Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March 24-April 7, 1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Well documented study according to OECD Guideline 403, Proposal for Updating Test Guideline 403 for Acute Inhalation Toxicity and EPA guideline OPPTS 870.1300. No influence of the test results is expected by the deviations.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report date:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
May 12, 1981
Deviations:
yes
Remarks:
No influence of the test results is expected by the deviations.
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Version / remarks:
June 1996
Deviations:
yes
Remarks:
No influence of the test results is expected by the deviations.
Qualifier:
according to guideline
Guideline:
other: Proposal for Updating Test Guideline 403 for Acute Inhalation Toxicity, OECD, Paris, August 1996.
Version / remarks:
August 1996
Deviations:
yes
Remarks:
No influence of the test results is expected by the deviations.
GLP compliance:
yes
Remarks:
OECD (Environmental Monograph No. 45, OECD, Paris 1992) and of the EPA (40 CFR, part 160).
Test type:
traditional method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
1-[2-(allyloxy)ethyl-2-(2,4-dichlorophenyl)-1H-imidazolium hydrogen sulphate
EC Number:
261-351-5
EC Name:
1-[2-(allyloxy)ethyl-2-(2,4-dichlorophenyl)-1H-imidazolium hydrogen sulphate
Cas Number:
58594-72-2
Molecular formula:
C14H14Cl2N2O.H2O4S
IUPAC Name:
1-[2-(2,4-DICHLOROPHENYL)-2-(PROP-2-EN-1-YLOXY)ETHYL]-1H-IMIDAZOLE SULFATE (1:1)
Test material form:
solid: particulate/powder
Details on test material:
- Product names: FUNGAFLOR® 75C / FUNGAFLOR® 75SP / FUNGAZIL® 750SP / FUNGAFLOR® 75PS / FUNGAZIL® 75SP / FUNGAFLOR® 750SP
- Reference number: JNJ-2634372-ABI / R027180 (product code)
- Physical state: solid
- Appearance: beige solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: ZR027180PUE131/Janssen code No. Imazalil sulphate: ZR027180
- Expiration date of the lot/batch: October 15, 2003
- Purity test date: 100% imazalil sulphate (75% as imazalil base equivalent)
- CAS No. 83918-57-4

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was stirred in the dust generator while pressing dry air through the powder to prevent agglomeration. It was trickled on to an adjustable vibratory metering system. From there it fell into the aerosol flask, was picked up by an air flow and transported to the lower center of the inhalation chamber. Larger particles sedimented within the inner chamber room and smaller ones followed the air stream to the outer chamber and reached the animals.


OTHER SPECIFICS:
Density: 1.46 g/ml at 20 °C
Melting point: 131.5 °C -132.8 °C

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
Him:OFA, SPF
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Forschungsinstitut fur Versuchstierzucht, A-2325 Himberg, Austria
- Females (if applicable) nulliparous and non-pregnant: no data
- Age at study initiation: Approximately 9 weeks
- Weight at study initiation: Males: 326-355 grams, Females 213-230 grams.
- Housing: Single caging in Makrolon cages type III (39 cm x 23 cm x 18 cm). Wire mesh lids, with bedding material for laboratory animals, type 4 HV (Finn Tapvie Ky, SF-73600 Kaavi), autoclaved. Bedding changed 1/week.
- Diet (e.g. ad libitum): Altromin No. 1314 forte, gamma irradiated with 25 kGy 60Co, ad libitum, except for the exposure period.
- Water (e.g. ad libitum): Tap water, acidified to pH 3 with HCl, from an automated watering system, ad libitum, except for the exposure period.
- Acclimation period: 6 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): average 22 °C
- Humidity (%): average 55 %
- Air changes (per hr): 12/hr
- Photoperiod (hrs dark / hrs light): Artificial light from 6 am to 6 pm.

Administration / exposure

Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
8.4 µm
Geometric standard deviation (GSD):
3.4
Remark on MMAD/GSD:
Original Test: 34% of the mass was in the fraction with a diameter of less than 5 µm. The size distribution did not exactly follow a log-normal distribution but had an additonal fraction of particles larger than 16 µm.

