Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
aug-oct, 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: MAFF in Japan (12-Nousan-No. 8147, 2-1-3, 2000)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): E-C104
- Physical state: blue powder
- Storage condition of test material: under dark condition at room temperature (actual range 20.5-22.3°C)

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Tsukuba Breeding Center (Ibaraki, Japan) of Charles River Japan, Inc.
- Age at study initiation: 8 weeks
- Weight at study initiation: 310-331 grams (males); 199-216 grams (females)
- Fasting period before study: not indicated
- Housing: wire-mesh stainless steel cage; during quarantine and acclimatization period, 5 or 2 animals of the same sex were housed in a cage. After selection of animals, 5 animals of the same sex were housed in a cage.
- Diet: pellet diet MF (Oriental Yeast Co., Ltd., Tokyo); ad libitum
- Water: local tap water; ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23±2°C
- Humidity (%): 60±20
- Air changes (per hr): 8
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: August 23, 2011 To: September 27, 2011

Administration / exposure

Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
Animals were individually held in animal holders attached to a snout-only exposure chamber (total volume 31.2 L). The dust was generated by a turn-table type dust feeder with compressed air. The compressed air was supplied to the dust feeder through an air filter. Airflow to the chamber was controlled by an ejector in the dust feeder at a rate of 20 L/min. The chamber air was exhausted through an air filter system consisting of a bag filter, a HEPA filter and an activated charcoal filter. The differential pressure in the chamber was adjusted to and maintained between -20 and -5 mm H2O using exhaust-adjusting valve. Aiflow rate in the dust feeder was continuously confirmed and recorded every 30 minutes during the exposure period.

Nominal concentration: the total amount of active ingredient of the test substance supplied to the chamber was determined gravimetrically with adjustment for the purity. The nominal concentration (mg/L) was calculated by dividing the total amount of active ingredient (mg) by total air volume (L) deliverd during exposure.
Actual concentration: the actual concentration was quantified at 1, 2 and 3 hours after the initiation of exposure. Six litters of chamber air was drawn from the chamber sampling port. The test substance dust was trapped on a pre-weighed glass fiber filter and the amount of the test substance trapped was determined gravimetrically using an electronic balance. The actual concentration (mg/L) was calculated by dividing the amount of the active ingredient (mg) with adjustment for the purity trapped on a glass fiber filter by air volume (L) sampled.

TEST ATMOSPHERE (if not tabulated)
high dose
- Particle size distribution (MMAD): 4.49 µm (4.42-4.60 µm)
- GSD (Geometric st. dev.): 2.34 (2.24-2.51)
- Mean value of actual concentration = >4.99 mg/L (>4.68->5.18)
low dose
- Particle size distribution (MMAD): 4.28 µm (4.12-4.38 µm)
- GSD (Geometric st. dev.): 2.11 (2.09-2.12)
- Mean value of actual concentration = >2.02 mg/L (>1.94->2.11)
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
5 mg/L (m, f), 2 mg/L (m)
No. of animals per sex per dose:
high dose: 5m, 5f
low dose: 5m
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observation for mortality and clinical signs at 2 hours after initiation of exposure, immediately, 1 and 4 hours after the exposure, and once daily thereafter until termination of the observation period. Each animal was weighed shortely before the exposure on the day of treatment and when found dead, on 1, 3, 7 and 14 days after exposure.
- Necropsy of survivors performed: yes
- Other examinations performed: none
Statistics:
The range of LC50 value was determined from the mortality.

Results and discussion

Preliminary study:
A dose-finding acute inhalation toxicity study with 1.0 and 5.0 mg/L (E-C104) was performed with 5 rats/sex (study no IET 11-0059). In the 1 mg/L group no mortality was observed during the 7-day observation period, whereas in the 5 mg/L group 1 male and 1 female died at 1 day after exposure. Based on these results, a limit test at 5 mg/L was considered for the main study.
Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
In the 5 mg/L group 1 male died 4 hours after exposure. No mortality was observed in the 2 mg/L group.
Clinical signs:
No clinical signs were observed in the low dose group. In the high dose group, sedation, deep respiration and labored respiration until one day after exposure, and disappeared on day 2 after exposure.
Body weight:
In the low dose group, body weight lossess were noted in all animals on day 1 after exposure, in 3 males on day 3 after exposure and in 1 male on day 7 after exposure (as compared with body weight before exposure).
In the high dose group, body weight losses were noted in all animals on day 1 after exposure and in 4 males on day 3 after exposure (as compared with body weight before exposure).
All animals gained their body weights at the end of the observation period (14 days after exposure).
Gross pathology:
No macroscopic abnormalities were noted in any surviving animal at necropsy at the end of the 14-day observation period.
Necropsy of the dead animal in the high dose group showed red color of the lung and foamy fluid in the trachea.
Other findings:
no

Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In the 5 mg/L group, 1 male died on day 1 after snout-only exposure to E-C104 during 4 hours. Based on these results, the LC50 of E-C104 for rats in this study was more than 5 mg/L, and E-C104 needs not to be classified for acute inhalation toxicity according to GHS.
Executive summary:

The acute inhalation toxicity of E-C104 was tested in Sprague-Dawley rats. Animals were exposed for 4 hours in a snout-only exposure chamber to 5 mg/L (5m, 5f) or 2 mg/L (5m). MMAD and GSD were 4.49 µm and 2.34, respectively (high dose), and 4.28 µm and 2.11, respectively (low dose). In the low dose group, no mortality was observed, whereas 1 male rat died in the high dose group the day after exposure. No clinical signs were observed in the low dose group. In the high dose group, sedation, deep respiration and labored respiration until one day after exposure, which disappeared on day 2 after exposure. In the low dose group, body weight losses were noted in all animals on day 1 after exposure, in 3 males on day 3 after exposure and in 1 male on day 7 after exposure (as compared with body weight before exposure). In the high dose group, body weight losses were noted in all animals on day 1 after exposure and in 4 males on day 3 after exposure (as compared with body weight before exposure). All animals gained their body weights at the end of the observation period (14 days after exposure). No macroscopic abnormalities were noted in any surviving animal at the end of the observation period. Based on these results, The LC50 of E-C104 for both sexes of Sprague-Dawley rats in this study is therefor more than 5 mg/L.