Registration Dossier

Administrative data

Description of key information

Acute Oral Toxicity:           

In Acute oral toxicity ,LD50 value for target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) was considered to be 3290 mg/kg bw;<5000 mg/kg bw;4700 mg/kg bw and approximately 4000 mg/kg bw in rats. All these studies concluded that the LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity:          

In Acute inhalation toxicity ,LC50 value for target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) was considered to be 11.71 mg/l and 182 mg/l in rats. All these studies concluded that the LC50 value is greater than 5 mg/L air. Thus, comparing this value with the criteria of CLP regulation, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) cannot be classified for acute inhalation toxicity.

Acute Dermal Toxicity:

In Acute dermal toxicity, LD50 value for target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) was considered to be >2000 mg/kg bw;>8000 mg/kg bw and >2000 mg/kg bw. All these studies concluded that the LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) cannot be classified for acute dermal toxicity.

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:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data is from peer-reviewed journal
Reference:
Composition 0
Composition 0
Composition 0
Composition 0
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
Acute Oral toxicity test was carried out to study the effects of Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) on rats.
GLP compliance:
not specified
Test type:
other: no data available
Limit test:
no
Test material information:
Composition 1
Specific details on test material used for the study:
- Name of test material (as cited in study report):Solantine Blue 10GL
- Molecular formula :C32H20Cu2N6Na4O16S4
- Molecular weight :1087.82 g/mol
- Substance type:organic
Species:
rat
Strain:
other: Albino SASCO
Sex:
male/female
Details on test animals and environmental conditions:
Details on test animal
TEST ANIMALS
- Weight at study initiation: 190 to 304 grams
- Fasting period before study: the animals were fasted overnight
- Housing: The animals were individually housed in metal, wire-bottomed cages elevated above the droppings.
- Diet (e.g. ad libitum): Purina Laboratory Chow was provided ad libitum
- Water (e.g. ad libitum): water was provided ad libitum
Route of administration:
oral: gavage
Vehicle:
other: deionized water
Details on oral exposure:
Details on exposure
VEHICLE
- Amount of vehicle (if gavage): 50% w/w
MAXIMUM DOSE VOLUME APPLIED: 6330 mg/kg bw

DOSAGE PREPARATION (if unusual):test substance was soluble in deionized water
Doses:
2520, 3170, 3990, 5020 and 6330 mg/kg bw
No. of animals per sex per dose:
Total: 25 animals
Control animals:
not specified
Details on study design:
Details on study design
- Duration of observation period following administration: 14 days
- Frequency of observations :
Gross pathology: The animals were observed for gross effect at regular intervals on the day of dosing and daily thereafter for 14 days.
- Frequency of weighing: Following the observation period, all surviving animals were weighed.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology
Statistics:
The acute oral LD50 calculated by the method of Weil, C.S.
Preliminary study:
Preliminary test:
the actual LD50 determination, exploratory doses were administered to six rats to estimate the order of toxicity of the test material. Based on the results of the preliminary assay, groups of five rats were dosed at levels designed to blanket the toxicity range as follows: 2520, 3170, 3990, 5020 and 6330 mg/kg bw.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
3 290 mg/kg bw
Based on:
test mat.
95% CL:
2 720 - 3 980
Remarks on result:
other: 50% mortality was observed
Mortality:
50% mortality was observed at dose 3290 mg/kg bw in treated animals
Clinical signs:
the following clinical signs of toxicity were observed Prostration, labored breathing, proneness, malaise, blue-black diarrhea, ptosis, morbidity, lethargy and weakness.
These appeared between 45 minutes and 6 hours after dosing .Surviving animals were normal within 6 days after dosing.
Body weight:
Final bodyweight records of survivors at termination showed weight gains within expected limits except in one animal at the 5020 mg/kg body weight level which showed no gain.
Gross pathology:
Gross necropsy of animals which succumbed showed generalized congestion of the lung,liver, kidneys and adrenal glands.
In all animals, tissues from all major organ systems and muscles were dyed blue to green by the compound. The color level was dose dependent. This dying effect was observed in muscle- and tendon.
Gross necropsy of animals sacrificed at termination showed no tissue damage from the administration of the compound except at the 5020 mg/kg body weight dosage. The two animals surviving at this level showed some gross indication of liver damage.
Other findings:
no data

