Sodium 1-amino-4-(cyclohexylamino)-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Endpoint summary

Description of key information

LD₅₀ oral = 9476.1 mg/kg bw
LD₅₀ dermal > 5020 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:

From May 26 to August 15, 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: TOP Velaz Ltd.
- Weight at study initiation: 150 g.
- Housing: plastic polypropylene cages T4, equipped with dry softwood chip which was sterilized in a hot air sterilizer HS 401 A/1 at 150°C for 90 minutes.
- Diet (e.g. ad libitum): standard commercially manufactured complete Mixed fodders ST-1, 10 g/animal/day.
- Water (e.g. ad libitum): drinking water according to CSN 757111 ad libitum.
- Acclimation period: one week.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3°C.
- Humidity: 50 ± 15%
- Photoperiod: 12 hrs cycle dark/light.
- Other: fluorescent light.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20% suspension

Doses:

7.943 g/kg, 8.913 g/kg, 10.00 g/kg, 12.59 g/kg

No. of animals per sex per dose:

5 per sex per dose

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations: the animals were observed for clinical signs of intoxication immediately after application (after 30 minutes), 3 hours after application and once a day for 14 days.
- Frequency of weighing: the animals were weighed before the oral administration and during the observation period.
- Necropsy of survivors performed: yes.
- Other examinations performed: clinical diagnosis was focused on observation of the appearance of skin, fur, visible mucous membranes, nutritional status, mental activity, somatomotor activity, responses to stimuli, focusing on sensibility and reactivity, lacrimation, assessment function respiratory, digestive, urogenital and circulatory system. Organs and muscles were examined macroscopically. After dissection internal organs were judged according to their color, size, consistency and structure. If the post-mortem bladder is filled with urine, the urine were carried out biochemical tests indicative indicator strips Heptaphan focusing on the detection of proteins, blood sugars, ketones, bilirubin, urobilinogen and pH.

Statistics:

LD50 is calculated by the probit method according to Bliss. Mortality data used and the frequency and level logarithmic doses were entered into a computer and analyzed by the program PROBIT from October 1991 VÚOS.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 9 476.1 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD0

Effect level:

ca. 7 943 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 6 633.27 mg/kg bw

Based on:

act. ingr.

Mortality:

Mortality is observed in doses higher than 7.943 g/kg.

Clinical signs:

other: After application of the logarithmic dose of 12.59 g / kg the following clinical symptoms of intoxication were observed: - appearance of skin and hair: blue coloration of the skin after application and smoot, shiny hair - nutritional status: good - appear

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 (Male/Female, Rats) = 9476.1 mg/kg b.w. (6633.27 mg/kg b.w. based on the active ingredient)

Executive summary:

The substance has been tested for acute toxicity by oral route accordimg to the OECD Guideline 401. Forty Wistar rats were tested with the following dose: 7.943 g/kg, 8.913 g/kg, 10.00 g/kg, 12.59 g/kg.

After 14 days of observation period the rats showed weight loss in all the application doses. The clinical signs observed are: conjuntivitis, locomotion disorders due to ataxia, diarrhea, hypergie, hyperaesthesia and blue coloration of the skin.

 

The substance shows an LD50 for male and female rats over the period of 14 days equal to 9476.1 mg/kg b.w. (6633.27 mg/kg b.w. based on the active ingredient).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

9 476.1 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:

From May 26 to August 15, 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: TOP Velaz Ltd. Praha.
- Weight at study initiation: 190-202 g.
- Housing: experimental animals were housed individually in cages made of plastic polypropylene T3, equipped with dry softwood chip which was sterilized in a hot air sterilizer HS 401 A/1 at 150°C for 90 minutes.
- Diet (e.g. ad libitum): standard commercially produced set-Mixed fodders ST-1, 20 g/animal/day.
- Water (e.g. ad libitum): drinking water according to CSN 757111 ad libitum.
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 3°C
- Humidity (%): 50% + 15%
- Photoperiod (hrs dark / hrs light): 12 hrs cycle dark/light
- Other: fluorescent light

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: 6 x 6 cm (36 cm^2) shaved skin.
- Type of wrap if used: animals were covered with gauze, aluminum foil and adhesive tape around the perimeter of the hull. This was accompanied by a fixation bandage. The dressing was covered with adhesive tape and clamped around the periphery of the fuselage.

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 24 hours.

Duration of exposure:

The duration of the exposure was 24 hours.

Doses:

logarithmic dose is 5.020 g/kg

No. of animals per sex per dose:

5 males per dose

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations: animals were observed for clinical signs of intoxication immediately after application (after 30 minutes), 3 hours after application and once a day for 14 days.
- Frequency of weighing: the animals were weighed at the beginning and at the end of the experiment.
- Necropsy of survivors performed: yes.
- Other examinations performed: clinical diagnosis was focused on observation of the appearance of hair, skin, visible mucous membranes, nutritional status, mental activity, somatomotor activity, responses to stimuli, focusing on sensibility and reactivity, lacrimation, assess the functioning of the respiratory, digestive and urogenital system Organs and muscles were examined macroscopically. After dissection internal organs were judged according to their color, size, consistency and structure. After macroscopic assessment of internal organ (heart, lungs, liver, spleen, kidneys, adrenal glands) biometric examination was performed. If the post mortem bladder is filled with urine, the urine was biochemical tested using Heptaphan indicative strips focusing on the detection of proteins, blood sugars, ketones, bilirubin, urobilinogen and pH.

