Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates

Endpoint summary

Description of key information

Skin irritation

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the skin irritation potential of the test chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates. The studies are as mentioned below:

 

An Acute Dermal Irritation/corrosion Study of read across chemical was conducted in Rabbits as per OECD guideline No. 404. Three healthy young adult female rabbits were used. Rabbits with good intact skin were selected for the study. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment. A dose of 0.5 ml of test item (as such) was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one side of rabbits. The other untreated side was kept as control area and0.5 ml of distilled water was applied at this site. At the end of 4 hours, the gauze patch was removed and test item application site was wiped with water without altering the integrity of the epidermis. Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch. The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method. The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 0.33, 1.00, 0.33 and 0.00, 0.33, 0.00, for erythema and oedema formation, respectively. Hence, under the experimental conditions tested it is concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits.

 

The above result was further supported by another experimental study conducted for another similar read across chemical that followed the same principle and procedure as mentioned in above study. During the study, in Animals Nos. 2 at 1 and 24 hours observation after post patch removal, revealed, slight erythema (barely perceptible) and no oedema which was recovered to normal at 48 hours observation. In Animals Nos. 3 at 1 hour observation after post patch removal, revealed, slight erythema (barely perceptible) and no oedema which was recovered to normal at 24 hours. At 48 and 72 hours observation no Erythema and oedema post patch removal in Animal nos. 2 and 3. The individual mean score at24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.33, 0.33, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.   Hence, under the experimental test conditions, it was concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits under the experimental conditions tested.

 

Based on the above summarized studies for target chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates and its structurally similar read across substances,it can be concluded that the testchemical is not able to cause skin irritation and considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

 

Eye irritation

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the ocular irritation potential of the test chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates . The studies are as mentioned below:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405. Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 0.1 mlof test itemwas instilled in the other (treated) eye of each rabbit.The eye was observed at 1, 24, 48, 72 hours and on day 7 after test item instillation.Ophthalmoscope was used for scoring of eye lesions. Untreated eye of all the three rabbits was normal throughout the experimental period of 7 days. The following grading scores were observed in treated eye of tested rabbits. Observation at 1 hour after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was observed in all 3 animals;Chemosis:Some swelling above normal (includes nictitating membranes) was seen in all 3 animals. Observation at 24 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity- Zero was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3;Chemosis:Some swelling above normal (includes nictitating membranes) was seen in all 3 animals. At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 30 %, 30 % and 35 % damage in animal no. 1, 2 and 3 respectively. Observation at 48 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals. Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3;Chemosis:No swelling (Normal) was seen in all 3 animals. Observation at 72 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 and 3 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2;Chemosis:No swelling (Normal) was seen in all the animals. Observation at day 7 after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Blood vessels normal in all 3 animals;Chemosis:No swelling (Normal) was seen in all 3 animals. The individual mean score for animal nos. 1, 2 and 3at 24, 48, 72 hoursfor corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 1.00, 0.33; 0.00, 0.00, 2.00, 0.33 and 0.00, 0.00, 1.67, 0.33, respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbit no. 2 and 3 was observed at 24 hours.  Hence under the experimental test conditions, the test chemical is “Mildly Irritating to Eyes” to New Zealand White female rabbit eyes. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within 7 days. Hence, the test chemical is “Mildly irritating to eyes” to New Zealand White Female rabbit eyes.

 

The above result was further supported by another eye irritation study conducted by peer reviewed journal for read across chemicalon New Zealand White rabbit to determine its irritation potential.Eye injury in rabbits is recorded in a 10- grade ordinal series and is based upon the degree of corneal necrosis that results from instillation of various volumes and concentrations of test chemical, as detailed by Carpenter and Smyth.The chemical caused eye injury of grade 8 that indicted severe damage to the treated eye.Based upon the score (8),the test chemical can be considered as irritating to the rabbits eye.

