Trisodium 5-[[4-chloro-6-(ethylphenylamino)-1,3,5-triazin-2-yl]amino]-4-hydroxy-3-[(1-sulphonato-2-naphthyl)azo]naphthalene-2,7-disulphonate

Endpoint summary

Description of key information

The acute oral and dermal LD50 of FAT 40147 is greater than 2000 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:

1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

None

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Principles of method if other than guideline:

None

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

None

Species:

rat

Strain:

other: Tif:RAIf (SPF)

Remarks:

F3-crosses of RII 1/Tif x RII 2/Tif

Sex:

male/female

Details on test animals or test system and environmental conditions:

Species and strain: Rat, Tif:RAIf(SPF), F3-crosses of RII 1/Tif x RII 2/Tif
Source: CIBA-GEIGY LTD. Tierfarm, 4334 Sisseln, Switzerland
Initial body weight range: 168-203 g
Initial age: 7-8 weeks
Individual identification: by colour code using picric acid
Husbandry: The animals were kept under conventional laboratory conditions. They were caged in groups of 5 in Macrolon cages type 4 with standardized soft wood bedding (Société Parisienne des sciures, Pantin).
The animal room was air conditionned: temperature 22±3° C, relative humidity 55±15%, 12 hours light/day, approximately 15 air changes/h.

Diet: Rat food, NAFAG No. 890, NAFAG AG, Gossau, SG (Switzerland), and water were provided ad libitum.

Route of administration:

oral: gavage

Vehicle:

other: Distilled water containing 0.5% carboxymethylcellulose and 0.1% polysorbate 80.

Details on oral exposure:

Prior to dosing, the animals were fasted overnight.
Administration: oral, by gastric intubation (gavage)
Volume (ml/kg body weight) applied: 10 and 20

Doses:

2000 and 5000 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 or until all symptoms have disappeared, whichever lasts longer,
- Mortality: daily; a.m. and p.m. on working days, a.m. on weekend days,
- Signs and symptoms: daily,
- Weighing: on days 1, 7, 14 and at death,
- Necropsy of survivors performed: yes,
Spontaneously dying animals were submitted to a gross necropsy as soon as possible; survivors at the end of the observation period.

Statistics:

From the body weights, the group means and their standard deviations were calculated.
Where feasible, the LD50 including the 95 % confidence limit were computed by the logit method (J. Berkson, J.Am. Stat. Ass. 39. 357-65, 1944).

Preliminary study:

None

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred at the dose of 2000 mg/kg bw. However, at 5000 mg/kg bw, 3 males and 5 females were found dead within day 1 after treatment.

Clinical signs:

other: Dyspnoea, ruffled fur, diarrhoea and curved body position were seen with the animals treated at 2000 mg/kg bw. Animals treated with 5000 mg/kg bw, also showed sedation, exophthalmos , ventral body position and lateral body position in addition to the prev

Gross pathology:

No gross lesions were found in the animals of the 2000 mg/kg group. With the exception of the two survivors, all animals of the 5000 mg/kg groups had blue stained bodies.

Other findings:

None

None

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance was determined to have a LD50 of >2000 mg/kg bw.

Executive summary:

The acute oral toxicity of the test substance was evaluated in a study conducted according to OECD Guideline 401. Two groups of 5 male and 5 female rats received the test item at the doses of 2000 and 5000 mg/kg bw by oral intubation. Observations of mortality and clinical signs were carried out after 1, 2, 3, 5 and 24 hrs of dosing, and daily thereafter for 14 days; while weighing was done on days 1, 7 and 14. Spontaneously dying animals were submitted to a gross necropsy as soon as possible; survivors at the end of the observation period. No mortality occurred at the dose of 2000 mg/kg bw. However, at 5000 mg/kg bw, 3 males and 5 females were found dead within day 1 after treatment. Dyspnoea, ruffled fur, diarrhoea and curved body position were seen with the animals treated at 2000 mg/kg bw. Animals treated with 5000 mg/kg bw, also showed sedation, exophthalmos, ventral body position and lateral body position in addition to the previously described signs. The surviving animals recovered within 9-10 days. No effect on body weight gains was seen. No gross lesions were found in the animals of the 2000 mg/kg group. With the exception of the two survivors, all animals of the 5000 mg/kg groups had blue stained bodies. Hence, based on the above findings, the test substance was determined to have a LD50 of >2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Good quality studies

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:

2010-01-05 till 2010-01-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See chapter 13 for detailed read across justification.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

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:

TEST ANIMALS RccHan: WIST(SPF)
- Source: Harlan Laboratories B.V. Kreuzelweg 53 5961 NM Horst / The Netherlands Postbus 6174 5960 AD Horst / The Netherlands
- Age at study initiation: Males: 9 weeks, Females: 11 weeks
- Weight at study initiation: Males 244.8 - 261.9 g ; Females: 191.8 - 210.3g
- Fasting period before study: no data
- Housing: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 30/09 (Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) ad libitum.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum.
Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): relative humidity between 30-70% (values above 70% during cleaning process possible)
- Air changes (per hr): Air-conditioned with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.

Type of coverage:

semiocclusive

Details on dermal exposure:

TEST SITE
- Area of exposure: no data
- % coverage: approx. 10% of the body surface
- Type of wrap if used: skin with a syringe and covered with a surgical gauze pad (ca. 5 x 5 cm) held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and an elastic adhesive restrainer bandage wrapped around the abdomen.


REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, flushed with lukewarm water and drapped off with disposable paper towels.
- Time after start of exposure: 24h

TEST MATERIAL AND VEHICLE
- Amount(s) applied (volume or weight with unit): 6mL/kg
- Concentration (if solution): dose: 2000mg/kg BW
- Constant volume or concentration used: yes
- test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle was added (weight:volume).

Duration of exposure:

24h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:

Viability / Mortality: Daily during the acclimatization period, within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15.

Clinical Signs: Daily during the acclimatization period, within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15.

Local Dermal Signs: Once daily during days 2 (following dressing removal) through day 15 using the numerical scoring system described in Appendix I.

Body Weights: On test days 1 (prior to administration), 8 and 15.

- Necropsy of survivors performed: yes
- Other examinations performed: macroscopic examinations were performed. An external examination and opening of the abdominal and thoracic cavities for examinations of major organs were performed. The appearance of any macroscopic abnormalities was recorded.No organs or tissues were retained.

Statistics:

No statistical analysis was used.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no statistical analysis was performed

Mortality:

No intercurrent deaths occurred during the course of the study.

Clinical signs:

other: No clinical signs were recorded throughout the complete observation period.

Gross pathology:

No macroscopic findings were recorded at necropsy.

Other findings:

In all animals, strong violet staining produced by the test item was noted on the treated skin area from test day 2 up to test day 6 (males) or 9 (females). However, this staining prevented the assessment of a possible erythema. When assessable, no local dermal signs were observed in all
animals.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The median lethal dose of FAT 40850/A TE after single dermal administration to rats of both sexes, observed over a period of 14 days, is: LD50 (rat): greater than 2000 mg/kg body weight

Based upon the classification criteria according to the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, FAT 40850/A TE does not have to be classified with respect to acute dermal toxicity in the rat.

Executive summary:

Five male and five female RccHan:WIST (SPF) rats were treated with FAT 40850/A TE at 2000 mg/kg by dermal application in accordance with EU Method B.3 and GLP. The test item was formulated in purified water at a concentration of 0.33 g/mL and administered at a volume dosage of 6 mL/kg. The application period was 24 hours. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2-15. Local signs were evaluated once daily after removal of the dressing on test day 2 up to test day 15. Mortality/viability was recorded within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

No intercurrent deaths occurred during the course of the study. No clinical signs were recorded throughout the complete observation period. In all animals, strong violet staining produced by the test item was noted on the treated skin area from test day 2 up to test day 6 or 9. However, this staining prevented the assessment of possible erythema. When assessable, no local dermal signs were observed in all animals. The body weight of all animals was considered to be within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy.

The median lethal dose of FAT 40850/A TE after single dermal administration to rats of both sexes, observed over a period of 14 days, is: LD50 (rat): greater than 2000 mg/kg body weight

Based upon the classification criteria according to the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, FAT 40850/A TE does not have to be classified with respect to acute dermal toxicity in the rat.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Study conducted on a similar substance FAT 40850 in accordance to OECD guideline 402 and GLP.

Additional information

Acute oral toxicity:

Four studies are available for the assessment of oral toxicity potential of the substance.

In a key study (1984), the acute oral toxicity of the test substance was evaluated in a study conducted according to OECD Guideline 401. Two groups of 5 male and 5 female rats received the test item at the doses of 2000 and 5000 mg/kg bw by oral intubation. Observations of mortality and clinical signs were carried out after 1, 2, 3, 5 and 24 hrs of dosing, and daily thereafter for 14 days; while weighing was done on days 1, 7 and 14. Spontaneously dying animals were submitted to a gross necropsy as soon as possible; survivors at the end of the observation period. No mortality occurred at the dose of 2000 mg/kg bw. However, at 5000 mg/kg bw, 3 males and 5 females were found dead within day 1 after treatment. Dyspnoea, ruffled fur, diarrhoea and curved body position were seen with the animals treated at 2000 mg/kg bw. Animals treated with 5000 mg/kg bw, also showed sedation, exophthalmus, ventral body position and lateral body position in addition to the previously described signs. The surviving animals recovered within 9-10 days. No effect on body weight gains was seen. No gross lesions were found in the animals of the 2000 mg/kg group. With the exception of the two survivors, all animals of the 5000 mg/kg groups had blue stained bodies. Hence, based on the above findings, the test substance was determined to have a LD50 of >2000 mg/kg bw.

