Reaction products of 4,4'-methylenebis(2,6-dimethylaniline) and 4-aminobenzenesulfonic acid with 2,4,6-trichloro-1,3,5-triazine, diazotised and subsequently coupled with N-(2-methoxyphenyl)-3-oxobutanamide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 February 2006 to 17 March 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction product of 4-{4-[4-(4-Amino-3,5-dimethyl-benzyl)-2,6-dimethyl-phenylamino]-6-chloro-[1,3,5]triazin-2-ylamino}-benzenesulfonic acid, sodium nitrite, hydrochloric acid and acetoacet-o-anisidin

Cas Number:

1165939-52-5

IUPAC Name:

Reaction product of 4-{4-[4-(4-Amino-3,5-dimethyl-benzyl)-2,6-dimethyl-phenylamino]-6-chloro-[1,3,5]triazin-2-ylamino}-benzenesulfonic acid, sodium nitrite, hydrochloric acid and acetoacet-o-anisidin

Test material form:

solid: particulate/powder

Remarks:

Yellow solid

Details on test material:

- Expiry date: 20 Dec 2007
- Storage conditions: At room temperature (ca 20°C)

Test animals

Species:

rat

Strain:

Wistar

Remarks:

HanRCC:WIST (SPF)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 13 - 14 weeks
- Weight at study initiation: 194.7 -203.1 g
- Fasting period before study: Fasted for approximately 18 hours. Access to water was permitted. Food was provided again approximately 3 hours after dosing.
- Housing: In gropus of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 % (values above 70 % possible during cleaning process).
- Air changes (per hr): 10 - 15 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 h light and 12 h dark.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

VEHICLE
- Justification for choice of vehicle: The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. This formulation trial is excluded from the GLP statement of compliance.
- Lot/batch no.: 11577905 110527

MAXIMUM DOSE VOLUME APPLIED
- 10 mL/kg body weight.

DOSAGE PREPARATION
- The dose formulations were made shortly before each dosing occasion using a magnetic stirrer and a spatula as homogenisers.
- The test material was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume).
- Homogeneity of the test material in the vehicle was maintained during administration using a magnetic stirrer.

Doses:

A single dose of 2000 mg/kg body weight.

No. of animals per sex per dose:

Two groups, each of three females.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality / viability: daily during the acclimatisation period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2 – 15.
Body weights: On test days 1 (prior to administration), 8 and 15.
Clinical signs: Daily during the acclimatisation period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2 - 15. All abnormalities were recorded.
- Necropsy of survivors performed: All animals were killed at the end of the observation by carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. No organs or tissues were retained.

Statistics:

No statistical analyses were used.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the study.

Clinical signs:

other: Slightly ruffled fur was noted in five out of six animals treated at 2000 mg/kg at the 2- and 3-hour reading and persisted in four of them up to the 5-hour reading. Yellow faeces were noted in the cage of the first treated group on test day 2.

Gross pathology:

Not macroscopic findings were recorded at necropsy.

Applicant's summary and conclusion

Interpretation of results:

other: Not classified according to EU criteria.

Conclusions:

Under the conditions of the study the acute oral LD50 of the test material in female rats was > 2 000 mg/kg body weight.

Executive summary:

The acute oral toxicity of the test material was assessed according to OECD Test Guideline 423 and EU Method B.1 tris according to the Acute Toxic Class method and in compliance with GLP.

During the study, two groups each of three female HanRcc:WIST (SPF) rats were treated with the test material by oral gavage administration at a dosage of 2000 mg/kg bodyweight. The test material was diluted in vehicle (PEG 300) at a concentration of 0.2 g/mL and administered at a volume dosage of 10 mL/kg.

The animals were examined daily during the acclimatisation period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2 – 15. Mortality/viability were recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2 – 15. Bodyweights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

All animals survived until the end of the study period.

Slightly ruffled fur was noted in five out of six animals treated at 2000 mg/kg at the 2- and 3-hour reading and persisted in four of them up to the 5-hour reading. Yellow faeces were noted in the cage of the first treated group on test day 2.

The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy.

The median lethal dose of the test material after a single oral administration to female rats, observed over a period of 14 days is > 2000 mg/kg bodyweight.