9,10-Anthracenedione, 1,4-diamino-, N,N'-mixed 2-ethylhexyl and hexyl and octyl derivs.

Acute Toxicity: inhalation

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 with acceptable restrictions

Justification for type of information:

Data is from experimental study report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Principles of method if other than guideline:

The study was conducted to find out the mortality(LC50), clinical effect and histopathological effect of test compound at different aerosol concentration in Wistar albino rats.

GLP compliance:

no

Test type:

other: not specified

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

9,10-Anthracenedione, 1,4-diamino-, N,N'-mixed 2-ethylhexyl and hexyl and octyl derivs.

EC Number:

297-755-3

EC Name:

9,10-Anthracenedione, 1,4-diamino-, N,N'-mixed 2-ethylhexyl and hexyl and octyl derivs.

Cas Number:

93762-42-6

Molecular formula:

C36H54N2O2

IUPAC Name:

9,10-Anthracenedione, 1,4-diamino-, N,N'-mixed 2-ethylhexyl and hexyl and octyl derivs.

Test material form:

liquid

Details on test material:

Name of test material : 9,10-Anthracenedione, 1,4-diamino-, N,N'-mixed 2-ethylhexyl and hexyl and octyl derivs.
IUPAC name: 9,10-Anthracenedione, 1,4-diamino-, N,N'-mixed 2-ethylhexyl and hexyl and octyl derivs.
Molecular Formula: C36H54N2O2
Molecular Weight: 546 g/mole
Smiles: CCCCCCCCNc1ccc(N(CCCCCC)CC(CC)CCCC)c2c1C(=O)c1ccccc1C2=O
Substance Type: Organic
Physical State: Liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Institute for Industrial Research and Toxicology.
- Age at study initiation: 7 to 9 weeks
- Weight at study initiation: 200±20 g
- Fasting period before study: No data available.
- Identification : By cage tag and corresponding colour body marking.
- Housing: 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 (e.g. ad libitum): Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India.
- Water (e.g. ad libitum): Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad-libitum.
- Acclimation period: Twenty healthy albino rats were selected and acclimatized for standard laboratory condition for period of one week in experimental room under vaterinary examination.
- Randomization : After acclimatization and veterinary examination all the selected rats randomly divided into two groups of five females and five males.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 degC
- Humidity (%): 30-60 %
- Air changes (per hr): Air conditioned rooms with 10-15 air changes per hour.
- Photoperiod (hrs dark / hrs light): Illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.

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 chamb er, 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 the system, as well as its dilution with outside air.
- System of generating particulates/aerosols: Nanotek aerosol generator
- Method of particle size determination: No data available
- 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: Group I : 22.15±1.14 and Group II: 22.37±1.41,
Relative humidity (%): Group I : 47.32±3.15 and Group II: 48.15±3.27

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

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:

Two groups
Group I - Limit test (5 mg/L)
Group II - Confirmatory test (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: daily
- Necropsy of survivors performed: yes
- Other examinations performed:
Body weight:
The body weight of all the animals was observed weekly on day 0 (Before treatment), 7th and 14th (post treatment).

Clinical Signs:
The treated animals were observed for signs of intoxication, at various intervals for first six hours after dosing and thereafter twice a day for 14 days. Any clinical signs in the treated group, if observed, recorded and mentioned in the report.

Mortality:
All the animals were observed for mortality at various intervals for first six hours on the day of dosing and thereafter twice a day for 14 days.

Necropsy:
Necropsy was carried out on all the animals that died during the study or surviving animals were sacrificed at the end of the study to observe any gross pathological changes.

Statistics:

No data available

Results and discussion

Preliminary study:

No data

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: no mortality was observed

Mortality:

The test compound did not elicit any mortality at the tested dose level of 5.0 mg/L throughout the period of observation after exposure.

Clinical signs:

other: The test compound did not elicit any clinical signs of intoxication at the tested aerosol concentration of 5 mg/L observed for the period of 14 days.

Body weight:

The body weight of all the animals recorded individually on day 0th and thereafter 7th and 14th (post treatment) showed normal increase as compared to day 0th.

Gross pathology:

Necropsy finding:
A.EXTRENAL:
i:skin: skin and hair coat was observed wet
ii:all external orifices: normal
B.INTERNAL:
i:subcutaneous: no change was observed
ii:superficial and deep lymph node: no change in mesenteric lymph node
ABDOMINAL CAVITY:
i. opening and general examination: in the abdominal cavity all the organs were present in normal position
ii. spleen:no changes were observed
iii. digestive sysytem: no gross changes were observed in stomach and intestine
iv. liver and biliary ducts: no gross pathological changes were observed
v. excretory sysytem: no gross pathological changes were observed
vi. adrenal: observed normal
vii. male/female genital organs: showed normal color,consistancy and no inflammartory changes
2. THORACIC CAVITY:
i. opening and general examination: thoracic cavity was found to be normal without any fluid,mucous or blood etc.
ii. lungs:no changes were observed
iii. heart: no changes were observed in color and consistancy. heart found normal
iv. thyroid: normal in shape,size and surface
3. CRANIAL CAVITY:
brain: normal in shape and size

