Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories Ltd., Kreuzelweg 53, 5961 NM Horst / Netherlands
- Age at study initiation: 10 weeks old (males), 11 weeks old (females)
- Weight at study initiation: 275.4-310.7 g (males), 195.3-215.4 g (females)
- Housing: in groups of five (same sex) in Makrolon type-IV cages
- Diet: Kliba-Nafag 3433 rat maintenance diet (Provimi Kliba AG, 4303 Kaiseraugst / Switzerland), ad libitum, except during exposure
- Water: community tap water, ad libitum
- Acclimation period: 14 days (performed under Harlan Laboratories Study B68308)

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30-70 %
- Air changes: 10-15/hour
- Photoperiod: 12 hours dark/12 hours light

IN-LIFE DATES: From: 23-Sep-2009 To: 07-Oct-2009

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: animals were confined separately in restraint tubes which were positioned radially around the flow-past, nose-only exposure chamber.
- Method of holding animals in test chamber: restraint tubes
- Rate of air: 1.0 L/min
- System of generating particulates/aerosols: nebulizer with test item reservoir (Hospitak no. 950 with stainless steel injector)
- Method of particle size determination: gravimetrically, using a 7-stage cascade Mercer impactor (Model 02-130. In-Tox. Products Inc., Albuquerque, New Mexico, U.S.A.)
- Oxygen concentration, temperature and humidity in air chamber (n = 9): mean oxygen concentration: 20.6 ± 0.1 %, mean temperature: 23.1 ± 0.2 °C, mean relative humidity: 4.0 ± 1.02

TEST ATMOSPHERE
- Brief description of analytical method used: aerosol concentrations were determined gravimetrically using Millipore durapore filters (type HVLP). The filters were weighed before and immediately after sampling. The test aerosol concentration was calculated from the amount of test item determined by gravimetry and the sample volume. The particle size distribution was measured using a 7-stage cascade Mercer impactor. Mass median aerodynamic diameters and geometric standard deviations were calculated on the basis of the results from the impactor.
- Samples taken from breathing zone: Yes

TEST ATMOSPHERE (if not tabulated)
- MMAD (mass median aerodynamic diameter) / GSD (Geometric st. dev.): 1.39-1.40 µm (n = 3) / 1.95-1.97 (n = 3)
Analytical verification of test atmosphere concentrations:
yes
Remarks:
from the analytical data and the volume of the inhalation atmosphere sample, the concentrations were calculated in mg/L.
Duration of exposure:
4 h
Concentrations:
5.4 ± 0.1 mg/L (n = 4)
No. of animals per sex per dose:
5 animals
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: animals were examined for clinical signs five times on day 1 and daily thereafter; body weight was recorded on days 1 (before exposure), 4, 8 and 15.
- Necropsy of survivors performed: on day 15
- Other examinations performed: macroscopic examination
Statistics:
No statistical analysis was performed as only one group was allocated to the study.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.4 mg/L air
Exp. duration:
4 h
Remarks on result:
other: None
Mortality:
There was no mortality.
Clinical signs:
All animals showed a slightly decreased spontaneous activity and moderately ruffled fur immediately after exposure and one hour afterwards. Slightly ruffled fur was still present in all animals up to test day 4 and for two further days in some animals. Additionally, three females had a red secretion from the nose on test days 2 and/or 3.
Body weight:
From day 1 to day 4, body weight loss (up to 7 %) or stagnation of body weight was noted in all animals. This observation was considered to be mainly related to treatment with the test item due to the incidence and the duration of the effect in some animals. However, the restraint procedure is likely to have added to the observed effect. Thereafter, all animals showed elevated body weights on day 15 as compared to the body weights on day 1.
Gross pathology:
There were no macroscopic findings at necropsy.

Applicant's summary and conclusion

Interpretation of results:
other: slightly toxic
Executive summary:

In an acute inhalation toxicity study (Schuler, 2010), a group of young adult Wistar rats (5 males, 5 females) were exposed by inhalation route (nose only) to an aerosol generated from undiluted Montanol 800 (>98 %, MMAD 1.39 -1.40 µm) for 4 hours at a single concentration of 5.4 mg/L. Animals then were observed for 14 days. No animal died and all gained body weight. The transient clinical findings noted during the observation period were confined to decreased activity, ruffled fur and red secretions from the nose. The LC50 was >5.4 mg/L after 4 hours.

Montanol 800 is classified as being of low toxicity based on the LC50 in males and females.