Aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex

Endpoint summary

Description of key information

Short term toxicity to fish:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex.

The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with 2 hrs continuous stirring for achieving test concentrations of 100mg/L,respectively.

The test was performed on the limit concentration i.e nominal concentration selected for the experiment were 100 mg/L and test fish were exposed to these concentration for 96 hours. After 96hrs of exposure LC0, LC50 and LC100 was observed. Effect on the symptoms and the normal activity was checked in the interval of 24rs. pH, tempterature and dissolved oxygen was also checked.

The lethal concentrations LC50 was determine to be >100 mg/L

LC0 (96 hours) (highest loading at which no mortality was observed) = 100 mg/L

LC50 (96 hours) Experimental = >100 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) =No mortality was observed

Thus based on the LC50 it can be concluded that the chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex was nontoxic and can be consider to be not classified as per the CLP classification criteria.

Long term toxicity to fish:

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the six closest read across substances, toxicity on Oncorhynchus mykiss (previous name: Salmo gairdneri) was predicted for aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]- 2-naphthalenesulfonic acid complex (15790-07-5). The LC50 value was estimated to be 3.638 mg/l when aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex exposed to Oncorhynchus mykiss (previous name: Salmo gairdneri) for 672hrs.  

Short term toxicity to aq. invertebrates:

Based on the prediction done using the OECD QSAR toolbox version 2.3 with log kow as the primary descriptor and considering the six closest read across substances, toxicity on Daphnia magna was predicted for aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]- 2-naphthalenesulfonic acid complex (15790-07-5). The EC50 value was estimated to be 128.230 mg/l when aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex exposed to Daphnia magna for 48hrs.   

 

Long term toxicity to aq. invertebrates:

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the six closest read across substances, toxicity on daphnia magna was predicted for aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]- 2-naphthalenesulfonic acid complex (15790-07-5). The EC50 value was estimated to be 83.527 mg/l when aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex exposed to daphnia magna for 504 hrs. Thus based on the EC50 it can be concluded that the chemical was nontoxic and can be consider to be not classified as per the CLP classification criteria.

Toxicity to aq. algae and cyanobacteria:

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. The microscopic observations were noted down in each of the control vessel. All the cells appeared healthy, round and green throughout the study duration in the control. Also, the drift in pH in the control vessels did not increase by >1.5 units when observed on 72 hours as compared to 0 hours. The average pH drift observed in the control vessels was 0 units. After 72 hours of exposure to aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) to various nominal test concentration, EC50 calculated from equation through probit analysis was determine to be > 200 mg/l. Based on the EC50, it can be concluded that the chemical was nontoxic and can be consider to be not classified as toxic as per the CLP classification criteria.

Toxicity to microorganisms:

Based on the toxicity on the (15790 -07 -5) was predicted by using prediction done using the OECD QSAR toolbox version 2.3 with log kow as the primary descriptor and considering the six closest read across substances, toxicity was measured. The inhibition growth concentration (IGC50) value of aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex in Tetrahymena pyriformis in a 48 hr study on growth inhibition effect was determine to be 88.5 mg/L.

 

Additional information

Short term toxicity to fish:

Based on the various experimental data for the target chemical and experimental data for read across chemicals study have been reviewed to determine the toxic nature of target chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) on the fishes. The studies are as mentioned below:

In the first key study for the target chemical (15790 -07 -5) from UERL report. Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with 2 hrs continuous stirring for achieving test concentrations of 100mg/L,respectively.

The test was performed on the limit concentration i.e nominal concentration selected for the experiment were 100 mg/L and test fish were exposed to these concentration for 96 hours. After 96hrs of exposure LC0, LC50 and LC100 was observed. Effect on the symptoms and the normal activity was checked in the interval of 24rs. pH, tempterature and dissolved oxygen was also checked. The lethal concentrations LC50 was determine to be >100 mg/L

LC0 (96 hours) (highest loading at which no mortality was observed) = 100 mg/L

LC50 (96 hours) Experimental = >100 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) =No mortality was observed

Thus based on the LC50 it can be concluded that the chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex was nontoxic and can be consider to be not classified as per the CLP classification criteria.

