Alkali salt of substituted alkyl amino sulfonyl triazinyl amino arylamido diazo aryl sulfonyl sulfonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 August to 24 August 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

During the animal exposures, the animal room temperature was in the range 24.3 – 26.2 °C,from 16 August 2011 to 17 August 2011. This was a deviation from the range stated in the guidelines (19.0 – 25.0 °C).

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

During the animal exposures, the animal room temperature was in the range 24.3 – 26.2 °C,from 16 August 2011 to 17 August 2011. This was a deviation from the range stated in the guidelines (19.0 – 25.0 °C).

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

During the animal exposures, the animal room temperature was in the range 24.3 – 26.2 °C,from 16 August 2011 to 17 August 2011. This was a deviation from the range stated in the guidelines (19.0 – 25.0 °C).

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Details on test material:

Substance name: Reactive Yellow F01-0555

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

EXPERIMENTAL ANIMALS

Species and strain: RjHan:WI rats
Source: Laboratoire Elevage Janvier, B.P. 4105, Route des Chênes Secs, 53940 Le Genest-St-Isle CEDEX FRANCE
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Number of animals: 6 animals, 3 animals/group
Sex: Female, nulliparous and non-pregnant.
Age of animals at dosing: Young healthy adult rats, 10 weeks old
Date of receipt: 28 July 2011
Body weight at treatment: 221 – 230 g
Acclimation period: At least 12 days

Husbandry

Animal health: Only healthy animals were used for the test. The veterinarian certified the health status.
Number of animal room: 522/10
Housing: 3 animals / cage
Cage type: Type II polypropylene/polycarbonate
Bedding: Lignocel Bedding for Laboratory Animals was available to animals during the study. A copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 21.4 – 26.2°C
Relative humidity: 30 - 70%
Ventilation: 15-20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.

The temperature and relative humidity were recorded twice daily during the study.

Food and Water Supply

Animals received ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle ad libitum. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

Animal Identification

Animals were individually identified using numbers written on the tail with an indelible marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.' s Master File, for each animal allocated to the treatment groups. The cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Distilled

Details on oral exposure:

Vehicle: Distilled water
Dispense Code: 0110111
Expiry Date: 31 January 2014
Dose volume: 10 mL/kg bw

Test item was freshly formulated at a concentration of 200 mg/mL in the vehicle (without correction for purity), in the Central Dispensary Unit of CiToxLAB Hungary Ltd. on the day of administration. The formulation container was stirred continuously during administration to ensure that the syringe was filled from a homogeneous suspension/solution.

Justification of the dose:

The initial dose level was selected on the basis of the information provided by the Sponsor. The LD50 value was expected to be above 2000 mg/kg bw.

Initially, three female animals were treated with 2000 mg/kg bw of Reactive Yellow F01-0555. As no mortality occurred within 24 hours after dosing, a second group of three animals received 2000 mg/kg bw of the test item approximately 24 hours after treatment of the first group. No mortality occurred in the second treatment group; hence, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris). The test was terminated on completion of the 14-day observation period.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Initially, three female animals were treated with 2000 mg/kg bw of Reactive Yellow F01-0555. As no mortality occurred within 24 hours after dosing, a second group of three animals received 2000 mg/kg bw of the test item approximately 24 hours after treatment of the first group.

Control animals:

no

Details on study design:

Procedure

A single oral dose was administered by gavage followed by a fourteen-day observation period. The animals were fasted for about 16 hours prior to treatment. The food but not water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral gavage in the morning. The food was returned 3 hours after the treatment. A constant treatment volume of 10 mL/kg bodyweight was applied.


OBSERVATIONS

Clinical Observations

Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement

The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0), Day 3, Day 7 and Day 14.

NECROPSY

Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthasol® 40 %; details are presented in 3.1.3.). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.

EVALUATION OF THE RESULTS

The method used was not intended to allow the calculation of a precise LD50 value.
The test item was ranked into categories of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423.
Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.

Statistics:

None

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Reactive Yellow F01-0555 did not cause mortality at a dose level of 2000 mg/kg bw.

Clinical signs:

other: Treatment with Reactive Yellow F01-0555 at a dose level of 2000 mg/kg bw caused orange urine and faeces on Day 1 in all Group 1 animals and at 6 hours after treatment in all Group 2 animals. All animals fully recovered and were free of any clinical signs

Gross pathology:

There were no test item related macroscopic findings in animals dosed at 2000 mg/kg bw at necropsy.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the acute oral LD50 value of the test item Reactive Yellow F01-0555 was found to be above 2000 mg/kg bw in female RjHan:WI rats.

Executive summary:

A single-dose oral toxicity of Reactive Yellow F01-0555 was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris) in RjHan:WI rats. Two groups of three female RjHan Wistar rats (10 weeks old) were treated with Reactive Yellow F01-0555 at a dose level of 2000 mg/kg bw (Group 1 and Group 2).

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. Rats were maintained without compound administration for a 2‑week observation period after the day of dosing. As no mortality was observed in this dose group within 24 hours after dosing, a confirmatory treatment was performed on 3 further females (Group 2) at the same dose level. As no mortality was observed in the second dose group, no further testing was required according to OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris.

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal (about 16 hours prior to treatment). Food was made available again 3 hours after the treatment. Reactive Yellow F01-0555 was administered formulated in distilled water at a concentration of 200 mg/mL at a dosing volume of 10 mL/kg bw. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0, 3 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

Reactive Yellow F01-0555 did not cause mortality at a dose level of 2000 mg/kg bw. Treatment with Reactive Yellow F01-0555 at a dose level of 2000 mg/kg bw caused orange urine and faeces in all animals (6/6). All animals fully recovered and were freeof any clinical signsfrom Day 2 until the end of the observation period. Body weights or bBody weight gains showed no indication of a treatment-related effect.

There were no test item related in animals dosed at 2000 mg/kg bw at necropsy.

Under the conditions of this study, the acute oral LD50 value of the test item Reactive Yellow F01-0555 was found to be above 2000 mg/kg bw in female RjHan:WI rats.