Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 June 2004 to 11 August 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Version / remarks:
Annex V
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agiruclture, Forestry and Fisheries, 12 NohSan No.8147 (2-1-3), 24 Nov 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Date of inspection: 2 December 2002 Date of Signature: 13 February 2003
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): TIPX
- Substance type: Colourless liquid.
- Physical state: Liquid.
- Purity test date: 99.3%.
- Lot/batch No.: 042028.
- Storage condition of test material: Stored cold at 4ºC.
- Others: Mass median aerodynamic diameter (for liq.+solid aerosol) - 3.15 micron.

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, Kent, UK.
- Age at study initiation: 8 to 12 weeks of age.
- Weight at study initiation: 200 - 350 g except one male weighed 352 g which considered not to affect the purpose or validity of the study.
- Fasting period before study: Not reported.
- Housing: The animals were housed in groups of 5 by sex in solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes and cardboard, provided with an environmental enrichment.
- Diet: EU Rodent Diet 5LF2 (BCM IPS Limited, London, UK) ad libitum. The diet was routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
- Water: Mains drinking water ad libitum. The water was routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Set to achieve limits of 19 to 23ºC.
- Humidity (%): Set to achieve limits of 40 to 70%.
- Air changes (per hr): At least 15 changes per hour.
- Photoperiod (hrs dark / hrs light): Twelve hours continuous light (06:00 to 18:00) and 12 hours darkness.

IN-LIFE DATES: From: Day 0 To: Day 14

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: Not applicable.
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Cylindrical exposure chamber.
- Exposure chamber volume: 30 litres. - -- Method of holding animals in test chamber: Each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the exposure chamber and sealed by means of a rubber 'O' ring.
- Rate of air: The chamber flow rate was maintained at 60 L/min providing 120 air changes per hour.
- System of generating particulates/aerosols: Glass concentric jet nebuliser (Radleys, Saffron Walden, Essex, United Kingdom).
- Method of particle size determination: Using a Marple Personal Cascade Impactor (Schaefer Instruments Ltd, Oxon, United Kingdom), determined 3 times during the exposure period.
- Treatment of exhaust air: With a high efficiency filter to a metered exhaust system.
- Temperature, humidity, pressure in air chamber: The temperature 19 - 21ºC and the humidity 78 - 89% during the exposure epriod. The test material was supplied under pressure but no specific values were reported.
TEST ATMOSPHERE
- Brief description of analytical method used: Two litres of test asmosphere was drawn through a glass fibre filter and the quality of the test substance retained by the filters was determined by GC using an external standard technique. The test filter samples were extracted with methanol at final concentration of approximately 0.1 mg/mL. As standards, aliquots (≤ 0.1g) of the test material at a concentration of 0.1 mg/mL was used. The standard and sample solutions were analysed by GC. The concentration found was in the range of 93 - 105% of the nominal concentrations.
- Samples taken from breathing zone: Yes.

VEHICLE
Not applicable.

TEST ATMOSPHERE
- MMAD (Mass median aerodynamic diameter): 3.15 μm.
- GSD (Geometric st. dev.): 2.10.

CLASS METHOD (if applicable)
Not applicable.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Linearity, specificity, stability and accuracy were tested. The analytical method was shown to be sufficiently accurate for the purpose of the study.
Duration of exposure:
4 h
Concentrations:
Mean of 5.99 ± 0.28 mg/L.
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:
Clinical signs: At hourly intervals during exposure, immediately on removal from the restraining tubes at the end of exposure, one hour after termination of exposure and subsequently once daily for 14 days.
Bodyweight: prior to the treatment on the day of exposure and on Days 7 and 14.
- Necropsy of survivors performed: Yes. Animals were killed by intravenous overdose of sodium pentobarbitone. All animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded. The respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity.
Statistics:
Not applicable.