Similarly in the second supporting study for the read across chemical toxicity was studied from the UERL lab, 2017. Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) aluminium 6-hydroxy-5-[(2-methoxy-5-methyl-4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex. The stock solution prepared as 400mg/4L, with the concentration of 100mg/L, and was kept for 24 hr stirring. After this filter the stock, give it for analytical detection and then stock taken for experiment. The test was performed on the limit concentration i.e nominal concentration selected for the experiment were 100 mg/L and test fish were exposed to these concentration for 96 hours. After 96hrs of exposure LC0, LC50 and LC100 was observed. Effect on the symptoms and the normal activity was checked in the interval of 24rs. pH, tempterature and dissolved oxygen was also checked. The lethal concentrations LC50 was determine to be >100 mg/L

LC0 (96 hours) (highest loading at which no mortality was observed) = 100 mg/L

LC50 (96 hours) Experimental = >100 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) =No mortality was observed

Thus based on the LC50 it can be concluded that the chemical aluminium 6-hydroxy-5-[(2-methoxy-5-methyl-4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex was nontoxic and can be consider to be not classified as per the CLP classification criteria.

Based on the data from various data, it can be concluded that the substance aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) is considered to be not toxic to aquatic environment (fishes) and cannot be classified as toxic as per the criteria mentioned in CLP regulation.

Long term toxicity to fish:

Based on the various predicted data for the target chemical and experimental data for read across chemicals study have been reviewed to determine the toxic nature of target chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) on the growth of fish. The studies are as mentioned below: 

In the first key study for the target chemical used based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the six closest read across substances, toxicity on Oncorhynchus mykiss (previous name: Salmo gairdneri) was predicted for aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]- 2-naphthalenesulfonic acid complex (15790-07-5). The LC50 value was estimated to be 3.638 mg/l when aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex exposed to Oncorhynchus mykiss (previous name: Salmo gairdneri) for 672hrs.   

 

Similarly in the second predicted report which was based on the prediction done using ECOSAR version 1.1, the long term toxicity on fish was predicted for test substance aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5). On the basis of effects observed in a static freshwater system, the NOEC value for the substance is estimated to be 107.088mg/l for fish for 28 days of exposure duration. Based on this value, it can be concluded that the test chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) can be considered as non-toxic to fish at environmentally relevant conentrations and can be considered not-classified as per the CLP classification criteria. 

Based on the predicted value, it can be concluded that the test chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) can be considered as non-toxic to fish at environmentally relevant conentrations and can be considered not-classified as per the CLP classification criteria. 

Short term toxicity to aq. invertebrates:

Based on the various predicted data for the target chemical and experimental data for read across chemicals study have been reviewed to determine the toxic nature of target chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5). The studies are as mentioned below:

In the first weight of evidence study for the target chemical (15790-07-5) toxicity was predicted based on the prediction done using the OECD QSAR toolbox version 2.3 with log kow as the primary descriptor and considering the six closest read across substances, toxicity on Daphnia magna was predicted for aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]- 2-naphthalenesulfonic acid complex (15790-07-5). The EC50 value was estimated to be 128.230 mg/l when aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex exposed to Daphnia magna for 48hrs.   

