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Acute Toxicity: inhalation

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Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
other information
Study period:
07 July 2010 to 13 July 2010
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study has been performed according to OECD and/or EC guidelines and according to GLP principles. Due to the particle size distribution of the test substance it was not possible to perform the inhalation toxicity test with the solid. Instead the test was conducted with an aqueous suspension. This study results are not used for the hazard assessment of the solid substance. Therefore the reliability of the study is set not assignable.

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
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes

Test material

Constituent 1
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study report): 202240/A
- Physical state: solid, white crystalline powder
- Expiration date of the lot/batch: 13 December 2010
- Storage condition of test material: At room temperature in the dark
- Stability under storage conditions: Stable

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany and Charles River France, L’Arbresle Cedex, France.
Two animal sources were used. Since the results of the animals obtained from both sources are interchangeable, the study integrity was not adversely affected by the deviation.
- Age at study initiation: Young adult animals (approximately 9 weeks old) were selected.
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean.
- Housing:
Before exposure: Group housing of five animals per sex per cage in labelled Macrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
After exposure : Group housing as described above, except that a paper sheet was introduced into the cage covering the bedding and cage enrichment to prevent suffocation in case of bad health condition. At the end of the Day of exposure the paper sheet was removed.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) except during exposure to the test substance.
- Water (e.g. ad libitum): Free access to tap water except during exposure to the test substance.
- Acclimatization period: at least 5 days before start of treatment under laboratory conditions.
- Health inspection: A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.7 – 20.3°C
- Humidity (%): 44 - 81%
Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70% for relative humidity. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

IN-LIFE DATES: From: 07 July 2010 to 13 July 2010

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: Water
Details on inhalation exposure:
The design of the exposure chamber is based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983). The chamber consisted of three animal sections with eight animal ports each (Appendix 1, Figure 1). Each animal port had its own atmosphere inlet and exhaust outlet. The animals were placed in restraining tubes and connected to the animal ports. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min, which ensures an adequate oxygen supply to the animals. The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.
The placement of the individual animals in the inhalation chamber is shown in Appendix 1 (Figure 2). All components of the exposure chamber in contact with the test material were made of stainless steel, glass, rubber or plastic. To avoid exposure of the personnel and contamination of the laboratory the exposure chamber was placed in a fume hood, which maintained at a slight negative pressure.
Analytical verification of test atmosphere concentrations:
no
Duration of exposure:
4 h
Concentrations:
3.6 and 1.2 mg/L
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
202240/A was formulated in water and administered as an aerosol by inhalation for 4 hours to two groups of five male and five female Wistar rats each. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15. Macroscopic examination was performed on the day of death or after terminal sacrifice (day 15).

For the highest exposure level, the mean actual test substance concentration was 3.6 ± 0.6 mg/L. The nominal concentration was 13.2 mg/L, resulting in a generation efficiency (ratio of actual and nominal concentration) of 17%. For the lowest exposure level, the mean actual test substance concentration was 1.2 ± 0.1 mg/L. The nominal concentration was 2.3 mg/L, resulting in a generation efficiency (ratio of actual and nominal concentration) of 52%. The concentration measurements distributed over time showed that the concentrations were sufficiently stable

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice for each exposure level. For 3.6 mg/L, the MMAD was 4.3 µm (gsd 1.8) and 3.9 µm (gsd 1.8) respectively. For 1.2 mg/L, the MMAD was 3.7 µm (gsd 1.7) and 3.8 µm (gsd 1.6) respectively.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 1 - < 5 other: mg/l aqueous suspension of 202240/A
Based on:
test mat.
Exp. duration:
4 h
Mortality:
All animals exposed to the highest level of 3.6 mg/L were found dead or sacrificed on the day of exposure (Day 1). At 1.2 mg/L, one male was found dead immediately after exposure. No further mortality occurred.
Clinical signs:
other: At 3.6 mg/L, laboured respiration was noted among the animals during exposure. After exposure, lethargy, hunched posture, laboured respiration and ventral lateral recumbency were noted among the animals that survived the exposure. Due to their very poor c
Body weight:
Overall body weight gain in surviving males and females was within the range expected for rats of this strain and age used in this type of study.
Gross pathology:
Macroscopic post mortem examination of the animals that were found dead or sacrificed for ethical reasons during the study revealed abnormalities of the lungs (many grey-white or reddish foci) and liver (enlarged in one male and one female). Also one surviving male showed abnormalities of the lungs (many grey-white foci). No abnormalities were found in the remaining surviving animals.

