Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013-05-13 to 2013-05-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: 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:
2009
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Version / remarks:
2008
Qualifier:
according to
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Version / remarks:
1998
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
Characteristics: clear, colourless, viscous
The density of the test item was 1.0953 g/cm3.

Test animals

Species:
rat
Strain:
other: Rattus norvegicus
Sex:
male/female
Details on test animals and environmental conditions:
TEST ORGANISMS: 
- Strain: Rattus norvegicus) / CD / Crl: CD(SD)
- Source: Charles River Laboratories, 97633 Sulzfeld, Germany
- Age: males: approx. 7 weeks, females: approx. 9 weeks
- Weight at study initiation: males: 233 - 251 g, females: 217 - 239 g
- Number of animals: 3 males and 3 females
- Fasting period before study: 16 hours
- Housing: groups of two or three
- Diet: ad libitum, ssniff R/m-H V 1534
- Water: ad libitum, tab water
- Acclimatisation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 3 °C
- Humidity (%): 55 + / - 15 %
- Photoperiod (hrs dark / hrs light): 12 hours darkness, 12 hours artifical light

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: compressed filtered air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Diluent: Dimethyl sulfoxide (DMSO), the test item is an emulsion with a high viscosity. Hence, a mixture of the test item with DMSO (4 + 1, m/m) was the highest possible concentration to obtain an appropriate aerosol.
- Type of exposure: nose-only, using a dynamic inhalation apparatus, air changes >/= 12/h
- Method of holding animals in test chamber (volume 40 l): separatley in Pyrex tubes at the edge of the chamber in radial position
- Source of air: compressed filtered air
- Method of conditioning air: manometer and air-flow meter (ROTA Yokogawa), flow rates checked once per hour and corrected if necessary
- Oxygen content: 21 %, determined at beginning and end of exposure, Carbon dioxide concentration did not exceed 1 %
- Type or preparation of particles: The aerosol of the test item was obtained using a spray-jet. The spray-jet was fed with compressed air (5.0 bar)
from a compressor and with the test item using a TSE infusion pump. At the bottom of the exposure chamber, the air was sucked off at a lower rate than created by the spray-jet in order to produce a homogenous distribution and a positive pressure in the exposure chamber (inflow 900 L/h,
outflow 800 L/h).
- Analysis of the aerosol concentration: The actual aerosol concentration in the inhalation chamber was measured gravimetrically with an air sample
filter (Minisart SM 17598 0.45 µm) and pump (Vacuubrand, MZ 2C ) controlled by a rotameter. Dust samples were taken once every hour during the exposure
- Method of particle size distribution: An analysis of the particle size distribution was carried out twice during the exposure period using a cascade
impactor according to MAY . The impactor is a device that classifies particles present in a sample of air or gas into known size ranges. It does this by drawing the air sample through a cascade of progressively finer nozzles. The air jets from these nozzles impact on pre-weighed plane sampling
surfaces (slides). The aerosol from the exposure chamber was drawn through the cascade impactor for 5 minutes at a constant flow rate of 5 L/min. The slides were removed from the impactor and weighed on an analytical balance (SARTORIUS, type 1601 004, precision 0.1 mg). The mass median aerodynamic diameter (MMAD) was estimated by means of non-linear regression analysis. The particulate size distribution was analysed using a
cascade impactor. The mass median aerodynamic diameter (MMAD) was determined as 2.598 μm. The Geometric Standard Deviation (GSD) of the
MMAD was calculated as 2.76.
- Temperature, humidity: T: 21.8 +/- 0.05°C, H: 60.3 % +/- 0.09 %, measured once every hour
TEST ATMOSPHERE
- Concentrations: 5.09 mg/l 
Air flow entrance (L/h): 900
Air flow exit (L/h): 800
Air change (changes per hour): 22.5

Analytical verification of test atmosphere concentrations:
yes
Remarks:
Gravimetric determination of aerosol concentrations with an air sample filter. Dust samples were taken once every hour during the exposure.
Duration of exposure:
4 h
Concentrations:
The gravimetric aerosol concentration of 5.09±0.03 mg test item/L air was measured at the animals’ nose.
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
EXAMINATIONS: 
- Post dose observation period: 14 days
- body weights: before,  7 and 14 days after treatment   
- mortality: once per hour during, and once after treatment on day of  exposure;  thereafter twice daily
- clinical signs: during and following exposure, observations were made and recorded systematicallye,  at least twice daily until all symptoms
subsided, thereafter each working day
- Necropsy: all animals (macroscopic) when found dead or at terminal  sacrifice. In addition, the weight of the lungs was determined.
Statistics:
not necessary

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.09 mg/L air
Based on:
act. ingr.
Exp. duration:
4 h
Mortality:
No animal died prematurely.
Clinical signs:
other: Slight ataxia and slight dyspnoea immediately until 30 minutes or 3 hours after end of exposure.
Body weight:
One of the 3 female animals appeared to be reduced in body weight gain since test day 8.
Gross pathology:
None findings
Other findings:
no other findings

Any other information on results incl. tables

The mean actual exposure concentration, MMAD and GSD of the test item were as follows:

 

nominal concentration

 

gravimetric concentration

(actual)

 

mass median aerodynamic diameter

(MMAD)

 

geometric standard deviation

(GSD)

 

[µL/L air]

 

[mg/L air]

 

[µm]

 

 

55.56

5.09

2.598

2.76

Applicant's summary and conclusion

Conclusions:
Under the present test conditions, LC50-value for CD rats following inhalation of the test item for 4 hours was determined as follows (actual concentration): LC50: exceeding 5.09 mg test item/L air.
Executive summary:

The aim of the present experiment was to obtain information on the acute toxicity and respiratory irritation, following a single 4-hour inhalation exposure of rats to the test item in an acute inhalation toxicity study.

Rats were exposed to the test item at a concentration of 5.09±0.03 mg test item/L air for 4 hours by inhalation using a dynamic nose-only exposure chamber. The aerosol was generated with the aid of a spray-jet.

Clinical signs/mortality:

Under the present test conditions, a 4-hour inhalation of 5.09 mg test item/L air revealed slight ataxia and slight dyspnoea immediately until 30 minutes or 3 hours after end of exposure. No animal died prematurely. No pathological findings were noted at necropsy.

Body weight:

One of the 3 female animals appeared to be reduced in body weight gain since test day 8.

Aerosol concentration and particle size distribution:

The gravimetric aerosol concentration of 5.09±0.03 mg test item/L air was measured at the animals’ nose. The particulate size distribution was analysed using a cascade impactor. The mass median aerodynamic diameter (MMAD) was determined as 2.598 μm. The Geometric Standard Deviation (GSD) of the MMAD was calculated as 2.76.

Conclusion:

Under the present test conditions, LC50-value for CD rats following inhalation of the test item for 4 hours was determined as follows (actual concentration):

LC50: exceeding 5.09 mg test item/L air.