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Administrative data

Description of key information

Theoral and dermal median lethal dose (LD50) of Reactive Orange 136 in rat was greater than 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:
N0V. 14 to DEC.19, 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
GLP compliance:
yes (incl. certificate)
Remarks:
German GLP
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: WP2/96
- Expiration date of the lot/batch: November 30, 2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature at about 20 °C; away from direct sunlight.
- Stability of the test substance in the solvent/vehicle: Stable in bi-distilled water for at least 48 h at 20 °C
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wölferstrasse 4, 4414 Füllinsdorf / Switzerland
- Females (if applicable) nulliparous and non-pregnant: not specified
- Age at study initiation: males: 8 weeks; females: 10 weeks
- Weight at study initiation: males: 197.1 - 218.5 g; females: 174.8 - 186.3 g
- Fasting period before study: 17 h
- Housing: Groups of five in Makrol on type-4 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz)
- Diet : Pelleted standard Kliba 343, Batch nos. 78/96 and 81/96 rat maintenance diet (Kliba Mühlen AG, CH-4303 Kaiseraugst) available ad libitum
- Water: Community tap water from Itingen, 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 +/- 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 h artificial fluorescent light/12 h dark

IN-LIFE DATES: From: 21-N0V-1996 to 05-DEC-1996
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
bi-distilled water
Details on oral exposure:
The animals received a single dose of the test article on a mg/kg body weight basis by oral gavage following fasting for approximately 17 h, but with free access to water. Food was provided again approximately 3 hours after dosing.
Doses:
2000 mg/kg body weight (10 mL/kg body weight)
No. of animals per sex per dose:
5 males and 5 females
Control animals:
no
Details on study design:
The test item was administration to a group of 5 male and 5 female rats by oral gavage, at the single dose of 2000 mg/kg body weight. The application volume was 10 mL/kg body weight. The animals were examined and mortality, viability and clinical signs were recorded. Body weights were taken on test day 1 (pre-administration), 8 and 15 for surviving animals. Necropsies were performed by experienced prosectors. At the end of the observation period all animals were sacrificed. The animals were examined macroscopicaliy. Thereafter, they were discarded.
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 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, except one female animal (no. 8) showed a slight loss of body weight during the second observation period.
Gross pathology:
No macroscopic findings were observed at necropsy.
Interpretation of results:
GHS criteria not met
Conclusions:
The median lethal dose (LD50) of FAT 45176/A by oral route in rat is greater than 2000 mg/kg body weight.
Executive summary:

An acute oral toxicity study was carried out in Wistar rats according to OECD 401 and EU B.1 guideline. A group of five male and five female HanIbm:WIST (SPF) rats was treated with FAT 45'176/A at 2000 mg/kg body weight 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 body weight. 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, except one female animal (no. 8) showed a slight loss of body weight during the second observation period. No macroscopic findings were observed at necropsy.

In conclusion, the median lethal dose (LD50) FAT 45176/A by oral route in rat 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 and OECD guideline compliance

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:
Nov. 26, 1996 - Jan. 10, 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
GLP compliance:
yes (incl. certificate)
Remarks:
German GLP
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: WP2/96
- Expiration date of the lot/batch: November 30,2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature at about 20 °C; away from direct sunlight
- Stability of the test substance in the solvent/vehicle: Stable in bi-distilled water for at least 48 h at 20 °C
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wolferstrasse 4, 4414 Füllinsdorf / Switzerland
- Females (if applicable) nulliparous and non-pregnant: Not Specified
- Age at study initiation: males: 8 weeks; females: 11 weeks
- Weight at study initiation: males: 233.5 to 258.7 g; females: 201.8 to 224.9 g
- Housing: individually in Makrolon type-3 cages with standard softwood bedding
- Diet: Pelleted standard Kliba 343, Batch no. 81/96 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 health
examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 h artificial fluo- rescent light/12-h dark

IN-LIFE DATES: From: 03-DEC-1996 to 17-DEC-1996
Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
bi-distilled 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 semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.
Application volume/kg body weight: 4 mL/kg
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.
Duration of exposure:
24 h
Doses:
2000 mg/kg body weight (4 mL/kg)
No. of animals per sex per dose:
5
Control animals:
no
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 deaths occured during the study.
Clinical signs:
No systemic signs of toxicity were observed.
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.
Other findings:
no data
Interpretation of results:
GHS criteria not met
Conclusions:
The dermal median lethal dose (LD50) of FAT 45176/A in rats was above 2000 mg/kg body weight.
Executive summary:

The acute dermal toxicity study was carried out on Wistar rats according to OECD 402 and EU B.3 guideline. The test item was applied to a group of 5 male and 5 female rats by dermal application, at the single dose of 2000 mg/kg body weight.

No death occurred during the study period. No clinical signs of systemic toxicity were observed during the observation period except

scales were noted in eight animals at different durations between test day 3 and 15. Orange 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.

There were no test item-related effects on body weight of the male and females animals during the observation period. No organ abnormalities were observed at necropsy.

Based on the study results, the median lethal dose (LD50) of FAT 45176/A was estimated to be 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 and OECD guideline compliance

Additional information

Acute Oral Toxicity:

An acute oral toxicity study was carried out in Wistar rats according to OECD 401 and EU B.1 guideline. A group of five male and five female HanIbm:WIST (SPF) rats was treated with FAT 45'176/A at 2000 mg/kg body weight 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 body weight. 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, except one female animal (no. 8) showed a slight loss of body weight during the second observation period. No macroscopic findings were observed at necropsy. In conclusion, the median lethal dose (LD50) FAT 45176/A by oral route in rat is greater than 2000 mg/kg body weight.

Acute Inhalation Toxicity:

Currently no study to assess the acute inhalation toxicity potential of Reactive Orange 136 is available. However, the vapour pressure for the substance can be considered low (5.75 X 10-20Pa) and owing to the high melting point (>400 °C). Hence, 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 vapour pressure and/or 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 liquid formulation, the exposure via inhalation is considered to be unlikely. Further, in case the substance is entering the respiratory tract, it will be trapped in the mucus and cleared, thereby further limiting the absorption. The chemical showed low toxicity potential in the available acute oral and dermal toxicity studies (LD50>2000 mg/kg body weight) with no mortality or systemic toxicity, hence it does not need to be classified STOT SE. Taking the above arguments into account, low toxicity potential is expected on acute exposure of Reactive Orange 136 via inhalation route and hence testing by the inhalation route was considered scientifically not necessary.

Acute Dermal Toxicity:

The acute dermal toxicity study was carried out on Wistar rats according to OECD 402 and EU B.3 guideline. The test item was applied to a group of 5 male and 5 female rats by dermal application, at the single dose of 2000 mg/kg body weight.

No death occurred during the study period.No clinical signs of systemic toxicity were observed during the observation period.There were no test item-related effects on body weight of the male and females animals during the observation period.No organ abnormalities were observed at necropsy. Based on the study results, the median lethal dose (LD50) of FAT 45176/A was estimated to be 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, Reactive Orange 136 does not considered to be classified according to according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.