main component 1 (isomer 1): 2-{6-fluoro-4-[3-(2,5-disulfo-phenylazo)-4-hydroxy-2-sulfonapht-7-ylamino]-1,3,5-triazin-2-ylamino}-3-{6-fluoro-4-[3-(1,5-disulfonaphth-2-ylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-propane sodium salt;main component 1 (isomer 2): 2-{6-fluoro-4-[3-(2,5-disulfo-phenylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-3-{6-fluoro-4-[3-(2,5-disulfo-phenylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-propane sodium salt;main component 2: 2,3-bis-{6-fluoro-4-[3-(2,5-disulfo-phenylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-propane sodium salt;main component 3: 2,3-bis-{6-fluoro-4-[3-(1,5-disulfonaphth-2-ylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-propane sodium salt

Endpoint summary

Description of key information

An acute median lethal dose (LD50) of test substance is >2000 mg/kg bw in the acute oral and acute dermal toxicity study.

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:

09 October 1996 to 14 November 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

No randomization of the animals was performed. They were distributed according to the sex into groups of five.

Qualifier:

according to guideline

Guideline:

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

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 40557/A
Batch-no.: PV1
Purity: 65 %
Appearance: Solid powder, red-brown
Stability in test article: Stable under storage condition; expiration date: 30-SEP-1996
Stability of Test Article Dilution: Stable in polyethylene glycol for at least 48 hours
Expiration date: 30 September, 2002
Storage conditions: In the original container at room temperature away from direct sunlight.

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wölferstrasse 4, 4414 Füllinsdorf / Switzerland
- Age at start of treatment: males: 8 weeks, females: 10 weeks
- Weight at start of treatment: males: 191.1 - 202.0 g, females 177-198.8 g
- Fasting period before study: 19.5 hours
- Housing: Groups of five in Makrolon type-4 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz).
- Diet: Pelleted standard Kliba 343, Batch no. 78/96 rat maintenance diet (Kliba Mühlen AG, CH-4303 Kaiseraugst) available ad libitum
- Water: Community tap water from Itingen, available ad libitum.
- Acclimation period: One week under laboratory conditions, after veterinary examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 40-70 (values above 70 % during cleaning process possible)
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Treatment: 09-OCT-1996 to 23-OCT-1996

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10mL/kg bw

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: Mortality / Viability: Four times during test day 1, and once daily during days 2-15, Body Weights: Test days 1 (pre-administration), 8 and 15, Symptoms: Each animal was examined for changes in appearance and behaviour four times during day 1, and daily during days 2-15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs (general behaviour, respiration, eye, nose, motility, body posture, motor susceptibility, skin), gross pathology

Statistics:

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

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured.

Clinical signs:

other: Diarrhea was noted in one male animal on test day 1, 5 hours after the test article administration.

Gross pathology:

No macroscopic findings were observed at necropsy.

The animals received a single dose of the test article on a mg/kg body weight basis by oral gavage following fasting for approximately 19.5 hours, but with free access to water. Food was provided again approximately 3 hours after dosing.

Rationale:

Oral administration was used as this is one possible route of human exposure during manufacture, handling and use of the test article.

Necropsy:

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, and all abnormalities recorded. Thereafter, they were discarded.

Interpretation of results:

GHS criteria not met

Conclusions:

The oral median lethal dose (LD50) of FAT 40557/A was estimated to be greater than 2000 mg/kg bw.

Executive summary:

In a GLP-compliant oral toxicity study, performed according to OECD guideline 401, Wistar rats (5/sex) were administered the test substance (2000 mg/kg bw in polyethylene glycol) by oral gavage followed by a 14-day observation period. Mortality did not occur, body weights were within the range of physiological variability known for rats of this strain and age, and no macroscopic findings were observed at necropsy. Diarrhoea was noted in one male animal on test day 1. Based on these observations, the oral median lethal dose (LD50) of FAT 40557/A in an 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.