Table 1: Time of Death Following Dosage

Dosage

Level

gm/kg

(Days)

%

mortality

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2.52

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0

3.17

0/5

1/5

1/5

1/5

1/5

1/5

1/5

1/5

1/5

1/5

2/5

2/5

2/5

2/5

40

3.99

0/5

2/5

4/5

4/5

4/5

4/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

100

5.02

0/5

2/5

3/5

3/5

3/5

3/5

3/5

3/5

3/5

3/5

3/5

3/5

3/5

3/5

60

6.33

0/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

100

*Specific gravity = 1.299 at 23° Centigrade

Interpretation of results:
other: not classified
Conclusions:
The lethal concentration (LD50) value for acute oral toxicity test was considered to be 3290 mg/kg bw,when male and female albino SASCO rats were treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) orally via gavage.
Executive summary:

Acute oral toxicity study was done in Male and female albino SASCO rats (SASCO strain) weighing 190 to 304 grams .Each animal was examined and only healthy animals were used on this study. The animals were individually housed in metal, wire-bottomed cages elevated above the droppings.Prior to dosing, the animals were fasted overnight.food and water was provided ad libitum.Prior to the actual LD50 determination, exploratory doses were administered to six rats to estimate the order of toxicity of the test material. Based on the results of the preliminary assay, groups of five rats were dosed at levels designed to blanket the toxicity range as follows: 2520, 3170, 3990, 5020 and 6330 mg/kg bw.The test material was prepared as a 50% weight/weight dilution indeionized water. The animals were observed for gross effect at regular intervals on the day of dosing and daily thereafter for 14 days. Animals which succumbed were necropsied. Following the observation period, all surviving animals were weighed, sacrificed and necropsied. The following clinical signs of toxicity were observed Prostration, labored breathing, proneness, malaise, blue-black diarrhea, ptosis, morbidity, lethargy and weakness. These appeared between 45 minutes and 6 hours after dosing .Surviving animals were normal within 6 days after dosing. Final bodyweight records of survivors at termination showed weight gains within expected limits except in one animal at the 5020 mg/kg body weight level which showed no gain. Gross necropsy of animals which succumbed showed generalized congestion of the lung,liver, kidneys and adrenal glands. In all animals, tissues from all major organ systems and muscles were dyed blue to green by the compound. The color level was dose dependent. This dying effect was observed in muscle- and tendon.Gross necropsy of animals sacrificed at termination showed no tissue damage from the administration of the compound except at the 5020 mg/kg body weight dosage. The two animals surviving at this level showed some gross indication of liver damage.Hence, Thelethal concentration (LD50) valuefor acute oral toxicity testwas considered to be3290mg/kg bw,when male and female albino SASCO rats were treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) orally via gavage.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
3 290 mg/kg bw
Quality of whole database:
Data is Klimisch 2 and from authoritative database

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
data is from secondary source
Reference:
Composition 0
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
Acute inhalation toxicity study of Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) in rats
GLP compliance:
not specified
Test type:
other: no data
Limit test:
no
Test material information:
Composition 1
Specific details on test material used for the study:
- Name of test material (as cited in study report):Solantine Blue 10GL
- Molecular formula :C32H20Cu2N6Na4O16S4
- Molecular weight :1087.82 g/mol
- Substance type:organic
Species:
rat
Strain:
other: albino SASCO
Sex:
not specified
Details on test animals and environmental conditions:
Details on test animal
TEST ANIMALS
- Weight at study initiation: 217 to 292 grams
- Diet (e.g. ad libitum): Purina Laboratory Chow (pelletized) was provided ad libitum
- Water (e.g. ad libitum):tap water was provided ad libitum
Route of administration:
inhalation: dust
Type of inhalation exposure:
not specified
Vehicle:
not specified
Details on inhalation exposure:
no data
Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
1 h
Concentrations:
182 mg/l in air
No. of animals per sex per dose:
Total: 10 animals
Control animals:
not specified
Details on study design:
Details on study design
- Duration of observation period following administration: 14 days
- Frequency of observations: The animals were observed frequently for gross effects during the exposure
- Necropsy of survivors performed: yes
Statistics:
no data
Preliminary study:
no data
Sex:
male/female
Dose descriptor:
LC50
Effect level:
182 mg/L air
Based on:
test mat.
Exp. duration:
1 h
Remarks on result:
other: No mortality was observed
Mortality:
No mortality was observed at dose 182 mg/l in air
Clinical signs:
No data
Body weight:
Changes in body weight were observed
Gross pathology:
No gross signs of toxicity were observed during the test period of the 14-day observation
period.
Gross necropsy of animals sacrificed at termination showed moderate congestion of lungs in 5 rats, two of these had lesions. Slight congestion of lungs in the other 5 rats and of these, one had lesions, Otherwise, the animals were not remarkable.
Other findings:
no data

