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:
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 and/or snouts (frequent instances), decreased respiratory rate and isolated occurences of ptosis and red fur staining of the stomach or leg (occasional instances), animals recoverd to appear normal from days 7 to 10 post-exposure.

Hunched posture, pilo-erection and red/browen staining around the eyes and snout are commonly seen in animals for short periods on removal from the chamber in inhalation studies. Wet fur is commonly recorded both during and for a short period after exposure. These observations are considered to be associated with the restraint procedure and, in isolation, are not indicative of toxicity.
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