF 1476

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:
Janvier Labs SAS, Le Genest St. Isle, 53941 Saint Berthevin Cedex, France
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation:
8 - 12 weeks
- Weight at study initiation:
204 - 246 g
- Fasting period before study:
overnight
- Housing:
groups of one to five rats
- Historical data:
available
- Diet (e.g. ad libitum):
ad libitum
- Water (e.g. ad libitum):
ad libitum
- Acclimation period:
at least 5 days prior to start of dosing

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
22 +/- 2°C
- Humidity (%):
45-65%
- Air changes (per hr): at least 8 per hour
- Photoperiod (hrs dark / hrs light):
12/12

IN-LIFE DATES: From: 2020-07-20 To: 2020-08-12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 and 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: stability in vehicle given

CLASS METHOD (if applicable)
- In the absence of data regarding the toxicity of the test item 300 mg/kg was chosen as the initial starting dose, since a severe toxicity which may necessitate humane euthanasia was not expected at this dose level.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

1 female for the pre-test with 300 mg/kg bw and 2000 mg/kg bw each
4 females for the main study with 2000 mg/kg bw

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations and inspections for morbidity / mortality were performed at least three times within the first six hours after application (i.e., 30 minutes and 1 hour, 2 hours and 4 hours after dosing), thereafter at least once daily for 14 days. Body weights were recorded on Day 0 (prior to dosing), Day 7, and 14.
- Necropsy of survivors performed: yes

Statistics:

not needed

Results and discussion

Preliminary study:

In the sighting test with one female and a single dose of 2000 mg/kg bw

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

One female (animal number 2) had to be humanely euthanised on day 1 after application due to deterioration of clinical signs.

Clinical signs:

other: Clinical signs included secretion of Harderian glands, fur stained by the test item, staining around the mouth, blue to black staining of the skin, staining of the urine and faeces. The euthanized animal showed the following additional symptoms: hunched p

Gross pathology:

Three animals showed dark kidneys, possibly due to test item residuals in the parenchyma.
For animal number 2 discoulouring of the following organs to blue was noted: fur, skin, orifices, tongue, pharynx, eyes, mucosa, small and large intestine, peritoneum, kidneys, and caecum. The skin of the urinary bladder was discoloured as well but the content was clear.
Additionally, fluffed-up cheeks and a thickened right masseter muscle, a filled stomach and a moniliform content of the small intestine and black content of the caecum was observed.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Executive summary:

The registered substance was tested at a starting dose of 300 mg/kg bw and at 2000 mg/kg bw in the main study of an acute oral toxicity study in female Wistar rats following OECD TG 420. One animal of the main test treated with 2000 mg/kg body weight was euthanized on day 1 after application due to deterioration of clinical signs that were, if compared to the symptoms observed in the four other animals dosed with 2000 mg/kg b.w., unexpectedly severe. Therefore, it is possible that these individual symptoms were incidental. Clinical signs in the other 4 animals included secretion of Harderian glands, and staining by the couloured test item of fur, mouth, skin, urine and faeces. The acute median lethal oral dose (LD50) was demonstrated to be > 2000 mg/kg bw.