In addition to the above key study, three supporting studies are available.

The acute oral toxicity of the test substance was evaluated in a study (1982) conducted according to OECD Guideline 401. Two groups of 5 male and 5 female rats received the test item at the doses of 2500 and 5000 mg/kg bw by oral intubation. Observations of mortality and clinical signs were carried out after 1, 3, 5 and 24 hrs of dosing, and daily thereafter for 14 days; while weighing was done on days 1, 7 and 14. Spontaneously dying animals were submitted to a gross necropsy as soon as possible; survivors at the end of the observation period. No mortality occurred at the dose of 2500 mg/kg bw. However, at 5000 mg/kg bw, 2 males and 3 females were found dead within 5 hours after treatment. Clinical signs like dyspnoea, exophthalmus, ruffled fur, diarrhoea and curved body position were seen with the animals treated at 2500 mg/kg bw. Animals treated with 5000 mg/kg bw, also showed sedation in addition to previously described signs. The surviving animals recovered within 10-12 days. No effect on body weight gains was seen. No substance related gross organ changes were seen during the gross pathological examination. Hence, based on the above findings, the test substance was determined to have a LD50 of >2500 mg/kg bw.

In another supporting study (1981), the acute toxicity of the test substance was evaluated in a study according to the methodology equivalent to OECD Guideline 401. Two groups of 5 male and 5 female rats received the test item at the doses of 2500 and 5000 mg/kg bw by oral intubation. Observations of mortality and clinical signs were carried out after 1, 2, 3, 5 and 24 h of dosing, and daily thereafter for 14 days; while weighing was done on days 1, 7 and 14. They were submitted at random for necropsy whenever they died, survivors at the end of the observation period. Two females died at 5000 mg/kg bw within 3 hours after treatment. At 2500 mg/kg bw, no mortality was seen. Dyspnoea, exophthalmos, curved body position, diarrhoea and ruffled fur were the clinical signs observed at both dose levels. The surviving animals recovered within 10-11 days. Two females died at 5000 mg/kg bw within 3 hours after treatment. At 2500 mg/kg bw, no mortality was seen. No effect on body weight gains was seen. No substance related gross organ changes were seen during the gross pathological examination. Hence, based on the above findings, the test substance was determined to have a LD50 of >5000 mg/kg bw.

Another supporting study (1981, oral-1), the acute toxicity of the test substance was evaluated according to the methodology equivalent to OECD Guideline 401. A group of 5 male and 5 female rats received the test item at the dose of 5000 mg/kg bw by oral intubation. Observations of mortality and clinical signs were carried out after 1, 2, 3, 5 and 24 h of dosing, and daily thereafter for 14 days; while weighing was done on days 1, 7 and 14. They were submitted at random for necropsy whenever they died, survivors at the end of the observation period. No mortality was seen throughout the duration of the observation period. Dyspnoea, curved body position and ruffled fur were the clinical signs observed. The animals recovered within 8 days. No effect on body weight gains was seen. No substance related gross organ changes were seen during the gross pathological examination. Hence, based on the above findings, the test substance was determined to have a LD50 of >5000 mg/kg bw.

Hence, based on the above information, FAT 40147 is considered to have an acute oral LD50 value of >2000 mg/kg bw.

Acute inhalation toxicity:

Currently no study to assess acute inhalation toxicity is available. But due to the intrinsic properties of the test item and the high values of the acute oral toxicity studies available no adverse effects are expected via the inhalation route. Experience with similar chemical structures demonstrated that it is very unlikely that toxicity related to intrinsic properties of the test item only show up via the inhalation route and not via the oral-gastric route of exposure.

 

Acute dermal toxicity:

Currently no study is available investigating acute dermal toxicity of the test item. Nevertheless, data from a Read across substance FAT 40850 is available. Five male and five female RccHan:WIST (SPF) rats were treated with FAT 40850/A TE at 2000 mg/kg by dermal application in accordance with EU Method B.3 and GLP. The test item was formulated in purified water at a concentration of 0.33 g/mL and administered at a volume dosage of 6 mL/kg. The application period was 24 hours. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. At the end of the observation period of 14 days all animals were necropsied and examined macroscopically.

 

No intercurrent deaths occurred during the course of the study. No clinical signs were recorded throughout the complete observation period. In all animals, strong violet staining produced by the test item was noted on the treated skin area from test day 2 up to test day 6 or 9. However, this staining prevented the assessment of possible erythema. When assessable, no local dermal signs were observed in all animals. The body weight of all animals was considered to be within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy.

 

The median lethal dose of FAT 40850/A TE after single dermal administration to rats of both sexes, observed over a period of 14 days, is: LD50 (rat): greater than 2000 mg/kg body weight. Taking these results into consideration no further studies to assess dermal toxicity will not be conducted.

Justification for classification or non-classification

Based on the results of available acute oral and dermal toxicity studies, the substance does not warrant classification according to the CLP (Regulation 1272/2008) criteria.