4-[(1-amino-9,10-dihydro-4-hydroxy-9,10-dioxo-2-anthryl)oxy]-N-(3-ethoxypropyl)benzenesulphonamide

Endpoint summary

Description of key information

The acute oral median lethal dose (LD₅₀) of Disperse Red 092 in the female Wistar rat was estimated to be greater than 2000 mg/kg body weight.

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:

20 July 2016 to 09 August 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

Identification: FAT 40444/B
Batch: PCR92X140707 (China)
Physical state/Appearance: dark red powder
Purity: 95.4 %
Expiry Date: 14 October 2019
Storage Conditions: approximately 4 °C in the dark

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Female Wistar (RccHan™:WIST) strain rats were supplied by Envigo RMS (UK) Limited, Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non- pregnant. After an acclimatization period of at least 5 days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card. At the start of the study the animals were 8 to 12 weeks of age. The body weight variation did not exceed ± 20 % of the mean body weight at the start of treatment.

Animal Care and Husbandry
The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet, drinking water, and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study. The temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70 % respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness. The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Route of administration:

oral: gavage

Vehicle:

other: Dimethyl sulphoxide

Remarks:

Dimethyl sulphoxide was used because the test item did not dissolve/suspend in distilled water or arachis oil BP

Details on oral exposure:

All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted body weight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each dose group to confirm the survival of the previously dosed animals.

Doses:

A single animal was treated with a dose level of 2000 mg/kg.
In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of four animals were treated at a dose level of 2000 mg/kg.

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

Test Item Preparation and Analysis
For the purpose of the study the test item was freshly prepared, as required, as a solution in dimethyl sulphoxide. Dimethyl sulphoxide was used because the test item did not dissolve/ suspend in distilled water or arachis oil BP. The test item was formulated within 2 hours of being applied to the test system. It is assumed that the formulation was stable for this duration. No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.

Study Design
Using available information on the toxicity of the test item, 2000 mg/kg was chosen as the starting dose.
A single animal was treated as follows:
Dose Level (mg/kg) Concentration (mg/mL) Dose Volume (mL/kg) Number of Rats
Female
2000 200 10 1

In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of animals was treated as follows:
Dose Level (mg/kg) Concentration (mg/mL) Dose Volume (mL/kg) Number of Rats
Female
2000 200 10 4

A total of five animals were therefore treated at a dose level of 2000 mg/kg in the study. All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted body weight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each dose group to confirm the survival of the previously dosed animals. Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for 14 days. Morbidity and mortality checks were made twice daily. Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14. At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Data Evaluation
The test item was classified according to Annex 3 of the OECD Guidelines for Testing of Chemicals No. 420 "Acute Oral Toxicity - Fixed Dose Method" (adopted 17 December 2001).
Evaluation of data included identification of the number of animals that died during the study (or that were killed for humane reasons), and determination of the nature, severity, onset and duration of the toxic effects. If possible, the signs of evident toxicity were described. Evident toxicity refers to the toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probable mortality. Effects on body weights and abnormalities noted at necropsy were also identified.
Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made.

Preliminary study:

There was no preliminary study.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths found.

Clinical signs:

other: Red colored staining of the feces was noted in the initial treated animal 2 to 4 days after dosing. No signs of systemic toxicity were noted during the observation period.

Gross pathology:

No abnormalities were noted at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral median lethal dose (LD50) of FAT 40444/B in the female Wistar rat was estimated to be greater than 2000 mg/kg body weight.

Executive summary:

An acute oral toxicity study was performed in female Wistar rat according to OECD 420 guideline. Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test item, as a solution in dimethyl sulphoxide, at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There were no deaths and signs of systemic toxicity. All animals showed expected gains in body weight. No abnormalities were noted at necropsy. Based on the study results, the acute oral median lethal dose (LD₅₀) of FAT 40444/A in the female Wistar rat was estimated to be greater than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Guideline and GLP compliance study

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

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute toxicity: oral

The acute oral toxicity of FAT 40444/B in the Wistar strain rat was evaluated using the fixed dose procedure (OECD 420 guideline). Following a combatability testing at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test item, as a solution in dimethyl sulphoxide, at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There were no deaths and no signs of systemic toxicity observed. All animals showed expected gains in body weight. No abnormalities were noted at necropsy. Hence, the acute oral median lethal dose (LD₅₀) of FAT 40444/B in the female Wistar strain rats was estimated to be greater than 2000 mg/kg body weight. 

 

Acute toxicity: inhalation

Currently no study to assess the acute inhalation toxicity potential of the target chemical, Disperse Red 092, is available. However, the vapour pressure for the substance is low (3.0 x 10^-13 Pa) and hence, the substance is considered to have low volatility. Synthesis and spray drying of this chemical is performed in a closed process and the final product consists of non-dusty granules. Hence, the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. Based on column 2, ‘Specific rules for adaptation from column 1’ of the table given in REACH Annex VIII, the study on acute inhalation toxicity only needs to be conducted if the 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 liquid formulation, the exposure via inhalation is considered to be unlikely. The chemical showed low toxicity potential in the available acute oral toxicity studies (LD₅₀>2000 mg/kg bw) with no mortality or systemic toxicity, hence it does not need to be classified for STOT SE. Taking the above arguments into account, low toxicity potential is expected on acute exposure of Disperse Red 092 via inhalation route and hence, testing by the inhalation route was considered scientifically not necessary.

 

Acute toxicity: dermal

Currently no study to assess the acute dermal toxicity potential of the target chemical, Disperse Red 092, is available. However, the molecular weight of the chemical is 496.53 g/mol, indicating limited dermal absorption. Synthesis and spray drying of this chemical is performed in a closed process without isolation of reaction products. Isolated products consist of either liquid formulations or dust free granules (non-dusty solid). In addition, the use of this substance will not result in aerosols, particles or droplets, so exposure to humans via the dermal route will be unlikely to occur. The water solubility of Disperse Red 092 was found to be low (8.1 mg/L), this indicates that the possibility of the substance partitioning from the stratum corneum into the epidermis will be low, thereby further limiting the absorption if any dermal exposure occurs. The chemical showed low toxicity potential in the available acute oral toxicity studies (LD₅₀>2000 mg/kg bw) with no mortality or systemic toxicity, hence, it does not need to be classified for STOT SE. Similarly, absence of systemic toxicity in skin sensitisation studies further supports the conclusion that low toxicity is expected for the chemical via the dermal route. Taking the above arguments into consideration, limited exposure and further reduced absorption is expected on acute exposure of Disperse Red 092 via dermal route. Hence, Disperse Red 092 is expected to have low toxicity potential on acute dermal exposure and testing by the dermal route was considered scientifically not necessary.

Justification for classification or non-classification

Based on the observed acute oral median lethal dose (LD₅₀) of >2000 mg/kg bw, Disperse Red 092 does not considered to be classified according to according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.