Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-06-03 to 2011-07-13
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study

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)
Version / remarks:
2009
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Version / remarks:
2008
Qualifier:
according to
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Version / remarks:
1998
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
CAS name: 1,4-Cyclohexanedicarboxamide, N1,N1,N4,N4-tetrakis(2-hydroxyethyl)-, trans
Chemical characterization: Trans-N,N,N',N'-Tetrakis(2-hydroxyethyl)-cyclohexyl-1,4-diamide
Characteristics: Whitish, solid, powder, hydroscopic
Contents:
91.53 % Trans-N,N,N',N'-Tetrakis(2-hydroxyethyl)-cyclohexyl-1,4-diamide

Test animals

Species:
rat
Strain:
other: Rattus norvegicus
Sex:
male/female
Details on test animals and environmental conditions:
TEST ORGANISMS: 
- Strain: Rattus norvegicus) / CD / Crl: CD(SD)
- Source: Charles River Laboratories, 97633 Sulzfeld, Germany
- Age: males: approx. 7 weeks, females: approx. 9 weeks
- Weight at study initiation: males: 260 - 288 g, females: 242 - 247 g
- Number of animals: 3 males and 3 females
- Fasting period before study: 16 hours
- Housing: groups of two or three
- Diet: ad libitum, ssniff R/m-H V 1534
- Water: ad libitum, tab water
- Acclimatisation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 3 °C
- Humidity (%): 55 + / - 15 %
- Photoperiod (hrs dark / hrs light): 12 hours darkness, 12 hours artifical light

Administration / exposure

Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
other: compressed filtered air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Type of exposure: nose-only, using a dynamic inhalation apparatus, air changes >/= 12/h
- Method of holding animals in test chamber (volume 40 l): separatley in Pyrex tubes at the edge of the chamber in radial position
- Source of air: compressed filtered air
- Method of conditioning air: manometer and air-flow meter (ROTA Yokogawa), flow rates checked once per hour and corrected if necessary
- Oxygen content: 21 %, determined at beginning and end of exposure, Carbon dioxide concentration did not exceed 1 %
- Type or preparation of particles: dust is generated with a rotating brush dust generator (RBG 1000, PALAS GmbH Partikel und Lasermesstechnik),
fed with compressed air (5.0 bar). At the bottom of the exposure chamber, the air was sucked off at a lower flow rate than it was created by the
dust generator in order to produce a homogenous distribution and a positive pressure in the exposure chamber (inflow 900 L/h, outflow 800 L/h).
- Method of particle size distribution: using a cascade impactor acording to MAY, analysis is carried out twice during inhaltion period.
Dust from the exposure chamber was drawn through the cascade impactor for 5 minutes at a constant flow rate of 5 L/min. The slides were
removed from the impactor and weighed on an analytical balance. The mass median aerodynamic diameter (MMAD) was determined as 3.615 µm.
The Geometric Standard Deviation (GSD) of the MMAD was calculated as 2.85.
- Temperature, humidity: T: 23 +/- 0.29°C, H: 63.4 % +/- 0.18 %, measured once every hour
TEST ATMOSPHERE
- Concentrations: 5.07 mg/l 
Air flow entrance (L/h): 900
Air flow exit (L/h): 800
Air change (changes per hour): 22.5

Analytical verification of test atmosphere concentrations:
yes
Remarks:
Gravimetric determination of aerosol concentrations with an air sample filter. Dust samples were taken once every hour during the exposure.
Duration of exposure:
4 h
Concentrations:
5.07 mg/l gravimetric concentration
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
EXAMINATIONS: 
- Post dose observation period: 14 days
- body weights: before,  7 and 14 days after treatment   
- mortality: once per hour during, and once after treatment on day of  exposure; 
thereafter twice daily
- clinical signs: during and following exposure, observations were made and recorded systematicallye,  at least twice daily until all symptoms
subsided, thereafter each working day
- Necropsy: all animals (macroscopic) when found dead or at terminal  sacrifice. In addition, the weight of the lungs was determined.
Statistics:
not necessary

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.07 mg/L air
Based on:
act. ingr.
Exp. duration:
4 h
Mortality:
No animal died prematurely.
Clinical signs:
other: Slight ataxia and slight dyspnoea in all 3 male and 3 female rats.
Body weight:
One of 3 female animals appeared to be reduced in body weight gain.
Gross pathology:
None findings
Other findings:
no other findings

Any other information on results incl. tables

no further remarks


Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: other: EC Regulation 1272/2008
Conclusions:
Under the present test conditions, LC50-value for CD rats following inhalation of trans-N1,N1,N4,N4-Tetrakis(2-hydroxyethyl)1,4-cyclohexanedicarboxamidefor 4 hours was determined as follows (actual concentration): LC50: exceeding 5.07 mg/L air. According to the EC Regulation 1272/2008 and subsequent regulations on classification, labelling and packaging of substances and mixtures, the substance does not require classification for acute inhalation toxicity.
Executive summary:

The aim of the present experiment was to obtain information on the acute toxicity and LC50, following a single 4-hour inhalation exposure in an acute inhalation study in rats.

3 male and 3 female rats were exposed to trans-N1,N1,N4,N4-Tetrakis(2-hydroxyethyl)1,4-cyclohexanedicarboxamide at a concentration of 5.07±0.01 mg test substance/L air for 4 hours by inhalation using a dynamic nose-only exposure chamber. The aerosol was generated with the aid of a dry, rotating brush dust generator. In the inhalation chamber, close to the animals' noses, the generated aerosol particulates had a mass median aerodynamic diameter (MMAD) of 3.615 µm as deter­mined with a cascade impactor. The Geometric Standard Deviation (GSD) of the MMAD was calculated as 2.85.

Under the present test conditions, a 4-hour inhalation LC50 of trans-N1,N1,N4,N4-Tetrakis(2-hydroxyethyl)1,4-cyclohexanedicarboxamide revealed slight ataxia and slight dyspnoea immediately until 60 minutes or 3 hours after end of exposure. No animal died prematurely. No pathological findings were noted at necropsy. One of the 3 female animals appeared to be reduced in body weight gain.

Therefore LC50 exceeding 5.07 mg/L air for both sexes combined.