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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

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 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

The study was conducted to find out the mortality, clinical sign of toxicity and histopathological effect of the given test chemical at different dose level in Wistar albino rats.

GLP compliance:

no

Test type:

other: Acute Dermal Toxicity

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: 8 to 10 weeks
- Weight at study initiation: 200±20 g
- Fasting period before study: Animals were fasted overnight prior to test and food was offered 3 hours after dosing.
- Identification : By cage tag and corresponding colour body marking.
- Housing: Groups of two animals of same 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): Fresh and clean water filtered through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad-libitum.
- Acclimation period: One week
- Randomization : After acclimatization and veterinary examination all the animals randomly divided into two groups and each group having five male and five female rats.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-25 degC
- Humidity (%): 40-60 %
- 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

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Back skin of total body surface area.
- % coverage: Approximate 10% area.
- Type of wrap if used: The test compound was held in contact with the skiin with impervious dressing secured in place with an adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The dressing was removed and the site of application was cleaned with lukewarm water wiping the test compound.
- Time after start of exposure: 24 hrs

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg b.wt

Duration of exposure:

24 hours

Doses:

Group - I : 2000 mg/kg b.wt
Group - II : 2000 mg/kg b.wt

No. of animals per sex per dose:

Group - I : 2000 mg/kg b.wt : 5 female and 5 male
Group - II : 2000 mg/kg b.wt : 5 female and 5 male

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily
- Necropsy of survivors performed: yes, necropsy was carried out on all animals that died during the study or surviving animals were sacrificed at the end of the study to observe any gross pathological changes.
- Other examinations performed:
- Body weight: The body weight of all the animals was observed weekly on day 0 (pre treatment), 7th and 14th (post treatment).
- Clinical signs : The treated animals were closely observed for clinical signs of intoxication, first 4 hours and thereafter for every 1 hrs interval for 24 hours after dosing and twice a day for 14 days. All the rats were observed at least twice daily with the purpose of recording any symptoms of ill-health or behavioral changes. These observations included changes in skin and fur in the eyes and mucous membranes, respiratory, circulatory, central nervous and autonomous systems, somatomotor activity and behavior changes. The following clinical signs were observed in rats to characterize with erythema, hypersensitivity, edema etc.
- Mortality: All the animals were observed for mortality at 30 minutes time interval for first six hours on the day of test compound administration and thereafter twice a day for 14 days.

Statistics:

No data

Results and discussion

Preliminary study:

no data

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality was observed

Mortality:

The test compound did not produce any mortality throughout the observation period of 14 days.

Clinical signs:

The test compound did not elicit any clinical signs at the dose level of 2000 mg/kg b.wt. in entire observation period.

Body weight:

All the animals treated with the test compound at the dose level of 2000 mg/kg b.wt. showed normal gain in body weight as compared to control group on day 0th (pre treatment).

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 chnage was observed
ii:superficial and deep lymph node: no chnage 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

SUMMARY OF BODY WEIGHT (GM)

Group	Animal ID	Day 0	Day 7	% Gain/loss	Day 14	% Gain/loss
Group-I 2000 mg/kg b. wt	20171-1	198.26	204.73	3.11	211.85	6.85
	20171 -2	200.43	206.05	2.80	213.26	6.40
	20171 -3	202.82	208.96	3.02	215.58	6.29
	20171 -4	201.20	207.61	3.18	212.07	5.40
	20171 -5	203.56	209.84	3.08	215.35	5.79
	20171 -6	200.78	206.39	2.94	213.22	6.19
	20171 -7	198.53	204.75	3.13	211.60	6.58
	20171 -8	199.38	205.62	3.12	212.54	6.60
	20171 -9	202.64	208.08	2.68	214.37	5.78
	20171 -10	201.95	207.33	2.66	212.28	5.11
Group-II 2000 mg/kg b. wt	20171 -11	199.03	205.85	3.42	210.56	5.79
	20171 -12	198.77	204.74	3.00	211.93	6.62
	20171 -13	199.35	205.82	3.24	209.52	5.10
	20171 -14	202.64	207.51	2.40	213.26	5.24
	20171 -15	200.25	206.83	3.28	212.40	6.06
	20171 -16	203.75	208.59	2.37	214.28	5.16
	20171 -17	200.37	206.64	3.12	211.19	5.40
	20171 -18	201.82	207.93	3.02	213.61	5.84
	20171 -19	200.45	206.31	2.92	210.58	5.05
	20171 -20	204.52	209.63	2.49	217.11	6.15

