Reaction mass of 2-ethyl-2-[[3-(2-methylaziridin-1-yl)propionyl]methyl]propane-1,3-diyl bis(2-methylaziridine-1-propionate) and 2,2-bis({[3-(2-methylaziridin-1-yl)propanoyl]oxy}methyl)butyl 3-[2,2-bis({[3-(2-methylaziridin-1-yl)propanoyl]oxy}methyl)butoxy]propanoate

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 25, 2001 - Jan 09, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The experiment was conducted according to current OECD guidelines in compliance with GLP standards.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

1981

Deviations:

no

Remarks:

, according to 2009 adopted guideline, traditional protocol was used

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

IONAC XAMA-220

IUPAC Name:

IONAC XAMA-220

Details on test material:

- Chemical name of test material: Polyfunctional aziridine
- CAS no.: 64265-57-2
- Appearance: amber liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan, Winkelmann GmbH, Borchen (Germany)
- Age at study initiation: appr. 2 months
- Weight at study initiation: all within 10% of the mean for each sex
- Fasting period before study: none
- Housing: individual housing in Makrolon cages.
- Diet: ad libitum, standard fixed-formula diet (NAFAG No. 9439 pellets maintenance diet for rats and mice)
- Water: ad libitum, tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±2
- Humidity (%):appr. 50%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: August 7, 2001 To: September 3, 2001

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: directed-flow nose-only exposure
- Exposure chamber volume: inner diameter = 14 cm, outer diameter = 35 cm (two chamber system), height = 25 cm (internal volume = about 3.8 l).
- Method of holding animals in test chamber: Rats were kept in place in plexiglas exposure tubes accommodated to fit the animals size (commercially available: TSE, 61348 Bad Homburg). The tail remained outside the tubes.
- Source and rate of air:>200 air exchanges per hour
- Method of conditioning air: Compressed air was supplied by Boge compressors and was conditioned (i.e. freed from water, dust, and oil) automatically by a VIA compressed air dryer. Adequate control devices were employed to control supply pressure.
- System of generating particulates/aerosols: Under dynamic conditions the test substance was nebulized neat into a baffle (pre-seperator) from which the substance was conveyed into the intake of the inhalation chamber. The substance was nebulized using a binary nozzle with conditioned compressed air (15 L/min; dispersion pressure approximately 600 kPa).
- Method of particle size determination: analyzed by a BERNER-TYPE AERAS low-pressure critical orifice cascade impactor.
- Treatment of exhaust air: purified via cotton-wool/ HEPA filters.
- Temperature, humidity, pressure in air chamber: 22-24C; <7%;

TEST ATMOSPHERE
- Brief description of analytical method used: Gravimetric analysis
- Samples taken from breathing zone: yes
- Integrity and stability of aerosol generation and exposure system was monitored by a RAS-2 real-time aerosol photometer (MIE, Bedford, Massachusetts, USA)

TEST ATMOSPHERE
- Particle size distribution: Aerosol mass <3µm= appr. 92%
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 1.36-1.44 µm/ 1.69-1.72

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetrically

Duration of exposure:

4 h

Concentrations:

100, 170, 300, 400, 460 mg/m3

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: several times on the day of exposure, at least daily thereafter
- Frequency of weighing: Day 0, 3, 7 and 14
- Necropsy of survivors performed: yes
- Other examinations performed: cage side observations including clinical signs,somatomotor activity and behavior pattern; reflexes were tested; rectal temperature was measured 30 min after exposure.

Statistics:

One-way ANOVA analysis was used for statistical evaluation of body weight gain, necropsy findings and rectal temperature.
Normal distribution of data was checked by comparing the median and the mean by analysis of variance (ANOVA).
LC50 was calculated according to the method of Rosiello et al (1977), modified by Pauluhn (1983), based on the maximum-likelihood nethod of Bliss (1938).

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

252 mg/m³ air

Based on:

test mat.

95% CL:

> 209 - < 304

Exp. duration:

4 h

Mortality:

Mortality occurred at exposure level equal to and above 105 mg/m3 (actual concentration). Rats succumbed on the exposure day up to day 8.
conc. M F
100 1/5 0/5
170 0/5 0/5
300 2/5 4/5
400 3/5 5/5
460 5/5 5/5

Clinical signs:

other: At lowest exposure (100mg/ m3) piloerection, ungroomed hair-coat, labored breathing pattern, bradypnea, breathing sounds (rales), dyspnea, limp, reduced motility, high-legged-gait, reddened nostrils with red crustrations, nasal discharge (serous), cyanosi

Body weight:

Comparisons between control animals revealed statistically significant effects on body weights in all groups exposed to the test compound.

Gross pathology:

Animals succumbed during the course of the study: Muzzle/nose: red encrustations. Discharge of clear liquid from the nose. Lung less collapsed, dark-red areas, trachea with foamy content. Hydrothorax. GI-tract: yellowish mucus.
Animals sacrificed at the end of the observation period: Rats exposed to the test compound displayed a somewhat increased incidence of macroscopic findings in the lung. These findings in the lung (discolorations) are suggestive of lung edema and associated damage in the respiratory tract.

Other findings:

Changes in reflexes were observed in all exposed groups on the first postexposure day.
Statistical comparisons between control animals with the groups exposed to the test substance revealed statistically significant differences, i.e., a hypothermic response.

Any other information on results incl. tables

Nominal conc.: 100, 170, 300, 400, 460 mg/m3

Grav. conc.: 105, 164.4, 280.8, 403.8, 478 mg/m3

Applicant's summary and conclusion

Interpretation of results:

toxic

Remarks:

Migrated information class 2 Criteria used for interpretation of results: EU

Conclusions:

In an acute inhalation study with male and female rats according to current EC/OECD guidelines and GLP principles, the 4-hour LC50 was found to be 0.252 mg/L air.

Executive summary:

An acute inhalation study was performed with 5 male and 5 female rats at several concentrations according to current EC/OECD guidelines and GLP principles. Mortality occurred at exposure levels equal to and above 105 mg/m3 (actual concentration). Rats succumbed on the exposure day up to day 8. Exposed rats showed several clinical signs (pilolerection, lung effects, limbness etc.). Animals exposed to the test substance showed hypothermic reactions shortly after exposure and weight gain was negatively affected in surviving animals (statistically significant). Animals succumbed during the course of the study showed red encrustations and discharge of clear liquid from the nose, lung less collapsed, dark-red areas, trachea with foamy content. Animals sacrificed at the end of the observation period displayed a somewhat increased incidence of macroscopic findings in the lung. These findings in the lung (discolorations) are suggestive of lung edema and associated damage in the respiratory tract. The 4-hour LC50 was found to be 0.252 mg/L air.