 

Based on the above summarized studies for target chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates and its structurally similar read across substances,it can be concluded that the testchemical is able to cause eye irritation and considered as irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Category 2 (irritating to eyes)”.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally and functionally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on 2 skin irritation studies as- WoE-2 and WoE-3. Skin irritation study of test chemical was conducted on rabbits to assess its skin irritating effects.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

1.TEST ANIMALS- Source: Procured from GENTOX Bio services., Hyderabad, India- Age at study initiation: 3 to 4 Months (Approximately)- Weight at study initiation: Minimum: 1.796 kg and Maximum: 1.998 kg (Prior to Treatment)- Housing: The animals were housed individually in stainless steel cages. - Diet (e.g. ad libitum): All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum.- Water (e.g. ad libitum): Aqua guard filtered tap water was provided ad libitum- Acclimation period: 5 days (Animal No.-1) and 7 days (Animal No.-2 and 3)ENVIRONMENTAL CONDITIONS- Temperature (°C): Minimum: 19.60 °C, Maximum: 22.30 °C- Humidity (%): Minimum: 51.10 % , Maximum: 67.40 %- Air changes (per hr): More than 12 changes per hour- Photoperiod (hrs dark / hrs light): 12:122.TEST ANIMALS- Source:Procured from RABBI ROOF, Hyderabad.- Age at study initiation:3.5 to 4.5 Months (Approximately)- Weight (Prior to Treatment):Minimum: 2.228 kg & Maximum: 2.502 kg - Health Status: Healthy young adults and nulliparous and non-pregnant Rabbits were used for the study.- Housing:The animals were housed individually in stainless steel cages.- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day. - Cages and water bottle:All the cages and water bottles were changed minimum twice a week.- Diet (e.g. ad libitum):All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum. - Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum. - Acclimation period:Rabbits were acclimatised to the test conditions for a period of 5 days (Animal No.-1) and 8 days (Animal No.-2 and 3) prior to the application of the test item.ENVIRONMENTAL CONDITIONS- Temperature (°C):Minimum: 20.00 °C and Maximum: 22.50 °C- Humidity (%):Minimum: 47.20 % and Maximum: 68.40 %- Air changes (per hr):More than 12 changes per hour- Photoperiod (hrs dark / hrs light):12:12

Type of coverage:

occlusive

Preparation of test site:

other: 1.shaved 2.clipped

Vehicle:

other: 1.unchanged (no vehicle) 2.not specified

Controls:

yes

Amount / concentration applied:

1. TEST MATERIAL- Amount(s) applied (volume or weight with unit): 0.5 ml2.- Amount(s) applied (volume or weight with unit):0.5 ml

Duration of treatment / exposure:

1.4 hours 2.4 hours

Observation period:

1.72 hours 2.72 hours

Number of animals:

1.3 female rabbits2.3 female rabbits

Details on study design:

1.TEST SITE- Area of exposure: Approximately 24 h prior to treatment, the fur coat of each rabbit was removed from dorsal lumbar region approximately 6 X 6 cm at contralateral sites on each rabbit using clipper, one as control and other site as treatment. Rabbits with healthy intact skin were selected for the study- % coverage: 6 X 6 cm- Type of wrap if used: porous gauze dressing and non-irritating tapeREMOVAL OF TEST SUBSTANCE- Washing (if done): cotton soaked in distilled water- Time after start of exposure: 4-hour exposure periodSCORING SYSTEM:Gradation as per draize method2.TEST SITE- Area of exposure:The dorsal lumbar region at contralateral sites. - % coverage:Approximately 6 X 6 cm. - Type of wrap if used:A porous gauze dressing and non-irritating tape (Micropore 3”). REMOVAL OF TEST SUBSTANCE- Washing (if done):The residual test item was removed by using cotton soaked in distilled water.- Time after start of exposure:4 hourSCORING SYSTEM:Draize Method

