Pentasodium 2,2'-((2,4-diamino-5-(4-methoxy-2-sulfophenylazo)-1,3-phenylene)bis(azo))bis(5-(2-(sulfonatooxyethyl)sulfonyl)benzenesulfonate

Endpoint summary

Description of key information

The test substance is considered to be of low acute oral and dermal toxicity with LD50 values >2,000 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 July 27, 2005 to August 12, 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

CRL:(WI) BR rats

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (Europe) laboratories INC.
- Hygienic level at arrival: SPF
- Age at study initiation: Young healthy adult rats, less than 10 weeks old
- Body weight range at treatment: 175-188g
- Housing: II type polypropylene/polycarbonate, 3 animals per cage
- Bedding: Laboratory bedding
- Diet: Ssniff RIM-Z+H, ad libitum
- Water: Tap water from 500 mL bottle, ad libitum
- Acclimation period: 14 d
-Animal identification: Numbers on the tail written with a marker pen
ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C
- Humidity: 30 - 70%
- Air changes: 8-12 air exchanges/hour by central air-condition system
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: July 27, 2005 to August 12, 2005

Route of administration:

oral: gavage

Vehicle:

other: 1 % aqueous methylcellulose

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Lot/batch no.: 05-07/01
- Expiry date: August 4, 2005
- Storage: In refrigerator

MAXIMUM DOSE VOLUME APPLIED: 1 mL/100 g bw (i.e., 10 mL/kg bw)

DOSAGE PREPARATION: For treatment the test substance was applied in a concentration of 200 mg/mL in 1 % aqueous methylcellulose. Formulations were prepared just before the treatment.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: No mortality was expected at 2,000 mg/kg bw dose level, so this was chosen as the starting dose.

Doses:

2,000 mg/kg bw

No. of animals per sex per dose:

6 animals (i.e., 3 animals/group)

Details on study design:

Test procedure
A single oral administration - followed by a 14 d observation period was performed by gavage. On the day before each treatment the animals were fasted. The food but not water was withheld during an overnight period. Animals were weighed just before the application. The test substance was administered by oral gavage in the morning hours. The food was given back 3 h after the treatment. A constant treatment volume of 1 mL/100 g bw was applied.

Three female animals were treated with a dose level of 2,000 mg/kg bw of test substance in the first step. All animals survived; so three further animals were dosed at 2,000 mg/kg bw dose level next day, as the second step. No mortality occurred in either step, so the test was finished meeting the stopping criteria ofguidelines.

Clinical Observations
Careful clinical observation was made once within the first 30 minutes, then 30 minutes, 1 h, 2 h, 3 h, 4 h, 6 h after the treatment and once each day for 14 d thereafter. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern as well. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma.

Body Weight Measurement

The body weight was recorded on Day 0 (shortly before the treatment), then on Days 7 and 14 with precision of 1g.

NECROPSY

All animals were subjected to gross pathology. Animals were exsanguinated under pentobarbital anesthesia. After examination of the external appearance the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. Abnormalities were recorded on post mortem data sheets.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

Test substance caused no mortality in female CRL:(Wl) BR rats after a single oral (i.e., by gavage) administration of 2,000 mg/kg bw.

Clinical signs:

other: Slight diarrhoea was observed in two animals from 2 h after the dosing to the end of the day. No clinical symptoms appeared on the day of the treatment on the other animals and the following 14 d observation period. The physical condition and behaviour of

Gross pathology:

No macroscopic alterations related to the toxic effect of test substance were found at necropsy. The pinprick-sized haemorrhages (3/6) observed in the lungs were attributed to the termination procedures and exsanguination.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the oral LD50 was found to be >2,000 mg/kg bw in rats.

Executive summary:

A study was conducted to assess the acute oral toxicity of the test substance in CRL:(WI) BR female rats according to OECD Guideline 423, EPA OPPTS 870.1100 and EU Method B.1 tris, in compliance with GLP.

Group of three female rats received a single oral (gavage) dose of 2,000 mg/kg bw in a first and second step, respectively. A concentration of 200 mg/mL of test substance was prepared in 1 % aqueous rnethylcellulose and administered at a volume of 1 mL/100 g bw (i.e., 10 mL/kg bw).

 

No mortality occurred, no effect on body weight was observed and no significant macroscopic abnormalities were seen at necropsy. Except for slight diarrhoea in two animals from 2 h after the dosing to the end of the day, no clinical symptoms were observed in other animals.

 

Under the study conditions, the oral LD50 was found to be >2,000 mg/kg bw in rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

High quality GLP study.

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, 16 2006 to May 30, 2006

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

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

CRL:(WI) BR Wistar rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (Europe) laboratories INC.
- Hygienic level at arrival: SPF
- Age at study initiation: Young adult rats
- Body weight range at treatment: Male: 243 - 271 g; Female: 236 - 245 g
- Sex: Male and female; female rats were nulliparous and non-pregnant
- Housing: Type II polypropylene/polycarbonate, individual caging
- Bedding: Laboratory bedding
- Diet: Ssniff RIM-Z+H complete diet, ad libitum
- Water: Tap water from 500 mL bottle, ad libitum
- Acclimation period: Female: 20 d; male: 6 d
-Animal identification: Numbers on the tail written with a permanent marker; The cages were marked by identity cards with information about study code, sex, dose group, cage number and individual animal number

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C
- Humidity: 30 - 70%
- Air changes: 8-12 air exchanges/hour by central air-condition system
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: May, 16 2006 to May 30, 2006

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
The back of animals was shaven (approximately 10% area of the total body surface) 24 h prior to the treatment. The test substance was applied in a single dose in original form uniformly over the shaved skin throughout a 24 h exposure period. Sterile gauze pads were placed on the skin of rats. These gauzes were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was wrapped with semi occlusive plastic wrap for 24 h.

