2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs.

Endpoint summary

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

GLP compliance:

yes

Test type:

acute toxic class method

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Age : 7 to 9 weeks
Sex : Female, nulliparous and non pregnant. It has been observed that females are generally more sensitive than males to toxic effects
Body weight range : 200±20g
Acclimation : One week in experimental room after veterinary examination.
Randomization : After acclimation and veterinary examination randomly selected in groups of three females.
Nutritional conditions : Fasted overnight prior to treatment. Food was offered three hours after dosing.
Environmental conditions :Air conditioned rooms with 10-15 air changes per hour, temperature between 22-25 0C, relative humidity 40-60% and illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.

Route of administration:

other: oral cannula

Vehicle:

corn oil

Details on oral exposure:

DOSE FORMULATION
Dose preparation of the test article was done freshly, few minutes prior to dosing. Test substance 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs. was mixed corn oil to obtain final concentration of 200 and 30mg/ml.

ADMINISTRATION OF TEST COMPOUND:
The test compound was administered by oral route by using of oral cannula at the dose volume of 10 ml/kg b.wt

Doses:

Two + one vehicle control
Group I : Dist. water, 10ml/kg body wt.
Group II : 2000 mg/kg body wt.
Group III : 300 mg/kg body wt.
Group IV : 300 mg/kg body wt.

No. of animals per sex per dose:

Three (3 females)/step

Control animals:

yes

Details on study design:

STUDY DESIGN:
The toxicity of the test compound following oral administration was assessed. Three female rats were used per step for each dose level. The rats were observed for incidence of mortality and signs of intoxication for 14 days after the administration of test article
OBSERVATION
Body weight:
The body weight of all the animals was observed weekly on day 0 (pre treatment), 7th and 14th (post treatment).
Mortality:
The test compound was administered at different dose level in wistar albino rats observed for mortality at the time interval of 30 minutes, 1hr, 2hr, 4 hr, and 6hr time interval on the day of test compound administration and thereafter twice a day for 14 days.
Clinical Signs
The treated animals were closely observed for clinical signs of intoxication, first 4 hours and every 1 hrs interval for 24 hrs after dosing and thereafter twice a day for 14 days. All the rats were observed at least twice daily to observe any clinical signs or behavioral changes. These observations included changes in skin and fur, in the eyes and mucous membranes, respiratory, circulatory, central nervous and autonomic nervous systems, somatomotor activity and behavioral changes. The following clinical signs were observed in female mice to characterize the various systemic studies: Salivation, lacrimation, pale mucous membrane, diarrheal feaces, hunched posture, scratching, polyuria, hypoactivity etc. The clinical sign will be graded as 0 = Normal, + = Mild, ++= Moderate, +++ =High and ++++ = Severe.
Necropsy:
Necropsy was carried out on all the animals which died during the study or surviving animals were sacrificed at the end of the study to observe any gross pathological changes.
Histopathological study
The organ which showed gross pathological change during necropsy subjected for histopathological study

Sex:

female

Dose descriptor:

LD50

Effect level:

300 mg/kg bw

Based on:

test mat.

Mortality:

Dose 2000 mg/kg body weight- Test compound produced two mortalities after 24 hrs of administration.
Dose 300 mg/kg body weight- Test compound did not elicit any mortality throughout the period of observation

Clinical signs:

other: Dose 2000 mg/kg body weight- The test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs administered at the dose 2000 mg/kg body weight to wistar albino rats (female) showed moderate to severe clinical signs of toxicity viz; decre

Gross pathology:

NECROPSY FINDING
EXTERNAL
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 nodes- No change in mesenteric lymph node.
1. ABDOMINAL CAVITY
i.Opening and general examination- In the abdominal cavity all the organs were present in normal position.
ii.Spleen- Normal upto highest tested dose level 2000 mg/kg b.wt.
iii.Digestive system- No gross changes were observed in stomach and intestine upto highest tested dose level 2000 mg/kg b.wt.
iv.Liver and biliary ducts- No gross pathological changes were observed
v.Excretory system- No gross pathological changes were observed upto highest tested dose level 2000 mg/kg b.wt.
vi.Adrenal- Observed normal.
vii.Male/female genital organs – Showed normal colour, consistency and no inflammatory changes upto highest tested dose level 2000 mg/kg b.wt.
2. THORACIC CAVITY
i.Opening and general examination- Thoracic cavity was found to be normal without any fluid, mucous or blood etc.
ii.Lungs- observed normal.
iii.Heart- No changes were observed in color and consistency. Heart found normal upto highest tested dose level 2000 mg/kg b.wt.
iv.Thyroid- Normal in shape, size and surface upto highest tested dose level 2000 mg/kg b.wt.
3. CRANIAL CAVITY
i.Brain- Normal in shape and size.

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

From the above results it is concluded that the test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs is non-toxic to Wistar albino rats at the tested dose level 300 mg/kg b.wt. According to Globally Harmonized Classification System for Chemical Substances, it comes under the Globally Harmonized Classification (GHS) Category-4 (>300-2000). The cut-off LD50 is estimated to be 1000 mg/kg body weight.

Executive summary:

From the above results it is concluded that the test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs is non-toxic to Wistar albino rats at the tested dose level 300 mg/kg b.wt. According to Globally Harmonized Classification System for Chemical Substances, it comes under the Globally Harmonized Classification (GHS) Category-4 (>300-2000). The cut-off LD50 is estimated to be 1000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Quality of whole database:

K1 level data

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Age : 8 to 10 weeks
Sex : Male and female
Body weight range : 200±20g
No. of animals per dose group : 10 (5male & 5 female)
Acclimatization : The healthy wistar albino rats selected for study acclimatized to standard laboratory condition for period of one week under close Veterinary supervision.
Nutritional conditions : Animals were fasted overnight prior to test and food was offered three hours after dosing.
Environmental conditions : Air conditioned rooms with 10-15 air changes per hour, temperature between 22-250C, relative humidity 40-60% and illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.