Irritation parameter:

other: overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

1.The following were observed in treated rabbits.The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method.Animal No. 1, very slight erythema (barely perceptible) and very slight oedema (barely perceptible) observed at 1 hour of observation. At 24 hour very slight erythema (barely perceptible) and no oedema was observed. At 48 and 72 hours observation no erythema and no oedema was observed in animal no 1.Animal No. 2, revealed very slight erythema (barely perceptible) and very slight oedema (barely perceptible) observed at 1 hour of observation post patch removal. At 24 hour well defined erythema and very slight oedema (barely perceptible) was observed. At 48 hour very slight erythema (barely perceptible) and no oedema was observed. At 72 hour no erythema and no oedema was observed. Animal No. 3, revealed very slight erythema (barely perceptible) and very slight oedema (barely perceptible) observed at 1 hour of observation post patch removal. At 24 hour slight erythema (barely perceptible) and no oedema was observed. At 48 and 72 hour no erythema and no oedema was observed. The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 0.33, 1.00, 0.33 and 0.00, 0.33, 0.00, for erythema and oedema formation, respectively2.The following were observed in treated rabbits.The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method.After 4 hours of exposure in Animal No. 1 and 2, there were very slight erythema (barely perceptible) and no oedema observed at 1 and 24 hour of observation which was recovered to normal at 48 hours observation. In Animals Nos. 3 at 1 hour observation after post patch removal, revealed, slight erythema (barely perceptible) and no oedema which was recovered to normal at 24 hours. The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.33, 0.33, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.

Other effects:

1.Clinical ObservationNo systemic toxicity was observed at treated rabbits during the experimental period. MortalityNo mortality was observed during the observation period.Body WeightsBody weights were increased as compared to day 0 in all the three animals at terminal sacrifice2.Clinical ObservationNo systemic toxicity was observed at treated rabbits during the experimental period. MortalityNo mortality was observed during the observation period.

1.

In Treated area Dose:0.5 ml of test item                                                              Sex:Female

 

Animal No.	Test	Treated  area*	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Right	1	1	0	0	1	0	0	0
2	Confirmatory	Right	1	2	1	0	1	1	0	0
3		Right	1	1	0	0	1	0	0	0

In Control area  Dose:0.5 ml of distilled water                                               Sex:Female

 

Animal No.	Test	Treated area*	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Left	0	0	0	0	0	0	0	0
2	Confirmatory	Left	0	0	0	0	0	0	0	0
3		Left	0	0	0	0	0	0	0	0

Key:h = Hour.

 

Erythema                                                                               Oedema

0 =No erythema                                                                  0 =No oedema

1 = Very slight erythema (barely perceptible)                       1 = Very slight oedema(barely perceptible)

2 = Well defined erythema

Mean Individual Animal Score at 24, 48 and 72 hours

 

Animal Number                   Observations	1	2	3
Erythema	0.33	1.00	0.33
Oedema	0.00	0.33	0.00

Individual Animal BodyWeight

Sex:Female

Animal No.	Body Weight (kg)
	Prior to Dosing	At termination
1	1.796	1.856
2	1.940	1.992
3	1.998	2.110

Key: kg = kilogram

Individual AnimalClinical Signs

Sex:Female

Animal No.	Days (Post dosing Observation)
	0	1	2	3
1	1	1	1	1
2	1	1	1	1
3	1	1	1	1

Key: ./. = Not Applicable. 1 = Normal.

2.

Table 1

Skin Reaction

 

In Treated area Dose:0.5 ml of test item                                                               Sex:Female

 

Animal No.	Test	Treated  area*	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Left	1	1	0	0	0	0	0	0
2	Confirmatory	Left	1	1	0	0	0	0	0	0
3		Left	1	0	0	0	0	0	0	0

 

 

 

In Control area                 Dose:0.5 ml of distilled water                              Sex:Female

 

Animal No.	Test	Treated area*	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Right	0	0	0	0	0	0	0	0
2	Confirmatory	Right	0	0	0	0	0	0	0	0
3		Right	0	0	0	0	0	0	0	0

Key: h = Hour.