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Remarks:
the solid test substance 202240/A was not tested
Conclusions:
Generation of the test atmosphere using the powdery test substance as delivered by the sponsor provided a Mass Median Aerodynamic Diameter (MMAD) between 5 and 7 µm. This MMAD exceeded the required maximum diameter of 4 µm as indicated in the test guidelines. Therefore the solid test substance was not tested. Instead a suspension in water was used to generate a test atmosphere that met the MMAD requirement.

The abnormalities of the lungs, seen in all non-surviving animals and one surviving male, might indicate that local toxicity was introduced in these animals. It can not be excluded that the local toxicity contributed to the death of the animals.
Based on the particle size the inhalatory toxicity test with 202240/A in Wistar rats was not performed.

Based on the results, the inhalatory LC50, 4h value of an aqueous suspension of 202240/A in Wistar rats was considered to be within the range of 1 – 5 mg/L.
Executive summary:

Assessment of acute inhalation toxicity with 202240/A in the rat

 

The study was carried out based on the guidelines described in:

-OECD Guidelines, Section 4, Health Effects. No.403, "Acute Inhalation Toxicity", September 2009.

- Commission Regulation (EC) No 440/2008,B.2. Acute Toxicity (inhalation),L142, May 2008.

- EPA OPPTS 870.1300, Acute inhalation Toxicity. EPA 712-C-98-193, August 1998.

- JMAFF, 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.

 

 

202240/A was formulated in water and administered as an aerosol by inhalation for 4 hours to two groups of five male and five female Wistar rats each. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15.Macroscopic examination was performed on the day of death or after terminal sacrifice (day 15).

 

For the highest exposure level, the mean actual test substance concentration was 3.6 ± 0.6 mg/L. The nominal concentration was 13.2 mg/L, resulting in a generation efficiency (ratio of actual and nominal concentration) of 17%. For the lowest exposure level, the mean actual test substance concentration was 1.2 ± 0.1 mg/L. The nominal concentration was 2.3 mg/L, resulting in a generation efficiency (ratio of actual and nominal concentration) of 52%.The concentration measurements distributed over time showed that the concentrations were sufficiently stable.

 

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice for each exposure level. For 3.6 mg/L, the MMAD was 4.3µm (gsd 1.8) and 3.9µm (gsd 1.8) respectively. For 1.2 mg/L, the MMAD was 3.7µm (gsd 1.7) and 3.8 µm (gsd 1.6) respectively.

  

All animals exposed to the highest level of 3.6 mg/L were found dead or sacrificed on the day of exposure (Day 1). At 1.2 mg/L, one male was found dead immediately after exposure. No further mortality occurred.

 

At 3.6 mg/L, laboured respiration was noted among the animals during exposure. After exposure, lethargy, hunched posture, laboured respiration and ventral lateral recumbency were noted among the animals that survived the exposure. Due to their very poor condition, the remaining animals were sacrificed at the end of Day 1.

At 1.2 mg/L, lethargy, hunched posture, laboured respiration and piloerection were noted in the animals on Days 1 and/or 2.

 

Overall body weight gain in surviving males and females was within the range expected for rats of this strain and age used in this type of study.

 

Macroscopic post mortem examination of the animals that were found dead or sacrificed for ethical reasons during the study revealed abnormalities of the lungs (many grey-white or reddish foci) and liver (enlarged in one male and one female). Also one surviving male showed abnormalities of the lungs (many grey-white foci). No abnormalities were found in the remaining surviving animals.

  

The abnormalities of the lungs, seen in all non-surviving animals and one surviving male, might indicate that local toxicity was introduced in these animals. It can not be excluded that the local toxicity contributed to the death of the animals. 

 

The inhalation toxicity test with the solid was not possible due to the MMAD exceeded the required maximum diameter of 4µm.

Generation of the test atmosphere using the powdery test substance as delivered by the sponsor provided a Mass Median Aerodynamic Diameter (MMAD) between 5 and 7µm. This MMAD exceeded the required maximum diameter of 4µm as indicated in the test guidelines.

Therefore the solid test substance was not tested.

Instead a suspension in water was used to generate a test atmosphere that met the MMAD requirement.

Based on the results, the inhalatory LC50, 4hvalue of a prepared aqueous suspension of 202240/A in Wistar rats was considered to be within the range of 1 – 5 mg/L. This result was not used for the inhalation hazard assessment fo the solid material.