Sodium 1-amino-9,10-dihydro-4-[5-[(2-hydroxyethyl)sulphamoyl]-3,4-xylidino]-9,10-dioxoanthracene-2-sulphonate

Endpoint summary

Description of key information

The acute lethal doses for Acid Blue 277 via oral, dermal and inhalation routes were >5000 mg/kg bw, >3000 mg/kg bw and >0.29 mg/L, respectively.

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:

supporting study

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

not specified

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Tif RAIf

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxicology/Pathology, Ciba-Geigy Limited, Basle, Switzerland
- Weight at study initiation: 102 - 136 g
- Fasting period before study: Overnight
- Housing: Macrolon cages (type 3) in group
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±1
- Humidity (%): 55±5
- Photoperiod (h dark / h light): 14 h light cycle

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

The test substance was administered as a single dose by gavage as solution in carboxymethylcellulose (CMC) 0.5 %

Doses:

1000, 3000, 10000 and 15000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 d
- At the end of the observation period, surviving animals were sacrificed and autopsy was performed.
- Other examinations performed: Mortality, clinical signs and symptoms, body weights (pre-test and Day 15)

Preliminary study:

not applicable

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 15 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: No clinical signs were observed in the animals treated at 1000, 3000 and 10000 mg/kg bw. In the animals treated at 15000 mg/kg bw, reduction in spontaneous motility was seen 30 min after gavage that lasted >6 h. Diarrohea was also seen 3 h post gavage. Th

Gross pathology:

At autopsy no gross organ changes were seen.

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 of the FAT 20033/D was found to be >15,000 mg/kg bw in male and female rats.

Executive summary:

A study was conducted to assess the acute oral toxicity of FAT 20033/D in Tif RAIf rats. Groups of rats, each with 5 males and 5 females, received the test substance in a single oral (gavage) dose of 1000, 3000, 10000 and 15000 mg/kg bw. Post gavage, the treated animals were observed for mortality, clinical symptoms and change in body weights. At the end of the observation period, surviving animals were sacrificed and autopsy was performed. No mortality was observed during the observation period. No clinical signs were observed in the animals treated at 1000, 3000 and 10000 mg/kg bw. In the animals treated at 15000 mg/kg bw, reduction in spontaneous motility was seen 30 min after gavage that lasted >6 h. Diarrohea was also seen 3 h post gavage. These symptoms were found to have been reversed by 24 h observation. No adverse effect on body weights observed. At autopsy no gross organ changes were seen. Based on the results of the study, the oral LD50 of FAT 20033/D was found to be >15,000 mg/kg bw male and female rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

yes

Remarks:

(Duration of exposure was 1 h without any justification)

GLP compliance:

not specified

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: E01A25-11 (DCT No. 70083)
Test material is dark blue powder

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: Males and Females 275 to 344 g
- Housing: Housed individually, in suspended, wire bottom cages
- Diet: Ad libitum
- Water: Ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature: Approx. 74±1 °F (ca. 23 °C)
- Humidity: 45-55 %
- Photoperiod: 12 h dark / 12 h light

Route of administration:

inhalation: dust

Type of inhalation exposure:

whole body

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

1.36 µm

Details on inhalation exposure:

- Generation of test atmosphere / chamber description: This test was conducted in a 40 L (36 x 36 x 31 cm) glass exposure chamber. The sides and the bottom of the chamber had centered holes (3-4 cm in diameter) to allow access to the chamber for testing and exhaust of the atmosphere. The port in the bottom of the chamber was centered over a 10 cm hole in a wooden platform. A funnel (8.5 cm in diameter) was brought from the underside of the platform through the hole and centered on the port in the bottom of the chamber. Dynamic air flow within the chamber was maintained by connecting the funnel to a vacuum pump for continuous changing of the chamber atmosphere.

A raised, tightly-fitting, wire mesh screen was placed over the bottom of the chamber and served as flooring for the test animals. This screen also served to raise the animals to a level such that the side ports in the chamber were at the breathing level of the animals. All measurements of particle size and concentration of the test substance were made from these side ports. Also, the delivery of the test substance into the chamber was done through one of the ports on the side of the chamber. The lid of the chamber also had a port which was used (if desired) for measuring the chamber atmosphere to insure even distribution of the test substance.

- Test substance generation: The test substance was generated as a dust using a 3-neck, round-bottom, 250 mL Pyrex flask. A stirring mechanism was placed through the center neck of the flask and an air line through one of the side necks. The third neck was connected by a glass elbow (which had an air vent to allow flushing) to the chamber. The dust was introduced into the chamber through a side port. A piece of rubber was taped over the outer area around the hole and a vertical slit made in the rubber to allow the entrance of the glass elbow from the dust generator. Constant flow of material was maintained by a calibrated flowmeter connected between the air line and the generating apparatus. In all instances, the air flow was maintained at or above 0.5 L of air/minute/rat.

