Registration Dossier

Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Principles of method if other than guideline:
Acute Inhalation Toxicity Study of 9,10-Anthracenedione, 1, 4-Diamino-N-N’-Bis (4-C7-17-Branched Alkyl phenyl) Derives in albino rat
GLP compliance:
no
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Details on test material:
-Substance Name: 9,10-Anthracenedione, 1,4-diamino-, N,N'-bis(4-C7-17-branched alkylphenyl) derivs
-Substance type: Organic
-Physical state: Liquid

Specific details on test material used for the study:
Name: 9, 10-Anthracenedione, 1, 4-Diamino-N-N’-Bis (4-C7-17-Branched Alkylphenyl) Derives
- Physical state: Dark blue liquid

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Source : Institute for Industrial Research & Toxicology
Age : 7 to 9 weeks
Sex : Male and Female
Body weight range :200 ±20g
Identification : By cage tag and corresponding colour body marking
No. of animals per dose group :10 (5 males and 5 females)
No. of dose group : Two :Group I – Limit test (5 mg/L) and Group II – Confirmatory test (5 mg/L)
Acclimatization :Twenty healthy albino rats were selected and acclimatized for standard laboratory condition for period of one week in experimental room under veterinary examination.
Randomization : After acclimatization and veterinary examination all the selected rats randomly divided into two groups of five females and five males.
Nutritional conditions :For feeding conventional Laboratory diets may be used with an unlimited supply of drinking water.
HUSBANDRY
Environmental conditions :Air conditioned rooms with 10-15 air changes per hour, temperature between 19-25 0C, relative humidity 30-60% and illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.
Accommodation :Groups of five animals of similar sex in polypropylene cages with stainless steel grill top, facilities for food and water bottle, and bedding of clean paddy husk.
Diet : Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India
Water : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad-libitum

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: 0.1% tween 80 in Distilled water
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:Nanotek aerosol generator
- Exposure chamber volume: 8 liters
- Method of holding animals in test chamber:For the inhalation purpose the rats were placed in polycarbonate holder tubes positioned radically around exposure chamber, so that only the snouts and nostrils of the animals were exposed to the aerosol.
- Source and rate of air: No data available
- Method of conditioning air:The chamber was maintained at a slightly negative pressure to prevent leakage of the test atmosphere from system, as well as its dilution with outside air.
- System of generating particulates/aerosols:Nanotek aerosol generator
- Method of particle size determination:
- Treatment of exhaust air: The exhaust air was decontaminated by subsequent passage through 1% NaOH solution, silica gel and activated charcoal filters.
- Temperature, humidity, pressure in air chamber: Temperature- 23.29±1.08 - 23.47±1.74, Relative humidity (%): 46.31±3.24 - 47.34±3.56

TEST ATMOSPHERE
- Brief description of analytical method used: No data available
- Samples taken from breathing zone: no

VEHICLE
- Composition of vehicle (if applicable):0.1% tween 80 in Distilled water
- Concentration of test material in vehicle (if applicable): 5 mg/L
- Justification of choice of vehicle: No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: less than 1 micron
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): No data available

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration:No data available
Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
4 h
Concentrations:
5 mg/L
No. of animals per sex per dose:
Total: 20
Group I – Limit test (5 mg/L) : 5 males and 5 females
Group II – Confirmatory test (5 mg/L): 5 males and 5 females
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:closely observed for any clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days.
- Necropsy of survivors performed: yes
- Other examinations performed: Mortality, clinical signs, body weight and gross pathology were examined.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 mg/L air (nominal)
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: No mortality observed
Mortality:
No mortality was recorded in any of the Wistar albino rats in both limit test and confirmatory test after 4 hours of exposure.
Clinical signs:
other: No clinical signs of intoxication at the tested aerosol concentration of 5 mg/L observed for the period of 14 days.
Body weight:
No significant increase or decrease in body weight on day 7th and 14th were observed when compared with day 0.
Gross pathology:
No gross pathological changes were observed in treated rats.

Any other information on results incl. tables

EXPOSURE ATMOSPHERE DATA 

Parameters

Group-I (5 mg/L)

(limit test)

Group-II (5 mg/L)

(confirmatory test)

Chamber temperature °C (Mean ± S.E.)