Table1: Inhalation Mortality Data

During Exposure

% Mortality

During 14-days observation period

% Mortality

Mort. /No. Animals

 

Mort. /No. Animals

 

0/10

0

0/10

0

Interpretation of results:
other: not classified
Conclusions:
The acute inhalation toxicity study in rats by using Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) for exposure of 1 hr was considered to be 18200 mg/m3 (182 mg/L).
Executive summary:

Ten albino rats (SASCO strain) weighing 217 to 292 grams exposed to a ambient concentration of approximately 182 mg./liter of air at a flow rate of 10 liters/minute survived the 1-hour exposure (plus a 6-minute equilibration period) and the 14-day observation period which followed.No mortality was observed was observed at dose 182 mg/l in air. Changes in body weight were observed. No gross signs of toxicity were observed during the test period of the 14-day observation period. Gross necropsy of animals sacrificed at termination showed moderate congestion of lungs in 5 rats, two of these had lesions. Slight congestion of lungs in the other 5 rats and of these, one had lesions, Otherwise, the animals were not remarkable.Hence, The acute inhalation toxicity study in rats by usingCopper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6)for exposure of 4 hr was considered to be 18200 mg/m3 (182 mg/L).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
11 710 mg/m³
Quality of whole database:
Data is Klimisch 2 and from authoritative database

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data is from secondary source
Reference:
Composition 0
Composition 0
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
Acute dermal toxicity test was carried out to study the effects of Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) on rabbits.
GLP compliance:
not specified
Test type:
other: no data
Limit test:
no
Test material information:
Composition 1
Specific details on test material used for the study:
- Name of test material (as cited in study report):Lumisol blue VG
- Molecular formula :C32H20Cu2N6Na4O16S4
- Molecular weight :1087.82 g/mol
- Substance type:organic
- Physical state:Dark Grey Powder
- Purity :100%
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
Details on test animal
TEST ANIMALS
- Age at study initiation: approximately 8 to 11 weeks
- Housing: The rabbits were individually housed in elevated wire mesh cages in temperature controlled rooms reserved exclusively for rabbits on acute tests
- Diet (e.g. ad libitum): Purina Rabbit Chow was provided ad libitum
- Water (e.g. ad libitum): water was provided ad libitum
- Acclimation period:7 days
Type of coverage:
occlusive
Vehicle:
other: distilled water
Details on dermal exposure:
TEST SITE
- Area of exposure: The test substance was applied to the the intact and abraded skin.
-Type of wrap if used: The test site was covered with gauze and the trunk was wrapped with impervious material for 24 hours.
REMOVAL OF TEST SUBSTANCE
- Washing (if done): the test site was washed with warm tap water

Details on exposure
VEHICLE
- Amount of vehicle (if gavage): 50% w/v

DOSAGE PREPARATION (if unusual):test substance was soluble in distilled water
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Total: 4 animals
2000 mg/kg bw:2 male and 2 female
Control animals:
not specified
Details on study design:
Details on study design
- Duration of observation period following administration: 14 days
- Frequency of observations : Skin sites were read again at 7 and 14 days. The rabbits were observed daily for 14 days for signs of toxicity and mortality.
-frequency of weighing: Body weights were recorded pretest and at 7 and 14 days.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights
Statistics:
no data
Preliminary study:
no data
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:
No mortality was observed at dose 2000 mg/kg bw
Clinical signs:
Animal 1 - Diarrhea, days 10 and 11; bloated abdomen, day 11:Few feces in pan, days 11 and 12.
Animal 2 - Lethargy, day 12.
Body weight:
Changes in body weight were observed
Gross pathology:
Necropsy findings concluded that all animals appeared normal.
Other findings:
no data

