Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 Jul - 06 Sep 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
The Department of Health of the Government of the United Kingdom
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 57033
- Expiration date of the lot/batch: 25 November 2014

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark.

Test animals

Species:
rat
Strain:
other: RccHan™:WIST
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: 8 to 12 weeks
- Weight at study initiation: 200-350 g
- Housing: animals were housed in groups of five by sex in solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes and provided with environmental enrichment items: wooden chew blocks and cardboard "fun tunnels" (Datesand Ltd., Cheshire, UK).
- Diet: Harlan 2014C Rodent Diet (Harlan Laboratories UK Ltd., Oxon, UK), ad libitum
- Water: main drinking water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 °C
- Humidity (%): 30-70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
clean air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: cylindrical exposure chamber
- Exposure chamber volume: 30 L (28 cm diameter x 50 cm high)
- Method of holding animals in test chamber: animals were individually held in a tapered, polycrbonate restraining tube fitted onto a single tier of the exposure chamber and sealed by means of a rubber "O" ring.
- Source and rate of air: compressed air was supplied by means of an oil free compressor and passed through a water trap and respiratory quality filters before it was introduced to the nebuliser. The chamber flow rate was maintained at 40 L/min providing 80 air changes per hour.
- System of generating aerosols: the test item was aerosolised using a glass concentric jet nebuliser (Radleys, Saffron Walden, UK).
- Method of particle size determination:
- Temperature, humidity, pressure in air chamber: temperature, relative humidity and oxygen levels were measured by an electronic thermometer/humidity meter (Hanna Instruments Ltd, Beds, UK) or oxygen analyser (Servomex Ltd, Crowborough, UK) located in a vacant port in the animals’ breathing zone of the chamber.

TEST ATMOSPHERE
- Brief description of analytical method used: the actual concentration of the test item was measured off-line by gas chromatography (GC).
- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- Particle size distribution: Inhalable fraction
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):
Analytical verification of test atmosphere concentrations:
yes
Remarks:
GC
Duration of exposure:
4 h
Concentrations:
12.3 mg/L (mean achieved concentration)
35.6 mg/L (nominal concentration)
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: bodyweights were recorded on arrival, prior to treatment on the day of exposure and on Days 1, 3, 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: animals were observed for clinical signs and the respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity.

Results and discussion

Preliminary study:
In a preliminary study, two rats (one male, one female) were exposed to an atmosphere of the test item at a mean achieved atmosphere concentration of 2.83 mg/L for approx. 4 hours. No significant effects were noted in either animal.
Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 12.3 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: No animal died at the limit dose of 12.3 mg/L.
Mortality:
No mortality occurred during the study period.
Clinical signs:
other: The following abnormalities were detected: (during exposure and directly after exposure): increased respiratory rate; (one day after exposure): hunched posture, pilo-erection (all animals), increased respiratory rate and red/brown staining around the eyes
Body weight:
5/5 males and 4/5 females exhibited bodyweight losses on the first day post-exposure. Further bodyweight losses or no bodyweight gain were noted in 4 males and 3 females from days 1 to 3 post-expossure. Reasonable bodyweight development was noted in all animals during the remainder of the recovery period.
Gross pathology:
No macroscopic abnormalities were detected in any animal at necropsy.

Any other information on results incl. tables

Table 1. Mean achieved atmosphere concentration

Atmosphere concentration

Mean achieved (mg/L)

Standard deviation

Nominal (mg/L)

12.3

0.15

35.6

 

Table 2. Results of the acute inhalation study: Mortality

Mean achieved atmosphere concentration (mg/L)

Deaths

Male

Female

Total

12.3

0/5

0/5

0/10

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified