Registration Dossier

Toxicological information

Acute Toxicity: inhalation

Currently viewing:

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From February 21, 1990 to April 30, 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report Date:
1990

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPP 81-3 (Acute inhalation toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Details on test material:
This is formulation of 49.2% of the test substance in water.
Specific details on test material used for the study:
- Batch no: RCD/VRE-55
- Date of receipt: 10 January 1990
- Storage: in dark at room temperature
- Expiry date: December 1990

Composition:
- cocobenzyldimethylammoniumchloride (CAS no. 68424-85-1; EC no. 270-325-2): 40% w/v
- dicocodimethylammoniumchloride (CAS no. 8001-54-3; EC no. 263-087-6): 37.5% w/v
- total amount of quarternary ammonium compounds: 77.3% w/v (according to COA, dated 21 December 1989)
- water: 7.7% (according to COA)

- pH (10% in water): 7.2
- Color Gardner: 2

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Test animals:
- Source: Charles River UK Ltd., Margate, UK
- Weight at study initiation: ca. 200 g on the d of exposure
- Housing: 5/sex in polypropylene cages with detachable wire mesh tops and floors
- Diet (e.g. ad libitum): ad lib
- Water (e.g. ad libitum): ad lib
- Acclimation period: at least 5 d

Environmental conditions:
- Temperature (°C): 18-24
- Humidity (%): 35-65

In-life dates: From 21 February, 1990 to 30 April, 1990

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
whole body
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
Generation of test atmosphere / chamber description
- Exposure apparatus: perspex whole body chamber (square section with pyramidal top)
- Exposure chamber volume: ca. 120L
- Method of holding animals in test chamber: wire mesh partitions to provide 10 separate animal compartments
- Source and rate of air: filtered and oil-free compressed air
- Method of conditioning air: dried
- System of generating particulates/aerosols: atomiser
- Method of particle size determination: cascade impaction (Andersen mini-sampler and Marple cascade impactor (model 296)
- Treatment of exhaust air: passage through a collection filter
- Temperature, humidity, pressure in air chamber: temperature measured at 30-min intervals (22-23°C), relative humidity not measured (reason: aqueous solution). However, as dried air was used and the test substance contained only 7.7% water, relative humidity could have been measured. Pressure in air chamber: not indicated. However, as the whole body chamber was placed in a hood, this is not as important.

Test atmosphere
- Brief description of analytical method used:
Five air samples (5 L for groups 2 and 4 (0.34 and 0.24 mg/L), 10L for group 3 (0.17 mg/L) were taken from the chamber during each exposure and the collected material was analysed to determine the concentration of the test substance in the chamber air. Each air sample was withdrawn, at 4 L per min, through a weighed glass fibre filter (Whatman GF/A) mounted in an open face filter holder. The volume of the air sample was measured with a wet-type gas meter. Two further air samples were taken using an Andersen mini-sampler or a series 290 Marple cascade impactor (Model 296), and the collected material was weighed to determine the particle size distribution of the test subtance. The samples were taken at approximately 1.5 and 3.5 h from the start of exposure. The filters from the open face sampler were transferred to extraction columns and compacted with a glass road. The test substance was eluted with five 2 mL portions of methanol into a 20 mL volumetric flask and diluted to volume with methanol.
The filters from the Andersen and Marple samplers were similarly treated t o give a final volume of 5 mL. The stages of the Andersenand Marple samplers were washed off with small amounts of methanol into 5 mL volurnentric flasks. The extracts were diluted with mobile phase to obtain solutions for HPLC-analysis with expected maximum concentrations of the test substance of 150 pg/mL.

- Samples taken from breathing zone: taken from the whole body chamber.