TABLE 1:AcuteDermalToxicity in Rabbits

Individual Body Weights and Skin Grades

Rabbit Number

Sex

Weights - kg

Redness

Edema

0

7

14

25H

7

14

25H

7

14

1

M

2.8

2.8

2.8

0

0

0

0

0

0

2ab

M

2.5

2.7

2.7

0

0

0

0

0

0

3

F

2.2

2.4

2.7

0

0

0

0

0

0

4ab

F

2.3

2.5

2.7

0

0

0

0

0

0

ab = abraded

Evaluation of Skin Reactions

value

Erythema & Eschar Formation:

     No erythema

     Very slight erythema (barely perceptible)

     Well defined erythema

     Moderate to severe erythema

     Severe erythema (beet redness) to slight eschar formation (injuries in depth)

 

0

1

2

3

4

Ederra Formation:

    No edema

    Very slight edema (barely perceptible)

    Slight edema (edges of area well defined by definite raising)

    Moderate edema (raised approximately 1 millimeter)

    Severe edema (raised more than 1 millimeter and extending beyond the area of exposure)

 

0

1

2

 

3

 

 

4

 

Interpretation of results:
other: not classified
Conclusions:
The LD50 value was considered to be >2000 mg/kg bw,when male and female New Zealand white rabbits were occlusively treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) by dermal application following 14 days of observation period.
Executive summary:

In acute dermal toxicity study, male and femaleNew Zealand white rabbitswere occlusively treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) in the concentration of 2000 mg/kg bw by dermal application.Distilled water was used as vehicle.Twenty-four hours prior to dosing the backs of the rabbits were clipped free of fur with an Oster ANG-RA clipper head designed specifically for clipping rabbits. The rabbits were returned to their cages overnight. Just prior to dosing, the backs of even-numbered rabbits were abraded with a 21 gauge bent tip needle. The abrasions, made every 2 to 3 cm longitudinally, scratched the stratum corneum but did not disturb the derma or produce bleeding.The test site was covered with gauze and the trunk was wrapped with impervious material for 24 hours.Following removal of the binder at 24 hours. The test site was washed with warm tap water. One hour after washing, the test sites were graded for skin irritation according to the attached scale. Skin sites were read again at 7 and 14 day.No mortality was observed in treated rabbits at dose 2000 mg/kg bw.Clinical signs such asDiarrhea, bloated abdomen, Few feces in pan and Lethargy was observed in treated rabbits. Changes in body weight were observed. Necropsyfindings concluded that all animals appeared normal.Hence,The LD50 value was considered to be >2000 mg/kg bw,when 2male and 2 female New Zealand white rabbitswere occlusively treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6)by dermal application following 14 days of observation period.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 2 and from authoritative database

Additional information

Acute Oral Toxicity: 

In different studies, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) has been investigated for acute oral toxicity to a greater or lesser extent. Often the studies are based on in vivo experiments data in rodents, i.e. most commonly in rats for target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). The different studies for target substance are summarized as below –

In experimental study conducted by U.S. National Library of Medicine(Chemidplus Database,U.S. National Library of Medicine,2017); S. Gangolli (The Dictionary of Substance and their effects,2nd edition,vol.2 (part 6)); National Technical Reports Library(National Technical Reports Library, Fiche No. OTS 215154,1972) and RTECS (RTECS (registry of toxic effect of chemical substance database ), 2018) for the target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). Acute oral toxicity study was done in Male and female albino SASCO rats (SASCO strain) weighing 190 to 304 grams .Each animal was examined and only healthy animals were used on this study. The animals were individually housed in metal, wire-bottomed cages elevated above the droppings.Prior to dosing, the animals were fasted overnight.food and water was provided ad libitum.Prior to the actual LD50 determination, exploratory doses were administered to six rats to estimate the order of toxicity of the test material. Based on the results of the preliminary assay, groups of five rats were dosed at levels designed to blanket the toxicity range as follows: 2520, 3170, 3990, 5020 and 6330 mg/kg bw.The test material was prepared as a 50% weight/weight dilution in deionized water. The animals were observed for gross effect at regular intervals on the day of dosing and daily thereafter for 14 days. Animals which succumbed were necropsied. Following the observation period, all surviving animals were weighed, sacrificed and necropsied. The following clinical signs of toxicity were observed Prostration, labored breathing, proneness, malaise, blue-black diarrhea, ptosis, morbidity, lethargy and weakness. These appeared between 45 minutes and 6 hours after dosing .Surviving animals were normal within 6 days after dosing. Final bodyweight records of survivors at termination showed weight gains within expected limits except in one animal at the 5020 mg/kg body weight level which showed no gain. Gross necropsy of animals which succumbed showed generalized congestion of the lung,liver, kidneys and adrenal glands. In all animals, tissues from all major organ systems and muscles were dyed blue to green by the compound. The color level was dose dependent. This dying effect was observed in muscle- and tendon.Gross necropsy of animals sacrificed at termination showed no tissue damage from the administration of the compound except at the 5020 mg/kg body weight dosage. The two animals surviving at this level showed some gross indication of liver damage.Hence, The lethal concentration (LD50) value for acute oral toxicity test was considered to be3290mg/kg bw,when male and female albino SASCO rats were treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) orally via gavage.

The above study was further supported by National Technical Reports Library (National Technical Reports Library, Fiche No. OTS 206237,1982) for the target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). Acute oral toxicity study was done in five Male and five female wistar rats.Distill water was used as vehicle.The rats were housed five per sex per cage in elevated wire mesh cages in a temperature controlled room reserved exclusively for rats on acute tests. Each rat was uniquely identified with picric acid dye.Purina Rat Chow and water from bottles was available ad libitum except for approximately 18 hours prior to dosing.After at least seven days equilibration. The rats were dosed at 5000 mg/kg orally by gavage(intubation).Following treatment the rats were observed at 1, 2 and 4 hours and daily for 14 days for signs of pharmacologic and/or toxicologic effects and mortality. Body weights were recorded prior to dosing and in the survivors at Day 14. Necropsies were performed on all rats.50% mortality was observed in treated rats. Clinical signs like blue Diarrhea, Chromorhinorrhea, chromodacryorrhea, ataxia,Piloerection, diarrhea, lethargy ,ptosis were observed. Both gain and loss of Bodyweight were observed in treated animals. In necropsy observations, exudate nose/mouth (blue), congenital blue intestine(contained blue fluid),stomach (contained blue fluid),lung areas purple),kidneys(greenish/blue),spleen (large) were observed.Hence, The lethal concentration (LD50) value for acute oral toxicity test was considered to be <5000mg/kg bw,when male and female wistar rats were treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6)orally via gavage.

This is further supported by National Technical Reports Library (National Technical Reports Library, Fiche No. OTS 206237,1982) for the for the target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). Acute oral toxicity study was done in four groups of five Male and female wistar rats.Distill water was used as vehicle. The animals were housed 5/cage in suspended wire mesh cages (20" x 10" x 7").Fresh Purina rat chow and water were freely available except for 16-20 hours prior to dosing when food was removed. The test material was given orally via gavage. The rats were observed 1, 2 and 4 hours after dosing and once daily for 14 days. Mortality, toxicity and pharmacological effects were recorded.Body weights were recorded pretest and in the survivors at 14 days. At 14 days the survivors were sacrificed. All animals were examined for gross pathology. No deaths occurred at the lowest level 3510 mg/kg. 25/30 animals died at the three highest levels:5000, 7120 and 10140 mg/kg. Significant predeath toxic signs included lethargy, ptosis, piloerection, ataxia, flaccid muscle tone, emaciation and diarrhea. In the surviving animals,weight gains were noted. Necropsy observations were non-specific.Hence, The lethal concentration (LD50) value for acute oral toxicity test was considered to be 4700 mg/kg bw (95% CI:3600 mg/kg bw-6100 mg/kg bw),when four groups of 5 male and 5 female wistar rats were treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6)orally via gavage.

