2-{4-[4-[4-fluoro-6-(2-(2-vinylsulfonylethoxy)ethylamino)-1,3,5-triazin-2-ylamino]phenylazo]phenylazo}naphthalene-4,6,8-trisulfonate, trisodium salt

Endpoint summary

Description of key information

The acute oral as well as dermal median lethal dose (LD50) of test substance in Wistar rats is >2000 mg/kg bw.

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:

04 May 2001 to 29 May 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

Identity: FAT 40800/A
Batch number: WP 2/01
Active ingredient: ca. 70 %
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Stability of test item: Stable under storage conditions
Stability of test item in vehicle: 24 hours at room temperature
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd., Biotechnology and Animal Breeding Division, CH-4414 Füllinsdorf / Switzerland
- Age at study initiation: Males: 8 weeks, Females: 10 weeks
- Weight at study initiation: Males: 194.1 - 198 g, Females: 168.5 - 174.2 g
- Fasting period before study: 16-20 hours
- Housing: In groups of three per sex in Makrolon type-4 cages with wire mesh tops and standardized softwood bedding ('Lignocel' Schill AG, CH-4132, Muttenz/Switzerland).
- Diet : Pelleted standard Provimi Kliba 3433 rat maintenance diet, batch no. 07/00 (Provimi Kliba AG, CH-4303, Kaiseraugst/Switzerland) ad libitum.
- Water : Community tap-water, from Itingen ad libitum.
- Acclimation period: Under laboratory conditions, after health examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12, music during the light period

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10mL/kg bw

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Treatment: The animals received a single oral dose of the test item by gavage at 2000 mg/kg body weight after being fasted for 16 to 20 hours (access to water was permitted). Food was provided again approximately 3 hours after dosing. The application volume was 10 mL/kg body weight.
- Observations:
Mortality/Viability: Daily during acclimatization and at least once daily during days 1-15.
Body weights: On test days 1 (pre-administration), 8 and 15.
Clinical signs: Daily during acclimatization and at least four times (target times: approximately one, two, three and five hours) on test day 1 after the test item administration, depending on the occurrence of clinical signs of toxicity. At least once daily during days 2-15, depending on the occurrence of clinical signs of toxicity. All abnormalities were recorded.
- Necropsy: All surviving animals were killed at the end of the observation period by an intraperitoneal injection of NARCOREN (Rhône Mérieux 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) and discarded after macroscopic examinations were performed. No organs or tissues were retained.

Statistics:

No statistical analysis was used.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No death occurred during the study.

Clinical signs:

other: No clinical signs were noted during the course of the study.

Gross pathology:

No macroscopic findings were observed at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute median lethal dose (LD50) of FAT 40'800/A after single oral administration to Wistar rats of both sexes, observed over a period of 14 days is >2000 mg/kg bw.

Executive summary:

In a GLP-compliant oral toxicity study, performed according to OECD guideline 423, Wistar rats (step 1: 3 females and step 2: 3 males) were administered the test substance (2000 mg/kg bw) by oral gavage followed by a 14-day observation period. All animals survived until the end of the study period. No clinical signs were evident during the course of the study. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy. Based on the observations, the acute median lethal dose (LD50) of FAT 40800/A after single oral administration to Wistar rats of both sexes, observed over a period of 14 days is >2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

GLP-compliant guideline study, Klimisch code 1

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

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:

09 May 2001 to 23 May 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

Identity: FAT 40800/A
Batch number: WP 2/01
Active ingredient: ca. 70 %
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Stability of test item: Stable under storage conditions
Stability of test item in vehicle: 24 hours at room temperature
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd., Biotechnology & Animal Breeding Division, CH-4414 Füllinsdorf
- Age at study initiation: Males: 9 weeks, Females: 12 weeks
- Weight at study initiation: Males: 230.6 - 246 g, Females: 187.7 - 210.4 g
- Housing: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz) during treatment and observation.
- Diet: Pelleted standard Kliba 3433, batch no. 07/00 rat maintenance diet (Provimi Kliba AG, CH-4303 Kaiseraugst) available ad libitum.
- Water: Community tap water from Itingen, available ad libitum.
- Acclimation period: Under laboratory conditions, after health examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12, music during light period.

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

One day before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10 % of the total body surface. Only those animals without injury or irritation on the skin were used in the test. On test day 1, the test item was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe and covered with a dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage. Application volume/kg body weight: 4 mL. Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels. Thereafter, the reaction sites were assessed.

