Reaction mass of Trisodium 4-({4-chloro-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)-2-{[1-ethyl-2-hydroxy-4-methyl-6-oxo-5-(sulfonatomethyl)-1,6-dihydropyridin-3-yl]diazenyl}benzenesulfonate and Trisodium 4-({4-chloro-6-[(3-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)-2-{[1-ethyl-2-hydroxy-4-methyl-6-oxo-5-(sulfonatomethyl)-1,6-dihydropyridin-3-yl]diazenyl}benzenesulfonate

Endpoint summary

Description of key information

The acute oral LD50 of FAT 40000/G in rats of both sexes observed over a period of 14 days is >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:

Study initiation date - 24 June 1992; Experiment start date - 01 July 1992; Experiment completion date - 15 July 1999; Study completion date - 20 August 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

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:

Code number: FAT 40000/G
Batch number: 67/92
Purity: 79.5%
Appearance: solid
Solubility: miscible
Storage: room temperature
Expiration date: 01 June 1993.

Species:

rat

Strain:

Wistar

Remarks:

SPF

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wölferstrasse 4, CH-4414 Füllinsdorf
- Age at study initiation: males: 8 weeks females: 10 weeks
- Weight at study initiation: males: 199 - 209 g females: 165 - 179 g
- Fasting: overnight
- Housing: Makrolon type-3 cages (size: 22 x 37.5 x 15 cm) with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard
- Water (e.g. ad libitum): Community tap water
- Acclimation period: One week under laboratory conditions, after veterinary examination. Only animals without any visual signs of illness were used for the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+-3 °C
- Humidity (%): 40-70 %,
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light (approx. 100 Lux) / 12 hours dark.

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

bidistilled

Details on oral exposure:

Application Volume/kg body weight: 10 ml at 2000 mg/kg

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 animals per dose

Control animals:

yes

Details on study design:

Mortality / Viability: Four times during test day 1 (according to the laboratories SOP's the last check was conducted 5 hours after application), and daily during days 2 - 1 5.
Body Weights: Test days 1 (pre-administration), 8 and 15.

Clinical Signs
Each animal was examined for changes in appearance and behaviour four times during day 1, and daily during days 2-15. All abnormalities were recorded. The animals were checked for the clinical signs listed below.
GENERAL BEHAVIOUR
aggressiveness
vocalization
restlessness / excitation
nervousness, fear
sedation
somnolence
sleep
coma

RESPIRATION
apnea
dyspnea
rales

EYE
chromodacryorrhea
exophthalmos
miosis
mydriasis
whitish discharge
lid adhesion
lacrimation
negative corneal reflex

NOSE
rhinorrhea
epistaxis

MOTILITY
akinesia
ataxia
dropped head
hyperkinesia
hypokinesia
paralysis, flaccid
paralysis, spastic
paddling movements
stiff gait
rolling movements

BODY POSTURE
ventral body position

MOTOR SUSCEPTIBILITY
spasms
tonic muscle spasms
clonic muscle spasms
opisthotonus
saltatory spasms
trismus
tremor
muscle-twitching
muscle-twitching
generalized

SKIN: erythema, edema, necrosis
OTHERS: Loss of weight, emaciation, diarrhea, ruflled fur, salivation, pallor, cyanosis.

Necropsy
Necropsies were performed by experienced prosectors. All animals were necropsied. All animals were euthanized by intraperitoneal injection of sodium pentobarbitone

Statistics:

The LOGIT-Model could not be applied to the observed rates of death. The toxicity was estimated without use of a statistical model.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No death occurred during the study period.

Clinical signs:

other: No clinical signs were noted in the animals.

Gross pathology:

No macroscopical organ findings were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral median lethal dose (LD50) of compound FAT 40000/G in rats is greater than 2000 mg/kg bw.

Executive summary:

The acute oral toxicity of FAT 40000/G was evaluated according to OECD test guideline 401 and Directive 84/449 EEC B.1 in a GLP certified laboratory.

The test article FAT 40000/G was administered to groups of 5 male and 5 female rats by oral gavage, at single dose of 2000 mg/kg. The following death rate was observed: 0 % at 2000 mg/kg. No clinical signs were noted in the animals. The body weight gain of the animals was not affected by the test article treatment throughout the entire study period. No macroscopical organ findings were observed.

The acute oral toxicity of FAT 40000/G in rats of both sexes, observed over a period of 15 days, was estimated to be greater than 2000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Good quality database with Klimisch rating 1.

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:

A key study was performed for FAT 40000/G which was administered to groups of 5 male and 5 female rats by oral gavage, at single dose of 2000 mg/kg. The following death rate was observed: 0 % at 2000 mg/kg. The acute oral toxicity of FAT 40000/G in rats of both sexes, observed over a period of 15 days, was estimated to be greater than 2000 mg/kg. Several other studies conducted with different batches of the test item more or less confirmed the non-toxic behaviour of the substance. Therefore effects of different batches of the test item can be ruled out.

Several other supporting studies were also performed with following results:

LD50 - >5000 mg/kg bw

LD50 - 14530 mg/kg bw

Acute inhalation toxicity waiver

Currently no study to assess the acute inhalation toxicity potential of Reactive yellow 095 is available. The vapour pressure is considered negligible since the melting point was determined to be 300 °C. Hence, the substance is considered to have low volatility. Synthesis and spray drying of this chemical is performed in a closed process; 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 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 chemical 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. Further, the chemical is found to have water solubility of 330 g/L, hence, in the case of dust of the substance entering the respiratory tract, it will be trapped in the mucus and cleared, thereby further limiting the absorption. The chemical showed low toxicity potential in the available acute oral toxicity studies (LD50: > 2000 mg/kg bw), with no systemic toxicity being seen even with doses >2000 mg/kg bw, hence it does not need to be classified STOT SE and low toxicity is expected for this chemical via the inhalation route. Taking into consideration the above arguments, low toxicity potential is expected on acute exposure of Reactive yellow 095 via inhalation route and hence testing by the inhalation route was considered scientifically not necessary.

Acute dermal toxicity waiver

Currently no study to assess the acute dose dermal toxicity of Reactive Yellow 095 is available. However, the molecular weight of the chemical is 797.1 g/mol, indicating it being too large for dermal absorption. It has water solubility of 330 g/L and n-octanol/water partition coefficient (log P) of -5.70, indicating it being too hydrophilic to cross the lipid rich environment of the stratum corneum. Hence, the dermal uptake for the substance will be low. The chemical showed low toxicity potential in the available acute oral toxicity studies (LD50: >2000 mg/kg bw). Similarly, absence of systemic toxicity in skin irritation studies, further supports the conclusion that low toxicity is expected for the chemical via the dermal route. Further experience with similar chemical substances has demonstrated that it is very unlikely that toxicity related to the intrinsic properties of the chemical only show up upon dermal exposure and not after systemic application. Taking the above arguments into consideration, low toxicity potential is expected on repeated exposure of Reactive Yellow 095 via dermal route and safety for human health can be estimated using the principles of route to route extrapolation. Hence, the conduct of repeated dose toxicity study via dermal route for Reactive Yellow 095 is considered to be scientifically not necessary.

Justification for classification or non-classification

Based on the available data from acute toxicity studies with FAT 40000, the test substance does not meet the criteria for classification for acute toxicity according to the CLP (1272/2008) Regulation.