Vehicle: not used

- Test atmosphere (if not tabulated)
Concentrations:
0.34 mg/L (± 6%); nominal: 2.21 mg/L
0.17 mg/L (± 13%); nominal: 0.52 mg/L
0.24 mg/L (± 17%); nominal: 0.88 mg/L

- MMAD (mass median aerodynamic diameter) / GSD (geometric standard deviation):
0.34 mg/L : MMAD 1.9 µm, gsd 3.5
0.17 mg/L : MMAD 1.1 µm, gsd 2.7
0.24 mg/L : MMAD 0.8 µm, gsd 0.8
Analytical verification of test atmosphere concentrations:
yes
Remarks:
HPLC
Duration of exposure:
4 h
Concentrations:
0, 0.17, 0.24 and 0.34 mg/L
No. of animals per sex per dose:
5 per sex per concentration
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 21d
- Frequency of observations and weighing: observations continuously duringe exposure and at least twice daily thereafter; BW daily
- Food and water intake: daily
- Necropsy of survivors performed: yes
- Other examinations performed: lung weight, histopathology of lungs, liver and kidneys
Statistics:
LC50 determination by log probit method of Miller LC and Tainter ML, Proc. Soc. Exp. Bio. Med., 57(2),1944:261-264.

Results and discussion

Effect levelsopen allclose all
Key result
Sex:
male
Dose descriptor:
LC50
Effect level:
0.22 mg/L air (analytical)
95% CL:
>= 0.17 - <= 0.27
Exp. duration:
4 h
Key result
Sex:
female
Dose descriptor:
LC50
Effect level:
0.28 mg/L air (analytical)
95% CL:
>= 0.21 - <= 0.35
Exp. duration:
4 h
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
0.25 mg/L air (analytical)
95% CL:
>= 0.22 - <= 0.28
Exp. duration:
4 h
Mortality:
control group: 0/10
0.17 mg/L: 1 male (1/10)
0.24 mg/L: 3 males and 1 female (4/10)
0.34 mg/L: 5 males and 4 females (9/10)
Clinical signs:
During exposure:
The signs seen during exposure were considered to be consistent with inhalation of an irritant aerosol. Closing or partial closing of the eyes and exaggerated respiratory movement were seen in all rats exposed to the test subtance. Additional signs observed in rats exposed at 0.34 mg/L were gasping, wetness around the snout and the death of 3 male rats.
During the observation period:
At removal from the chamber, exaggerated respiratory movements, noisy respiration, a reduced respiration rate, brown staining around the snout and jaws, a discharge from the eyes and wet fur were seen. The exaggerated respiratory movements and the noisy respiration persisted in most rats during an extended observation period (21d). Signs that regressed during the observation period included brown staining around the snout and jaws, a coloured discharge from the eyes, wet fur and gasping. Other signs indicative of general decline in condition (swollen abdomen and lethargy) were seen in a small number of exposed rats.
Body weight:
There were moderate to marked decreases of body weight or reductions in the rate of body weight gain for up to 8d in male rats and for up to 14d in female rats following exposure at 0.17 mg/L or 0.24 mg/L. Subsequently weight gain for rats that survived exposure to the test substance was similar to that of the control rats.
Gross pathology:
The findings for rats that died as a result of exposure to the test substance were typified by congestion of the lungs, fluid in the trachea and gas-filled stomach. Macroscopic abnormalities in a proportion of rats that survived exposure to the test substance were a swollen appearance of the lungs and gas-filled stomachs and intestines.
Other findings:
The food and water consumption for the rat that survived exposure at 0.34 mg/L was variable and reduced. Food consumption was reduced for up to 12 d following exposure to the test substance at concentrations of 0.24 or 0.17 mg/L. The water consumption for these groups was reduced for up to 14d following exposure.

The lung weight to body weight ratio was increased, due to a high lung weight, for most rats that died as a result of exposure to ARMOBLEN 400 for a few decedents and the majority of the surviving rats was increased because of low body weight.

Histopathology
Group 2 (0.34 mg/L test substance)
Decedents
Treatment-related changes were seen in the 5 male and 4 female decedents. The distribution of these lesions was as follows:
focal alveolar wall necrosis in 3 males; diffuse congestion in 3 males and 4 females; eosinophilic material in alveoli in 5 males and 4
females; alveolitis in 2 males and 4 females; focal alveolar oedema in 1 male; perivascular oedema in 4 males.
Survivors
Treatment-related changes were seen in the single female rat surviving to the end of the observation period. These were as
follows: focal alveolitis; focal bronchiolitis; prominent bronchiolar goblet cells.