Erythema                                                                                                       Oedema

0 =No erythema                                                                                           0 =No oedema

Table 1 Continued…

Mean Individual Animal Score at 24, 48 and 72 hours

 

Animal Number                   Observations	1	2	3
Erythema	0.33	0.33	0.00
Oedema	0.00	0.00	0.00

 

 

 

Table 2

Individual Animal BodyWeight

Sex:Female

Animal No.	Body Weight (kg)
	Prior to Dosing	At termination
1	2.502	2.540
2	2.228	2.352
3	2.474	2.522

Key: kg = Kilogram

Table 3

Individual AnimalClinicalSigns

Sex:Female

Animal No.	Days (Post dosing Observation)
	0	1	2	3
1	1	1	1	1
2	1	1	1	1
3	1	1	1	1

Key:1 = Normal.

Interpretation of results:

other: not irritating

Conclusions:

The test material Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates was considered to be irritating to the skin of rabbits.

Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the skin irritation potential of the test chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates. The studies are as mentioned below:

 

An Acute Dermal Irritation/corrosion Study of read across chemical was conducted in Rabbits as per OECD guideline No. 404. Three healthy young adult female rabbits were used. Rabbits with good intact skin were selected for the study. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment. A dose of 0.5 ml of test item (as such) was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one side of rabbits. The other untreated side was kept as control area and0.5 ml of distilled water was applied at this site. At the end of 4 hours, the gauze patch was removed and test item application site was wiped with water without altering the integrity of the epidermis. Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch. The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method. The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 0.33, 1.00, 0.33 and 0.00, 0.33, 0.00, for erythema and oedema formation, respectively. Hence, under the experimental conditions tested it is concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits.

 

The above result was further supported by another experimental study conducted for another similar read across chemical that followed the same principle and procedure as mentioned in above study. During the study, in Animals Nos. 2 at 1 and 24 hours observation after post patch removal, revealed, slight erythema (barely perceptible) and no oedema which was recovered to normal at 48 hours observation. In Animals Nos. 3 at 1 hour observation after post patch removal, revealed, slight erythema (barely perceptible) and no oedema which was recovered to normal at 24 hours. At 48 and 72 hours observation no Erythema and oedema post patch removal in Animal nos. 2 and 3. The individual mean score at24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.33, 0.33, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.   Hence, under the experimental test conditions, it was concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits under the experimental conditions tested.

 

Based on the above summarized studies for target chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates and its structurally similar read across substances,it can be concluded that the testchemical is not able to cause skin irritation and considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on 2 eye irritation studies as- WoE-2 and WoE-3. An eye irritation study of test chemical was conducted on rabbits to assess its eye irritating effects.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

1.TEST ANIMALS- Source:Procured from Rabbiroof, Hyderabad, India.- Age at study initiation:4.5 to 5.0 Months (Approximately)- Weight at study initiation:Minimum: 2.042 kg and Maximum: 2.392 kg (Prior to Treatment)- Health Status:Healthy young adult, Females were nulliparous and non pregnant.- Housing:The animals were housed individually in stainless steel cages.- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day. - Cages and water bottle:All the cages and water bottles were changed minimum twice a week.- Diet (e.g. ad libitum):All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum. - Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum. - Acclimation period:Rabbits were acclimatised to the test conditions for a period of 6 days (Animal No.-1) and 10 days (Animal No. 2 and 3) prior to the application of the test item.ENVIRONMENTAL CONDITIONS- Temperature (°C):Minimum: 19.50 °C and Maximum: 22.70 °C- Humidity (%):Minimum: 41.20 % and Maximum: 67.30 %- Air changes (per hr): More than 12 changes per hour- Photoperiod (hrs dark / hrs light): 12:12

Vehicle:

other: 1.unchanged (no vehicle) 2.water or propylene glycol

Controls:

yes

Amount / concentration applied:

1.TEST MATERIAL- Amount(s) applied (volume or weight with unit):0.1 ml2.concentration applied: 0.5 ml of undiluted chemical 0.005 ml and 0.5 ml of a 1% solution in water or propylene glycol.

