N-[2-[(2,6-dicyano-4-nitrophenyl)azo]-5-(diethylamino)phenyl]acetamide

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 6, 1979 to March 26, 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Inhalation toxicity was tested according to the method of Sachsse et al. (1973, 1976).

Deviations:

no

Principles of method if other than guideline:

K. Sachsse, L. Ullmann, G. Voss and R. Hess: Measurement of inhalation toxicity of aerosols in small laboratory animals. In: Proceedings of the Europ. Soc. for the Study of Drug Toxicity. Vol XV, pp. 239-251, Zurich, June 1973.
K. Sachsse, L. Ullmann, K. Zbinden: Toxikologische Prüfungen von Aerosolen im Tierexperiment: Aus "Chemische Rundschau" 29 (1976), Nr. 38 Seite 1-4.

GLP compliance:

no

Remarks:

Pre-dates GLP

Test type:

traditional method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N-[2-[(2,6-dicyano-4-nitrophenyl)azo]-5-(diethylamino)phenyl]acetamide

EC Number:

255-473-8

EC Name:

N-[2-[(2,6-dicyano-4-nitrophenyl)azo]-5-(diethylamino)phenyl]acetamide

Cas Number:

41642-51-7

Molecular formula:

C20H19N7O3

IUPAC Name:

N-[2-[(2,6-dicyano-4-nitrophenyl)azo]-5-(diethylamino)phenyl]acetamide

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

other: Tif: RAIf

Sex:

male/female

Details on test animals or test system and environmental conditions:

The test was performed with 30 male and 30 female young adult rats of the Tif: RAIf (SPF) strain, raised on the premises. An additional group of 20 (10 males/10 females) rats served as a control.
Before and after the inhalation the animals were kept at a room temperature of 22 + 2° C, at a relative humidity of 55 + 10 % and on a 10 hours light cycle day. They received ad libitum rat food - NAFAG, Gossau SG - and water. Prior to treatment they were adapted to our laboratories for a minimum of 4 days. Bodyweights were recorded immediately prior to exposure (control weights) and at day 7 and 14 (Table 2). The males and females were segregated and kept in Macrolon cages, type 4, (10 animals to a cage), individually marked with picric acid.

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

Preparation of aerosol
The aerosol was generated by injecting the solid test material with the help of a Grafix Exaktomat Injector (Cerutti AG, Bern, Switzerland) into an air stream which was discharged into the exposure chamber at a rate of 20 l/min. The control animals were exposed to filtered air under the same conditions as described above.
The concentration and the particle size distribution of the aerosol in the vicinity of the animals were monitored at regular intervals throughout the aerosol exposure. The concentration was determined 5 times gravimetrically by sampling the test atmosphere through a selectron filter of 50 mm diameter and with a pore size of 0.2 μm (Schleicher and Schuell, Feldbach, Switzerland) at an air flow rate of 10 1/min. The size distribution of the particles was measured twice with a 4 stage Cascade Impactor with selectron filters of 25 mm diameter and with a pore size of 0.2 μm (Schleicher and Schuell) at an air flow rate of 17.5 l/min.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

107 ± 6 mg/m3, 194 ± 8 mg/m3 and 317 ± 22 mg/m3

No. of animals per sex per dose:

10 male/10 female per dose

Control animals:

yes

Details on study design:

Testing procedures
For inhalation the rats were kept in separate PVC tubes positioned radially around the exposure chamber such that snout and nostrils of the animals only were exposed to the aerosol.
During the exposure period the following parameters were controlled once at half time of the study inside the inhalation cylinder: temperature (with a Therm 2104 contact thermometer, Ahlborn Messund Regeltechnik, 815 Holzkirchen, Germany), relative humidity (with a VASALA Humidity Indicator HMI 11, Kelag AG, 8057 Zurich, Switzerland) and oxygen content (with a DRAEGER E 15 stationary control system, Draegerwerk AG, Lübeck, Germany).
After a 4 hour inhalation the rats were returned to their cages. Physical condition and incidence of death were monitored throughout an observation period of 14 days.

Statistics:

Not specified in the study report.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 300 mg/m³ air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality

Clinical signs:

other: The animals exposed to the test material recovered within 3 to 4 days. They were submitted to a necropsy at the end of the observation period. The control rats failed to show any symptoms, during the exposure and observation period.

Body weight:

Reported in table form - see Any other information

Gross pathology:

Partially congested organs were observed.