1st Amendment: The MMAD was 5.18 µm with a mean gometric standard deviation of 2.52.
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Nose-only inhalation apparatus (TSE, Technical & Scientific Equipment GmbH, Kronberg, Germany, Article No. 504101)
- Exposure chamber volume: The chamber was 30 cm in diameter and 27 cm in height, resulting in a total volume of 19 litres.
- Method of holding animals in test chamber: no data
- Source and rate of air: Central pressure pump (type GA22 from Atlas Copco, Vienna, Austria). The relative humidity was reduced to about 10% in a freeze-dryer (type FD23 from Atlas Copco). The air was filtered oil-free (DEL-Tech filter type 150, DELtech Engineering Ltd, England) and distributed within the Research Center. In the Toxicology Department the pressure was reduced to 3 bar.
- System of generating particulates/aerosols: The test item was stirred in the dust generator (Technical & Scientific Equipment GmbH, Kronberg, Germany, article number 594203) while pressing dry air through the powder to prevent agglomeration. It was trickled on to an adjustable vibratory metering system. From there it fell into the aerosol flask, was picked up by an air flow and transported to the lower center of the inhalation chamber. Larger particles sedimented within the inner chamber room and smaller ones followed the air stream to the outer chamber and reached the animals.
- Administration procedure: Vibratory metering system was adjusted to about 1 g of test item per minute. The air flow was 700 l per hour. Larger particles, about 99% of the test item, were caught in the inner chamber by sedimentation. Smaller particles, about 1% of the test item, reached the outer chamber and the inhalation tubes.
- Method of particle size determination: The size of the dust particles was analysed with a cascade impactor. It contains nine steps with cut-off-diameters from 0.06 um to 16 um. The cut-off diameters were obtained from the manufacturer. About 18 liters of the dust were passed through the impactor within 180 seconds and the amount sedimented in the individual steps was determined gravimetrically after drying. The site of collection was the same as for the analysis. For calculation of the mean particle size, the probit of the fraction of mass smaller than the cut-off diameters was plotted against the logarithm of the cut-off diameters and the liner regresssion of this graph was calculated, preferring the data points around 50%. The diameter, where the regression gives a probit of 5 is the mass mediated aerodynamic diameter (MMAD). The proportion of the diameter at a probit of 6 to the mass mediated diameter is the geometric standard deviation (GSD).
- Treatment of exhaust air: no data
- Temperature, humidity, pressure in air chamber: The temperature inside the chamber was 20.2 to 22.5 °C. The relative humidity ranged from 35.5 % to 42.3%. Air-flow, humidity and temperature: Recording ever half hour.

TEST ATMOSPHERE
- Brief description of analytical method used:
The humidity of the air inside the chamber was measured with a hygrometer (Hygrotest serie 55, type 0555 6020, Testoterm Ges.m.b.H., Vienna, Austria), the temperature with a glass-mercury thermometer. The air flow on the low pressure side (after the generator) was measured with a rotameter (Rota Apparatebau GmbH&Co KG, Wehr, Germany, type L 63/2400-9048, ranging till 2000 l/h). The air flow on the high pressure side (before the generator) was monitored by a rotameter (Fischer & Porter PrazisionsmeBrohr Nr. FP 1/4-10-6-5-81) and by recording the pressure of the air.
Concentration of the test item: At least 6 times during the 4-hour exposure period.
Size distribution of the test item: Once during the 4-hour exposure period.
- Samples taken from breathing zone: yes

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: The target concentration was set according to the Sponsor. 0.5 mg/l is the minimum LC50 for a substance for a classification in toxicity class III.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Gravimetric analysis. The dust was collected 9 times during the exposure period.
Duration of exposure:
4 h
Remarks on duration:
37 air changes per hour
Concentrations:
Actual concentration: The mean was 655 mg/m3 or 0.66 mg/l.
Nominal concentration: The mean nomimal was 103 g per m3 air (103 mg/l). The recovery of respirable dust was 0.6%
No. of animals per sex per dose:
5 males and 5 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations: 1, 2, 3, 4, 5 and 6 hours after exposure and then at least once a days for a total of 14 days. Body weights: Before exposure and 7 and 14 days after expsoure.
- Necropsy of survivors performed: Yes. All surviving animals were sacrified by CO2 14 days p.a. and examined macroscopically in an attempt to detect possible lesions.
- Other examinations performed: behavior, reactions, physical signs, and body weight.
Statistics:
Statistical evaluation of body weight changes versus the test item concentration.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 0.66 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: General malaise and to signs of damage to the respiratory system. Not life threatening and the animals recovered within a few days.
Mortality:
All animals survived till the end of the study.
Clinical signs:
other: One male and one female animal were normal at each observation term. In the other animals, piloerection, eyelid closure, chromodacryorrhoea, hunched posture and sedation were the most prominent findings, indicating general malaise of the animals. With the
Body weight:
The mean body weights at the day of exposure were 343 g for males and 223 g for females. The mean body weight gain in the first week after the exposure was 15 g for males and 16 g for females. In the second week males gained 42 g, and females 16 g.

One of the females lost weight in the first week after the exposure. In the second week all animals gained weight.
Gross pathology:
Nothing abnormal was seen in any of the animals.
Other findings:
- Other observations: Some observations in life were only seen in males. This may however be an accidental event. As most findings were seen in males and females and as there were not life threatening signs in any of the sexes, no clear sex differences can be established from the results of this study.

Any other information on results incl. tables

The gravimetric determination gave concentrations from 0.45 to 0.85 mg of test item per litre. About 2 hours (1 h 48 min) after the start of the exposure a concentration of less than 0.5 mg per litre was found. Therefore the input of test item was increased until the actual concentration was again higher than 0.5 mg/l. The reason for this lower output remained unclear.

The 1st Amendment was issued on 2000-06-15 because the test item aerodynamic diameter was 8.4 µm which outside the limits of 1-4 µm required by the EPA guideline.

It was therefore requested to prove experimentally if grinding of the test item and using the ground product for preparing the dust would lower the diameter so that it conforms with the limits of 1-4µm.

The grinding of the test item reduced the aerodynamic diameter to some extent but it still remained outside the range of 1-4µm.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The inhalative exposure of rats to the test item at a concentration of 0.66 mg/l leads to signs of general malaise and to signs of damages to the respiratory system. None of these signs were life threatening and the animals recovered within a few days. At terminal necropsy no abnormal findings were noted. The LC50 (inhalative), four hour exposure of test item for male and female rats is therefore >0.66 mg/l.