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

Endpoint summary

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted February 24, 1987

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

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

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.

Preliminary study:

Not performed.

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.

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

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Klimisch code 1

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Acclimatization: October 15, 1996 - Report: November 26, 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

February 24, 1987

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Version / remarks:

July 31, 1992

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

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 FOllinsdorf I Switzerland
- Females (if applicable) nulliparous and non-pregnant: no
- Age when treated: 8 weeks (males), 11 weeks (females)
- Weight when treated: males 231.0-243.6 g, females 203.3-222.9 g
- Housing: During acclimatization in groups of five in Makrolon type-4 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz). During treatment and observation individually in Makrolon type-3 cages with standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standard Kliba 343, Batch no. 78/96 rat maintenance diet (Kliba Muhlen AG, CH-4303 Kaiseraugst) available ad libitum.
- Water (e.g. ad libitum): Community tap water from Itingen, available ad libitum.
- 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°C ± 3°C
- Humidity (%): 40-70% (values above 70% during cleaning process possible)
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12 (music was played during the daytime light period)

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

bio-distilled water

Details on dermal exposure:

TEST SITE
- Area of exposure: back
- % coverage: 10%
- Type of wrap if used: the dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): twenty-four hours after the application the dressing was removed and the skin was washed with lukewarm tap water and dried with disposable paper towels.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg body weight
- Concentration (if solution): 0.5 g/ mL vehicle
- Constant volume or concentration used: yes

VEHICLE: bi-distilled water
- Amount(s) applied (volume or weight with unit): 4 mL (vehicle + test item)/kg
- Concentration (if solution): 0.5 g/ mL vehicle

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/viability: Four times during test day 1 and once daily for surviving animals during days 2-15.

- 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 NARCDREN (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 and all abnormalities recorded. Thereafter, they were discarded.

- Other examinations performed: clinical signs, body weight, organ weights, histopathology, ...

Body weights: On test days 1 (pre-administration), 8 and 15 for surviving animals.
Clinical signs: Each animal was examined for changes in behaviour and appearance (with special emphasis on the application area, except for the time when the semi-occlusive dressing was in place) four times during day 1, and once daily during days 2-15. All abnormalities were recorded.

Statistics:

No statistical analysis was performed.

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.
Red discoloration of the skin at the application site was evident in all animals after the removal of the dressing on test day 2 and persisted until study termination. Residual test article was noted in three males and one female.

Body weight:

The body weight of the animals was within the range of physiological variability known for rats of this strain and age. For results, please refer to the table in "Any other information on results incl. tables" hereafter.

Gross pathology:

No macroscopic organ findings were observed at necropsy.

Evolution of body weight (in grams) :

Sex/dose	Animal No.	Day 1 (= day of treatment)	Day 8	Day 15
Male / 2000 mg/kg	1	231.0	250.3	270.4
	2	240.2	267.1	294.8
	3	243.6	269.6	296.9
	4	239.1	262.0	280.6
	5	232.4	261.5	293.5
Female / 2000 mg/kg	6	208.9	228.6	250.6
	7	203.3	214.6	222.1
	8	222.9	224.9	232.6
	9	208.6	221.0	237.0
	10	209.9	221.9	235.4

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LDso (rat) of SCARLET RN 1165 is > 2000 mg/kg bw in test performed according to OECD TG 402 and following GLP.

Executive summary:

A group of five male and 5 female Hanibm:WIST (SPF) rats was treated with SCARLET RN 1165 at 2000 mg/kg by dermal application. The test article was suspended in vehicle (bi-distilled water) at a concentration of 0.5 g/ml and administered at a volume of 4 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 period.

Red skin (back) and residual test article (back) were observed on the test sites.

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

No macroscopic organ findings were observed at necropsy.

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Klimisch code 1

Justification for classification or non-classification

For each route of exposure, the LD50/LC50 is > threshold dose leading to classification according to CLP criteria.