Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)

Endpoint summary

Description of key information

Short-term toxicity to fish:

Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) was used as a test material to evaluate its acute toxicity effect on Fathead Minnow (Pimephales promelas)

 10 Fathead Minnow (Pimephales promelas) were acclimatized in 5-gallon wide mouth glass jar for 10 days before the test in the test temperature 17°C. The test organisms were pre-treated with tetracycline .

 Test material was subjected in the test vessel upto the concentration of 180 mg/l . No toxic effect were observed after 96 hrs. Hence the effect concentration was considered to be >180g/l

By considering LC50 value it is concluded that test chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) is not toxic tofish and can be considered as “Not classified” according to CLP classification criteria.

Short-term toxicity to aquatic invertebrate :

Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) when used as a test material to evaluate aquatic toxicity on Daphnia magna .The neonatal 24 hrs old Daphnis were used. Ten neonates were placed in individual 50 mL containers, with 40 mL of the sample solution, Two sample solutions were prepared for each concentration.

Concentrations selected were 0.8,1.3, 2.2, 3.6, 6.0, and 10.0 mg/L. D. magna were placed in upright incubator (cycle 16 h on/8 off) and covered loosely with parafilm to prevent evaporation.

The mortality of daphnids was observed at 24 and 48 h from initiation of the test. The results indicate that the LC50 of Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)  for D. magna is about 6.0 mg/l at 24-h and 3.6-6.0 mg/L at 48-h tests. No mortality or lethal effects were observed at 0.8-2.2 mg/L and 100% mortality was observed at 48-h in 10.0 mg/L .

This value indicates that the substance is likely to be hazardous to aquatic invertebrates can can be classified as Aquatic chronic category 1 as per the CLP criteria.

Toxicity to aquatic algae and cyanobacteria:

Toxicity of test chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) (CAS no. 28407 -37 -6) was estimated by using OECD QSAR tool box v3.3 with log Kow as primary descriptor and considering the seven closest read across substances. When test chemical was exposed for 72 hrs on Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) alga, EC50 value was estimated to be 105.1 mg/L. By considering EC50 value it is concluded that test chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) is not toxic to aquatic algae and can be considered as ''Not classified"according to CLP classification criteria.

Additional information

Short-term toxicity to fish:

Experimental study for short-term toxicity to fish was summarized in Handbook of Environment and Waste Management air and water pollution control , Yung-Tse Hung , Lawrence K Wang, Nazih K Shammas 2012 and Ecology committee American Dye Manufacturers Institute, INC. Sept. 1972 for target chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)) (CAS 28407-37-6).

Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) was used as a test material to evaluate its acute toxicity effect on Fathead Minnow (Pimephales promelas)

  10 Fathead Minnow (Pimephales promelas) were acclimatized in 5-gallon wide mouth glass jar for 10 days before the test in the test temperature 17°C. The test organisms were pre-treated with tetracycline .

  Test material was subjected in the test vessel upto the concentration of 180 mg/l . No toxic effect were observed after 96 hrs. Hence the effect concentration was considered to be >180g/l

By considering LC50 value it is concluded that test chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) is not toxic tofish and can be considered as “Not classified” according to CLP classification criteria.

Short-term toxicity to aquatic invertebrate :

Various experimental studies describing the short term toxicity to aquatic invertebrate 1972 for target chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)) (CAS 28407-37-6) are summarized below.

An experimental study conducted by Jin-Seok Bae , Harold S. Freeman summarized in Dyes and Pigments 73 (2007) 126-132.

Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) when used as a test material to evaluate aquatic toxicity on Daphnia magna .The neonatal 24 hrs old Daphnis were used. Ten neonates were placed in individual 50 mL containers, with 40 mL of the sample solution, Two sample solutions were prepared for each concentration.

Concentrations selected were 0.8,1.3, 2.2, 3.6, 6.0, and 10.0 mg/L. D. magna were placed in upright incubator (cycle 16 h on/8 off) and covered loosely with parafilm to prevent evaporation.

