disodium 7-(4,6-difluoropyrimidin-2-ylamino)-4-hydroxy-3-(4-methoxy-2-sulfonatophenylazo)naphthalene-2-sulfonate; reaction mass of: disodium 7-(2,4-difluoropyrimidin-6-ylamino)-4-hydroxy-3-(4-methoxy-2-sulfonatophenylazo)naphthalene-2-sulfonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Acclimatization: October 15, 1996 - Termination: November 05, 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted February 24, 1987

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Version / remarks:

July 31, 1992

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:

-

EC Number:

426-840-1

EC Name:

-

IUPAC Name:

tetrasodium 7-[(2,6-difluoropyrimidin-4-yl)amino]-4-hydroxy-3-[2-(4-methoxy-2-sulfophenyl)diazen-1-yl]naphthalene-2-sulfonate 7-[(4,6-difluoropyrimidin-2-yl)amino]-4-hydroxy-3-[2-(4-methoxy-2-sulfophenyl)diazen-1-yl]naphthalene-2-sulfonate

Test material form:

solid

Details on test material:

Identity: Scarlet RN 1165
Appearance : Solid powder, dark-red

Test animals

Species:

rat

Strain:

other: HanIbm: WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wolferstrasse 4, 4414 Fullinsdorf I Switzerland
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation (age when treated): males 8 weeks, females 10 weeks
- Weight at study initiation (body weight range when treated): males 199.8-207.0 g, females 169.8-193.3 g
- Fasting period before study: overnight fasting period prior to application
- Housing: Groups of five in Makrolon type-4 cages with standard softwood bedding ("LignocelN, Schill AG, CH-4132 Muttenz).
- Diet (e.g. ad libitum): Pelleted standard Kliba 343, Batch no. 78/96 rat maintenance diet (Kliba MOhlen AG, CH-4303 Kaiseraugst) available ad libitum (except for the overnight fasting period prior to application). Results of analyses for contaminants are included in the report.
- Water (e.g. ad libitum):Community tap water from Itingen, available ad libitum. Results of bacteriological, chemical and contaminant analyses are included in the report.
- Acclimation period:One week 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 (%): between 40-70% (values above 70% during cleaning process possible)
- Air changes (per hr): Air-conditioned with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/12 hours dark, music during the light period

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

bi-distilled water

Details on oral exposure:

VEHICLE: bi-distilled water
- Concentration in vehicle: 0.2g test item/ml vehicle
- Amount of vehicle (if gavage): 10 ml vehicle/kg bw
- Justification for choice of vehicle: not detailed
- Lot/batch no. (if required): not detailed
- Purity: not detailed

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg body weight

DOSAGE PREPARATION (if unusual):
The test article was placed into a glass beaker on a tared Mettler PM 460 balance and the vehicle (bi-distilled water) was added. A weight by volume dilution was prepared using a glass rod and a magnetic stirrer (Janke & Kunkel,
0-79219 Staufen) as homogenizers. Homogeneity of the test article in the vehicle was maintained during treatment using the same magnetic stirrer (Janke & Kunkel, 0-79219 Staufen). The preparation was made shortly before dosing.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: limit concentration of 2000 mg/kg bw

TREATMENT
The animals received a single dose of the test article on a mg/kg body weight basis by oral gavage following fasting for approximately 18 hours, but with free access to water. Food was provided again approximately 3 hours after dosing.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing:
MORTALITY/VIABILITY: four times during test day 1 and once daily for surviving animals during days 2-15.
CLINICAL SIGNS: each animal was examined for changes in appearance and behaviour four times during day l, and once daily for surviving animals during days 2-15. All abnormalities were recorded.
BODY WEIGHTS: on test day 1 (pre-administration), 8 and 15 for surviving animals.
- Necropsy of survivors performed: yes
Necropsies were performed by experienced prosectors. At the end of the observation period all animals were sacrificed by intraperitoneal injection of NARCOREN (Rhone Merieux GmbH, D-88471 Laupheim) at a dose of at least 2.0 ml/kg body weight (equivalent to at least 320 mg sodium pentobarbitone/kg body weight). The animals were examined macroscopically. Thereafter, they were discarded.

- Other examinations performed: -

Statistics:

The LOGIT-Model could not be used as no deaths occured.

Results and discussion

Preliminary study:

Not performed.

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the study.

Clinical signs:

No clinical signs of toxicity were observed during the study period.

Body weight:

The body weight of the animals was within the range of physiological variability known for rats of this strain and age.

Gross pathology:

No macroscopic findings were observed at necropsy.

Any other information on results incl. tables

Evolution of body weight (in grams):

Sex/dose	Animal No.	Day 1 (= day of treatment)	Day 8	Day 15
Male / 2000 mg/kg	1	204.6	272.1	305.3
	2	207.0	265.1	278.6
	3	201.9	253.3	277.0
	4	199.8	260.7	286.1
	5	200.2	253.9	286.1
Female/ 2000 mg/kg	6	193.1	221.1	232.2
	7	189.5	209.4	215.4
	8	169.8	192.5	209.2
	9	186.6	215.4	222.8
	10	193.3	232.9	238.7

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The mean lethal dose of SCARLET RN 1165 after single oral administration to rats of both sexes, observed over a period of 14 days, could not be estimated because LD50: greater than 2000 mg/kg.

Executive summary:

A group of five male and 5 female Han!bm:WIST (SPF) rats was treated with SCARLET RN 1165 at 2000 mg/kg by oral gavage. The test article was suspended in vehicle (bi-distilled water) at a concentration of 0.2 g/ml and administered at a volume of 10 ml/kg. Four times during day 1 and once daily during days 2-15 the animals were examined for clinical signs. Mortality/viability were recorded together with clinical signs at the same time intervals. Body weights were recorded on day 1 before administration and on days 8 and 15. All animals were necropsied and examined macroscopically.

No deaths occurred during the study.

No clinical signs of toxicity were observed during the observation period.

The body weight of the animals was within the range of physiological variability known for rats of this strain and age.

No macroscopic findings were observed at necropsy.

CONCLUSION

The mean lethal dose of SCARLET RN 1165 after single oral administration to rats of both sexes, observed over a period of 14 days, could not be estimated, because LD50: greater than 2000 mg/kg