Results and discussion

Preliminary study:
Not applicable.
Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.99 mg/L air
Exp. duration:
4 h
Mortality:
Male: 5.99 mg/L; Number of animals: 5; Number of deaths: 0
Female: 5.99 mg/L; Number of animals: 5; Number of deaths: 0
Clinical signs:
Increased respiratory was noted in all animals during exposure, on removal and one hour post exposure.
One day after exposure, increased respiratory rate and hunched posture was noted in all animals. Animals recovered quickly to appear normal on Day 2 or Day 3 post exposure, no further clinical signs were noted throughout the remainder of the recovery period.

Signs of hunched posture, pilo-erection and red/brown staining to the eyes were commonly seen in animals for short periods on removal from the chamber following 4-hour inhalation studies. Wet fur was recorded both during and for a short period after exposure. These observations were
considered to be associated with the restraint procedure, and were not indicative of toxicity.


Body weight:
No treatment-related effects were observed during the study.
Gross pathology:
Apart from two instances of dark patches on the lungs, no macroscopic abnormalities were detected amongst animals at necropsy.
Other findings:
Not reported.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
No deaths occurred in a group of 10 rats exposed to a mean achieved atmosphere concentration of 5.99 mg/L for 4 hours. It was therefore considered that the acute inhalation median lethal concentration (4 h LC50) of TIPX, in the Sprague-Dawley Crl:CD® (SD) IGS BR strain rat, was greater than 5.99 mg/L. The test material does not meet the criteria for classification according to EU classification system (Council Directive 67/548/EEC and Regulation (EC) No 1272/2008).
Executive summary:

Introduction.A study was performed to assess the acute inhalation toxicity of the test material. The method used followed that described in the US Environmental Protection Agency (EPA) health Effects test Guidelines, OPPTS 870.1300, Acute Inhalation Toxicity, August 1998. The method was also designed to meet OECD guideline 404 (may 1981) and EC Guideline (Method B2 of Commission Directive 92/69/EEC, Annex V) and to comply with the requirements of the Japanese Ministry of Agriculture, Forestry and Fisheries (MAFF), testing Guidelines for Toxicology Studies, 12 NohSan No.8147 (2-1-3), 24 Nov 2000 (partially received on 26 June 2001).

 

Method. A group of 10 Sprague-Dawley Crl:CD® (SD) IGS BR strain rats (5 males and 5 females) was exposed to an aerosol atmosphere. The animals were exposed for 4 hours using a nose only exposure system, followed by a 14-day observation period.

 

Results.

The mean achieved atmosphere concentration was as follows:

Mean Achieved: 5.99 mg/L

Standard Deviation: 0.28

Nominal: 12.9 mg/L.

 

The characteristics of the achieved atmosphere were as follows:

Mean Achieved Atmosphere Concentration: 5.99 mg/L

Mean mass Median Aerodynamic Diameter: 3.15 um

Inhalable Fraction (< 4 um): 62.6%

Geometric Standard Deviation: 2.10.

 

Mortality. There were no deaths in both sexes during the study.

Clinical Observations. Common abnormalities noted during the study included increased respiratory rate, hunched posture, pilo-erection and wet fur. There was an isolated instance of red/brown staining to the eyes. All animals recovered quickly to appear normal by Day 2 or Day 3 post-exposure.

Bodyweight. Normal bodyweight development was noted during the study.

Necropsy. Apart from two instances of dark patches on the lungs, no macroscopic abnormalities were detected amongst animals at necropsy.

 

Conclusion. No deaths occurred in a group of 10 rats exposed to a mean achieved atmosphere concentration of 5.99 mg/L for 4 hours. It was therefore considered that the acute inhalation median lethal concentration (4 h LC50) of TIPX, in the Sprague-Dawley Crl:CD® (SD) IGS BR strain rat, was greater than 5.99 mg/L. The test material does not meet the criteria for classification according to EU classification system (Council Directive 67/548/EEC and Regulation (EC) No 1272/2008).