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Toxicological information

Acute Toxicity: inhalation

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Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (analytical purity of test substance not specified, only 4 days acclimatisation time, particle size of 5.2 µm MMAD in accordance to former guideline)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1987
Report Date:
1987

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
yes
Remarks:
analytical purity of test substance not specified, only 4 days acclimatisation time, particle size of 5.2 µm MMAD in accordance to former guideline
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
not specified
Details on test material:
- Name of test material (as cited in study report): Acrawax C
- Analytical purity: no data

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc.
- Age at study initiation: young adult
- Weight at study initiation: 180-300 g
- Housing: All animal housing and care conformed to AAALAC standards and to those published in the "Guide for the CareI and Use of Laboratory Animals," NIH Publication N0. 85-23. The animals were individually housed in suspended stainless steel wire mesh bottom cages.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 4 days


ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12/12


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: Rochester type inhalation chamber
- Exposure chamber volume: 270 L
- Method of holding animals in test chamber: The animals were individually housed during the exposure in wire mesh cages without access to food or water.
- Source and rate of air: High pressure external air source, 75 L / min
- System of generating particulates/aerosols: Particle generator (Model FD-100. Unifab Corporation, Kalamazoo. Michigan)
- Method of particle size determination: Particle size analysis was performed once per hour during the exposure using an Anderson 2000 impactor (Model 20-800). Stages one to eight of the impactor, and the final filter stage were fitted with pre-weighed glass fiber filters. A known volume of chamber air (30 L) was drawn through the impactor and the change in weight of each filter was then determined and recorded.
- Treatment of exhaust air: Air treatment system which consisted of a HEPA filter, a charcoal filter and a water scrubber
- Temperature, humidity, pressure in air chamber: The test atmosphere temperature, relative humidity and percent oxygen content, and the air flow rate to the chamber were recorded at approximately 30 minute intervals during the exposure. Average temperature, relative humidity, and oxygen content of the test atmosphere were 21°C, 56.8% and 21.0%, respectively.

TEST ATMOSPHERE
The inhalation chamber was maintained at a slightly negative pressure at all times during operation. Air flow rate to the chamber was monitored continuously during the exposure using calibrated Dwyer air flow meters (Dwyer Instruments, Inc.).
- Brief description of analytical method used: The average actual concentration of the test atmosphere was determined by gravimetric sampling. At the time of theoretical chamber equilibration a test atmosphere sample was drawn from the breathing zone of the chamber (5 L) through a pre·weighed glass fiber filter. The change in weight of the filter (mg) was determined and this value was divided by the volume of test atmosphere sampled (5 L) to yield the actual test atmosphere concentration. Additional gravimetric samples were obtained at approximately 30 minute intervals during the exposure. The average actual concentration of the test atmosphere was calculated for the exposure based on the initial and subsequent concentration analyses.
- Samples taken from breathing zone: yes
- MMAD: The mass median aerodynamic diameter of the generated particles was 5.2 µm and the standard geometric deviation was 1.7. Approximately 87% of the particles were less than 10 µm in size.



Analytical verification of test atmosphere concentrations:
yes
Remarks:
by gravimetric sampling
Duration of exposure:
240 min
Concentrations:
The average actual test atmosphere concentration was determined to be 6.3 mg/L. The calculated nominal concentration was 27 mg/L. Thus, the average actual concentration was approximately 23.3% of the calculated nominal test atmosphere 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: Mortality checks were performed twice daily, a minimum of 5 hours apart. The animals were observed for outward signs of toxicity 3 times on study day 1 (post exposure) and once daily thereafter for the duration of the study (day 15). Due to the density of the test atmosphere, an accurate observation of the study animals could not be performed during the actual exposure. Individual body weights were determined and recorded on study days 1, 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology
Statistics:
not determined

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 6.3 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
240 min
Mortality:
1 male, necropsy findings suggest that death was caused by asphyxiation
Clinical signs:
other: Labored breathing and/or rales, dark material around nose or mouth, decreased activity, urine stain, trashing (in cage), for details see table 1 in remarks
Body weight:
Decreased body weight gain and/or weight loss were observed in both the male and female animals. No net change in mean body weight was observed in the male rats between days 1 and 15. In the female animals, mean body weight was decreased approximately 5% during the period of the study.
Gross pathology:
Necropsy examinations of the surviving animals revealed yellow material in the stomach (4 females), pale lungs (one male) and
multifocal dark red foci on the lungs (one male). The significance of the above findings, if any was not determined in this study.
Other findings:
- Histopathology: No microscopic lesions were observed in the kidneys or liver of any animal.

Any other information on results incl. tables

Table 1: Clinical signs during the 4 day observation period after exposure; because of the death of one male, 4 males and 5 females (9 animals) were examined

 

Incidence of animals exhibiting finding / number of total animals on day

Finding

1

2

3

4

Labored breathing and/or rales

7 / 9

7 / 9

3 / 9

2 / 9

Dark material around nose or mouth

3 / 9

4 / 9

1 / 9

0 / 9

Decreased activity

0 / 9

3 / 9

0 / 9

0 / 9

Urine stain

0 / 9

1 / 9

0 / 9

0 / 9

Trashing (in cage)

1 / 9

0 / 9

0 / 9

0 / 9

Based on the results of this study, the acute inhalation LC50 of ACRAWAX C in rats is estimated to be greater than 6.3 mg/L.

According to the criteria laid down in Regulation (EC) 1272/2008 and Directive 67/548/EEC ACRAWAX C does not have to be classified for acute toxicity via the inhalation route.

Applicant's summary and conclusion