Registration Dossier

Administrative data

Description of key information

The oral and dermal LD50 for Reactive Orange 35 was considered to be >2000 mg/kg bw in rats. 

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:
1975
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
not specified
Principles of method if other than guideline:
The test substance was administered orally by gavage at single dose to a group of experimental animals. Subsequently observations of effects and deaths were made. At the end of the observation period the surviving animals were sacrificed and autopsied.
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
Name: FAT 40075/A
Purity: 74.5 %
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 6-7 weeks
- Weight at study initiation: 234 g (males), 160 g (females)
- Fasting period before study: 18 h
- Housing: Rats were caged singly
- Diet: Commercial pelleted diet (ad libitum)
- Water: Ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2 °C.
- Photoperiod (hrs dark / hrs light): 12 h/12 h
Route of administration:
oral: gavage
Vehicle:
other: deionised water
Details on oral exposure:
The test substance was administered as a single dose by gavage as a 25 % w/v solution in deionised water at a rate of 20 mL/kg (equivalent to 5 g/kg bw of test substance) to rats which had been fasted for 18 h.
Doses:
5000 mg/kg bw
No. of animals per sex per dose:
5/sex/dose
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 d.
- At the end of the observation period, surviving animals were killed by exsanguinations under other anaesthesia and an autopsy performed.
- Other examinations performed: Mortality, clinical symptoms.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 3 725 mg/kg bw
Based on:
act. ingr.
Mortality:
No death occurred during the study
Clinical signs:
No clinical signs were observed during the study period.
Gross pathology:
No changes in organs or tissues were observed at autopsy.
Interpretation of results:
GHS criteria not met
Conclusions:
The oral LD50 of the test substance was found to be >5000 mg/kg bw (i.e. ca. >3725 mg a.i./kg bw) in rats.
Executive summary:

A study was conducted to assess the acute oral toxicity of the test substance (of ca. 74.5 % purity) in Sprague-Dawley rats. This test was conducted in accordance to guideline similar to OECD 401. Group of 10 fasted animals (5/sex/dose) received a single oral (gavage) dose of 5000 mg/kg bw. Parameters assessed included mortality, clinical symptoms and autopsy findings after a 14 d observation period. No mortality and no clinical symptoms were observed throughout the observation period and no significant changes in organs or tissues were seen at autopsy. Under the study conditions, the oral LD50 of the test substance was found to be >5000 mg/kg bw (i.e. ca. >3725 mg a.i./kg bw) in rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Qualifier:
according to
Guideline:
EPA OTS 798.1100 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
other: Testing Methods for New Chemical Substances according to the Revised Japanese Chemical Substance Law (March 31, 1987).
GLP compliance:
yes (incl. certificate)
Remarks:
RCC, Holding Company Ltd. 4452 Itinqen Switzerland
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
None
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
- Age at study initiation: males: 9 weeks, females: 11 weeks
- Weight at study initiation: males: 253.6 - 286.2 g; females: 196.6 - 212.8 g
- Housing: during acclimatization in groups of five in Makrolon type-4 cages (size: 33 x 55 x 20 cm) with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz). During treatment and observation individually in Makrolon type-3 cages (size: 22 x 37.5 x 15 cm) with standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standard Kliba 343, Batch no.73/94 rat maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst) available ad libitum.
- Water (e.g. ad libitum): community tap water from Füllinsdorf, available ad libitum.
- Acclimation period: one week under laboratory conditions, after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 36-64
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12, music during light period
Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
bi-distilled water
Details on dermal exposure:
TREATMENT
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. Only those animals without injury or irritation on the skin were used in the test.
On test day 1, the test article 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: 4.0 mL/kg. 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.

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 daily during days 2-15.
Body Weights: On test days 1 (pre-administration), 8 and 15.
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, (general behaviour, respiration, eyes, motor susceptibility, nose, motility, body posture, skin)

* The computerised system does not show that the mortality/viability checks were recorded at the same time as the clinical signs.