- Exposure monitoring: Measurement of the atmospheric concentration of test substance in the chamber was achieved using a Gelman Model 1235 stainless steel filter holder containing a pre-weighed glass fiber filter (Gelman type AE-47 mm). The filter holder was attached to a vacuum pump which was regulated to exhaust 1 L of air/min from the chamber through the filter. Subsequent weighing of the filter after a designated period permitted the calculation of the atmospheric concentration of the test substance in the chamber during the exposure. Particle size determinations were made using a Cascade Impactor.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

1 h

Concentrations:

Mean achieved: 0.29±0.12 mg/L

No. of animals per sex per dose:

5/sex/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Frequency of observations:
Mortality and clinical signs- Four times during the exposure (15, 30, 45 and 60 minutes), twice after the exposure (3 and 6 h) on same day and daily once thereafter.
Body weights- pre-test, Day 1, 7 and 14
- Necropsy of survivors performed: Yes

Statistics:

no data

Preliminary study:

none

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 0.29 mg/L air

Based on:

test mat.

Exp. duration:

1 h

Mortality:

No mortality was observed.

Clinical signs:

other: No adverse effects were noted during the 1 h exposure or during the 14 d post-exposure period.

Body weight:

All animals displayed a normal pattern of weight gain.

Gross pathology:

All organs examined were normal.

Other findings:

none

Table: Method of calculating analytical concentration

	SAMPLE 1	SAMPLE 2	SAMPLE 3	SAMPLE 4
Post Weight (mg)	1495.5	1500.3	1500.3	1495.8
Pre-Weight (mg)	1493.5	1495.9	1495.9	1493.3
Weight Difference (mg)	2.0	4.4	4.4	2.5
Flow Rate (L/min)	1	1	1	1
Sample Time (min)	10	10	10	10
Air Volume (L)	10	10	10	10
Analytical Concentration (mg/L)	0.20	0.44	0.44	0.25

 

The average mass median particle diameter was 1.36 ± 0.26 µ. The 1 h LC50 value is >290 mg/mm³ (>0.29 mg/L).

Interpretation of results:

GHS criteria not met

Conclusions:

The 1 h inhalation LC50 value of the FAT 20033/D was considered to be >290 mg/mm³ (>0.29 mg/L).

Executive summary:

A study was conducted to evaluate the acute inhalation toxicity of FAT 20033/D (ca. 46 % purity) following a method comparable to EPA OPPTS 870.1300 with deviations. A single group of 10 rats (5/sex) were exposed (whole body) for 1 h to the dust of the test substance at a mean achieved concentration of 0.29 mg/L (i.e. 290 mg/m³). A 14 d observation period followed. No mortality, change in body weight, clinical signs or gross pathological organ changes were observed during the study period. Hence, the 1 h inhalation LC50 value of the test substance was considered to be >290 mg/mm³ (>0.29 mg/L).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

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:

1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

not applicable

GLP compliance:

no

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: E0T425-11 (OCT No. 70083)

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ace Animals, Boyertown, Pennsylvania
- Weight at study initiation: 1.6 - 2.4 kg
- Housing: in accordance with standard laboratory procedures
- Diet: ad libitum
- Water: ad libitum

Type of coverage:

occlusive

Details on dermal exposure:

TEST SITE
- Area of exposure: Back side
- % coverage: 30%
- Type of wrap if used: elastic sleeve

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Warm water
- Time after start of exposure: 24 hr

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 3g/kg

Duration of exposure:

24 h

Doses:

3 g/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

Twenty-four hours prior to dosing, the animals were immobilized in an animal holder and their backs clipped free of hair with an Oster animal clipper, exposing approximately 30 % of each animal's skin surface. Immediately prior to dosing, one-half of the animals were further prepared by making epidermal abrasions longitudinally over the area of exposure. The abrasions were made sufficiently deep to penetrate the stratum corneum, but not deep enough to disturb the derma. The test material was applied, in the quantity of 3 g/kg of body weight, to the test area and held in contact with the skin by means of an elastic sleeve for a period of 24 hours at which time the sleeve was removed and the treated areas washed clean of the remaining excess test material with warm water. The animals were then returned to their individual cages and observed for toxic signs and survival.