 

In the second weight of evidence study for the read across chemical from ABITEC report, study on determination of the inhibition of the mobility of daphnids was carried out with the substance Aluminium,6-hydroxy-5- [(2-methoxy-5-methyl-4-sul fophenyl)azo ]-2-naphthalenesulfonic acid complex according to OECD Guideline 202. A limit test at sample concentration of 100 mg/L was performed. Effects on immobilisation were observed for 48 hours. At 100 mg/l only 8% inhibition was observed, thus it can be concluded that the EC50 was >100 mg/l. The median effective concentration (EC50) for the test substance, Aluminium, 6-hydroxy-5- [(2-methoxy-5-methyl-4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex, in Daphnia magna was determined to be >100 mg/L for immobilisation effects. Based on this EC50 value and after comparing with CLP criteria for aquatic classification of the substance it is concluded that the substance, Aluminium, 6-hydroxy-5- [(2-methoxy-5-methyl-4-sulfophenyl)azo ]-2-naphthalenesulfonic acid complex does not exhibit short term toxicity to aquatic invertebrate (Daphnia Magna).

 

Similarly in the third weight of evidence study for RA chemical 2,7-Naphthalenedisulfonic acid, 3-hydroxy- 4-[(4-sulfo-1-naphthalenyl), sodium salt (Amaranth dye) (915-67-3), ABITEC lab report, 2016. Determination of the inhibition of the mobility of daphnids was carried out with the substance 2,7-Naphthalenedisulfonic acid, 3-hydroxy- 4-[(4-sulfo- 1-naphthalenyl), sodium salt; Amaranth dye according to OECD Guideline 202. The limit test was performed at 100 mg/l. Effects on immobilisation were observed for 48 hours. The effective concentration (EC8) for the test substance, 2,7-Naphthalenedisulfonic acid, 3-hydroxy-4- [(4-sulfo-1 -naphthalenyl), sodium salt (Amaranth dye), in Daphnia magna was determined to be 100 mg/L on the basis of mobility inhibition effects in a 48 hour study. This value indicates that the substance is likely to be non-hazardous to aquatic invertebrates and cannot be classified as toxic as per the CLP criteria.

 

Study was conducted on the read across chemical RA chemical 2,7-Naphthalenedisulfonic acid, 3-hydroxy- 4-[(4-sulfo-1-naphthalenyl), sodium salt (Amaranth dye) (915-67-3), (from The Journal of Toxicological Sciences, 1997). The toxic effects of Amaranth were studied on Artemia salina larvae. Artemia salina (A. salina eggs) a crustacean, commonly known as brine shrimp eggs, are commercially available, and are easily cultured in the laboratory because they are resistant to environmental stresses. Active larvae can be obtained within 1 to 2 days and no live culture is required for a few days thereafter. A. salina eggs (encysted dried gastrulae) were commercially obtained, and were stored at -200°C. Eggs used in experiments were washed and stored at room temperature in a desiccators over anhydrous granular CaCl2. Larvae were obtained by incubating eggs in petri dishes containing muslin-filtered sea water at 30°C for 24 hours. The larvae were separated from shells, dead larvae and unhatched eggs by their phototactic movements towards a light source. Amaranth at concentrations of 6044.7mg/l and 604.47 mg/l were placed in a petri dish, and sea water containing 20 to 30 larvae was added. After this was incubated at 30°C for 24 hours and 48 hours, larvae surviving were measured by direct count. The same method was tested from 5 to 6 times for each concentration, and the death rate was calculated. Death was assumed to have occurred when there was no movement. The death rate was defined as the average of the percentage of deaths observed for 24 hours and 48 hours. 100% death rate was noted after 48 hours when 6044.7 mg/l of Amaranth was exposed to the test organism and 0% death rate after 24 hours in case of exposure to 604.47 mg/l of test chemical.

 

Based on the data from various data, it can be concluded that the substance aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) is considered to be not toxic to aquatic environment and cannot be classified as toxic as per the criteria mentioned in CLP regulation.

Long term toxicity to aq. invertebrates:

Based on the various predicted data for the target chemical and experimental data for read across chemicals study have been reviewed to determine the toxic nature of target chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5). The studies are as mentioned below: 

 

In the first weight of evidence study for the target chemical (15790-07-5) toxicity was predicted based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the six closest read across substances, toxicity on daphnia magna was predicted for aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]- 2-naphthalenesulfonic acid complex (15790-07-5). The EC50 value was estimated to be 83.527 mg/l when aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex exposed to daphnia magna for 504 hrs. Thus based on the EC50 it can be concluded that the chemical was nontoxic and can be consider to be not classified as per the CLP classification criteria.

