Trisodium 5,5'-[(2-sulphonato-1,4-phenylene)bis(azo)]bis(salicylate)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-08-03 to 2017-10-24

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

December 2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

May 2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 5,5'-[(2-sulphonato-1,4-phenylene)bis(azo)]bis(salicylate)

EC Number:

276-501-5

EC Name:

Trisodium 5,5'-[(2-sulphonato-1,4-phenylene)bis(azo)]bis(salicylate)

Cas Number:

72230-95-6

Molecular formula:

C20H14N4O9S.3Na

IUPAC Name:

trisodium 3,3'-[(2-sulfonato-1,4-phenylene)didiazene-2,1-diyl]bis(6-hydroxybenzoate)

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

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:

other: aqua ad injectionem (sterile water)

Remarks:

(Deltamedica, lot no. 612118, expiry date: 30/11/2019)

Details on oral exposure:

The animals were marked for individual identification by tail painting.
Prior to the administration a detailed clinical observation was made of all animals. Only healthy animals were used.
Prior to the administration food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing.
The test item was administered at a single dose by gavage using a feeding tube.
The test item was administered at a dose volume of 10 mL/kg body weight.

Doses:

The starting dose was selected to be 2000 mg a.i./kg body weight. No compound-related mortality was recorded for any animal of step 1. Based on these results and according to the acute toxic class method regime a second step was required. A second step was performed at a dose of 2000 mg a.i./kg body weight. No compound related mortality was recorded for any animal of step 2. Based on these results and according to the acute toxic class method regime no further testing was required.

No. of animals per sex per dose:

3 per step (2 steps performed)

Control animals:

no

Details on study design:

All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.
Weight Assessment
The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.
Clinical Examination
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.

Pathology
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.

Evaluation of Results
Results were interpreted according to OECD Guideline 423, Annex 2.
Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by dose level.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
Necropsy findings were described.
With few exceptions, data were captured using the validated departmental computerised system E WorkBook (version 10.1.2, ID Business Solutions Ltd.).

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the
results is not regarded as necessary.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

> 5 000 mg/kg bw

Based on:

act. ingr.

Remarks on result:

other: Limit dose tested: 2000 mg/kg body weight

Mortality:

All animals survived until the end of the study without showing signs of toxicity.

Clinical signs:

other: All animals survived until the end of the study without showing signs of toxicity.

Gross pathology:

At necropsy, no macroscopic findings were observed in any animal of any step.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the present study, a single oral application of the test item to rats at a dose of 2000 mg a.i./kg body weight was not associated with signs of toxicity or mortality.
The median lethal dose of the test item after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat): > 5000 mg a.i./ kg bw

Executive summary:

Summary Results

Two groups, each of three female WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg a.i./kg body weight. The test item was dissolved in sterile water at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

All animals used in the study after their entrance at BSL were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

Results per Step

Step	Sex / No.	Starting Dose (mg/kg bw)	Number of Animals	Number of Intercurrent Deaths
1	Female / 1 - 3	2000	3	0
2	Female / 4 - 6	2000	3	0

bw = body weight

All animals survived until the end of the study without showing signs of toxicity.

Throughout the 14-day observation period, the weight gain of the animals was within the normal range of variation for this strain.

Macroscopic findings of surviving animals:

At necropsy, no macroscopic findings were observed in any animal of any step.

LD₅₀cut-off (rat):                   > 5000mg a.i./kg bw

Species/strain:                      WISTAR Crl: WI(Han) rats

Vehicle:                                  Aqua ad injectionem

Number of animals:                3 per step / 2 steps performed

Method:                              OECD 423, EC 440/2008, Method B.1 tris, OPPTS 870.1100

Conclusion

Under the conditions of the present study, a single oral application of the test item Aluminium Orange RL pc-w to rats at a dose of 2000 mg a.i./kg body weight was not associated with signs of toxicity or mortality.

The median lethal dose of Aluminium Orange RL pc-w after a single oral administration to female rats, observed over a period of 14 days is:

LD₅₀cut-off (rat): > 5000 mg a.i./ kg bw