6,13-dichloro-3,10-bis-{2-[4-fluoro-6-(2-sulfo-phenylamino)-1,3,5-triazin-2-ylamino]-propylamino}-benzo[5,6][1,4]oxazino[2,3-.b.]phenoxazine-4,11-disulphonic acid, lithium, sodium salt.

Endpoint summary

Description of key information

A LD50 of >2000mg/kg bw was observed in the acute oral and acute dermal toxicity study.

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Italia, Calco Como ltaly
- Age at study initiation: 5 to 6 weeks
- Weight at study initiation: 126 - 150g
- Fasting period before study: Overnight
- Housing: Group of five of one sex in clear polycarbonate cages measuring 59x20x39 cm with a stainless steel mesh lid and floor. Each cage tray held absorbent material which was inspected daily and changed as necessary.
- Diet (e.g. ad libitum): Commercially available laboratory rodent diet (Altromin MT, A. Rieper S.p.A., Bolzano, Italy) ad libitum
- Water (e.g. ad libitum): Drinking water supplied to each cage via a water bottle, ad libitum
- Acclimation period: Seven days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 45-65
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5%

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10mL/kg bw

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: throughout the study all animals were checked twice daily for mortality. All animals were weighed at allocation to the study (day -1), immediately prior to dosing (Day 1) and at weekly intervals thereafter (Days 8 and 15). Animals were observed for clinical signs immediately upon dosing, approximately one, two and four hours after dosing and daily thereafter for a total of fourteen days.
- Necropsy of survivors performed: AII animaIs were killed by carbon dioxide narcosis on terminatlon of the defined post-dose observation perlod. Animals were subjected to a gross examination for external abnormalities. The cranial, thoracic and abdominal cavities were opened to allow examination of their contents and larger organs were sectioned. The stomach and representitive sections of the gastrointestinal tract were opened for examinatlon of the mucosal surfaces.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured.

Clinical signs:

other: No significant clinical signs were observed following dosing. Mucoid and blue stained faeces were observed in the litter tray under the cages of animals of both sexes. This was apparent approximately four hours after dosing and was no longer present from

Gross pathology:

No abnormalities were noted on necropsy examination of animals on termination of the study. Findings were limited to blue staining on the tail of all female animals. This staining would have arisen from the coloured test substance but was not a symptom of toxicity or intolerance.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the observations the toxicity of the test substance was estimated to be greater than 2000 mg/kg bw.

Executive summary:

In a GLP compliant oral toxicity study, performed according to OECD guideline 401, Sprague-Dawley rats (5/sex/) were administered the test substance (2000 mg/kg bw) by oral gavage followed by a 14-day observation period. Mortality did not occur. No significant clinical signs were observed and necropsy did not reveal any significant abnormality. Based on these observations, the acute oral toxicity of the test substance in rats of both sexes observed for a period of 14 days was greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

GLP compliant guideline study, klimisch 1

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia S.p.A., Calco (Como), ltaly
- Age at study initiation: 6 to 8 weeks
- Weight at study initiation: 176 - 200g
- Housing: Individually in polycarbonate cages measuring 59x39x20 cm with a stainless steel mesh lid and floor. Cages were suspended over trays and each tray held an absorbent material which was inspected daily and changed as necessary.
- Diet (e.g. ad libitum): Commercially available laboratory rodent diet (Altromin MT, A. Rieper S.p.A., Bolzano, Italy) ad libitum
- Water (e.g. ad libitum): Drinking water supplied to each cage via a water bottle, ad libitum
- Acclimation period: Seven days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 45-65
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

The fur was removed from the dorsaI surfaces of the trunk over an area estimated to be at least 10% of the total body surface of each animal. An electric cllpper with suitable blade was used and care was taken to avoid any irritation or damage to the skin.
The next day (Day 1), the amount of supplied test substance to be administered, at a dose level of 2000 mg/kg body weight was calculated for each animal according to body weight. This was mixed to a paste with sterile water and spread evenly over the prepared skin of each animal. The mixing with water and volume of water used was not recorded due to oversight. The dosed site was then covered by a porous gauze dressing, held in place by encircling the trunk of the animal with an elastic adhesive bandage overlying aluminium foil. After a period of twenty four hours, the adhesive bandage and gauze dressings were removed. The treated skin was washed gently with warm water to remove residual test substance.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Mortality/viability were measured throughout the study twice daily
- Body weights were measured at allocation of the study (Day -1), immediately prior to dosing (Day 1) and at weekly intervals thereafter (Days 8 and 15).
- Animals were observed for clinical signs immediately upon dosing, approximately two and four hours after dosing and daily thereafter for a total of fourteen days.
- Necropsy of survivors performed: AII animaIs were killed by carbon dioxide narcosis on terminatlon of the defined post-dose observation perlod. Animals were subjected to a gross examination for external abnormalities. The cranial, thoracic and abdominal cavities were opened to allow examination of their contents and larger organs were sectioned. Particular attention was paid to the treated site.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured during the fourteen day observation period following dosing.

Clinical signs:

other: No clinical signs were observed during this post-dose observation period that could be attributed to an effect of the test substance. The only finding was hair loss from the ventral surfaces of one female animal, observed on day 15. Such hair loss may be

Gross pathology:

No abnormalities were found on necropsy.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The toxicity of the test substance was estimated to be greater than 2000 mg/kg bw

Executive summary:

In a GLP compliant dermal toxicity study, performed according to OECD guideline 402, Sprague-Dawley rats (5/sex) were administered the test substance (2000 mg/kg bw). The test substance was dissolved in water, applied on the skin and covered with an occlusive dressing for 24 hours. The treated skin was washed after 24 hours and a 14-day observation period followed. No mortality was observed during this period. Also no clinical signs and significant changes in body weight were observed. Furthermore, no abnormalities were found on necropsy. Therefore, the toxicity of the test substance was estimated to be >2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

GLP compliant guideline study, klimisch 1

Additional information

Acute oral toxicity:

In a GLP compliant oral toxicity study, performed according to OECD guideline 401, Sprague-Dawley rats (5/sex/) were administered the test substance (2000 mg/kg bw) by oral gavage followed by a 14-day observation period (RTC 1995). Mortality did not occur. No significant clinical signs were observed and necropsy did not reveal any significant abnormality. Based on these observations, the acute oral toxicity of the test substance in rats of both sexes observed for a period of 14 days was greater than 2000 mg/kg bw.

Acute dermal toxicity

In a GLP compliant dermal toxicity study, performed according to OECD guideline 402, Sprague-Dawley rats (5/sex) were administered the test substance (2000 mg/kg bw) (RTC 1995). The test substance was dissolved in water, applied on the skin and covered with an occlusive dressing for 24 hours. The treated skin was washed after 24 hours and a 14-day observation period followed. No mortality was observed during this period. Also no clinical signs and significant changes in body weight were observed. Furthermore, no abnormalities were found on necropsy.Therefore, the toxicity of the test substance was estimated to be >2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
Only study available

Justification for selection of acute toxicity – dermal endpoint
Only study available

Justification for classification or non-classification

Based on the observed LD50 of >2000 mg/kg bw in the acute oral and dermal toxicity study, the test substance does not need to be classified according to Directive 67/548/EEC and according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.