 

In the second weight of evidence study for the read across chemical (657-84-1) from J-check study on determination of the inhibition of the mobility of daphnids was carried out with the substance Aluminium,6-hydroxy-5- [(2-methoxy-5-methyl-4-sul fophenyl)azo ]-2-naphthalenesulfonic acid complex according to OECD Guideline 211. Acute Immobilization Test of 4-methylbenzenesulfonic acid (Sodium p-toluenesulfonate) to Daphnia magna was studied for 21 days. Test was performed according to the OECD Guideline 211 (Daphnia magna Reproduction Test). Test performed in the semi-static system. Control, 100, 32, 10 mg/L nominal concentration was added in the test vessel filled with 80 ml of test solution. Test conducted in the ten replicates, 10 daphnia magna per concentration (1 per vessel was added). Based on the effect of chemical 4-methylbenzenesulfonic acid (Sodium p-toluenesulfonate) on the normal activity of daphnia magna for the 21 days, the EC50 was ≥ 100 mg/l and NOEC was 100 mg/l. Thus based on the EC50, it can be concluded that the 4-methylbenzenesulfonic acid was nontoxic and can be consider to be not classified as per the CLP classification criteria. 

 

Similarly third weight of evidence study supports the first predicted study for RA chemical (5281-04-9) from OECD SIDS report, 1994 similar to the target chemical. Study was conducted to determine the long term toxicity of calcium 3-hydroxy-4-[(4-methyl-2-sulfonatophenyl)diazenyl]-2-naphthoate on the mortality rate of daphnia magna. Test conducted in the open system by semi-static method. 4 replicates were used in which 10 organisms were used in each replicates. Control of DMSO: HCO-40 was 9:1 (100 mg/kg). Test conducted on the nominal concentration for 7 days, 14 days and 21days. After the exposure of chemical lethal concentration was measured. Based on the mortality rate of daphnia magna due to the long period exposure with chemical calcium 3-hydroxy-4-[(4-methyl-2 sulfonatophenyl) diazenyl]-2-naphthoate, the LC50 value was 13 mg/l, 10 mg/l and 9.7 mg/l for 7 days, 14 days and 21 days of exposure. The EC50 was 4.4 and 9.1 after 14 and 21 days of exposure. Based on the LC50, it can be concluded that the chemical was nontoxic and not classified as per the CLP classification criteria.

 

Based on the data from various data, it can be concluded that the substance aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) is considered to be not toxic to aquatic environment and cannot be classified as toxic as per the criteria mentioned in CLP regulation.

Toxicity to aq. algae and cyanobacteria:

Based on the various experimental data for the target chemical and read across chemicals study have been reviewed to determine the toxic nature of target chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5). The studies are as mentioned below:

In the first key study for the aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) from UERL lab, 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. The microscopic observations were noted down in each of the control vessel. All the cells appeared healthy, round and green throughout the study duration in the control. Also, the drift in pH in the control vessels did not increase by >1.5 units when observed on 72 hours as compared to 0 hours. The average pH drift observed in the control vessels was 0 units. After 72 hours of exposure to aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) to various nominal test concentration, EC50 calculated from equation through probit analysis was determine to be > 200 mg/l. Based on the EC50, it can be concluded that the chemical was nontoxic and can be consider to be not classified as toxic as per the CLP classification criteria.