The mortality of daphnids was observed at 24 and 48 h from initiation of the test. The results indicate that the LC50 of Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)  for D. magna is about 6.0 mg/l at 24-h and 3.6-6.0 mg/L at 48-h tests. No mortality or lethal effects were observed at 0.8-2.2 mg/L and 100% mortality was observed at 48-h in 10.0 mg/L .

This value indicates that the substance is likely to be hazardous to aquatic invertebrates can can be classified as Aquatic chronic category 1 as per the CLP criteria.

The above experimental data was further supported by experimental study conducted by Y Verma summarized in Toxicology and Industrial Health 2008; 24: 491–500 for the target chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)).

Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) when used as a test material to evaluate its aquatic toxicity on Daphnia magna. The test animal was maintained. Twenty-four hours before the test, adult daphnids (gravid females) were sorted and the young one (neonates) produced from these adults were used in the experiment.

Initially, a 24-h preliminary test was carried out at exposure concentrations of 100, 10, and 1.0 mg/LThe selected concentration for the test was 100, 80, 60, 40, 20, and 10 mg/L

The LC 50 value for the test chemical was observed to be 24-h LC50 6.0 mg/L;48-h LC50 3.6–6.0 mg/L

The above experimental data was further supported by experimental data summarized in Journal of Developing Drugs Volume 5 , Issue 1 2016.

Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) when subjected on Daphnia magna for 48 hrs produced high toxicity effect with the LC50 range between 1.0-10.0 mg/l

Based on the available data of target Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)) (CAS 28407-37-6) it can be considered that the substance is likely to be hazardous to aquatic invertebrates can can be classified as Aquatic chronic category 1 as per the CLP criteria.

Toxicity to aquatic algae and cyanobacteria:

Toxicity of test chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) (CAS no. 28407 -37 -6) was estimated by using OECD QSAR tool box v3.3 with log Kow as primary descriptor and considering the seven closest read across substances. When test chemical was exposed for 72 hrs on Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) alga, EC50 value was estimated to be 105.1 mg/L. By considering EC50 value it is concluded that test chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) is not toxic to aquatic algae and can be considered as ''Not classified"according to CLP classification criteria.

The above prediction was supported by experimental study conducted by Sustainability Support Services (Europe) AB for the structurally similar read across substance N-benzyl-2-[(2,6-dimethylphenyl)amino]-N,N-diethyl-2-oxoethanaminium benzoate hydrate (CAS: 3734-33-6)

The effect of test substance Denatonium Benzoate (granular form), CAS No. 3734-33-6 was studied on the growth of fresh water green alga Chlorella vulgaris.The study was conducted following OECD guideline 201- Alga growth inhibition test. The test concentration chosen for the study were 6.25 mg/l, 12.5 mg/l, 25mg/l, 50 mg/l, 100mg/l and 200 mg/l. The test concentrations were prepared using stock solution of the test substance using mineral media. The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of test substance. EC50 calculated graphically through probit analysis was observed to be 281.556mg/l.

The experimental data was further supported by experimental study conducted by Sustainability Support Services (Europe) AB for the structurally similar read across substance aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (CAS:15790-07-5)

The effect of test item aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) was studied on the growth of fresh water green alga Chlorella vulgaris. The study was conducted following OECD guideline 201- Alga, growth inhibition test. The test concentration chosen for the study were 6.25mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L, 200mg/L. The test solution was prepared in aseptic condition. The test item Aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200 mg/L. This stock solution was kept for stirring/sonication for 30 minutes to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The concentrations chosen were set up to the water solubility limit. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment. Test was performed in static manner at proper requirement of pH and temperature.

By considering EC50 value it is concluded that test chemical Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) and its structurally similar read across substances it can be considered that the target chemical is not toxic to aquatic algae and can be considered as ''Not classified"according to CLP classification criteria.

Based on the experimental data of target Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-) for short term toxicity to aquatic invertebrates the overall classification of aquatic toxicity can be considered as

Aquatic chronic category 1 as per the CLP criteria.