Duration of treatment / exposure:

1.24 hrs

Observation period (in vivo):

1.All the animals were observed at 1, 24, 48, 72 hours and on day 7 after instillation of test item.

Number of animals or in vitro replicates:

1.3 female rabbits

Details on study design:

1.REMOVAL OF TEST SUBSTANCE- Washing (if done):The treated eye of rabbit was washed with normal saline.- Time after start of exposure:24 hoursSCORING SYSTEM:Draize MethodTOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescent strips.2.Eye injury in rabbits is recorded in a 10- grade ordinal series and is based upon the degree of corneal necrosis that results from instillation of various volumes and concentrations of chemical, as detailed by Carpenter and Smyth

Irritation parameter:

other: 1.overall irritation score

Basis:

mean

Time point:

24/48/72 h

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: Not specified

Score:

8

Max. score:

10

Reversibility:

not reversible

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

1.The following were observed in treated rabbits.In the initial test, 0.1 ml of test item was applied into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 ml of test item was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. Ocular lesions were seen in animal no. 2 and 3 at 1 hour observation which recovered at day 7.Untreated eye of all the three rabbits was normal throughout the experimental period of 7 days.2.The chemical caused eye injury of grade 8 that indicted severe damage to the treated eye.The following grading scores were observed in treated eye of tested rabbits.Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was seen in all 3 animals.Observation at 24 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- Zero was seen in all 3 animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3; Chemosis: Some swelling above normal (includes nictitating membranes) was seen in all the animals.At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 30 %, 30 % and 35 % damage in animal no. 1, 2 and 3 resectively. Observation at 48 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- No opacity was seen in all 3 animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3; Chemosis: No swelling (Normal) was seen in all the animals.Observation at 72 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- No opacity was seen in all 3 animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 and 3 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2; Chemosis: No swelling (Normal) was seen in all the animals.Observation at day 7 after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- No opacity was seen in all 3 animals; Iris: Normal in all the animals. Conjunctivae - Blood vessels normal in all 3 animals; Chemosis: No swelling (Normal) was seen in all 3 animals.The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 1.00, 0.33; 0.00, 0.00, 2.00, 0.33 and 0.00, 0.00, 1.67, 0.33, respectively.2.

Other effects:

1.Clinical ObservationNo systemic toxicity was observed in treated rabbits during the experimental period. 2.The chemical caused eye injury of grade 8 that indicted severe damage to the treated eye.MortalityNo mortality was observed during the observation period.Body weightAll the rabbits were observed with body weight gain on day 7 as compared to day 0.

1.

Table 1 : Individual Animal Eye Irritation Scores

 

In Treated area Dose:0.1 ml of test item                                                               Sex:Female

Animal Numbers		1							2							3
Application Side		Right							Right							Right
Eye Reactions		At hour							At hour							At hour
		*		1		24		48		72		Day 7		*		1		24		48		72		Day 7		*		1		24		48		72		Day 7
Corneal Opacity		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0
Area of Opacity		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0
Iris		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0		0
Conjunctiva		0		1		1		1		1		0		0		1		2		2		2		0		0		1		2		2		1		0
Chemosis		0		1		1		0		0		0		0		1		1		0		0		0		0		1		1		0		0		0
Corneal Damage%		30							30							35

Dose:Untreated (Control Eye)                                                                       Sex:Female

Animal Numbers	1						2						3
Application Side	Left						Left						Left
Eye Reactions	At hour						At hour						At hour
	*	1	24	48	72	Day 7	*	1	24	48	72	Day 7	*	1	24	48	72	Day 7
Corneal Opacity	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Area of Opacity	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctiva	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Chemosis	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Corneal Damage%	0						0						0

Key:*= Pre-exposure eye examination. 