Group 3 (0.17 mg/L test substance)
Decedents
Treatment-related changes were seen in the single decedent male. These were as follows: diffuse congestion; eosinophilic materialin alveoli; alveolitis; focal alveolar oedema.
Survivors
Treatment-related changes were seen in one of the 4 males and in one of the 5 females surviving to the end of the observation
period. The distribution of these lesions was as follows: focal bronchiolitis in 1male; prominent bronchiolar goblet cells in 1 male;
foreign body giant cells in 1 male; focal alveolar haemorrhage in 1 female. A focus of emphysema was also seen in one surviving
female. The significance of this finding in a single animal is unclear, but possibility that it is related to treatment cannot be excluded.
No abnormalities were detected in 3 males and 3 females surviving to the end of the observation period.

Group 4 (0.24 mg/L test substance)
Decedents
Treatment-related changes were seen in the 3 male decedents and 1 female decedent. The distribution of these lesions was as
follows: diffuse or focal congestion in 2 males and 1 female; eosinophilic material in alveoli in 2 males and 1 female; alveolitis in 1
male and 1 female; focal alveolar oedema in 1 male; focal alveolar haemorrhage in 1 female; perivascular oedema in 1 female.
Survivors
Treatment-related change was seen in one of the two males and one of the 4 females surviving to the end of the observation
period. This lesion was: foreign body giant cells. No abnormalities were detected in 1 male and 3 females surviving to the end of theobservation period.

Comment from the authors
Treatment-related changes were found in the survivors from all groups. It is not possible to predict with certainty the progression of these lesions, but the changes were minimal and of a focal nature and it is possible that they would be resolved in the course of time leaving no permanent lesions.

Applicant's summary and conclusion

Conclusions:
Under the study conditions, the 4hr-LC50 of the mixture was calculated to be 0.25 mg/L (0.22 -0.28 mg/L), or based on 77.3% active material 0.32 mg QAC/L.
Executive summary:

A study was conducted to determine the inhalation acute inhalation toxicity of the test substance according to OECD Guideline 403 and US EPA OPP 81 -3, in compliance with GLP. The experiment was performed in Sprague-Dawley rats. Four groups of ten rats (five males and five females) were given a single, 4 h whole body exposure at concentration levels of 0, 0.17, 0.24 and 0.34 mg/L. The animals were observed for 21 d after exposure and were then culled for gross and histopathological examination of the lungs. Body weight, food and water intake and lung weight were also determined. There were no deaths in the control group; one animal (male) died at 0.17 mg/L, four animals died at 0.24 mg/L (3 males, 1 female), and nine animals died at 0.34 mg/L (5 males, 4 females). Clinical signs of toxicity noted were (partial) closing of the eyes and exaggerated respiratory movement during exposure in all test groups, gasping and wetness around the mouth during exposure at 0.34 mg/L. Clinical signs were noted in survivors throughout the 21 day observation period. Decrease of body weight, reduced weight gain, and reduced food and water intake was generally seen up to Day 14. Abnormalities noted at necropsy in survivors were increased relative lung weight, swollen appearance of the lungs and gas-filled stomach and intestines. Animals that died showed congestion of the lungs, fluid in the trachea, and gas-filled stomach. Histopathological lung changes in survivors generally consisted of focal alveolitis and bronchiolitis; changes in deceased animals generally consisted of focal alveolar wall necrosis, diffuse congestion, focal alveolar wall oedema and focal alveolar wall haemorrhage. Under the study conditions, the 4 h LC50 of the mixture was calculated to be 0.25 mg/L (0.22 -0.28 mg/L), or based on 77.3% active substance 0.32 mg QAC/L (Jackson, 1990).