2-[2-[4-[(2-cyanoethyl)methylamino]phenyl]vinyl]-1,3,3-trimethyl-3H-indolium hydrogen sulphate

Endpoint summary

Description of key information

 The LD50 for acute oral toxicity of the substance was determined to be between 300 and 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:

31 May 2016 to 29 June 2016

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)

Version / remarks:

17 December 2001

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

30 May 2008

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guideline of 12 Nousan No. 8147

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Test item No.: 16/0122-1
- Batch identification: Lot 4030158820
- Expiry date: March 2018
- Content: 100 g/100 g

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

OTHER SPECIFICS
- Physical state / color: Solid / violet to sparkling dark blue

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han) SPF

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approx. 10 weeks
- Weight at study initiation: Animals were of comparable weight (± 20% of the mean weight, experiment 1: 178.7 g, experiment 2: 176.7, experiment 3: 176.3 g)
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Housing: Makrolon cage, type III, Single housing
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: Tap water ad libitum
- Bedding: H 15005-29; Ssniff, Spezialdiäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)
- Enrichment: Wooden gnawing blocks (Type NGM E-022) ; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria
- Acclimation period: Acclimatization period of at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 (fully air-conditioned rooms)
- Humidity (%): 30 – 70
- Air changes (per hr): Approx. 10
- Photoperiod (hrs dark / hrs light): 12/12 (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)

Route of administration:

oral: gavage

Vehicle:

other: 0.5% solution of CMC (sodium carboxymethylcellulose, Dow Wolff Cellulosics GmbH) in deionized water

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: A good homogeneity in water could not be guaranteed, because the test item preparation was a suspension. Therefore a 0.5% solution of CMC in deionized water was applied.

DOSAGE PREPARATION
- Form of administration: Suspension
- Test item preparation (for all doses): The test item preparation for each test group was produced shortly before administration by stirring with a high speed homogenizer (Ultra-Turrax) and a magnetic stirrer.
- Homogenization until end of each administration period: The homogeneity of the test item preparation during administration was ensured by stirring with a magnetic stirrer.

FORM AS APPLIED IN THE TEST
Suspension

Doses:

EXPERIMENT 1
2000 mg/kg bw

EXPERIMENT 2 and Experiment 3
300 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Observation period: 14 days
- Body weight determination: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation.
- Clinical observations: Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter.
- Mortality: A check for any dead or moribund animals was made at least once each workday; these records are archived by Bioassay.
- Pathology: Necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations. Necropsy of all animals that died as early as possible after death.
- Histology: No histological examinations were performed.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

EXPERIMENT 1
All animals died at hour 3 after administration

EXPERIMENT 2 and 3
No mortality occurred

Clinical signs:

other: EXPERIMENT 1 - Impaired general state was noted in two animals from hour 0 until hour 1 after administration. Thereafter poor general state was noted in these animals at hour 2. Piloerection was seen in these animals from hour 0 until hour 2. - In the th

Gross pathology:

EXPERIMENT 1
The following macroscopic pathologic findings were observed in the animals that died: Red discoloration of the stomach contents, red discoloration of the small intestine contents and dark spotted liver.

EXPERIMENT 2 and 3
There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period.

Interpretation of results:

Category 4 based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

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 toxicity: oral

In a GLP compliant acute oral toxicity study performed according to OECD guideline 423 (Acute Toxic Class method), doses of 2000 and 300 mg/kg bw of the test substance (preparations in 0.5% sodium carboxymethylcellulose-solution) were administered by gavage to three test groups of three fasted Wistar rats each (2000 mg/kg bw in 3 females, 300 mg/kg bw in two times 3 females). All animals died at hour 3 after administration of 2000 mg/kg bw. In this test group, test-substance related clinical signs included impaired general state, piloerection, staggering and red discoloured defecation. Macroscopic pathologic observations in these animals were red discoloration of the stomach contents and of the small intestine contents, and dark spotted liver. No mortality occurred in the 300 mg/kg bw test groups. Red discoloured defecation and red discoloured urine were noted from study day 1 till study day 3. There were no macroscopic pathological findings in the surviving animals sacrificed at the end of the observation period. The LD50 was determined to be between 300 and 2000 mg/kg bw.

Justification for classification or non-classification

Based on the available information the substance needs to be classified as acute toxic 4, H302: Harmful if swallowed in accordance with EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation No. (EC) 1272/2008.