[No public or meaningful name is available]

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 04 - June 19, 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study has been performed according to OECD guideline 403 and GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

(1981 and draft document 1996)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(1992)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

(1998)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanes Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

453-140-3

EC Name:

-

Cas Number:

667465-46-5

Molecular formula:

C20H38N6O13 (not dissociated) C20H44N6O13 (dissociated in water)

IUPAC Name:

3,6,9-tris(2-{[(2-hydroxyethyl)amino]oxy}-2-oxoethyl)-3,6,9-triazaundecan-1,11-dioic acid

Constituent 2

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

Water

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): DHX2
- Physical state: clear yellow liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: young adult animals (13 weeks old)
- Weight at study initiation: +/- 20% of sex mean
- Housing: group housing (5/cage) in labelled Macrolon cages containing sterilised sawdust as bedding material and paper as cage-enrichment
- Diet: free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) except during exposure to the test substance
- Water: free access to tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3-21.4
- Humidity (%): 41-78
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: 4-19 June 2008

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: The chamber consisted of 3 animal sections with 8 animal ports each. The inlet of the test atmosphere was located in the top section and the outlet was located in the bottom section.
- Exposure chamber volume: not indicated
- Method of holding animals in test chamber: restraining tube
- Source and rate of air: pressurized air; 2.0 L/min
- System of generating aerosols: generated by two nebulizers (type 950, Hospitak Inc. Lindenhurst, NY, USA)
- Method of particle size determination: gravimetrically, samples collected with an 8-stage Marple personal cascade impactor fiber glass filters (FG-033450-GA, AE Fiber glass, SKC Omega Specialty Division, Chelmsford, MA, USA) and a fiber glass back-up filter (SEC-290-F1, Westech, Upper Stondon, Bedfordshire, England)
- Treatment of exhaust air: passed through a filter to determine the nominal concentration
- Temperature, humidity, pressure in air chamber: 21.0-21.4 °C; relative humidity was not measured; pressure was not reported

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetrical analysis: Samples were drawn through a glass fiber filter (type APFC04700, Millipore, Billerica, MA, USA). The filters were dried in a kiln for 5 minutes before weighing to allow the volatile part of the test substance to evaporate. The collected amount of the non-volatile part of the test substance in the air sample was measured gravimetrically. Sample volumes were measured by means of a dry gas meter (type G 1.6, Actaris Meterfabriek B.V., Dordrecht, The Netherlands).
- Samples taken from breathing zone: yes, from one of the free animal ports

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: determined twice
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 1.7 µm/1.9 at both occasions

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetrically, 8 times 3L sample during exposure

Duration of exposure:

4 h

Concentrations:

nominal 14.1 mg/L; actual 5.2 +/- 0.3 mg/L

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: during exposure, 3 times for mortality, behavioural signs of distress and effects on respiration; twice daily for mortality/viability; twice on the day of dosing and once daily thereafter for clinical signs; days 1 (pre-administration), 8 and 15 for body weights
- Necropsy of survivors performed: yes

Statistics:

None

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.2 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality occurred.

Clinical signs:

other: During exposure irregular breathing was noted in all animals at 1 hour after initiation of exposure. No symptoms were noted at 2 and 3 hours after initiation of the exposure. After exposure, between Days 1 and 5, chromodacryorrhoea on the snout, hunched p

Body weight:

During the first week after exposure the animals showed body weight loss or reduced body weight gain, The body weight gain during the second week after exposure shown by the animals was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The inhalatory LC50, 4h value of DHX2 in Wistar rats was established to exceed 5 mg/L.

Executive summary:

Rats (5/sex) were exposed to 5.2 mg/L of the test substance by inhalation according to OECD 403 and GLP principles.

No mortality occurred. During exposure irregular breathing was noted in all animals. Day 1 to 5 after exposure chromodacryorrhoea on the snout, hunched posture, laboured respiration, lethargy and rales were noted in the majority of the animals and piloerection in females. Body weight loss/reduced body weight gain was seen during the first week of exposure only. No abnormalities were found at macroscopic examination.

The inhalation LC50 (4 -hours) of the test substance in rats was established to exceed 5.2 mg/L.