Registration Dossier

Administrative data

acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to OECD guidelines and GLP

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guidelineopen allclose all
according to guideline
OECD Guideline 403 (Acute Inhalation Toxicity)
according to guideline
EU Method B.2 (Acute Toxicity (Inhalation))
according to guideline
EPA OPPTS 870.1300 (Acute inhalation toxicity)
according to guideline
other: JMAFF, 12 Nousan, Notification No 8147, November 2000; including the most recent partial revisions
GLP compliance:
Test type:
acute toxic class method
Limit test:

Test material

Constituent 1
Chemical structure
Reference substance name:
[2-(4-aminophenyl)ethyl][(2R)-2-hydroxy-2-phenylethyl]azanium chloride
EC Number:
Cas Number:
Molecular formula:
[2-(4-aminophenyl)ethyl][(2R)-2-hydroxy-2-phenylethyl]azanium chloride
Test material form:
solid: particulate/powder
migrated information: powder

Test animals

Details on test animals or test system and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approximately 9 - 12 weeks old
- Weight at study initiation: Bodyweight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study: No
- Housing: Group housing of 5 animals per cage in labelled makrolon cages containing sawdust as bedding material and paper as cage-enrichment.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet except during exposure.
- Water (e.g. ad libitum): Free access to tap water except during exposure.
- Acclimation period: At least 5 days before the start of treatment under laboratory conditions.

- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%
- Air changes (per hr): At least 10 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hour light, 12 hour dark cycle.

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
clean air
Details on inhalation exposure:
- Exposure apparatus: The design of the chamber is based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983).
- Exposure chamber volume: Approximately 150 mL
- Method of holding animals in test chamber: Animals were placed in restraining tubes.
- Source and rate of air: Pressurised air at a rate of 22 L/min for the initial exposure, and 17 L/min for the second exposure.
- Method of conditioning air: None, pressurised air used to supply an adequate oxygen supply.
- System of generating particulates/aerosols: For the initial exposure (ca. 1 mg.L), the test substance was administered in a stream
- Method of particle size determination: Samples collected with an 8 stage Marple personal cascade impactor containing fibre glass filters, and a fibre glass back up filter. Amounts of test substance collected were measure gravimetrically and the MMAD and GSD determined.
- Treatment of exhaust air: Air passed through a filter before being released into the exhaust of the fume hood.
- Temperature, humidity, pressure in air chamber: For ca. 1 mg/L the temperature ranged between 21.6 and 22.4°C with humidity between 11 and 21% and at ca. 7 mg/L temperature was 21°C and humidity was at 5%. These conditions were considered appropriate for the relatively short exposure duration.

- Brief description of analytical method used: A total of 13 and 4 representative samples were taken for determination of the actual concentration during exposure at ca. 1 and ca. 7 mg/L, respectively. Samples were drawn from the test atmosphere through a tube mounted in a free animal port and through a glass fibre filter. The collected amount of test material was measured gravimetrically, and measured by means of a dry gas meter. The time-weighted means and standard deviations were calculated.

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: At ca. 1 mg/L the MMAD and GSD were determined twice. The MMAD was 3.9 μm (gsd 1.6) and 4.5 μm (gsd 1.6). The MMAD values are at teh upper end of 1 - 4 μm. As one measurement was above the upper limit, and based on the effects on the animals, it can be assumed that sufficient deposition in the lower respiratory tract occured. At ca. 7 mg/L, the MMAD and gsd were not determined during the exposure, since the exposure was terminated after 32 minutes. Since the test substance was grinded in the same way as for the ca. 1 mg/L exposure and taken into account the effects on the animals, sufficient lung deposition can be expected. There was no evidence of test substance deposits in the upper air ways.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: Based on effects observed in the acute oral study, a starting concentration of ca. 1 mg/L was considered appropriate.
Analytical verification of test atmosphere concentrations:
Duration of exposure:
ca. 4 h
Remarks on duration:
At 1 mg/L, at ca. 7 mg/L exposure duration was 32 minutes due to observed toxicity
For the 1 mg/L group, the time-weighted mean actual concentration was 1.1 +/-0.04 mg/L. This was considered sufficiently stable to represent a 4 hour exposure to 1 mg/L.

For the second exposure targeted at 5 mg/L, the actual time-weighted mean concentration was 6.8 +/- 0.88 mg/L. Due to the mortality and signs of the animals already observed after 32 minutes, the exposure was terminated after 32 minutes. The concentration was measured four times during this exposure and showed a variation between 2 and 12 mg/L. Based on the early manifestation of the mortality and clinical signs, within 32 minutes after exposure and the variability in concentration range of 2 and 12 mg/L, it was not considered possible to determine whether mortality was due to a higher than expected initial exposure or whether the 4-hour LC50 would fall above or below 5 mg/L. There was no evidence of test substance overload of the respiratory tract.
No. of animals per sex per dose:
5 males and 5 females per dose.
Control animals:
Details on study design:
- Duration of observation period following administration: Up to Day 15 for surviving animals.
- Frequency of observations and weighing: Clinical signs were observed three times during exposure, shortly after exposure and then once daily until day 15. Bodyweights were recorded pre-exposure, and on days 2, 4, 8 and 15 and at death.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, bodyweights and macroscopic pathology.
No statistical analysis was performed, as it not a requirement on studies on this type.

Results and discussion

Effect levels
Dose descriptor:
Effect level:
> 1 mg/L air (nominal)
Based on:
test mat.
Exp. duration:
4 h
At ca. 1 mg/L no mortality occured.

At ca. 7 mg/L, three males and two females were found dead and one female was sacrificed for ethical reasons between 34 and 44 minutes after the start of exposure. One males was sacrificed for ethical reasons on Day 2. No further mortality occured for the remaining male and two females.
Clinical signs:
other: At ca. 1 mg/L, during exposure, no abnormalities were seen. After exposure, lethargy (1 male), flat posture (1 male), hunched posture (all animals), laboured respiration (2/5 males, 1/5 females), gasping (1 male), chromodacryorrhoea (1 male) and/or yello
Body weight:
At ca. 1 mg/L, overall body weight gain in four males and all females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity. One male showed body weight loss up to Day 8 and regained weight during the second week post-exposure.

At ca. 7 mg/L, overall body weight gain for the two surviving females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity. Body weight loss was noted in the surviving male during the first week post-exposure and the male regained weight during the second week post-exposure.
Gross pathology:
At ca. 1 mg/L, no abnormalities were found at macroscopic post mortem examination of the animals.

At ca. 7 mg/L, macroscopic examination for the animals that died or were sacrificed for ethical reasons during the study revealed several dark red foci for the lungs (one male), pelvic dilation of the kidneys (three males), dark red discolouration of the mandibular lymph nodes (one male and one female), enlarged thymus (one male) and diaphragmatic hernia of the right medial lobe of the liver (one female). No abnormalities were found for the three surviving animals.

Beginning autolysis was found for one animal found dead which was considered not related to treatment.

Applicant's summary and conclusion

Interpretation of results:
Migrated information Criteria used for interpretation of results: EU
The LC50 (4 hour) is normally ranked within the following ranges (0-0.05, 0.05-0.5, 0.5-1, 1-5 or >5 mg/L). However, given the actual concentrations obtained at target concentrations of 1 and 5 mg/L (1.1 and 6.8 mg/L) it was considered appropriate to rank the LC50 (4 hour) as > 1 mg/L.