 

Table 1 (Continued):Eye Irritation Scores - Mean Values after 24, 48, 72 Hours (Treated eye)

Animal No.  Eye Reaction	1	2	3
Corneal Opacity	0.00	0.00	0.00
Iris	0.00	0.00	0.00
Conjunctiva	1.00	2.00	1.67
Chemosis	0.33	0.33	0.33

 

 

Formula :

 

Mean Eye Irritation Score =

                          

Sum of the Individual Animal Score for eye reactionat24, 48 and 72 hours

Number of the Observations (3)

 

Table 2: Individual AnimalClinicalSigns

 

Sex:Female

Animal No.	Days (Post application observation)
	0	1	2	3	4	5	6	7
1	1	1	1	1	1	1	1	1
2	1	1	1	1	1	1	1	1
3	1	1	1	1	1	1	1	1

Key:1 = Normal

Table 3: Individual Animal Body Weight

Sex :Female

Animal No.	Animal Body Weight (kg)
	Prior to application	At termination
1	2.392	2.510
2	2.158	2.396
3	2.042	2.316

Key:kg = Kilogram

 

2. Not specified

 

 

 

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The test material [Benzenamine, N,N-dimethyl-, oxidized, molybdatetungstatephosphates was considered to be irritating to the skin of rabbits.

Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the ocular irritation potential of the test chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates . The studies are as mentioned below:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405. Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 0.1 mlof test itemwas instilled in the other (treated) eye of each rabbit.The eye was observed at 1, 24, 48, 72 hours and on day 7 after test item instillation.Ophthalmoscope was used for scoring of eye lesions. Untreated eye of all the three rabbits was normal throughout the experimental period of 7 days. The following grading scores were observed in treated eye of tested rabbits. Observation at 1 hour after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was observed in all 3 animals;Chemosis:Some swelling above normal (includes nictitating membranes) was seen in all 3 animals. Observation at 24 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity- Zero was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3;Chemosis:Some swelling above normal (includes nictitating membranes) was seen in all 3 animals. At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 30 %, 30 % and 35 % damage in animal no. 1, 2 and 3 respectively. Observation at 48 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals. Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3;Chemosis:No swelling (Normal) was seen in all 3 animals. Observation at 72 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 and 3 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2;Chemosis:No swelling (Normal) was seen in all the animals. Observation at day 7 after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Blood vessels normal in all 3 animals;Chemosis:No swelling (Normal) was seen in all 3 animals. The individual mean score for animal nos. 1, 2 and 3at 24, 48, 72 hoursfor corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 1.00, 0.33; 0.00, 0.00, 2.00, 0.33 and 0.00, 0.00, 1.67, 0.33, respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbit no. 2 and 3 was observed at 24 hours.  Hence under the experimental test conditions, the test chemical is “Mildly Irritating to Eyes” to New Zealand White female rabbit eyes. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within 7 days. Hence, the test chemical is “Mildly irritating to eyes” to New Zealand White Female rabbit eyes.

 

The above result was further supported by another eye irritation study conducted by peer reviewed journal for read across chemicalon New Zealand White rabbit to determine its irritation potential.Eye injury in rabbits is recorded in a 10- grade ordinal series and is based upon the degree of corneal necrosis that results from instillation of various volumes and concentrations of test chemical, as detailed by Carpenter and Smyth.The chemical caused eye injury of grade 8 that indicted severe damage to the treated eye.Based upon the score (8),the test chemical can be considered as irritating to the rabbits eye.

 

Based on the above summarized studies for target chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates and its structurally similar read across substances,it can be concluded that the testchemical is able to cause eye irritation and considered as irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Category 2 (irritating to eyes)”.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the skin irritation potential of the test chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates. The studies are as mentioned below:

 

An Acute Dermal Irritation/corrosion Study of read across chemical was conducted in Rabbits as per OECD guideline No. 404. Three healthy young adult female rabbits were used. Rabbits with good intact skin were selected for the study. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment. A dose of 0.5 ml of test item (as such) was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one side of rabbits. The other untreated side was kept as control area and0.5 ml of distilled water was applied at this site. At the end of 4 hours, the gauze patch was removed and test item application site was wiped with water without altering the integrity of the epidermis. Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch. The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method. The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 0.33, 1.00, 0.33 and 0.00, 0.33, 0.00, for erythema and oedema formation, respectively. Hence, under the experimental conditions tested it is concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits.