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 020 mg/kg bw

Based on:

test mat.

Sex:

male

Dose descriptor:

LD50

Effect level:

> 3 514 mg/kg bw

Based on:

act. ingr.

Gross pathology:

After application of the logarithmic dose of 5.020 g/Kg the following pathological/anatomical symptoms of intoxication were observed:
hair, skin and visible: normal appearance;
subcutaneous tissue and muscle: without macroscopic pathomorpholoical change;
nutritional status: very good;
head and neck: without macroscopic pathomorpholoical change;
heart: maroon (brownish-red), stiff consistency, without pathomorpholoical changes;
lung: a pinkish color, spongy, airy, without macroscopic pathomorfological changes;
stomach: filled with mushy food, without macroscopic pathomorfological changes;
intestine: filled with sparse food, without macroscopic pathomorfological changes;
liver: dark brownish color, stiff consistency, without pathomorfological changes;
spleen: reddish-brown color, stiff consistency, without pathomorfological changes;
kidneys: brownish red color, firmer texture, smooth surface, pathomorphological changes;
bladder: no macroscopic pathomorfological changes;
peritoneum: without pathomorfological changes.

Other findings:

Organ weights: the relative weight of the organs (heart, lungs, kidneys, liver, spleen, adrenal lang, testicles) of the animals assigned to the experimental group was higher than the relative weight of the organs of the animals assigned to the control group.

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 (male rats) > 5020 mg/kg b.w. (> 3514 mg/kg b.w. based on the active ingredient)

Executive summary:

The substance has been tested for acute toxicity by dermal route according to the OECD Guideline 402. Five Wistar rats were tested with the dose: 5.020 g/Kg. After 14 days of the observation period, rats showed increase of body weight. At the end of the experiment the relative weight of the organs (heart, lungs, kidneys, liver, spleen, adrenal lang, testicles) of the animals assigned to the experimental group was higher than the relative weight of the organs of the animals assigned to the control group.

The clinical signs observed after the application of the test material was the pinkish coloration of the skin which derived from physiological standards. After the application of the test material and the autopsy no macroscopic pathomorfological changes were observed.

 

The LD50 of the substance for male rats over the period of 14 days is greater than to 5020 mg/kg b.w. (3514 mg/kg b.w. based on the active ingredient).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 020 mg/kg bw

Additional information

The test substance was evaluated for its potential to cause acute toxicity in rats.

 

Acute Toxicity: Oral

The study was performed according to the OECD guideline 401, following oral administration of multiples doses to the rats. Mortality is observed in doses higher than 7.943 g/Kg. After 14 days of observation, rats showed weight loss in all the application doses. Clinical signs observed were : conjuntivitis, locomotion disorders due to ataxia, diarrhea, hypergie, hyperaesthesia and blue coloration of the skin.

The acute oral LD₅₀ was determined to be 9476.1 mg/kg bw.

 

Acute Toxicity: Dermal

The evaluation of acute dermal toxicity has been performed according to the OECD guideline 402. No mortality occurred during the study.

After 14 days of the observation period, rats showed increase of body weight. At the end of the experiment the relative weight of the organs (heart, lungs, kidneys, liver, spleen, adrenal lang, testicles) of the animals assigned to the experimental group was higher than the relative weight of the organs of the animals assigned to the control group. The clinical sign observed after the application of the test material was the pinkish coloration of the skin which derived from physiological standards. After the application of the test material and the autopsy no macroscopic pathomorfological changes were observed.

Justification for classification or non-classification

According to the CLP Regulation (EC) No. 1272/2008, 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to numeric criteria. Acute toxicity values are expressed as (approximate) LD₅₀ (oral, dermal) or LC₅₀ (inhalation) values or as acute toxicity estimates (ATE).

 

The oral LD₅₀ value was established to be 9476.1 mg/kg body weight, therefore the test substance is out of any classification limit for acute oral toxicity (oral acute toxicity Category 4: 300 < ATE ≤ 2000 mg/kg bw).

The dermal LD₅₀ value was established to be higher than 5020 mg/kg body weight, which exceeded the highest CLP limit for classification (dermal acute toxicity Category 4: 1000 < ATE ≤ 2000 mg/kg bw).

 

In conclusion, using the data obtained from oral and dermal acute toxicity, it is possible to conclude that the test substance is not classified for acute oral toxicity and acute dermal toxicity, according to the CLP Regulation (EC) No. 1272/2008.