OBSERVATIONS:
The animals were observed daily for adverse reaction and survival for a period of 14 days. In addition, local skin reactions were evaluated at 1, 7 and 14 days. Body weights were recorded prior to dosing and at days 7 and 14. Gross necropsies are performed on all animals.

Statistics:

None

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 3 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was seen throughout the observation period.

Gross pathology:

All gross necropsies were within normal limits.

Other findings:

Edema and erythema were present on all sites for 72 hours and on five sites for four days. The remaining site, at this time, displayed erythema. On the fifth day of scoring, four animals demonstrated erythema and two demonstrated both edema and erythema. On days six, seven and eight, irritation grading showed three animals displaying erythema. After this time, all animals returned to normal.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of FAT 20033/D in New Zealand albino rabbits is >3 g/kg bw.

Executive summary:

A study was conducted to assess the acute dermal toxicity of FAT 20033/D in New Zealand albino rabbits. A group of 6 animals (3/sex/dose) were used for the study. Twenty-four hours prior to dosing, the animals were immobilized in an animal holder and their backs clipped free of hair with an Oster animal clipper, exposing approximately 30% of each animal's skin surface. Immediately prior to dosing, one-half of the animals were further prepared by making epidermal abrasions longitudinally over the area of exposure. The abrasions were made sufficiently deep to penetrate the stratum corneum, but not deep enough to disturb the derma. The test material was applied, in the quantity of 3 g/kg of body weight, to the test area and held in contact with the skin by means of an elastic sleeve for a period of 24 hours at which time the sleeve was removed and the treated areas washed clean of the remaining excess test material with warm water. No mortalities were observed throughout the study. Oedema and erythema were present on all sites for 72 hours and on five sites for four days. The remaining site, at this time, displayed erythema. On the fifth day of scoring, four animals demonstrated erythema and two demonstrated both edema and erythema. On days six, seven and eight, irritation grading showed three animals displaying erythema. After this time, all animals returned to normal. All gross necropsies were within normal limits. Based on the study results, the dermal LD50 of the test substance was found to be >3,000 mg/kg bw in NZ albino rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

Oral:

An acute oral toxicity study was conducted using 5 Sprague Dawley rats/sex, which were gavaged with 5000 mg/kg bw of FAT 20033/C. Mortality and clinical signs were observed during 14 days. At the end of the observation period surviving animals were killed by exsanguination under ether anaesthesia and an autopsy performed. No mortality or clinical signs were seen during the observation period. At autopsy no changes in organs or tissues caused by the administration of the test compound were seen. On the basis of the study results, the median lethal dose (LD50) for FAT 20033/C derived is >5000 mg/kg bw. An acute oral toxicity study conducted with FAT 20033/D further supports the conclusion that Acid Blue 277 has low acute toxicity, with LD50 being >5000 mg/kh bw.

 

Inhalation:

A study was conducted to evaluate the acute inhalation toxicity of FAT 20033/D (ca. 46 % purity) following a method comparable to EPA OPPTS 870.1300 with deviations. A single group of 10 rats (5/sex) were exposed (whole body) for 1 h to the dust of the test substance at a mean achieved concentration of 0.29 mg/L (i.e. 290 mg/m³). A 14 d observation period followed. No mortality, change in body weight, clinical signs or gross pathological organ changes were observed during the study period. Hence, the 1 h inhalation LC50 value of the test substance was considered to be >290 mg/mm³ (>0.29 mg/L).

 

Dermal:

A study was conducted to assess the acute dermal toxicity of FAT 20033/D in New Zealand albino rabbits. A group of 6 animals (3/sex/dose) were used for the study. The test material was applied, in the quantity of 3 g/kg of body weight, to the shaved (3 animals) and abraded (3 animals) test area and held in contact with the skin by means of an elastic sleeve for a period of 24 hours. No mortalities were observed throughout the study. Edema and erythema were present on all sites for 72 hours and on five sites for four days. The remaining site, at this time, displayed erythema. On the fifth day of scoring, four animals demonstrated erythema and two demonstrated both edema and erythema. On days six, seven and eight, irritation grading showed three animals displaying erythema. After this time, all animals returned to normal. All gross necropsies were within normal limits. Based on the study results, the dermal LD50 of the test substance was found to be >3,000 mg/kg bw in NZ albino rabbits.

Justification for classification or non-classification

Based on the available information 1-amino-4-({3-[(2-hydroxyethyl)sulfamoyl]-4,5-dimethylphenyl}amino)-9,10- dioxo-9,10-dihydroanthracene-2-sulfonic acid, sodium salt does not need to be classified for acute toxicity according to CLP (Regulation (EC) No 1272/2008 Of The European Parliament And Of The Council) as implementation of UN-GHS in the EU.