REMOVAL OF TEST SUBSTANCE
At the end of the exposure period, residual test item was removed, using body temperature water.

PREPARATION OF THE TEST SUBSTANCE
For the treatment the test substance was applied in original form moistened with 0.5 mL of distilled water.

Duration of exposure:

24 h

Doses:

2,000 mg/kg bw

No. of animals per sex per dose:

Five/sex/dose

Control animals:

not required

Details on study design:

Experimental design

Dose Group Number of animals
Male Group (2,000 mg/kg bw) 5
Female Group (2,000 mg/kg bw ) 5

A single administration was performed by dermal route and was followed by a 14 d observation period.

OBSERVATIONS

Clinical Observations

Careful clinical observation was made at the following intervals: 1h, 5h after the treatment and once each day for 14 d thereafter. Individual observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern as well. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement

The body weight of each animal was recorded on Day 0 (shortly before the treatment), then on Days 7 and 14 with a precision of 1 g.

NECROPSY

At the end of the observation period gross pathology was conducted in every experimental animal. Animals were exsanguinated under pentobarbital anaesthesia. After examination of the external appearance, the cranial, thoracic, and abdominal cavities were opened and tissues, organs were observed. Gross pathological changes were recorded for each animal on the post mortem record sheets.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No mortality occurred after the 24 h dermal exposure to test substance in CRL(WI)BR rats.

Clinical signs:

other: No behavioural changes or general toxic symptoms were noted during the study. The skin remained brownish coloured after careful washing off of test substance, thus it was not feasible to evaluate erythema. In three cases (1/5 male, 2/5 female) some small

Gross pathology:

Test substance related macroscopic alterations were not found at the gross pathology. Pinprick-sized haemorrhages (5/5 male, 2/5 female) were observed in the lungs, which were caused by exsanguination procedures. Hydrometra (1/5) due to the sexual cycle is a common alteration in experimental rats.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the dermal LD50 of the test substance was found to be >2,000 mg/kg bw in rats.

Executive summary:

A study was conducted to assess the acute dermal toxicity of the test substance in CRL:(WI) BR Wistar rats according to OECD Guideline 402, EU Method B.3. and EPA OPPTS 870.1200, in compliance with GLP.

 

Groups of five female and five male rats received a single dermal dose of 2,000 mg/kg bw. Test substance moistened with 0.5 mL of distilled water was applied topically under semi-occlusive conditions for 24 h.

 

No mortality occurred, no clinical symptoms were observed and no significant macroscopic abnormalities were seen at necropsy. Body weight was also not impaired. Although, the skin of the animals was brownish coloured by the test substance and some small wounds were observed on the treated skin surface in three animals, skin recovered between Days 4 and 6. 

Under the study conditions, the dermal LD50 of the test substance was found to be >2,000 mg/kg bw in rats

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

High quality GLP study.

Additional information

Oral route:

A study was conducted to assess the acute oral toxicity of the test substance in CRL:(WI) BR female rats according to OECD Guideline 423, EPA OPPTS 870.1100 and EU Method B.1 tris, in compliance with GLP. Group of three female rats received a single oral (gavage) dose of 2,000 mg/kg bw in a first and second step, respectively. A concentration of 200 mg/mL of test substance was prepared in 1 % aqueous rnethylcellulose and administered at a volume of 1 mL/100 g bw (i.e., 10 mL/kg bw). No mortality occurred, no effect on body weight was observed and no significant macroscopic abnormalities were seen at necropsy. Except for slight diarrhoea in two animals from 2 h after the dosing to the end of the day, no clinical symptoms were observed in other animals. Under the study conditions, the oral LD50 was found to be >2,000 mg/kg bw in rats (Szakonyi, 2005).

Dermal route:

A study was conducted to assess the acute dermal toxicity of the test substance in CRL:(WI) BR Wistar rats according to OECD Guideline 402, EU Method B.3. and EPA OPPTS 870.1200, in compliance with GLP. Groups of five female and five male rats received a single dermal dose of 2,000 mg/kg bw. Test substance moistened with 0.5 mL of distilled water was applied topically under semi-occlusive conditions for 24 h. No mortality occurred, no clinical symptoms were observed and no significant macroscopic abnormalities were seen at necropsy. Body weight was also not impaired. Although, the skin of the animals was brownish coloured by the test substance and some small wounds were observed on the treated skin surface in three animals, skin recovered between Days 4 and 6. Under the study conditions, the dermal LD50 of the test substance was found to be >2,000 mg/kg bw in rats (Szakonyi, 2006).

Justification for classification or non-classification

Oral route:

Based on the results of an acute oral toxicity study, the test substance does not need to be classified for acute toxicity according to the EU CLP criteria (EC 1272/2008).

Dermal route:

Based on the results of an acute dermal toxicity study, the test substance does not need to be classified for acute toxicity according to the EU CLP criteria (EC 1272/2008).