Benzenemethanaminium, N,N,N-trimethyl-, 2-hydroxy-2-methylpropanoate (1:1)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 May 2012 to 31 June 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

benzyltrimethylazanium 2-hydroxy-2-methylpropanoate

EC Number:

805-580-6

Cas Number:

1431696-36-4

Molecular formula:

C14 H23 O3 N

IUPAC Name:

benzyltrimethylazanium 2-hydroxy-2-methylpropanoate

Test material form:

other: solid

Details on test material:

- Storage conditions: Room temperature; under N2; dry; protect against humidity
- Physical state/ color: solid/ beige
- Homogeneity: The test item was homogeneous by visual inspection

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: Young adult animals (female animals approx. 10 – 11 weeks)
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Weight at study initiation: Animals of comparable weight

HOUSING and DIET
- Room temperature/relative humidity: The animals were housed in fully air-conditioned rooms. Central air-conditioning guaranteed a range of 22°C ± 3°C for temperature and of 30 – 70% for relative humidity. There were no deviations from these ranges, which influenced the results of the study.
- Air changes per hour: approx. 10
- Day / night rhythm: 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)
- Type of cage: Makrolon cage, type III
- Number of animals per cage: Single housing
- Feeding: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany)
- Drinking water: Tap water ad libitum
- Bedding: H 15005-29; Ssniff, Spezialitäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)
- Enrichment: NGM E-022; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 2000 mg/kg bw: 40 g/100 mL; 300 mg/kg bw: 6 g/100 mL; 50 mg/kg bw: 1 g/100 mL
- Amount of vehicle (if gavage): 5.00 mL/kg bw
- Justification for choice of vehicle: Good homogeneity in olive oil Ph.Eur.

DOSAGE PREPARATION: For better handling the test item was ground with a pestle and mortar. The test item preparation was produced for
each application group shortly before application by stirring with a high speed homogenizer (Ultra-Turrax) and a magnetic stirrer. Additionally the homogeneity of the test item preparation during application was ensured by stirring with a magnetic stirrer.

CLASS METHOD
- Rationale for the selection of the starting dose: By request of the sponsor a starting dose of 2000 mg/kg bw was chosen in the first step with 3 female animals. As all animals died, 300 mg/kg bw were administered to 3 female rats in the second step. After administration all animals died in the second step. Therefore 50 mg/kg were given to a further group of three fasted females in the third step. Because no mortality occurred, 50 mg/kg bw were administered to another group of 3 female animals in the fourth step.

Doses:

2000, 300, 50 mg/kg bw

No. of animals per sex per dose:

2000 mg/kg bw: 3 females
300 mg/kg bw: 3 females
50 mg/kg bw: 6 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation.
- Necropsy of survivors performed: yes
- Pathology: Necropsy with gross-pathology examination on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with increasing concentrations over time. Necropsy of all animals that died before as early as possible after death.
- Clinical signs: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals.
- Mortality: A check for any dead or moribund animals was made at least once each workday, these records are archived by Bioassay.
- Histology: No histological examinations were performed.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 50 - < 300 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality occurred at 50 mg/kg bw. No animal survived dosing with 300 mg/kg bw.

Mortality:

All animals died in the 2000 mg/kg bw and 300 mg/kg bw test groups within 1 hour after application.
No mortality occurred in the 50 mg/kg bw test groups

Clinical signs:

other: All animals of the 2000 mg/kg bw test group and 2 animals of the 300 mg/kg bw died immediately after administration. Therefore no clinical signs could be determined. One animal of the 300 mg/kg bw test group showed poor general state, dyspnoea, piloerecti

Gross pathology:

The three animals that died in the 2000 mg/kg bw test group showed liquid content in the stomach and black discoloration of liver. In two animals of the 300 mg/kg bw test group dark red spotted discoloration of the lung was noted at necropsy. One animal of this test group didn’t show any findings. In the 50 mg/kg bw test group no macroscopic pathological findings were noted in the animals sacrificed at the end of the observation period.

Applicant's summary and conclusion

Interpretation of results:

toxic

Remarks:

Migrated information Criteria used for interpretation of results: EU