7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(6-sulpho-2-naphthyl)azo]naphthalene-2-sulphonic] acid, sodium salt, compound with 2,2',2''-nitrilotriethanol

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

Adopted in December 2001

Deviations:

not applicable

Remarks:

No influence to the quality or integrity of the study

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(6-sulpho-2-naphthyl)azo]naphthalene-2-sulphonic] acid, sodium salt, compound with 2,2',2''-nitrilotriethanol

EC Number:

279-255-7

EC Name:

7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(6-sulpho-2-naphthyl)azo]naphthalene-2-sulphonic] acid, sodium salt, compound with 2,2',2''-nitrilotriethanol

Cas Number:

79770-29-9

Molecular formula:

C41H28N6O15S4.xC6H15NO3.xNa

IUPAC Name:

2-[bis(2-hydroxyethyl)amino]ethan-1-ol 4-hydroxy-7-[({5-hydroxy-7-sulfo-6-[(E)-2-(6-sulfonaphthalen-2-yl)diazen-1-yl]naphthalen-2-yl}carbamoyl)amino]-3-[(E)-2-(6-sulfonaphthalen-2-yl)diazen-1-yl]naphthalene-2-sulfonic acid sodium hydride

Test material form:

liquid

Specific details on test material used for the study:

To consider the purity of the test item a correction factor of 1.1 was applied.

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
Species/strain: WISTAR rats Crl: WI(Han)
Source: Charles River, 97633 Sulzfeld, Germany
Sex: female (non-pregnant and nulliparous)
Number of animals: 3 per step
Age at the
beginning of the study: 8 – 10 weeks
Body weight on the
day of administration: Step 1: 163 – 177 g, Step 2: 154 – 173 g
Housing and Feeding Conditions
- Full barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding
- Certificates of food, water and bedding are filed for two years at BSL Munich and afterwards archived at Eurofins Munich
- Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

VEHICLE
-No vehicle used
MAXIMUM DOSE VOLUME APPLIED:
2000mg/kg bw
CLASS METHOD
Number of animals: 3 per step / 2 steps performed
Method: OECD 423
EC 440/2008, Method B.1 tris
OPPTS 870.1100

Doses:

2000 mg/kg bw for both steps

No. of animals per sex per dose:

3 females

Control animals:

no

Details on study design:

All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.
The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.
A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
At the end of the observation period the animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally at a dosage of 250-400 mg/kg bw
All animals were subjected to gross necropsy and examined macroscopically for gross pathological changes. In the absence of gross pathological changes no tissues were preserved for a possible histopathological evaluation.

Statistics:

None required

Results and discussion

Preliminary study:

LD50 in rats >2000mg/kg bw

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

None

Clinical signs:

other: Slight piloerection and hunched posture

Gross pathology:

No specific gross pathological changes were recorded for any animal.

Other findings:

None

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose of Direct Red 236 after a single oral administration to female rats, observed over a period of 14 days is:
LD50 (rat): > 2000 mg/ kg bw

Executive summary:

Under the conditions of the present study, a single oral application of the test item Direct Red 236 to rats at a dose of 2000 mg/kg body weight was associated with slight signs of toxicity but not mortality.

The median lethal dose of Direct Red 236 after a single oral administration to female rats, observed over a period of 14 days is:

LD50 (rat): > 2000 mg/ kg bw