Salts of 9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethylxanthenium chloride and 4-{[1-hydroxy-6-({[5-hydroxy-6-(phenyldiazo)-7-sulfonaphthalen-2-yl]carbamoyl}amino)-3-sulfonaphthalen-2-yl]diazo}benzoic acid (2:1)

Endpoint summary

Description of key information

In an acute oral toxicity study with rats the LD50-value was determined > 2000 mg/kg bw (limit dose). An acute dermal toxicity study revealed a LD50-value of > 2000 mg/kg bw (limit dose). The LC50 (acute worker)inhalation = 3526 mg/m3 was calculated assuming 70 kg per person and exposure for 8 h during light activity. For the general population the LD50 (acute general population)inhalation = 1500 mg/m3 was calculated assuming equal breathing volume (20 m3), a body weight of 60 kg/person and exposure over 24 h at basal caloric demand.

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 2008-03-20 to 2008-04-04

Reliability:

1 (reliable without restriction)

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

rat, Wistar Crl:(WI)BR (outbred, SPF-Quality)
nulliparous and non-pregnant

Route of administration:

oral: gavage

Vehicle:

other: 1 % aqueous methylcellulose

Doses:

2000 mg/kg (10 mL/kg) bw (limit dose)

No. of animals per sex per dose:

3 females (nulliparous, non pregnant)/group
(2 treatment groups, single oral administration at limit dose 2000 mg/kg bw tested)

Control animals:

no

Details on study design:

The acute toxic class method according to OECD 423 and Directive 2004/73/EC B.1. tris was performed with F 213 Red in rats. Two groups of 3 female CRL:(WI) BR Wistar rats were treated with F 213 Red by single oral gavage at dose levels of 2000 mg/kg bw (Treatment groups 1 and 2) in 1 % aqueous methylcellulose in two independent experiments. Based on the results of the study and according to the criteria of the relevant test guidelines no further testing was performed. The concentration of the formulations was 200 mg/mL in both steps to ensure the constant treatment volume of 10 mL/kg bw.

Statistics:

no data

Preliminary study:

not applicable

Sex:

female

Dose descriptor:

LD50

Effect level:

2 000 mg/kg bw

Mortality:

In both treatment groups one female CRL:(WI) BR rat died after single oral administration of F213 Red at 2000 mg/kg bw dose level.

Clinical signs:

Treatment group 1 (2000 mg/kg bw): Decreased activity (2/3), squatting position (2/3), swollen abdomen (1/3), piloerection (1/3), dyspnoea (1/3), noisy breathing (1/3) and emaciation (1/3) were observed in the first treatment group. The first clinical signs appeared from 1st and 3rd hours after the treatment and ceased on day 1. After a transient (4 days) recovery, clinical signs appeared again, and significant loss of the body weight was noted for animal found dead on day 14. In the bedding material, reddish coloured urine and stool were observed from the 2nd hour, which disappeared on day 1 and day 4, respectively.

Treatment group 2 (2000 mg/kg bw): Slight activity decrease and squatting position were observed on animal No.: 9883 3 hours after the treatment. One day thereafter, marked activity decrease, squatting position, piloerection and dyspnoea were noted and this animal was found dead on day 2. Two animals of this group were symptom-free during the day of the treatment and following 14-day observation period. In the bedding material, reddish coloured urine and stool were observed from the 2nd hour, which disappeared on day 1 and day 4, respectively.

Body weight:

Treatment group 1 (2000 mg/kg bw): A significant body weight loss was noted for animal No.: 9844 (died on day 14) on weeks 1 and 2. The body weight development of surviving animals was considered to be normal.

Treatment group 2 (2000 mg/kg bw ): There was no test item influence on the body weight and body weight gain of these animals.

Gross pathology:

Treatment group 1 (2000 mg/kg bw): In dead animal (No.: 9844), under nourishment, dark red coloured left lobe of lungs, congestive liver and empty, gas filled stomach and intestines were observed. In surviving animals (No.: 9833 and 9856) pale raised areas were found in the lungs.

Treatment group 2 (2000 mg/kg bw): Animal found dead (No.: 9883) showed dark red lungs, congestive liver and reddish content in the stomach and coecum. In surviving animals (No.: 9868 and 9870), pinprick sized haemorrhages were observed in the lungs. In summary, reddish content in the stomach and coecum was indicative of the presence of the test item in the gastro-intestinal tract in dead animal of Treatment group 2. The pulmonary alterations (dark red colour) and congestive liver of dead animals were signs of circulatory disturbance developed during the death. The pale raised areas and haemorrhages in the lungs were due to the method of anaesthesia and exsanguination, which are also observable in untreated animals after anaesthesia.

Other findings:

not applicable

no remarks

Interpretation of results:

Category 5 based on GHS criteria

Remarks:

Migrated information

Conclusions:

Under the conditions of the present study, a single oral administration of the test item F 213 Red caused death of female CRL:(WI)BR rats at 2000 mg/kg bw (2/6). According to the Globally Harmonised Classification System, the acute oral LD50 value of F 213 Red was greater than 2000 mg/kg body weight in female CRL:(WI) BR rats and it was ranked into Category 5 with a cut off value of 2500 mg/kg bw. According to directive 2001/59/EC, classification of F 213 Red by the oral route is not required based on the results of this study.