23.29±1.08

23.47±1.74

Relative humidity (%)

 (Mean ± S.E.)

46.31±3.24

47.34±3.56

Oxygen content (%)

(Mean ± S.E.)

20.21±1.16

21.34±1.52

 

Mean Body Weight (gm)

S.No.

Groups

BODY WEIGHT (gm)

DAY 0

DAY 7th

% gain or loss

DAY14th

% gain or loss

1.

Group-I (5.0 mg/L)

200.37

205.69

2.65

211.71

5.65

2.

Group-II (5.0 mg/L)

203.11

209.82

3.30

214.24

5.47

CLINICAL SIGNS AND MORTALITY

Group: I Limit test                                                                         Dose: 5.0 mg/L

 

Parameters

Incidence of Clinical Signs Observed after Dosing on

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

%

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0/10

 

0

Clinical Signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0         =   No clinical sign (Normal)

+         =   Mild

++        =   Moderate

+++      =   High

++++  =         Severe

CLINICAL SIGNS AND MORTALITY

Group: II Confirmatory test                                                                    Dose: 5.0 mg/L

 

Parameters

Incidence of Clinical Signs Observed after Dosing on

 

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

%

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0/10

 

0

Clinical Signs

0

0

0

0

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 0         =   No clinical sign (Normal)

+         =   Mild

++        =   Moderate

+++      =   High

++++    =   Severe

SUMMARY OF NECROPSY FINDING

 

S. No.

 

Fate

 

Wistar albino rats

Dose (mg/l)

5.0

(limit test)

5.0

(confirmatory test)

1

Terminal sacrifice

10/10

10/10

2

Found Dead

0/10

0/10

3

Abnormalities detected

0/10

0/10

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: I Limit test                                                                          Dose: 5.0 mg/L       

Animal ID

Fate

Time

Gross Findings

20172-1

TS

Day 14

NAD

20172-2

TS

Day 14

NAD

20172-3

TS

Day 14

NAD

20172-4

TS

Day 14

NAD

20172-5

TS

Day 14

NAD

20172-6

TS

Day 14

NAD

20172-7

TS

Day 14

NAD

20172-8

TS

Day 14

NAD

20172-9

TS

Day 14

NAD

20172-10

TS

Day 14

NAD

Day 0 is the day of exposure

TS=Terminal Sacrifice

NAD=No Abnormality Detected

FD=Found Dead

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: II Confirmatory test                                                                    Dose:5.0 mg/L       

Animal ID

Fate

Time

Gross Findings

20172-11

TS

Day 14

NAD

20172-12

TS

Day 14

NAD

20172-13

TS

Day 14

NAD

20172-14

TS

Day 14

NAD

20172-15

TS

Day 14

NAD

20172-16

TS

Day 14

NAD

20172-17

TS

Day 14

NAD

20172-18

TS

Day 14

NAD

20172-19

TS

Day 14

NAD

20172-20

TS

Day 14

NAD

Day 0 is the day of exposure

TS=Terminal Sacrifice

NAD=No Abnormality Detected

FD=Found Dead

Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
Acute lethal Concentration (LC50) of test compound was found be more than 5 mg/L. when Wistar Albino Male and Female rats were inhaled for 4 hours.
Executive summary:

In a acute inhalation toxicity study,Wistar Albino Male and Female rats were exposed with test chemical in the concentration of 5 mg/L in a dynamic nose-only cylindrical chamber built from stainless steel and glass. The chamber had a volume of 8 liters with inner and outer chamber to minimize the fluctuation in concentration and temperature for 4 hours. No mortality was recorded in any of the Wistar albino rats in both limit test and confirmatory test after 4 hours of exposure. No clinical signs of intoxication at the tested aerosol concentration of 5 mg/L observed for the period of 14 days. No significant increase or decrease in body weight on day 7thand 14thwere observed when compared with day 0. No gross pathological changes were observed in treated rats. After 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration. No effects were observed in exposed rats at 5 mg/L. Therefore, Acute lethal Concentration (LC50) of test compound was found be more than 5 mg/L. when Wistar Albino Male and Female rats were inhaled for 4 hours.