 

The above result was further supported by another experimental study conducted for another similar read across chemical that followed the same principle and procedure as mentioned in above study. During the study, in Animals Nos. 2 at 1 and 24 hours observation after post patch removal, revealed, slight erythema (barely perceptible) and no oedema which was recovered to normal at 48 hours observation. In Animals Nos. 3 at 1 hour observation after post patch removal, revealed, slight erythema (barely perceptible) and no oedema which was recovered to normal at 24 hours. At 48 and 72 hours observation no Erythema and oedema post patch removal in Animal nos. 2 and 3. The individual mean score at24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.33, 0.33, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.   Hence, under the experimental test conditions, it was concluded that the test chemical was Non-Irritating to the skin of female New Zealand White rabbits under the experimental conditions tested.

 

Based on the above summarized studies for target chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates and its structurally similar read across substances,it can be concluded that the testchemical is not able to cause skin irritation and considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

 

Eye irritation

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the ocular irritation potential of the test chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates . The studies are as mentioned below:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits was performed as per OECD guideline no. 405. Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 0.1 mlof test itemwas instilled in the other (treated) eye of each rabbit.The eye was observed at 1, 24, 48, 72 hours and on day 7 after test item instillation.Ophthalmoscope was used for scoring of eye lesions. Untreated eye of all the three rabbits was normal throughout the experimental period of 7 days. The following grading scores were observed in treated eye of tested rabbits. Observation at 1 hour after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was observed in all 3 animals;Chemosis:Some swelling above normal (includes nictitating membranes) was seen in all 3 animals. Observation at 24 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity- Zero was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3;Chemosis:Some swelling above normal (includes nictitating membranes) was seen in all 3 animals. At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 30 %, 30 % and 35 % damage in animal no. 1, 2 and 3 respectively. Observation at 48 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals. Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3;Chemosis:No swelling (Normal) was seen in all 3 animals. Observation at 72 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no.1 and 3 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2;Chemosis:No swelling (Normal) was seen in all the animals. Observation at day 7 after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-No opacity was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae -Blood vessels normal in all 3 animals;Chemosis:No swelling (Normal) was seen in all 3 animals. The individual mean score for animal nos. 1, 2 and 3at 24, 48, 72 hoursfor corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 1.00, 0.33; 0.00, 0.00, 2.00, 0.33 and 0.00, 0.00, 1.67, 0.33, respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbit no. 2 and 3 was observed at 24 hours.  Hence under the experimental test conditions, the test chemical is “Mildly Irritating to Eyes” to New Zealand White female rabbit eyes. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within 7 days. Hence, the test chemical is “Mildly irritating to eyes” to New Zealand White Female rabbit eyes.

 

The above result was further supported by another eye irritation study conducted by peer reviewed journal for read across chemicalon New Zealand White rabbit to determine its irritation potential.Eye injury in rabbits is recorded in a 10- grade ordinal series and is based upon the degree of corneal necrosis that results from instillation of various volumes and concentrations of test chemical, as detailed by Carpenter and Smyth.The chemical caused eye injury of grade 8 that indicted severe damage to the treated eye.Based upon the score (8),the test chemical can be considered as irritating to the rabbits eye.

 

Based on the above summarized studies for target chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates and its structurally similar read across substances,it can be concluded that the testchemical is able to cause eye irritation and considered as irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Category 2 (irritating to eyes)”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates   and its structurally and functionally similar read across substanceswere observed in various studies. The results obtained from these studies indicate that the chemical  is unlikely to cause skin irritation but can cause severe eye damage. Hence Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates   can be classified under the category “Not Classified” for skin and “Category 2 (irritating to eyes)” for eye as per CLP.