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:

exposure considerations

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:

10 October 1996 to 14 November 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

See overall remarks attachment section

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 40557/A
Batch-no.: PV1
Purity: 65 %
Appearance: Solid powder, red-brown
Stability of Test Article: Stable under storage condition; expiration date: 30-SEP-2002
Stability of Test Article Dilution: Stable in polyethylene glycol for at least 48 hours
Expiration date: 30 September, 2002
Storage conditions: In the original container at room temperature away from direct sunlight

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wölferstrasse 4, CH-4414 Füllinsdorf
- Age at start of treatment: males: 8 weeks, females, 11 weeks
- Weight at start of treatment: males: 215.1 - 245.4 g, females: 200.5 - 209.1 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: Pelleted standard Kliba 343, Batch 78/96 rat maintenance diet ("Kliba" Mühlen AG, CH-4303 Kaiseraugst), available ad libitum
- Water: Community water from Itingen, available ad libitum
- Acclimation period: One week under laboratory conditions, after veterinary examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 40-70 (values above 70 % during cleaning process possible)
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Treatment: 10-0CT-1996 to 24-0CT-1996

Type of coverage:

semiocclusive

Vehicle:

polyethylene glycol

Details on dermal exposure:

Approximately 24 hours before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10 % of the total body surface. On test day 1, 4 mL at 2000 mg/kg bw test article was applied evenly on the skin with a syringe and covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage. Twenty-four hours after the application, the dressing was removed and the treated skin was washed with lukewarm tap water and dried with disposable paper towels and the skin reaction was 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
- Frequency of observations and weighing: Mortality / Viability: Four times during test day 1, and once daily during days 2-15, Body Weights: Test days 1 (pre-administration), 8 and 15, Symptoms: Each animal was examined for changes in appearance and behaviuor four times during day 1, and daily during days 2-15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs (general behaviour, respiration, eye, nose, motility, body posture, motor susceptibility, skin), gross pathology

Statistics:

Since no mortality was recorded, the LOGIT-Model could not be applied.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occured during the study.

Clinical signs:

other: All animals showed orange discoloration while scaling was observed in one male.

Gross pathology:

No macroscopic organ findings were observed in the animals.

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal median lethal dose (LD50) of FAT 40557/A was estimated to be >2000 mg/kg bw.

Executive summary:

In a GLP-compliant dermal toxicity study, performed according to OECD guideline 402, Wistar rats (5/sex) were administered the test substance (2000 mg/kg bw). The test substance was dissolved in polyethylene glycol and applied on the skin with a syringe and covered with a semi-occlusive dressing for 24 hours. The treated skin was washed after 24 hours and a 14-day observation period followed. No mortality was observed during this period. Discoloration of the skin was observed in all animals while one male rat showed scaling. Slight loss of body weight was observed in three animals of which two recovered before the end of the observation period. No macroscopic findings were noted. Based on the study results, the dermal median lethal dose (LD50) of FAT 40557/A was estimated to be >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 401, Wistar rats (5/sex) were administered the test substance (2000 mg/kg bw in polyethylene glycol) by oral gavage followed by a 14-day observation period. Mortality did not occur, body weights were within the range of physiological variability known for rats of this strain and age, and no macroscopic findings were observed at necropsy. Diarrhoea was noted in one male animal on test day 1. Based on these observations, the oral median lethal dose (LD50) of FAT 40557/A in an 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 toxicity:

Currently no study to assess acute inhalation toxicity of Reactive Orange 135 is available. However, with the low vapour pressure (7.25 x 10^-37Pa) 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 Orange 135 was found to be miscible in water (water solubility 246 g/L) and have low log partition coefficient (-3.22), 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. Considering above arguments, low toxicity potential is expected on acute exposure of Reactive Orange 135 via inhalation routes 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, Wistar rats (5/sex) were administered the test substance (2000 mg/kg bw). The test substance was dissolved in polyethylene glycol and applied on the skin with a syringe and covered with a semi-occlusive dressing for 24 hours. The treated skin was washed after 24 hours and a 14-day observation period followed. No mortality was observed during this period. Discoloration of the skin was observed in all animals while one male rat showed scaling. Slight loss of body weight was observed in three animals of which two recovered before the end of the observation period. No macroscopic findings were noted. Based on the study results, the dermal median lethal dose (LD50) of FAT 40557/A 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 is not-classified according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.