Type of coverage:

open

Details on dermal exposure:

DOSE FORMULATION
The test compound was applied at the dose level of 2000 mg/kg b.w.t as such (as the compound was in liquid form).
APPLICATION OF TEST COMPOUND:
The test substance was applied uniformly over an exposed area of skin. The test compound was held in contact with the skin with an impervious dressing secured in place with an adhesive tape. The animals were then housed individually in cages with a collar around the neck in order to avoid the ingestion of the test compound. After 24 hours, the dressing was removed and the site of application was cleaned with lukewarm water wiping the test compound.

APPLICATION OF TEST COMPOUND:
The test substance was applied uniformly over an exposed area of skin. The test compound was held in contact with the skin with an impervious dressing secured in place with an adhesive tape. The animals were then housed individually in cages with a collar around the neck in order to avoid the ingestion of the test compound. After 24 hours, the dressing was removed and the site of application was cleaned with lukewarm water wiping the test compound.

Duration of exposure:

14 days

Doses:

No. of dose group : Two
Group-I: 2000 mg/kg b.wt (limit test)
Group-II: 2000 mg/kg b.wt (confirmatory test)

No. of animals per sex per dose:

10 (5male & 5 female)

Control animals:

no

Details on study design:

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 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs 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 (pre treatment), 7th and 14th (post treatment). The necropsy was performed on all animals which was died during the study or were sacrificed at termination of the study.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was recorded in Wistar albino rats after administration of test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs at the dose level of 2000 mg/kg b.wt. throughout observation period of 14 days.

Clinical signs:

other: Wistar albino rats treated with the test compound at the dose level of 2000 mg/kg b.wt. did not elicit any clinical sign of toxicity throughout the observation period of 14 days.

Gross pathology:

NECROPSY FINDING
EXTERNAL
i.Skin- Skin and hair coat was observed wet.
ii.All external orifices- Normal
B. INTERNAL
i. Subcutaneous- No changes were observed.
ii. Superficial and deep lymph nodes- 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 recorded.
iii.Digestive system- No gross changes were observed in stomach and intestine.
iv.Liver and biliary ducts- No gross pathological changes were observed
v.Excretory system- No gross pathological changes were observed.
vi.Adrenal- Observed normal.
vii.Male/female genital organs – Showed normal colour, consistency and no inflammatory 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 recorded.
iii.Heart- No changes were observed in color and consistency. Heart found normal.
iv.Thyroid- Normal in shape, size and surface.
3. CRANIAL CAVITY
i.Brain- Normal in shape and size.

CLINICAL SIGNS AND MORTALITY

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	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0		0		0
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	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

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute dermal LD50 value of test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs in Wistar albino rats when applied by dermal route was found to be more than 2000 mg/kg b.wt. (> 2000 mg/kg b.wt.). From this it is concluded that the substance 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs is non toxic by dermal route.

Executive summary:

The acute dermal LD50 value of test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs in Wistar albino rats when applied by dermal route was found to be more than 2000 mg/kg b.wt. (> 2000 mg/kg b.wt.). From this it is concluded that the substance 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs is non toxic by dermal route.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

K1 level data

Additional information

Acute oral-

The test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs is non-toxic to Wistar albino rats at the tested dose level 300 mg/kg b.wt. According to Globally Harmonized Classification System for Chemical Substances, it comes under the Globally Harmonized Classification (GHS) Category-4 (>300-2000). The cut-off LD50 is estimated to be 1000 mg/kg body weight.

Acute inhalation-

In accordance with column 2 of Annex VIII, this end point was considered for waiver since exposure of humans via inhalation is highly unlikely taking into account the low vapour pressure of 0.00000000061 Pa, estimated for 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs. Acute dermal-

Based on studies of target substance 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs. and readacross substances reviewed for acute toxicity dermal from reliable sources having Klimisch rating 1 and 4 considering the weight of evidence approach.

 The summary of the results are presented below

Sr. No	End point	Value	Species	Remark
1.	LD50	>2000 mg/L	Rat	IIRT Results
2.	LD50	>2000 mg/L	Rabbit	RA 1229-55-6(Study report)

 

Based on above table, endpoint value was found to be higher than 2000 mg/kg bw/d which is the criteria for classification towards acute toxicity, it is concluded that 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs. does not exhibits acute toxicity by the dermal route.

Justification for selection of acute toxicity – oral endpoint

From the above results it is concluded that the test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs is non-toxic to Wistar albino rats at the tested dose level 300 mg/kg b.wt. According to Globally Harmonized Classification System for Chemical Substances, it comes under the Globally Harmonized Classification (GHS) Category-4 (>300-2000). The cut-off LD50 is estimated to be 1000 mg/kg body weight.

Justification for selection of acute toxicity – dermal endpoint

The acute dermal LD50 value of test compound 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs in Wistar albino rats when applied by dermal route was  found to be more than 2000 mg/kg b.wt. (> 2000 mg/kg b.wt.). From this it is concluded that the substance 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs is non toxic by dermal route.

Justification for classification or non-classification

The test substance 2-Naphthalenol, 1-[[4-(phenylazo)phenyl]azo]-, ar',ar''-Me derivs.is toxic via oral route but non toxic via inhalation and dermal route.