Also these results are further supported by the experimental study conducted by National Technical Reports Library (National Technical Reports Library, Fiche No. OTS 215154,1972) for the for the target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). Acute oral toxicity (MLD) study was done in Male and female young adult New Zealand albino Rabbits weighing 2.0 to 2.5 kilograms.Each animal was examined and only healthy animals were used on this study. Each animal was examined and only healthy animals were used on this study. The animals were individually housed in metal, wire-bottomed cages elevated above the droppings. Prior to dosing, the animals were fasted overnight .Purina Laboratory Chow and water were freely available at all times.The test compound was administered as a 50% w/w aqueous dilution using deionized water.50% mortality was observed in treated animals. The final body weights of survivors show gains within expected limits at the low dose, but a severe weight loss in the animal at the M.L.D. dose. Gross necropsy of animal which succumbed showed intense blue staining of all tissues of the major organ systems and connective tissue. The staining was to such a degree as to mask congestion. Severe congestion was detected in the lungs.Hence, The median lethal dose (MLD ) value for acute oral toxicity test was considered to be approximately 4000 mg/kg bw,when male and female young adult New Zealand albino Rabbits were treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6)orally via gavage.

Thus, based on the above studies on Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6), it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity: 

In different studies, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) has been investigated for acute inhalation toxicity to a greater or lesser extent. Often the studies are based on in vivo experiments data in rodents, i.e. most commonly in rats for target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). The different studies for target substance are summarized as below –

In experimental study conducted by U.S. National Library of Medicine (HSDB (Hazardous Substances Data Bank); US national Library of Medicine,2017) andNational Technical Reports Library (National Technical Reports Library,Fiche No. OTS0206237,1979) for thetarget substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). Ten Haltzman Sprague-Dawley rats (5 male, 5 female) were exposed for 4 hours in an inhalation chamber to the maximum obtainable concentration of Lumisol Blue VG (TR No. 78-735) powder. The powder was dispersed by an NBS dust generator into the stainless steel-glass chamber in which the 10 animals were housed individually in wire mesh cages. The nominal concentration of the chemical in the exposure atmosphere was calculated from the weighed quantity of chemical dispersed into the chamber and the total volume of air passed through the chamber.The animals were observed frequently during exposure and during the 14-day post-exposure period for pharmaco-toxic signs. Body weights were obtained prior to exposure and at 1, 7 and 14 days after exposure. At the end of the 14-day post-exposure period, all surviving animals were necropsied and major organs were examined for macroscopic abnormalities.No mortality was observed at dose 11.71 mg/l. No clinical signs occurred during the exposure. Body weights of males were normal, but 2 females exhibited slight depressed weight gain. Necropsyfindings found macroscopic lesions in the lungs of all animals.Thesefindings included hemorrhagic spots and foci, a purplish discoloration and rough texture of lobes or portions of lobes, and nodular masses or growths with a shrunken appearance. Thesefindings were not considered treatment-related because it was concluded that thesefindings were typical of chronic murine pneumonia complex. Other prominent abnormalities were nodular masses or growths, shrunken appearance and rough texture of lobes and white patches or areas.Hence, The acute inhalation toxicity study in rats by using Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6)for exposure of 4 hr was considered to be 11710 mg/m3 (11.71 mg/L).

 

This is further supported by National Technical Reports Library (National Technical Reports Library, Fiche No. OTS 215154,1972)for thetarget substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). Ten albino rats (SASCO strain) weighing 217 to 292 grams exposed to a ambient concentration of approximately 182 mg./liter of air at a flow rate of 10 liters/minute survived the 1-hour exposure (plus a 6-minute equilibration period) and the 14-day observation period which followed.No mortality was observed was observed at dose 182 mg/l in air. Changes in body weight were observed. No gross signs of toxicity were observed during the test period of the 14-day observation period. Gross necropsy of animals sacrificed at termination showed moderate congestion of lungs in 5 rats, two of these had lesions. Slight congestion of lungs in the other 5 rats and of these, one had lesions, Otherwise, the animals were not remarkable.Hence, The acute inhalation toxicity study in rats by using Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) for exposure of 4 hr was considered to be 18200 mg/m3 (182 mg/L).

Thus, based on the above studies on Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) ,it can be concluded that LC50 value is greater than 5 mg/L air. Thus, comparing this value with the criteria of CLP regulation, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) cannot be classified for acute inhalation toxicity.