TABLE - 2

CLINICAL SIGNS AND MORTALITY

Group: I Limit test                                                 Dose: 2000 mg/kg b.wt

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- Local
Redness	-	-	-	-	-	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Pain	-	-	-	-	-	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Swelling	-	-	-	-	-	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Systemic signs
Clinical signs	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

-          =Observed after 24 hrs

0        = No clinical signs

+        = Mild

++      = Moderate

+++    = High

++++  = Severe

TABLE - 2 Contd...

CLINICAL SIGNS AND MORTALITY

Group: II Confirmatory test                                                        Dose: 2000 mg/kg b.wt

                                                                                   

                                                           

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- Local
Redness	-	-	-	-	-	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Pain	-	-	-	-	-	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Swelling	-	-	-	-	-	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Systemic signs
Clinical signs	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

-          =Observed after 24 hrs

0        = No clinical signs

+        = Mild

++      = Moderate

+++    = High

++++  = Severe

TABLE – 3

SUMMARY OF NECROPSY FINDINGS

S.No.	Fate	Wistar albino rats
		Dose (mg/kg b.wt)
		2000 (limit test)	2000 (confirmatory test)
1	Terminal sacrifice	10/10	10/10
2	Found dead	0/10	0/10
3	Abnormalities detected	0/10	0/10

Table 4: individual animal fate and necropsy findings

Group:I  (limit test)                                                                            2000 mg/kg bw

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-4 contd……………….

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

GROUP:II (confirmatory test)                                                   2000 mg/kg bw

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 LD50 value was considered to be >2000 mg/kg bw, when Wistar albino rats were occlusively treated with the given test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

Executive summary:

The acute dermal toxicity study was conducted by using the given test chemical on Wistar albino rats under OECD guideline-402 Guideline for Testing of Chemicals.

LIMIT TEST (2000 mg/kg b.wt): Ten healthy wistar albino rats of both sex (ranging b.wt 200±20 gm) selected for study after acclimatization. Approximate 10 percent back skin of total body surface area was prepared 24 hrs prior to application of test compound. The test drug was applied dermally at the dose level of 2000 mg/kg b.wt for each animal. The treated animals were observed for clinical signs of intoxication and mortality at different time interval for a period of 14 days. The body weight of each rat was observed on day 0 (pretreatment), 7thand 14th(post treatment). The necropsy was performed on all animals at the termination of the study. The test compound applied at the dose level of 2000 mg/kg b.wt in Wistar albino rats did not show any clinical signs of toxicity throughout the observation period of 14 days.  Furthermore, no mortality was observed throughout the period of observation (14 days). The necropsy was performed on all animals at the termination of the study did not show any gross pathological changes.

CONFIRMATORY TEST: After 72 hrs, a confirmatory test was conducted in same species of animals to confirm the limit test of test compound (OECD-402 guidelines). Ten healthy Wistar albino rats of both sex (ranging b.wt 200±20 gm) selected for study after acclimatization. Approximate 10 percent back skin of total body surface area was prepared 24 hrs prior to application of test compound. The test drug was applied dermally at the dose level of 2000 mg/kg b.wt for each animal. The treated animals were obser ved for clinical signs of intoxication and mortality at different time interval for a period of 14 days. The body weight of each rat was observed on day 0 (pretre atment), 7thand 14th(post treatment). The necropsy was performed on all animals at the termination of the study. The test compound did not produce any mortality throughout the observation period of 14 days. Furthermore, the test compound did not elicit any clinical signs at the dose level of 2000 mg/kg b.wt. in entire observation period. All the animals treated with the test compound at the dose level of 2000 mg/kg b.wt. showed normal gain in body weight as compared to control group on day 0th (pre treatment). Necropsy was conducted on day 15th (end of study) did not reveal any significant gross pathological changes related to compound toxicity.

The results obtained from present investigation can be concluded that, the LD50 value was considered to be >2000 mg/kg bw, when Wistar albino rats were occlusively treated with the given test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).