Reaction products of disodium 3,5-diamino-2-[(2-sulfonatophenyl)diazenyl]benzoate and  diazotised sodium 2-[(4-aminophenyl)sulfonyl]ethyl sulfate

Endpoint summary

Description of key information

FAT40810/A was assessed for oral and dermal acute toxicity according to OECD guideline study protocols. In both studies, no mortality was observed at a single dose of 2000 mg/kg bw and thus the LD50 dermal and LD50 oral in both studies were >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:

Experiment start date - 06 February 2003; Experiment completion date - 04 March 2003; Study completion date - 13 March 2003.

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

Identity: FAT 40810/A
Batch: WP 6/02
Purity: approx. 75 %
Appearance: Solid, dark brownish powder
Expiration date: 12 December 2010
Storage: At room temperature at about 20 °C

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test system: Rat, HanBrl: Wist (SPF)
Rationale: Recognized by the international guidelines as a recommended test system.
Source: RCC Ltd, Biotechnology and Animal Breeding, CH-4414 Füllinsdorf / Switzerland
Number of animals per group 3 males and 3 females
Total number of animals 3 males and 3 females
Age when treated: Males: 8 weeks, Females: 12 weeks
Identification: Unique cage number and corresponding color-coded spots on the tail. The animals were marked immediately prior to treatment.
Randomization: Selected by hand at time of delivery. No computer generated randomization program.
Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.
Husbandry: Room no. 104/ RCC Ltd, Füllinsdorf
Conditions: Standard Laboratory Conditions. Air-conditioned with 10 – 15 air changes per hour, and continuously monitored environment with target ranges for temperature 22 ± 3 °C and for relative humidity between 30 - 70 % (values above 70 % during cleaning process possible), 12 hours fluorescent light 12 hours dark, music during the light period.
Accommodation: In groups of three per sex in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).
Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 67/02 (Provimi Kliba AG, CH-4303 Kaiseraugst Switzerland) ad Iibitum. Results of analyses for contaminants are archived at RCC Ltd, ltingen.
Water: Community tap water from Füllinsdorf ad Iibitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd, ltingen.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

The animals received a single dose of the test item by oral gavage administration at 2000 mg/kg body weight after being fasted for 17 to 19 hours (access to water was permitted). Food was provided again 3 hours after dosing. The application volume was 10 ml/kg body weight.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3 females and 3 males

Control animals:

no

Details on study design:

The study strictly followed the OECD 423 toxic class methodology and in lack of mortality and other effects at 2000 mg/kg bw no further doses were tested. Animals were observed for 15 days following dosing for mortality/viability (daily), body weight (prior to administration, on days 8 and 15) and clinical signs (daily). Following the observation period, the animals were killed by intraperitoneal injection and macroscopic examinations were performed.

Statistics:

No statistics was applied in lack of symptoms/mortality.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Male: 2000 mg/kg bw; Number of animals: 3; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 3; Number of deaths: 0

Clinical signs:

other: No clinical signs were observed during the course of the study. No deaths occurred during the study. CLINICAL SIGNS: No clinical signs were observed in any animals. BODY WEIGHTS: The body weight of all animals was within the range commonly

Gross pathology:

Effects on organs: No macroscopic findings were observed at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

Acute LD50 (oral, rat) of FAT 40810/A: >2000 mg/kg bw

An acute oral toxicity study was conducted in accordance with OECD test guideline 423 and EEC B.1 to assess toxicity of FAT 40810/A upon single administration.

Executive summary:

An acute oral toxicity study was conducted in accordance with OECD test guideline 423 and EEC B.1 to assess toxicity of FAT 40810/A upon single administration. Three male and three female HanBrl: WIST (SPF) rats were treated with FAT 40810/A by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (purified water) at a concentration of 0.2 g/ml and administered at a volume dosage of 10 ml/kg. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded twice daily during test days 1-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically. All animals survived until the end of the study period. No clinical signs were observed during the course of the study. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy. The median lethal dose of FAT 40810/A after single oral administration to rats of both sexes, observed over a period of 14 days is: LD50 (rat): greater than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

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:

Experiment start date - 06 February 2003; Experiment end date - 27 February 2003; Study completion date - 13 March 2003.

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

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

Identity: FAT 40810/A
Batch: WP 6/02
Purity: approx. 75 %
Appearance: Solid, dark brownish powder
Expiration date: 12 December 2010
Storage: At room temperature at about 20 °C

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test system: Rat, HanBrl: Wist (SPF)
Rationale: Recognized by the international guidelines as a recommended test system.
Source: RCC Ltd, Biotechnology and Animal Breeding, CH-4414 Füllinsdorf / Switzerland
Number of animals per group 3 males and 3 females
Total number of animals 3 males and 3 females
Age when treated: Males: 8 weeks, Females: 12 weeks
Identification: Unique cage number and corresponding color-coded spots on the tail. The animals were marked immediately prior to treatment.
Randomization: Selected by hand at time of delivery. No computer generated randomization program.
Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.
Husbandry: Room no. 104/ RCC Ltd, Füillinsdorf
Conditions: Standard Laboratory Conditions. Air-conditioned with 10 – 15 air changes per hour, and continuously monitored environment with target ranges for temperature 22 ± 3 °C and for relative humidity between 30 - 70 % (values above 70 % during cleaning process possible), 12 hours fluorescent light 12 hours dark, music during the light period.
Accommodation: In groups of three per sex in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).
Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 67/02 (Provimi Kliba AG, CH-4303 Kaiseraugst Switzerland) ad Iibitum. Results of analyses for contaminants are archived at RCC Ltd, ltingen.
Water: Community tap water from Füllinsdorf ad Iibitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd, ltingen.

