Registration Dossier

Administrative data

Description of key information

The oral and dermal LD50s of Reactive Blue 72 were 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:
1987
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)
GLP compliance:
not specified
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
Test substance: FAT 400045/C
Batch No.: EN 19609.62
Species:
rat
Strain:
other: albino
Sex:
male/female
Details on test animals and environmental conditions:
not available
Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
no data
Doses:
2000 and 5000 mg/kg bw
No. of animals per sex per dose:
3

Control animals:
not specified
Statistics:
no data
Sex:
male/female
Dose descriptor:
approximate LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred at the dose of 2000 mg/kg bw. In the rats administered 5000 mg/kg bw, 1/3 males and 1/3 females were found dead during the observation period.
Clinical signs:
Dyspnoea, exophthalmos, ruffled fur, and curved body position were seen, being common symptoms in acute toxicity testing. The surviving animals recovered within 10-12 d.
Body weight:
No data
Gross pathology:
At autopsy, no deviations from normal morphology were found in the animals of the 2000 mg/kg bw dose group.
The gastrointestinal tract was dilated in the male and the female of the 5000 mg/kg bw dose group which died on day 1.
Other findings:
no data
Interpretation of results:
GHS criteria not met
Conclusions:
The oral LD50 of the test substance was >5000 mg/kg bw in rats.
Executive summary:

A study was conducted to assess the acute oral toxicity of the test substance in albino rats following a method comparable to OECD Guideline 401. Two groups of rats (3/sex/dose) received a single oral (gavage) dose of either 2000 or 5000 mg/kg bw of the test substance. No mortality occurred at 2000 mg/kg bw. However, in the rats administered 5000 mg/kg bw, 1/3 males and 1/3 females were found dead during the observation period. Dyspnoea, exophthalmos, ruffled fur and curved body position, being common symptoms in acute toxicity studies, were seen in treated animals. Surviving animals recovered within 10-12 d. At autopsy, no deviations from normal morphology were found at 2000 mg/kg bw. The gastrointestinal tract was dilated in the male and the female of the 5000 mg/kg bw dose group which died on Day 1. Based on these findings, the approximate oral LD50 of the test substance was considered to be >5000 mg/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

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:
1989
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))
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, CH 4414 Fuellinsdorf/Switzerland
- Age at study initiation: males: 12 weeks; females: 12 weeks
- Weight at study initiation: males: 297 - 337 g; females: 224 - 260 g
- Housing: Individually 1n Makrolon type-2 cages with standard softwood bedding ("Lignocel", Schill AG, 4132 Muttenz/Switzerland).
- Diet: Pelleted standard Kliba 343, Batch 31/88 and 34/88 rat maintenance diet ("Kliba", Klingentalmuehle AG, 4303 Kaiseraugst/Switzerland), ad libitum
- Water: ad libitum
- Acclimation period: At least one week under laboratory conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature: 22+/-3 °C
- Humidity: 40-70 %
- Air changes: 10-15 air changes per hour
- Photoperiod: 12 hours artificial fluorescent Iight/12 hours dark, music/light period.
Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
Approximately 24 hours before treatment, the backs of the animals were shaved with an electric clipper, exposing an area of approximately 10 % of the total body surface. On test day 1 the test article was applied evenly on the skin with a syringe and covered with an 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. The treated skin was washed with lukewarm tap water, dried with disposable paper towels and the skin reaction was assessed according to the method of Noakes and Sanderson (Br J Ind Med; 1969; 26:59-64).

Application volume/kg body weight: 4 ml at 2000 mg/kg

Rationale:
The dermal administration was used because this is one possible route of human exposure during manufacture, handling and use of the test article.

Duration of exposure:
24 hours
Doses:
Application volume/kg body weight: 4 ml at 2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations: Four times during test day 1, and daily during days 2 - 15.
- Frequency of weighing: Test days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes
Statistics:
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 mortality was seen throughout the observation period.
Clinical signs:
The following local findings were observed: erythema (females), blue discolored application area.
The female rats had recovered from erythema within 12 observation days, whereas blue discolored application area was observed until termination of test in animals of both sexes.
Body weight:
no adverse effect was seen.
Gross pathology:
No macroscopic organ changes were observed.
Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD50 of FAT 40336/B was estimated to be >2000 mg/kg bw.
Executive summary:

FAT 40336/B was assessed for toxicological potential when administered to rats by a single occlusive dermal application, with an observation period of 15 days. The test article FAT 40336/B was applied to the shaved skin of rats of both sexes for 24 hours at a dose of 2000 mg/kg bw. No mortality was observed. Therefore, the dermal LD50 of FAT 40336/B 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

Additional information

Oral:

Acute oral toxicity of Reactive Blue 72 was evaluated in two studies that were conducted according to OECD Guideline 401. In the study (1986) considered as key, the LD50was determined to be >5000 mg/kg bw. In this study, no mortality was observed at 2000 mg/kg bwHowever, in the rats administered 5000 mg/kg bw, 1/3 males and 1/3 females were found dead during the observation period. In the supporting study, conducted as a limit test, no mortality was seen at 5000 mg/kg bw.


Inhalation:

Currently no study to assess the acute inhalation toxicity potential of Reactive Blue 72 is available. However, the vapour pressure for the substance can be considered low owing to the high melting point (>350 °C). Hence the substance is considered to have low volatility. 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 a dust-free powder or as a granulate, the exposure via inhalation is considered to be unlikely. Reactive Blue 72 was found to have water solubility of 376 g/L, hence vapours of this very hydrophilic substance may be retained within the mucus and cleared through mucociliary system. Further, the chemical was found to have low acute toxicity when tested via oral route with no mortality upto 2000 mg/kg bw. Hence, considering all the above arguments, it is considered that Reactive Blue 72 has a low toxicity potential via inhalation route and thus the study on acute inhalation toxicity is considered scientifically not necessary.


Dermal

Currently no study to assess acute dermal toxicity of Reactive Blue 72 is available. However, a structurally similar substance, FAT 40336/B was assessed for toxicological potential when administered to rats by a single occlusive dermal application, with an observation period of 15 days. FAT 40336/B was applied to the shaved skin of rats of both sexes for 24 hours at a dose of 2000 mg/kg bw. No mortality was observed. Therefore, the dermal LD50 of FAT 40336/B was estimated to be >2000 mg/kg bw.

Justification for classification or non-classification

Based on the available information, Reactive Blue is considered to have low acute toxicity and does not warrant classification as per the Regulation (EC) No. 1272/2008 (CLP).