Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 May - 18 Jun 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
adopted in 2009
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Version / remarks:
adopted in 1998
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Version / remarks:
adopted in 2008
GLP compliance:
yes (incl. certificate)
Remarks:
GYEMSZI National Institute for Quality- and Organizational Development in Healthcare and Medicines, Budapest, Hungary
Test type:
traditional method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
other: CRL:(WI) rats
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 - 11 weeks
- Weight at study initiation: 324 - 435 g (males), 213 - 234 g (females)
- Housing: 5 animals of the same sex per cage (individual in case of sighting group) in type III solid floor cages with stainless stell mesh lids, deep wood sawdust bedding (Lignocel® Hygienic Animal Bedding, J. Rettenmaier & Söhne GmbH+Co. KG, Rosenberg, Germany)
- Diet: ssniff SM R/M-Z+H (ssniff Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
3.64 µm
Geometric standard deviation (GSD):
2.4
Remark on MMAD/GSD:
The mean mass median aerodynamic diameter was 3.72 µm in the sighting group and 3.64 µm in the main study. The geometric standard deviation was 2.47 in the sighting group and 2.40 in the main study.
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: concentric anodised aluminium chambers with a computer control system incorporating pressure detectors and mass flow controllers (TSE Rodent Exposure System; TSE Systems GmbH, Bad Homburg, Germany)
- Exposure chamber volume: 3.85 L (Theoretical Volume)
- Method of holding animals in test chamber: Each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the chamber.
- Source and rate of air: filtered air at 0.7 L/min
- System of generating particulates/aerosols: generation of an aerosol using a Dust Generator Budiman (TSE Systems GmbH, Bad Homburg, Germany) at the top of the chamber under dynamic conditions
- Method of particle size determination: The particle size of the test atmosphere was determined three times during the exposure period using a 7-stage impactor of Mercer style (TSE Systems GmbH, Bad Homburg, Germany) isolating particles in the discrete aerodynamic size ranges. Samples were taken from an unoccupied exposure port and the weight of the test item was determined gravimetrically.
- Treatment of exhaust air: Used aerosol entered the outer cylinder from where it was exhausted through a suitable filter system.
- Temperature, humidity in air chamber: 25.0 - 27.1°C, 25.0 - 50.0%,

TEST ATMOSPHERE
- Brief description of analytical method used: The test substance concentration was determined by gravimetric analysis (glass-fibre filter, Whatman GmbH, Dassel, Germany)
- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- Particle size distribution: During the study the test atmosphere was respirable to the animals. see table 1
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): see table 1
Analytical verification of test atmosphere concentrations:
yes
Remarks:
gravimetric
Duration of exposure:
4 h
Concentrations:
4.83 mg/L (sighting group, analytical)
5.01 mg/L (main study, analytical, maximum attainable concentration)
No. of animals per sex per dose:
1 (sighting group)
5 (main study)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: animals were observed hourly during exposure, 1 h after exposure and twice daily thereafter for mortality/ moribundity; hourly during exposure, twice on the day of exposure and once daily thereafter for clinical signs. Bodyweights were determined prior to treatment and on Days 1, 3, 7 and 14.
- Necropsy of survivors performed: yes

Results and discussion

Preliminary study:
In the preliminary sighting study 1 male and 1 female were exposed to test substance target concentration of 5.0 mg/L. No mortality occured during the study.
Effect levels
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.01 other: mg/L air (gravimetrical)
Based on:
test mat.
Remarks:
maximum attainable concentration
Exp. duration:
4 h
Mortality:
1/5 male died shortly after the exposure
Clinical signs:
other: Wet fur and fur staining were observed in most animals on the day of exposure and one day after exposure. These observations were considered to be related to the restraint and exposure procedures and therefore, not considered to be treatment-related. Slig
Body weight:
Bodyweight loss was observed in the majority of animals, which is due to the restrained procedure during the exposure. However, the mean bodyweight and bodyweight gain of the animals at termination of the study was in the normal range, compared to untreated animals of the same age and strain.
Gross pathology:
No macroscopic changes were noted. However, dark/red discoloration of the non-collapsed lungs was macroscopically found in the dead animal.

Any other information on results incl. tables

Table 2 Mortality and clinical signs

Group

Mortality

Clinical Signs

Males

Sighting group (4.83 mg/L)

0/1

1/1

Main study group (5.01 mg/L)

1/5

5/5

Females

Sighting group (4.83 mg/L)

0/1

1/1

Main study group (5.01 mg/L)

0/5

5/5

Applicant's summary and conclusion

Interpretation of results:
other: CLP/GHS criteria not met; no classification required according to Regulation (EC) No. 1272/2008
Conclusions:
CLP: not classified