Other findings:

No data

Any other information on results incl. tables

TABLE - 1

EXPOSURE ATMOSPHERE DATA

Parameters	Group-I (5 mg/L) (limit test)	Group-II (5 mg/L) (Confirmatory test)
Chamber temperature °C (Mean ± S.E.)	22.15±1.14	22.37±1.41
Relative humidity (%) (Mean ± S.E.)	47.32±3.15	48.15±3.27
Oxygen content (%) (Mean ± S.E.)	19.54±1.14	20.27±1.41

TABLE - 2

MEAN BODY WEIGHT (gm)

S. No.	Groups	Body Weight (gm)
		Day 0	Day 7th	% gain or loss	Day 14th	% gain or loss
1.	Group – I (5.0 mg/L)	202.43	208.21	2.85	213.51	5.47
2.	Group – II (5.0 mg/L)	200.46	206.61	3.06	213.12	6.31

TABLE – 3

CLINICAL SIGNS AND MORTALITY

GROUP – I                                                                                                                                 DOSE: 5.0 mg/L

WISTAR ALBINO RATS

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 signs

+        = Mild

++      = Moderate

+++    = High

++++  = Severe 

 

TABLE – 3 Contd…..

CLINICAL SIGNS AND MORTALITY

GROUP – II                                                                                                                        DOSE: 5.0 mg/L

WISTAR ALBINO RATS

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 signs

+        = Mild

++      = Moderate

+++    = High

++++  = Severe

TABLE – 4

SUMMARY OF NECROPSY FINDINGS

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

Table 5: individual animal fate and necropsy findings

Group:I                                                                               5.0 mg/l

WISTAR ALBINO RATS

Animal ID	Fate	Time	Gross findings
20171-1	TS	Day 15	NAD
20171-2	TS	Day 15	NAD
20171-3	TS	Day 15	NAD
20171-4	TS	Day 15	NAD
20171-5	TS	Day 15	NAD
20171-6	TS	Day 15	NAD
20171-7	TS	Day 15	NAD
20171-8	TS	Day 15	NAD
20171-9	TS	Day 15	NAD
20171-10	TS	Day 15	NAD

DAY 0 is the day of exposure

TS= terminal sacrifice

NAD=no abnormality

FD=found dead

 

Table-5 contd……………….

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

GROUP:II                                                       5.0 mg/l

Animal ID	Fate	Time	Gross findings
20171-11	TS	Day 15	NAD
20171-12	TS	Day 15	NAD
20171-13	TS	Day 15	NAD
20171-14	TS	Day 15	NAD
20171-15	TS	Day 15	NAD
20171-16	TS	Day 15	NAD
20171-17	TS	Day 15	NAD
20171-18	TS	Day 15	NAD
20171-19	TS	Day 15	NAD
20171-20	TS	Day 15	NAD

DAY 0 is the day of exposure

TS= terminal sacrifice

NAD=no abnormality

FD=found dead

Applicant's summary and conclusion

Interpretation of results:

other: Not classified

Conclusions:

The LC50 value for acute inhalation toxicity test was considered to be >5 mg/l, when wistar albino rats were exposed with the given test chemical by inhalation route for 4 hour according to OECD guideline 403(Acute inhalation toxicity).

Executive summary:

The acute inhalation study was conducted by using the given test chemical performed according to OECD-Guideline – 403 for testing of chemicals in albino rat.

LIMIT TEST: Ten healthy Wistar albino rats of both sexes (5 male and 5 female) of body weight 200±20 gm were selected for study after acclimatization. The test group of animals was exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various intervals such as 1hr, 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. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol of the test compound at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation. The necroscopy was performed on all the animals at the termination of study did not show any gross pathological changes.

CONFIRMATORY TEST: After 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration following same guideline. Ten healthy Wistar albino rats of both sex (body weight 200±20 gm) were selected for study after acclimatization. The test group of animals were exposed to aerosol of the test compound at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were 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 a experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5 mg/L did not show any clinical signs of intoxication. Again there was no mortality recorded during the entire observation period. The body weight of animals exposed to test compound, observed on day 0th(pretreatment) and day 7th(post treatment) showed normal gain as compared to control group. The necroscopy was performed on all the animals at the termination of study did not show any gross pathological changes.

The result obtained from present investigation can be concluded that, the LC50 value for acute inhalation toxicity test was considered to be >5 mg/l, when wistar albino rats were exposed with the given test chemical by inhalation route for 4 hour according to OECD guideline 403(Acute inhalation toxicity).