Reaction mass of N, N'-hexane-1,6-diylbis [12-hydroxyoctadecanamide] and 12-hydroxy-N-[6-[1-oxoalkyl)amino] hexyl ] octadecanamide

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7 November - 21 November 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to OECD and EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese 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:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): AD-1000
- Substance type: white powder
- Physical state: powder
- Storage condition of test material: at room temperature in the dark

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 were selected (10 weeks old).
- Weight at study initiation: Animals used within the study were of approx. the same age and body weight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study: not applicable
- Housing: Before exposure: Group housing of 5 animals per sex per cage in labelled Macrolon cages (type IV; height 18 cm.) containing sterilised sawdust as bedding material and paper as cage-enrichment. After exposure: Group housing, maximally 5 animals per sex per cage in labelled stainless steel wire mesh cages. Two days after exposure the animals were housed as described above.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet except during exposure to the test substance.
- Water (e.g. ad libitum): Free access to tap water except during exposure to the test substance.
- Acclimation period: Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 – 22.7
- Humidity (%): 37 - 60
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

Nose-only
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: restraining tubes, which were connected to the exposure chamber. The design of the exposure chamber was based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983). The chamber consisted of 4 animal sections with 8 animal ports each. The number of open animal ports was adapted to the air flow in such a way that at each animal port the theoretical air flow was approximately 2.0 l/min, which ensures an adequate oxygen supply to the test animals. The inlet of the test atmosphere was located in the top section and the outlet was located in the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal. The placement of the individual animals in the inhalation chamber is shown in figure 2. All components of the exposure chamber which could come in contact with the test material were made of stainless steel, glass, rubber or plastic. To avoid exposure of the personnel and contamination of the laboratory the exposure chamber was placed in a fume hood, which maintained a slight negative pressure.

- System of generating particulates/aerosols: A dry powder aerosol was generated by administering the test substance to a stream of pressurized air (mean air flow 38 l/min) by means of a combination of a spiral feeder. The aerosol was led to a cyclone where larger particles were allowed to settle. Subsequently the aerosol was passed through the exposure chamber. From the exposure chamber the test atmosphere was passed through a set of filters before it was released to the exhaust of the fume hood.

- Method of particle size determination: The droplet size distribution was characterized twice during the exposure period. The samples were drawn (2 l/min) from the test atmosphere through a tube mounted in one of the free animal ports of one of the middle sections of the exposure chamber. The samples were collected with an 8 stage Marple personal cascade impactor, fiber glass filters and a fiber glass back-up filter. Amounts of test substance collected were determined gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation were determined.

- Temperature, humidity, pressure in air chamber: 20.1-20.7oC; 30.5-37.5%;

TEST ATMOSPHERE
Nominal concentration: The nominal concentration was calculated by dividing the amount of test substance used by the volume of pressurized air (average air flow times exposure time) entering the exposure chamber used for exposure of the animals.
Actual concentration: The actual concentration was determined 7 times during the exposure period. Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of one of the middle sections of the exposure chamber. Samples were drawn through a glass fiber filter. The collected amount of test substance in the air sample was measured gravimetrically. Sample volumes were measured by means of a dry gas meter. Subsequently the mean concentrations and the standard deviations were calculated.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

The mean actual concentration was 5.2 ± 0.4 mg/l. The nominal concentration was 14.6 mg/l. The generation efficiency (ratio of actual and nominal concentration) was 35.6%.

No. of animals per sex per dose:

one group of five male and five female Wistar rats

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Necropsy of survivors performed: yes/no
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Statistics:

No statistical analysis was performed (the method used was not intended to allow the calculation of a precise LC50 value).

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5 mg/L air

Exp. duration:

4 h

Mortality:

Male: 5.2 mg/L; Number of animals: 5; Number of deaths: 0
Female: 5.2 mg/L; Number of animals: 5; Number of deaths: 1

Clinical signs:

other: During exposure: No clinical signs were noted. After exposure: Hunched posture, piloerection, ptosis and slow breathing were noted among the majority of animals. Laboured respiration was noted among the majority of the males and rales were noted in one f

Body weight:

The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

Effects on organs:
Granular, gray-white contents in the trachea, gray-white discolouration of the tongue and many, dark red
foci on the lungs were found at macroscopic post mortem examination of the animal that died during the study
Macroscopic post mortem examination of the surviving animals at termination did not reveal any abnormalities.

Any other information on results incl. tables

The mean ' standard deviation of the temperature and relative humidity during exposure were 20.6 ' 0.1°C and 34.8
' 2.5%.
The mean actual concentration was 5.2 ' 0.4 mg/l. The nominal concentration was 14.6 mg/l. The generation
efficiency (ratio of actual and nominal concentration) was 35.6%.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The inhalatory LC50, 4h value of AD-1000 in Wistar rats was established to exceed 5 mg/l.