Other findings:

Particle size distribution analysis of the chamber airborne particles showed that 20 to 60 % were smaller than 7 μm in diameter.

Any other information on results incl. tables

Temperature, relative humidity and oxygen control

The following values were obtained during the exposure period.

	Control group 0 mg/m^3	107 ± 6 mg/m^3	194 ± 8 mg/m^3	317 ± 22 mg/m^3
Temperature °C	23	24	24	24
Relative Humidity %	58	40	42	55
Oxygen content Vol. %	20	20	20	20

 

RATE OF DEATHS

Dose mg/m^3	Sex	Total Number animals in group	Total Number animals dead	Death rate percentage
0 Control	Male	10	0	0
107 ± 6		10	0	0
194 ± 8		10	0	0
317 ± 22		10	0	0
0 Control	Female	10	0	0
107 ± 6		10	0	0
194 ± 8		10	0	0
317 ± 22		10	0	0

 

BODYWEIGHT CHANGE

		Concentration mg/m^3
		0 Control	107 ± 6	194 ± 8	317 ± 22
Day 1 Male	MEAN BODYWEIGHT (g)	237	230	238	187
Day 1 Female		205	194	191	181
Day 7 Male		260	255	247	219
Day 7 Female		212	204	191	189
Day 14 Male		290	303	292	273
Day 14 Female		221	224	210	209

 

Signs and symptoms

Concentration (mg/m^3): 107 ± 6

Signs and symptoms

Exposure Hours

Days

1

2

4

6

24

2

3

4

5

6

7

8

9

10

11

12

13

14

Sedation

Dyspnoea

+

+

Dacryorrhoea

Chromodacryorrhoea

Rinorrhoera

Epistaxis

Exophthalmos

+

+

+

+

Salivation

Ruffled fur

+

+

+

Pallor

Cyanosis

Diarrhoea

Body position (ventral)

Body position (lateral)

Body position (curved)

+

Ataxia

Trismus

Tremor

Tonic clonic muscle spasms

Convulsions

Key:       + = slight,             ++ = moderate,                  +++ = severe

 

Concentration (mg/m^3): 194 ± 8

Signs and symptoms

Exposure Hours

Days

1

2

4

6

24

2

3

4

5

6

7

8

9

10

11

12

13

14

Sedation

Dyspnoea

+

+

Dacryorrhoea

Chromodacryorrhoea

Rinorrhoera

Epistaxis

Exophthalmos

+

+

+

+

Salivation

Ruffled fur

+

+

+

Pallor

Cyanosis

Diarrhoea

Body position (ventral)

Body position (lateral)

Body position (curved)

+

+

+

Ataxia

Trismus

Tremor

Tonic clonic muscle spasms

Convulsions

Key:       + = slight,             ++ = moderate,                  +++ = severe

 

Concentration (mg/m^3): 317 ± 22

Signs and symptoms

Exposure Hours

Days

1

2

4

6

24

2

3

4

5

6

7

8

9

10

11

12

13

14

Sedation

Dyspnoea

+

+

Dacryorrhoea

Chromodacryorrhoea

Rinorrhoera

Epistaxis

Exophthalmos

+

+

+

+

Salivation

Ruffled fur

+

+

+

Pallor

Cyanosis

Diarrhoea

Body position (ventral)

Body position (lateral)

Body position (curved)

+

+

+

Ataxia

Trismus

Tremor

Tonic clonic muscle spasms

Convulsions

Key:       + = slight,             ++ = moderate,                  +++ = severe

Applicant's summary and conclusion

Interpretation of results:

other: LC50 is greater than the highest dose tested in this study.

Conclusions:

The LC50 of a 4 hour aerosol exposure for rats of both sexes is greater than 300 mg/cubic-meter air, when evaluated for a 14 day post treatment observation period.

Executive summary:

Inhalation toxicity was tested according to the method of Sachsse et al. (1973, 1976).

 

For inhalation the rats were kept in separate PVC tubes positioned radially around the exposure chamber such that snout and nostrils of the animals only were exposed to the aerosol.

 

Results

Particle size distribution analysis of the chamber airborne particles showed that 20 to 60 % were smaller than 7 μm in diameter.

The animals exposed to the test material recovered within 3 to 4 days. They were submitted to a necropsy at the end of the observation period.

The control rats failed to show any symptoms, during the exposure and observation period.

 

The LC50 of a 4 hour aerosol exposure for rats of both sexes is greater than 300 mg/cubic-meter air, when evaluated for a 14 day post treatment observation period.