Reaction mass of pentasodium 2-{[4-chloro-6-(ethyl{3-[(2-sulfonatoethyl)sulfonyl]phenyl}amino)-1,3,5-triazin-2-yl]amino}-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate and tetrasodium 2-[(4-chloro-6-{ethyl[3-(vinylsulfonyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate

Endpoint summary

Description of key information

The acute median lethal dose ( LD₅₀) of test substance in the acute oral and dermal toxicity study was >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:

27 December 2005 to 03 Febuary 2006

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 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

Identity: FAT 40824/A
Batch: Red ROE 805 BOP 04/05
Appearance: dark red powder
Purity: Organic part (Na-salt): approx. 82 %; Main component 1 : approx. 36.2 %; Main component 2: approx. 27.5 %; Oligomers: 10%
Expiration date: 01 October 2010
Stability in water: Max. 7 days at room temperature
Storage: At room temperature at about 20 °C, in a desiccator because test substance is hygroscopic, away from direct sunlight

Species:

rat

Strain:

other: Rat, HanRcc:WIST (SPF)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services CH-4414 Füllinsdorf / Switzerland
- Age at study initiation: 12-13 weeks
- Housing: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding
- Diet: Pelleted standard Provimi Kliba 3433 rat/moitse maintenance diet, batch no. 63/05 ad libitum.
- Water: Community tap water from Fiillinsdorf ad libitum.
- Fasting period before study: 18-19 hours

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30-70
- Air changes (per hr): Air-conditioned with 10-15 air changes per hour.
- Photoperiod (hrs dark / hrs light): automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

The dose formulations were made shortly before each dosing occasion using a magnetic stirrer and a spatula as homogenizers.The test item was weighed into a tared glass beaker on a surtable precision balance and the vehicle added (weight/volume). Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

3 females per group and two groups per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 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.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights.

Statistics:

No statistical analysis was used.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality observed at highest dose group tested (2000 mg/kg bw)

Mortality:

No deaths occurred during the study.

Clinical signs:

other: Slightly ruffled fur was observed in two of 6 animals from the 2- to 3- hour reading. Dark red feces were noted in both cages on test 2 and 3.

Gross pathology:

No macroscopic findlings were recorded at necropsy.

Other findings:

No data

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral median lethal dose ( LD50) of FAT 40824/A to female rat was greater than 2000 mg/kg bw.

Executive summary:

In a GLP compliant acute toxicity study conducted according to OECD TG 423, two groups, each of three female Wistar rats, were treated with FAT 40824/A by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (purified water) 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 acclimatization 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 was 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. Body weights 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 observed in two out 6 animals from the 2- to the 3-hour reading. Dark red feces were noted in both cages on test day 2 and 3.

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 FAT 40824/A after single oral administration to female rats, observed over a period of 14 days (LD50) is greater than 2000 mg/kg body weight

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

GLP compliant and guideline study

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

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:

28 December 2005 to 03 Febuary 2006

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))

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

Identity: FAT 40824/A
Batch: Red ROE 805 BOP 04/05
Appearance: dark red powder
Purity: Organic part (Na-salt): approx. 82 %; Main component 1 : approx. 36.2 %; Main component 2: approx. 27.5 %; Oligomers: 10%
Expiration date: 01 October 2010
Stability in water: Max. 7 days at room temperature
Storage: At room temperature at about 20 °C, in a desiccator because test substance is hygroscopic, away from direct sunlight

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services CH-4414 Füllinsdorf / Switzerland.
- Age at study initiation: 12-13 weeks for females; 8-9 weeks for males.
- Housing: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding.
- Diet:: Pelleted standard Provimi Kliba 3433 rat/moitse maintenance diet, batch no. 63/05 ad libitum.
- Water: Community tap water from Fiillinsdorf ad libitum.
- Fasting period before study: 18-19 hours

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ±3
- Humidity (%): 30-70
- Air changes (per hr): Air-conditioned with 10-15 air changes per hour.
- Photoperiod (hrs dark / hrs light): automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: backs
- % coverage: 10% od the total body surface
- Type of wrap if used: gauze patch

REMOVAL OF TEST SUBSTANCE
- Washing (if done): lukewarm tap water
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 6mL

Duration of exposure:

24h

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
- Frequency of observations and weighing: 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.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights and local signs.

Statistics:

No statistics analysis was used.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality at highest dose group (2000 mg/kg bw)

Mortality:

No deaths occurred during the study

Clinical signs:

other: No systemic signs of toxicity were observed during the study period. A possible erythemateous reaction could not be assessed due lo a red discoloration produced by the test item in all male and all female animals from test day 2 to test day 5, 6, 7, 8 or

Gross pathology:

No macroscopic findings were observed at necropsy.

Other findings:

No data

Interpretation of results:

GHS criteria not met

Conclusions:

The acute dermal median lethal dose (LD50) to rat was greater than 2000 mg/kg bw.

Executive summary:

In a GLP compliant acute toxicity study conducted according to OECD TG 402,Wistar rats (5/sex) were treated with FAT 40824/A at 2000 mg/kg by dermal application. The test item was diluted in vehicle (purified water) at a concentration of 0.33 g/mL and administered at a volume dosage of 6 ml/kg. The application period was 24 hours. The animals were examined daily during the acclimatization 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. Local signs were noted once daily from test day 2 to 15. Mortality/viability was 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. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

No deaths occurred during the study. No clinical signs were observed throughout the whole observation period. A possible erythemateous reaction could not be assessed due to a red discoloration produced by the test item in all male and all female animals from test day 2 to test day 5, 6, 7, 8 or 13. Slight crusts were noted in one female from test day 8 to 10. 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 acute dermal median lethal dose (LD50) of FAT 40824/A to rat was greater than 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 and guideline study

Additional information

Acute oral toxicity:

In a GLP compliant acute toxicity study conducted according to OECD TG 423, two groups, each of three female Wistar rats, were treated with FAT 40824/A by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (purified water) 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 acclimatization period and mortality, viability and clinical signs were recorded.

All animals survived until the end of the study period. Slightly ruffled fur was observed in two out 6 animals from the 2- to the 3-hour reading. Dark red feces were noted in both cages on test day 2 and 3. 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 acute median lethal dose of FAT 40824/A after single oral administration to female rats, observed over a period of 14 days (LD50) is greater than 2000 mg/kg body weight.

Acute inhalation toxicity:

Currently no study to assess acute inhalation toxicity of Reactive Red 282 is available. However, with the low vapour pressure (1.8 x 10^-29 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 Red 282 was found to be miscible in water (water solubility 373 g/L) and have low log partition coefficient (-5.8), 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 Red 282 via inhalation routes and hence acute toxicity testing by the inhalation route was considered scientifically not necessary.

Acute dermal toxicity:

In a GLP compliant acute toxicity study conducted according to OECD TG 402,Wistar rats (5/sex) were treated with FAT 40824/A at 2000 mg/kg by dermal application. The test item was diluted in vehicle (purified water) at a concentration of 0.33 g/mL and administered at a volume dosage of 6 ml/kg. The application period was 24 hours. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded.

No deaths occurred during the study. No clinical signs were observed throughout the whole observation period. A possible erythemateous reaction could not be assessed due to a red discoloration produced by the test item in all male and all female animals from test day 2 to test day 5, 6, 7, 8 or 13. Slight crusts were noted in one female from test day 8 to 10. 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 acute dermal median lethal dose (LD50) of FAT 40824/A to rat was greater than 2000 mg/kg bw.

Justification for classification or non-classification

Based on the observed LD₅₀of >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.