cis-4-[3-(p-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03. - 05.2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

07 Sep 2009

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

certified by Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht Rheinland-Pfalz

Test type:

traditional method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

cis-4-[3-(p-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

EC Number:

266-719-9

EC Name:

cis-4-[3-(p-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

Cas Number:

67564-91-4

Molecular formula:

C20H33NO

IUPAC Name:

(2R,6S)-4-[3-(4-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

Test animals

Species:

rat

Strain:

Wistar

Remarks:

RccHan:WIST

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories B.V., Kreuzelweg 53, 5961 NM Horst, Netherlands
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Young adult animals (male animals approx. 8 weeks, female animals approx. 10 weeks)
- Weight at study initiation: Group 1 (2.036mg/L): 265.66 g mean bw of males and 210.70 g mean bw of females. Group 2 (5.036 mg/L): 271.88 g mean bw of males and 214.52 g mean bw of females.
- Fasting period before study: not specified
- Housing: Single housing or up to 5 animals
- Diet: Kliba-Labordiät, ad libitum
- Water: Tap water. ad libitum
- Acclimation period: at least 5 days before exposure
- Method of randomisation in assigning animals to test and control groups : Animals were randomly selected from a pool of animals. Five male and five female rats were used for each test group.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 – 24°C
- Humidity: 30 – 70%
- Air changes:1 5 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)

IN-LIFE DATES:
- From: 2011-03-17
- To: 2011-04-13

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose/head only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

>= 1 - <= 1.7 µm

Geometric standard deviation (GSD):

>= 3 - <= 5.3

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Head-nose inhalation system INA 20 (glass-steel construction)
- Exposure chamber volume: approx. 55 L
- Method of holding animals in test chamber: the animals were restrained in glass tubes and their snouts projected into the inhalation system
- Source and rate of air (airflow): Compressed air was produced by an oil-free compressor. Conditioned air was provided from a central air-conditioning system (1.5 m³/h).
- Method of conditioning air: Cold air passed through an activated charcoal filter, was adjusted to room temperature of 20 to 24°C and passed through a second particle filter.
- System of generating particulates/aerosols: Diaphragm metering pump (KNF, Switzerland), Two-component atomizer Mod. 970 (stainless steel, Schlick)
- Method of particle size determination: Cascade impactor measurement
- Treatment of exhaust air: The exhaust air was filtered and conducted into the exhaust air of the building (airflow: 1.35 m³/h).
- Temperature: The temperatures in the inhalation system were measured continuously with a digital thermometer.
- Humidity: The humidities in the inhalation system were measured with a dielectric probe (Testo).
- Pressure in air chamber: not specified

TEST ATMOSPHERE
- Brief description of analytical method and equipment used: For the quantitative determination of the aerosol an air sampler GS 312 (DESAGA) and high-performance-liquid chromatographic method was used.
- Samples taken from breathing zone: yes
- Time needed for equilibrium of exposure concentration before animal exposure : approx. 10 min.

CLASS METHOD
- Rationale for the selection of the starting concentration: The selection of the concentration of test group 1 was based on available information about the toxicity of the test substance.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

ca. 4 h

Concentrations:

Mean concentration of 2.036 mg/L (analytical concentration) for test group 1 and 5.036 mg/L (analytical concentration, limit concentration) for test group 2

No. of animals per sex per dose:

Five male and five female rats were used for each test group.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of clinical observations: Detailed clinical observations were recorded for each animal separately several times during exposure and at least once daily on the pre-exposure day and during the observation period.
- Frequency of weighing: Individual body weights once during the acclimatization period, shortly before exposure (day 0) and at least on days 1, 3 and 7, and before the sacrifice of the animals at the end of the observation period.
- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes
- Other examinations performed: check for mortality twice each work day and once on saturdays, sundays and on public holidays

Statistics:

Binomial test

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.036 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

Test group 1 and 2:
- No lethality was observed in male and female animals during the study period of 14 days. Therefore, the study satisfies the criteria of a limit test.

Clinical signs:

other:

Remarks:

see "Any other information on results incl. tables"

Body weight:

Test group 1 and 2:
- lower than 10% body weight loss
- other body weight observations: The mean body weights of the male animals decreased during the first post exposure observations day and increased from study day 3 onward. The mean body weights of the female animals decreased during the first few post exposure observations days and increased from study day 7 onward.

Gross pathology:

Test group 1 and 2:
- No gross pathological abnormalities were detected during the necropsy in the animals at the termination of the post exposure periods.

Any other information on results incl. tables

Clinical observations:

- Test group 1 (2.036 mg/L): Substance contamined fur, abdominal and accelerated respiration with sounds, piloerection from day 0 to day 3, but no clinical signs and findings were observed from study day 4 onwards.

- Test group 2 (5.036 mg/L): Colorless eye and nose discharge, substance contamined fur, red encrusted nose, accelerated respiration with sounds, piloerection from day 0 to day 4, but no clinical signs and findings were observed from study day 5 onwards.