Sodium 1-amino-9,10-dihydro-9,10-dioxo-4-[(2,2,4-trimethylcyclohexyl)amino]anthracene-2-sulphonate

Endpoint summary

Description of key information

Oral LD50 (male and female): 9476.1 mg/kg bw (6633.27 mg/kg bw based on the active ingredient)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From May 26th to August 15th, 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

The read across approach is detailed into the document attached to the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

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: 150g.
- Housing: 5 rats were housed in separated menagerie of 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: standard commercially manufactured complete Mixed fodders ST-1, 10 g /animal/ day.
- Water: 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:

7943, 8913, 10000 and 12590 mg/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.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

9 476.1 mg/kg bw

Based on:

test mat.

Remarks:

(ca 6633.27 a.i. mg/kg bw)

Mortality:

Mortality is observed in doses higher than 7943 mg/kg.

Clinical signs:

After application of the logarithmic dose of 12590 mg/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
- appearance of visible mucous membranes: blue color of the lining of the nose and conjunctivitis
- mental activity: within the physiological standards
- somatomotor activity: locomotion disorders due to ataxia
- reactivity and sensibility: hypergie, Hyperaesthesia
- functions of the digestive system: diarrhea

Body weight:

Body weight loss was observed for male and female rats.

Dose (mg/kg)	Mortality
	Male	Female	Total percentage
7943	0	0	0
8913	1	2	30
10000	2	5	70
12590	5	5	100

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No1272/2008.

Conclusions:

LD50 (male and female): 9476.1 mg/kg bw (6633.27 mg/kg bw based on the active ingredient)

Executive summary:

The substance has been tested for acute toxicity by oral route according to the OECD Guideline 401. 5 male and 5 female Wistar rats per group were tested with the following dose: 0, 7943, 8913, 10000 and 12590 mg/kg.

After 14 days of observation period the rats showed weight loss in all the application doses.

Mortality was observed in doses higher than 7943 mg/kg. After application of the logarithmic dose of 12590 mg/kg the following clinical symptoms of intoxication were observed: coloration of the skin after application and smoot, shiny hair, blue color of the lining of the nose and conjunctivitis, locomotion disorders due to ataxia, hypergie, hyperaesthesia and diarrhea.

Conclusion

LD50 (male and female): 9476.1 mg/kg bw (6633.27 mg/kg bw based on the active ingredient)

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

9 476 mg/kg bw

Additional information

ACUTE TOXICITY BY ORAL ROUTE

A sheet reporting the test results of an old experiment conducted on Acid Blue 278 is available; however, due to the fact that details on testing procedures and results are lacking, a reliability cannot be assigned. The result is here mentioned for completeness sake. The oral LD50 in rat was indicated to be higher than 5000 mg/kg bw (Sheet of test results, 1977).

In order to assess the oral acute potential toxicity of Acid Blue 278, the available information on the structural analogous Similar Substance 01 have been taken into consideration; the read across approach can be considered as appropriate and suitable to assess the property under investigation (details about the approach are reported into the IUCLID section 13).

The Similar Substance 01 tested for acute toxicity by oral route, according to the OECD Guideline 401. Mortality was observed in doses higher than 7943 mg/kg. After application of the logarithmic dose of 12590 mg/kg the following clinical symptoms of intoxication were observed: coloration of the skin after application and smoot, shiny hair, blue color of the lining of the nose and conjunctivitis, locomotion disorders due to ataxia, hypergie, hyperaesthesia and diarrhea.

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 the 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 greater than 2000 mg/kg body weight, therefore the test substance is expected to be out of any classification limit for acute oral toxicity (oral acute toxicity category 4: 300 < ATE ≤ 2000 mg/kg bw).