2-[[3-acetamido-4-[(2-chloro-4-nitrophenyl)azo]phenyl](2-cyanoethyl)amino]ethyl acetate

Endpoint summary

Description of key information

Oral LD50 (male and female) > 5350 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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

The read across approach is detailed into the document attached into the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

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
- Source: animals were purchased from KFM, CH-4414 Fülllinsdorf, Switzerland.
- Age at study initiation: young adult, age not specified.
- Weight at study initiation: ranged between 140 and 180 g.
- Fasting period before study: access of food only was prevented approximately 18 hours prior and four hours after the dosing. The water bottles were withdrawn two hours prior and four hours after dosing.
- Housing: housed individualry in Macroron cages.
- Diet: standard laboratory pelleted diet (KLIBA no. 24-343-7 from Klingentatmühle AG, Basle). The batch of diet used for the study was analysed for chernical and microbiological contaminants.
- Water: ad libitum.
- Acclimation period: animals were acclimatised to the experimental environment for a period of about five days prior to the start of the study.

ENVIRONMENTAL CONDITIONS
- Temperature: 23 ± 2 °C
- Humidity: 50 ± 10 %
- Air changes: approximately 15 changes/hour.
- Photoperiod: the light/dark cycle was 12 hours.

Route of administration:

oral: gavage

Vehicle:

DMSO

Details on oral exposure:

- Test fomulation: test item was dissolved in DMSO; the test sample was prepared immediately before dosing.
- Dose volume: 15 mI per kg body weight.

Doses:

5350 mg/kg bw, higher dosages were not applicable.

No. of animals per sex per dose:

Five male and five female.

Details on study design:

- Duration of observation period following administration: the animals, on both the preliminary and the main studies, were observed for 14 days after dosing.
- Frequency of observations: animals were observed soon after dosing, then at hourly intervals for the remainder day 1. On the subsequent days the animals were observed once in the morning and once in the late afternoon. Clinical signs were recorded at each observation.
- Frequency of weighing: individual body weights were recorded on day 1, 7 and 14.
- Necropsy of survivors performed: surviving animals were killed after two weeks. All animals which died during the study and those killed after two weeks were subjected to a macroscopic postmortem examination. The macroscopic appearance of the abnormal organs was recorded.

PRELIMINARY TEST
A preliminary study was carried out to establish a dosing regimen, using groups of two males and two females at two dosage, using 10-15 ml/kg body weight.

Statistics:

The acute oral LD5O was determined using the probit method of L.C. Miller and M.L. Tainter (Proc. Soc. exper. Biol. Med. 57 (1944) p. 26) or calculated by computer using the method of D.J. Finney (3rd. ed., 1971). The given doses aiways referred to the test material.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 350 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No death occurred during the range-finding experiment.
During the main experiment, one female died 59 hours after dosing.

Clinical signs:

No overt symptoms of poisoning were observed.

Gross pathology:

No special findings could be detected.

Mortality ratio and group mean bodyweight (g) of rats

Sex	Dosage	Bodyweight (g) at			Mortality ratio
		Dosing	1 wk	2wks
Male	Test item, 5350 mg/kg bw	177	207	246	0/5
Female	Test item, 5350 mg/kg bw	166	163	183	1/5

Interpretation of results:

other: not classified, according to CLP Regulation (EC 1272/2008)

Conclusions:

LD50 > 5350 mg/kg bw (males and females)

Executive summary:

Test item was dissolved in DMSO; the test sample was prepared immediately before dosing. A preliminary study was carried out to establish a dosing regimen, using groups of two males and two females at two dosage, using 10-15 ml/kg body weight. In the main experiment, groups of five male and five female rats were dosed at levels selected using the results of the preliminary study. The animals were dosed by oral gavage by means of a stomach tube and the dosed volume was 15 ml per kg body weight. The animals on both, the preliminary and the main studies were observed for 14 days after dosing.

Surviving animals were killed after two weeks. All animals which died during the study and those killed after two weeks were subjected to a macroscopic postmortem examination. The macroscopic appearance of the abnormal organs was recorded.

One female died 59 hours after dosing. No overt symptoms of poisoning were observed. No special findings could be detected.

Conclusion

LD50 > 5350 mg/kg bw (males and females)

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 350 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

ACUTE ORAL TOXICITY

There are no experiments on the oral acute toxicity potential of Disperse Red 202, thus, available data on the structural analogue Similar Substance 02 has been taken into consideration. The read across approach can be considered as suitable and appropriate to investigate the propriety (details on the read across are given into the document attached into the IUCLID section 13).

A study was performed to assess the acute oral toxicity of the Similar Substance 02 in the young adult male and female Han-wistar strain rat. Test item was dissolved in DMSO. A preliminary study was carried out to establish a dosing regimen, using groups of two males and two females at two dosage, using 10-15 ml/kg body weight. In the main experiment, the animals were dosed with 5350 mg/kg bw, by oral gavage. One female died 59 hours after dosing; no overt symptoms of poisoning were observed and no special findings could be detected.

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to the numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

The oral LD50 value was established to be greater than 5000 mg/kg bw, therefore the test substance is out of any classification limit for acute oral toxicity (oral acute toxicity category 4: 300 < ATE ≤ 2000 mg/kg bw).

In conclusion, Disperse Red 202 does not meet the criteria to be classified for oral acute toxicity, according to the CLP Regulation (EC 1272/2008).