Acute Dermal Toxicity:

In different studies, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) has been investigated for acute dermal toxicity to a greater or lesser extent. Often the studies are based on in vivo experiments and estimated data in rodents, i.e. most commonly in rabbits for Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) along with the study available on structurally similar read across substance Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate (CAS No. 2519-30-4). The studies are summarized as below –

In experimental study conducted by US national Library of Medicine (HSDB (Hazardous Substances Data Bank); US national Library of Medicine,2017) andNational Technical Reports Library(National Technical Reports Library, Fiche No. OTS 206237,1982)for thetarget substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). In acute dermal toxicity study, male and female New Zealand white rabbits were occlusively treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) in the concentration of 2000 mg/kg bw by dermal application.Distilled water was used as vehicle.Twenty-four hours prior to dosing the backs of the rabbits were clipped free of fur with an Oster ANG-RA clipper head designed specifically for clipping rabbits. The rabbits were returned to their cages overnight. Just prior to dosing, the backs of even-numbered rabbits were abraded with a 21 gauge bent tip needle. The abrasions, made every 2 to 3 cm longitudinally, scratched the stratum corneum but did not disturb the derma or produce bleeding.The test site was covered with gauze and the trunk was wrapped with impervious material for 24 hours.Following removal of the binder at 24 hours. The test site was washed with warm tap water. One hour after washing, the test sites were graded for skin irritation according to the attached scale. Skin sites were read again at 7 and 14 day.No mortality was observed in treated rabbits at dose 2000 mg/kg bw.Clinical signs such as Diarrhea, bloated abdomen, Few feces in pan and Lethargy was observed in treated rabbits. Changes in body weight were observed. Necropsy findings concluded that all animals appeared normal.Hence,The LD50 value was considered to be >2000 mg/kg bw,when 2male and 2 female New Zealand white rabbits were occlusively treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) by dermal application following 14 days of observation period.

The above study was further supported by National Technical Reports Library (National Technical Reports Library, Fiche No. OTS 215154,1973) for the target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6). In acute dermal toxicity study, male and female New Zealand white rabbits were occlusively treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) in the concentration of 8000 mg/kg bw by dermal application. Distilled water was used as vehicle.Prior to exposure, the animals were prepared by clipping the skin of the trunk free of hair and only those animals without observable skin defects or irritation were used.Solantine Blue 10GL,as a 50% w/w aqueous dilution was introduced under the plastic binder; the binder was then fastened tightly to keep the preparation in close contact with the skin for 24 hours.No mortality was observed at dose 8000 mg/kg bw. No signs of local toxicity were observed during the study. material tends to obscure local toxicity signs.No signs of systemic toxicity related to application of the material were observed. Final body weight of surviving animals at termination (14 days),showed 3/4 at the low level gaining weight within expected limits and 1/4 at the low level losing weight. At the high level, 1/3 lost weight, 1/3 remained steady and 1/3 gained weight. Gross necropsy of animal which succumbed showed a number(approximately 10) haemolytic lesions of the stomach and a thickening of the stomach wall. Staining occurred in the liver and stomach and intestine.Hence,The LD50 value was considered to be >8000 mg/kg bw,when male and female New Zealand white rabbits were occlusively treated with Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) by dermal application following 14 days of observation period.

Also these results are further supported by Sustainability Support Services (Europe) AB (study number: 18818, 2016-10-24) for the structurally similar read across substance Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate (CAS No. 2519-30-4). The acute dermal toxicity profile of Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulpho natophenylazo)-7-sulphonato-1-naphthyl azo))naphthalene-4,6-disulphonate in Sprague Dawley rats.water was used as vehicle.The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days.Animals exhibited normal body weight gain through the study period of 14 days.Gross pathological examination did not reveal any abnormalities attributable to the treatment.Hence,The LD50 value was considered to be >2000 mg/kg bw,when male and female Sprague Dawley rats were semiocclusively treated with Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6- disulphonate (CAS No. 2519-30-4) by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

Thus, based on the above studies on Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) and it’s structurally similar read across substances Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6- disulphonate (CAS No. 2519-30-4) , it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above experimental studies on target substance Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) and it’s structurally similar read across substances Tetra sodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6- disulphonate (CAS No. 2519-30-4) , it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Copper,[tetrahydrogen-3,3'-[(3,3'-dihydroxy-4,4'-biphenylylene)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphthalenedisulfonato](4-)]di-,tetrasodium salt (7CI) (28407-37-6) cannot be classified for acute oral ,dermal and inhalation toxicity.