Duration of exposure:

24 hours

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: 14 days
- Observations: Mortality/Viability: Daily during acclimatization and at least one, two, three and approximately five hours after the test item administration, depending on the occurrence of clinical signs of toxicity. At least once daily during days 2-15, depending on the occurrence of clinical signs of toxicity.
Body weights: On test days 1 (pre-administration), 8 and 15.
Clinical signs: Daily during acclimatization and at least one, two, three and approximately five hours or more on test day 1 after the test item administration, depending on the occurrence of clinical signs of toxicity. At least once daily during days 2-15, depending on the occurrence of clinical signs of toxicity. All abnormalities were recorded.
- Necropsy: At the end of the observation period all animals were sacrificed by intraperitoneal injection of NARCOREN (Rhône Mérieux 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.

Statistics:

No statistical analysis was used.

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:

other: In all animals slight discoloration and test item residuals were noted from the second to the sixth/seventh observation day. Those findings were considered to be test item-related non toxic effects.

Gross pathology:

No macroscopic findings were observed at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose (LD50) of FAT 40800/A after single dermal exposure to Wistar rats of both sexes, observed over a period of 14 days is >2000 mg/kg bw.

Executive summary:

In a GLP-compliant dermal toxicity study, performed according to OECD guideline 402, a group of Wistar rats (5/sex/dose) were exposed to the test substance (2000 mg/kg bw). The test substance was dissolved in water and applied on the skin with a syringe and covered with an dressing for 24 hours. The treated skin was washed after 24 hours and a 14-day observation period followed. No deaths occurred during the study. In all animals, slight discoloration and test substance residuals were noted from the second to the sixth/seventh observation day. Those findings were considered to be test substance-related non toxic effects. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy. Based on the study results, the median lethal dose (LD50) of FAT 40800/A after single dermal administration to Wistar rats of both sexes, observed over a period of 14 days is >2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

GLP-compliant guideline study, Klimisch code 1

Additional information

Acute oral toxicity:

In a GLP-compliant oral toxicity study, performed according to OECD guideline 423, Wistar rats (step 1: 3 females and step 2: 3 males) were administered the test substance (2000 mg/kg bw) by oral gavage followed by a 14-day observation period (RCC 2001). All animals survived until the end of the study period. No clinical signs were evident during the course of the study. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy. Based on the observations, the acute oral toxicity of the test substance in rats of both sexes observed for a period of 14 days was greater than 2000 mg/kg bw.

 

Acute inhalation study:

Currently no study to assess acute inhalation toxicity of Reactive Yellow 210 is available. However, with the low vapour pressure (7.23 E-25 Pa) and the high melting point (>400 °C), the substance is considered to have low volatility. Synthesis and formulation of this chemical is performed in a closed process; the final product consists of liquid formulations only. Hence, the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. Based on column 2, ‘Specific rules for adaptation from column 1’ of the table given in REACH Annex VII, the study on acute inhalation toxicity only needs to be conducted if an exposure via inhalation is to be expected, based on the likelihood of an exposure to aerosols, particles or droplets. Referring to the expected low volatility of the substance, the fact that the substance is imported into the EU in a formulated form as a liquid formulation, the exposure via inhalation is unlikely. Further, Reactive Yellow 210 was found to be miscible in water (water solubility 275 g/L) and have low log partition coefficient (log Pow <-5.9), hence in the case of dust of the substance entering the respiratory tract, it will be trapped in the mucus and cleared, thereby further limiting the absorption. The chemical showed low toxicity potential (LD50 >2000 mg/kg bw) in the available acute oral and dermal toxicity studies with no mortality or systemic toxicity, hence it does not need to be classified as STOT SE. Taking into consideration the above arguments, low toxicity potential is expected on acute exposure of Reactive Yellow 210 via inhalation route and hence acute toxicity testing by the inhalation route was considered scientifically not necessary.

 

Acute dermal toxicity:

In a GLP-compliant dermal toxicity study, performed according to OECD guideline 402, a group of Wistar rats (5/sex/dose) were exposed to the test substance (2000 mg/kg bw) (RCC 2001). The test substance was dissolved in water and applied on the skin with a syringe and covered with an dressing for 24 hours. The treated skin was washed after 24 hours and a 14-day observation period followed. No deaths occurred during the study. In all animals slight discoloration and test substance residuals were noted from the second to the sixth/seventh observation day. Those findings were considered to be test substance-related nontoxic effects. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy. Therefore, the toxicity of the test substance was estimated to be >2000 mg/kg bw.

Justification for classification or non-classification

Based on the observed LD₅₀ of >2000 mg/kg bw in the acute oral and dermal toxicity study, the test substance does not considered to be classified according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.