Executive summary:

he acute toxic class method according to OECD 423 and Directive 2004/73/EC B.1. tris was performed with F 213 Red in rats. Two groups of 3 female CRL:(WI) BR Wistar rats were treated with F 213 Red by single oral gavage at dose levels of 2000 mg/kg bw (Treatment groups 1 and 2) in 1 % aqueous methylcellulose in two independent experiments. Based on the results of the study and according to the criteria of the relevant test guidelines no further testing was performed. The concentration of the formulations was 200 mg/mL in both steps to ensure the constant treatment volume of 10 mL/kg bw.

Clinical signs:

Treatment group 1 (2000 mg/kg bw): One animal of the first treatment group was found dead on day 14. Clinical signs observed from 1st and 3rd hour after the treatment (dyspnoea, activity decrease, squatting position and piloerection) ceased on day 1. After a transient (4 days) recovery, decreased activity, squatting position, swollen abdomen, piloerection, dyspnoea, noisy breathing and significant loss of the body weight were found. Gross pathology revealed no test item related macroscopic findings. In one of surviving animals, decreased activity and squatting position were observed between the third and six hours after the treatment. There were no body weight changes and necropsy findings related to treatment with F 213 Red. Reddish coloured urine and stool were observed in the bedding material from 2nd hour, which disappeared on day 1 and day 4, respectively.

Treatment group 2 (2000 mg/kg bw): Decreased activity, squatting position, piloerection and dyspnoea were noted for animal found dead on day 2 of study. Residue of the test item was noted in the stomach and coecum at the necropsy. Remaining two animals of this group were symptom-free during the entire observation period. The body weight development was considered to be normal. No macroscopic findings related to the test item effect were observed. Reddish coloured urine and stool were observed in the bedding material from 2nd hour, which disappeared on day 1 and day 4, respectively.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Dose descriptor:

LC50

Value:

1 500 mg/m³

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 2008-03-20 to 2008-04-03

Reliability:

1 (reliable without restriction)

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

rat, Wistar Crl:(WI)BR (SPF-Quality)
Male and female (nulliparous, non pregnant)
Number of animals: 5 animals/sex
Acclimatisation time: 8 days

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

The test item was applied as a single dose uniformly over an area of approximately 10 % of the total body surface for a 24-hour exposure period. Test item was administered in its original form, it was pulverised and was moistened sufficiently with water to ensure good contact with the skin. Sterile gauze pads were placed on the skin of the rats and kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semiocclusive plastic wrap for 24 hours. At the end of the exposure period, residual test item was removed using water at body temperature.

Duration of exposure:

24 hours

Doses:

The test item was administered in a single dose of 2000 mg/kg bw.

No. of animals per sex per dose:

5 animals

Control animals:

not required

Details on study design:

not applicable

Statistics:

not applicable

Preliminary study:

not applicable

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No mortality observed.

Clinical signs:

No clinical signs observed. Behaviour and physical condition of animals were considered to be normal.

Body weight:

The mean body weight and the body weight gain of the male and female animals were considered to be normal during the two-week observation period, similar to the expected values in untreated animals of the same age and strain.

Gross pathology:

No macroscopic alterations due to the effects of the test item were found. Gross necropsy revealed pale raised areas (2/5 male; 2/5 female) and pinprick-sized haemorrhages (3/5 male, 3/5 female) in the lungs due to the method of anaesthesia and exsanguination, which are also observable in untreated animals after anaesthesia. Two animals showed hydrometra related to the sexual cycle and which is a frequent observation in experimental rats.

Other findings:

Due to the colour of F 213 Red, 2000 mg/kg bw of the test item caused red colouration of the skin. Skin recovered 11 days after the patch removal. The alteration thus being temporary, therefore considered not toxicologically relevant.

no remarks

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute dermal LD50 value of F 213 Red is greater than 2000 mg/kg bw in male and female CRL: (WI) BR rats. F 213 Red does not meet the criteria for classification according to EU labelling regulations Commission Directive 2001/59/EC for classification and labelling of dangerous substances.

Executive summary:

Under the conditions of the present study, a single 24-hour dermal administration of 2000 mg/kg bw of the test item, F 213 Red caused no toxic changes on the treated skin. Mortalities were not observed, neither in male nor in female CRL:(WI)BR rats. This acute toxicity study by the dermal route did not indicate skin irritation at the limit dose level of 2000 mg/kg body weight tested. The acute dermal LD50 value of F 213 Red is greater than 2000 mg/kg bw in male and female CRL: (WI) BR rats. F 213 Red does not meet the criteria for classification according to EU labelling regulations Commission Directive 2001/59/EC for classification and labelling of dangerous substances.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Justification for classification or non-classification

Based on the acute toxicity results obtained for F 213 Red, no classification is warranted.