- Necropsy of survivors performed:
Necropsies were performed by experienced prosectors. At the end of the observation period all animals were anesthetized by intraperitoneal injection of NARCOREN (Rhone Merieux GmbH, D-88417 Laupheim) at a dose of at least 2.0 mL/kg body weight (equivalent to at least 320 mg/kg sodium pentobarbitone/kg body weight) and sacrificed by exsanguination. The animals were examined macroscopically.
Statistics:
The LOGIT-Model could not be used as no deaths occurred.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths as a result of treatment with the test article.
Clinical signs:
No clinical signs of systemic toxicity were observed during the observation period.
Discoloration of the skin at the application site was evident in all animals after the removal of the dressing on test day two and persisted until study termination.
Body weight:
Slight loss of body weight (2.2 and 2.0 g) was noted in female animals no. 7 and 9 respectively during the first observation week. The other animals gained weight over the observation period.
Gross pathology:
No organ abnormalities were observed at necropsy.
Interpretation of results:
GHS criteria not met
Conclusions:
The toxicity of the read across substance was estimated to be greater than 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 read across substance (2000 mg/kg bw). The test substance was dissolved in water 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, clinical signs of systemic toxicity, or organ abnormalities were observed during this period. The pigment in the test article caused a discolouration of the skin at the application site which persisted throughout the observation period. There were no test article-related effects on the body weight of the animals during the observation period. The minimal to slight loss of body weight in two female animals during the first observation week was considered to be a consequence of the semi-occlusive dressing. Commonly female animals prove to be more sensitive in relation to effects on body weight caused by semi-occlusive dressing than males. The mean lethal dose of test substance after single dermal administration to rats of both sexes, observed over a period of 14 days, could not be estimated, because no deaths occurred at the maximal dose of 2000 mg/kg bw.

Endpoint:
acute toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Qualifier:
according to
Guideline:
EPA OTS 798.1100 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
other: Testing Methods for New Chemical Substances according to the Revised Japanese Chemical Substance Law (March 31, 1987).
Test type:
standard acute method
Limit test:
yes
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths as a result of treatment with the test article.
Clinical signs:
No clinical signs of systemic toxicity were observed during the observation period.
Discoloration of the skin at the application site was evident in all animals after the removal of the dressing on test day two and persisted until study termination.
Body weight:
Slight loss of body weight (2.2 and 2.0 g) was noted in female animals no. 7 and 9 respectively during the first observation week. The other animals gained weight over the observation period.
Gross pathology:
No organ abnormalities were observed at necropsy.
Interpretation of results:
GHS criteria not met
Conclusions:
The toxicity of the read across substance was estimated to be greater than 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 read across substance (2000 mg/kg bw). The test substance was dissolved in water 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, clinical signs of systemic toxicity, or organ abnormalities were observed during this period. The pigment in the test article caused a discolouration of the skin at the application site which persisted throughout the observation period. There were no test article-related effects on the body weight of the animals during the observation period. The minimal to slight loss of body weight in two female animals during the first observation week was considered to be a consequence of the semi-occlusive dressing. Commonly female animals prove to be more sensitive in relation to effects on body weight caused by semi-occlusive dressing than males. The mean lethal dose of test substance after single dermal administration to rats of both sexes, observed over a period of 14 days, could not be estimated, because no deaths occurred at the maximal dose of 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Acute oral toxicity

Acute oral toxicity of Reactive Orange 035 was evaluated in a study (1975) conducted using a methodology similar to OECD Guideline 401. In this study, a group of 10 fasted animals (5/sex/dose) received a single oral (gavage) dose of 5000 mg/kg bw. No mortality and no clinical symptoms were observed throughout the observation period and no significant changes in organs or tissues were seen at autopsy. Hence, the oral LD50 of Reactive Orange 35 was found to be >5000 mg/kg bw (i.e. ca. >3725 mg a.i./kg bw) in rats.

Acute inhalation toxicity

Currently no study to assess the acute inhalation toxicity potential of Reactive Orange 035 is available. However, the vapour pressure for the substance can be considered low owing to the high melting point (>270 °C). Hence the substance is considered to have low volatility. Synthesis and spray drying of this chemical is performed in a closed process; the final product consists of non-dusty granules. 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 chemical is imported into the EU in a formulated form as a dust-free powder or as a granulate, the exposure via inhalation is considered to be unlikely. Further the chemical is found to have water solubility of 334 g/L, 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 in the available acute oral toxicity study (LD50: >5000 mg/kg bw), with no mortality or systemic toxicity being seen, hence it does not need to be classified STOT SE and low toxicity is expected for this chemical via the inhalation route. Taking into consideration the above arguments, low toxicity potential is expected on acute exposure of Reactive Orange 035 via inhalation route and hence testing by the inhalation route was considered scientifically not necessary.

Acute dermal toxicity

Currently no study to assess the acute dermal toxicity of Reactive Orange 035 is available. However, the read-across substance,lithium sodium salt of Reactive Black039 was tested for acute dermal toxicity in a GLP compliant study, performed according to OECD Guideline 402. In this study, Wistar rats (5/sex) were exposed to the test substance (2000 mg/kg bw) dermally. No mortality was reported throughout the study. Hence, the lithium sodium salt of Reactive Black 039 was expected to pose no acute dermal hazard.

Justification for classification or non-classification

The available data indicate that Reactive Orange 035 is not acutely toxic and hence it does not need to be classified for acute toxicity as per the criteria of Regulation (EC) No. 1272/2008.