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

Bi-distilled

Details on dermal exposure:

One day before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10 % of the total body surface. Only those animals without injury or irritation on the skin were used in the test. On test day 1, the test item was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe and covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage (application volume/kg body weight: 6 ml). Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels. Thereafter, the reaction sites were assessed.

Duration of exposure:

24 h

Doses:

2000 mg /kg bw

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

Observations:
Mortality / Viability: Daily during acclimatization and twice daily during days 1-15.
Body weights: On test days 1 (prior to administration), 8 and 15.
Clinical signs: Daily during acclimatization and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15.
Necropsy: All animals were killed at the end of the observation period by an intraperitoneal injection of Vetanarcol at a dose of at least 2.0 ml/kg body weight (equivalent to at least 324 mg sodium pentobarbitone/kg body weight) and discarded after macroscopic examinations were performed. 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.

Mortality:

Male: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0

Clinical signs:

other: No clinical signs were observed during the observation period. Slight red staining of the treated skin area was observed in all animals immediately after removal of the dressing and persisted up to the end of the observation period.

Gross pathology:

Effects on organs: No macroscopic findings observed at necropsy.

Other findings:

Slight red staining of the treated skin area was observed in all animals immediately after removal of the dressing and persisted up to the end of the observation period.

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 (dermal, rat): >2000 mg/kg bw

Executive summary:

In a GLP-study conducted according to OECD test guideline 402, FAT 40810/A was assessed for its toxicity potential upon single application to skin. Five male and five female HanBrI: WIST (SPF) rats were treated with FAT 40810/A at 2000 mg/kg by dermal application. The test item was diluted in vehicle (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. No deaths occurred during the study. No clinical signs were observed during the observation period. Slight red staining of the treated skin area was observed in all animals immediately after removal of the dressing and persisted up to the end of the observation period. Three female animals showed a loss of body weight (0.3 % to 6.2%) one week after treatment whereas one other female animal did not gain body weight during the same period. They recovered at the end of the observation period. The body weight of all other animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy except the slight red staining of the treated skin area which was still present. In conclusion the median lethal dose of FAT 40810/A after single dermal administration to rats of both sexes, observed over a period of 14 days is LD50 (rat):greater than 2000 mglkg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Acute oral toxicity:

An acute oral toxicity study was conducted in accordance with OECD test guideline 423 and EEC B.1 to assess toxicity of FAT 40810/A upon single administration. Three male and three female HanBrl: WIST (SPF) rats were treated with FAT 40810/A by oral gavage administration at a dosage of 2000 mg/kg body weight. All animals survived until the end of the study period. No clinical signs were observed during the course of the study. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy. The median lethal dose of FAT 40810/A after single oral administration to rats of both sexes, observed over a period of 14 days is: LD50 (rat): greater than 2000 mg/kg body weight.

Acute dermal toxicity:

In a GLP-study conducted according to OECD test guideline 402, FAT 40810/A was assessed for its toxicity potential upon single application to skin. Five male and five female HanBrI: WIST (SPF) rats were treated with FAT 40810/A at 2000 mg/kg by dermal application. The application period was 24 hours. No deaths occurred during the study. No clinical signs were observed during the observation period. Slight red staining of the treated skin area was observed in all animals immediately after removal of the dressing and persisted up to the end of the observation period. Three female animals showed a loss of body weight (0.3 % to 6.2 %) one week after treatment whereas one other female animal did not gain body weight during the same period. They recovered at the end of the observation period. The body weight of all other animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy except the slight red staining of the treated skin area which was still present. In conclusion the median lethal dose of FAT 40810/A 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. 

Acute inhalation toxicity:

Currently no study to assess the acute inhalation toxicity potential of Reactive Orange 140 is available. However, the vapour pressure for the substance was found to be 6.6E-25 to 3.8E-26 Pa which is considered to be negligible. Hence, the substance is considered to have low volatility. Based on column 2, ‘Specific rules for adaptation from column 1’ of the table given in REACH Annex VII, the study on acute inhalation toxicity only needs to be conducted if an exposure via inhalation is to be expected, based on vapour pressure and/or the likelihood of an exposure to aerosols, particles or droplets. Referring to the expected low volatility of the substance, the fact that the substance is imported into the EU in a formulated form as a dust-free powder or as a granulate, the exposure via inhalation is considered to be unlikely. Reactive Orange 140 was found to have water solubility of 224 g/L, hence, vapours of this very hydrophilic substance may be retained within the mucus and cleared through mucociliary system. Further, the chemical was found to have low acute toxicity when tested via oral route with no mortality up to 2000 mg/kg bw. Hence, considering all the above arguments, it is considered that Reactive Orange 140 has a low toxicity potential via inhalation route and thus the study on acute inhalation toxicity is considered scientifically not necessary.

Justification for classification or non-classification

Based on the results of an oral and a dermal acute toxicity study the substance FAT40810/A is not classifiable for acute oral and dermal toxicity according to CLP (Regulation EC No 1272/2008).