 

 

In the second supporting study for the read across chemical from UERL lab, 2016 toxicity was measured on algae. The effect of test item aluminium, 6-hydroxy-5-[(2-methoxy-5-methyl-4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex, CAS No. 68583-95-9 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-[(2-methoxy-5-methyl-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 for 24 hours and filter it 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. The microscopic observations were noted down in each of the control vessel. All the cells appeared healthy, round and green throughout the study duration in the control. Also, the drift in pH in the control vessels did not increase by >1.5 units when observed on 72 hours as compared to 0 hours. The average pH drift observed in the control vessels was 0.1 units. 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 the test item. EC50 calculated graphically through probit analysis was observed to be 136.57 mg/L. Based on the EC50, it can be concluded that the chemical was nontoxic and can be consider to be not classified as toxic as per the CLP classification criteria.

 

 

Similarly in the third supporting study for RA chemical 2,7-Naphthalenedisulfonic acid, 3-hydroxy- 4-[(4-sulfo-1-naphthalenyl), sodium salt (Amaranth dye) (915-67-3), ABITEC lab report, 2016. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance 2,7-Naphthalenedi- sulfonic acid, 3-hydroxy-4-[(4-sulfo- 1-naphthal - enyl), sodium salt (Amaranth dye) according to OECD Guideline 201. The test substance was dissolved in OECD growth medium and tested at the concentrations 0, 12.5, 25, 50, 100, 200 mg/L. Effects on the growth rate of the organism were studied. The effective concentration (ErC50) for the test substance, 2,7-Naphthalen -edisulfonic acid, 3-hydroxy-4-[(4-sulfo-1- -naphthalenyl), sodium salt, in Desmodesmus subspicatus was determined to be 356.2 mg/L. This value indicates that the substance is likely to be non-hazardous to aquatic algae and cannot be classified as toxic as per the CLP criteria.

 

Similarly in the fourth supporting study for RA chemical 2,7-Naphthalenedisulfonic acid, 3-hydroxy- 4-[(4-sulfo-1-naphthalenyl), sodium salt (Amaranth dye) (915-67-3),UERL report, 2016, The effect of test substance Trisodium 3-hydroxy-4-(4’-sulphonatonaphthylazo) naphthalene-2,7-disulphonate (CAS No. 915-67-3) 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, 12.5mg/l, 25mg/l, 50mg/l, 100mg/l and 200mg/l were prepared using stock solution of the test substance using de-ionized water. 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 from equation through probit analysis was observed to be > 200 mg/L. Thus based on the EC50 it was concluded that the chemical was nontoxic and can be consider to be not classified as toxic to aquatic environment as per the CLP classification criteria.

 Based on the data from various data, it can be concluded that the substance aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) is considered to be not toxic to aquatic environment and cannot be classified as toxic as per the criteria mentioned in CLP regulation.

 

Thus based on the data for the toxicity to fish, aquatic invertebrates and algae from various data source, it can be concluded that the substance aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) is considered to be not toxic to aquatic environment and cannot be classified as toxic as per the criteria mentioned in CLP regulation.

Toxicity to microorganisms:

Based on the various experimental data for the target chemical and read across chemicals study have been reviewed to determine the toxic nature of target chemical aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5). The studies are as mentioned below:

 

In the first weight of evidence study for the target chemical(15790-07-5) from QSAR based on the toxicity on the (15790 -07 -5) was predicted by using prediction done using the OECD QSAR toolbox version 2.3 with log kow as the primary descriptor and considering the six closest read across substances, toxicity was measured. The inhibition growth concentration (IGC50) value of aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex in Tetrahymena pyriformis in a 48 hr study on growth inhibition effect was determine to be 88.5 mg/L.

 

 

In the second study for read across chemical (915-67-3) Biotechnology Letters, 2002. The Microtox acute toxicity assay was performed by using a modified strain of Vibrio fischeri . Frozen samples were brought to room temperature, and centrifuged. The pH of the samples was adjusted where necessary to 6 by adding 0.5 ml 0.58 M KH2 PO4 and 70μl 1 M NaOH. Colour correction was done at 490 nm. The Microtox acute toxicity assay was performed in a Microtox 500 Analyzer on samples before and after decoloration according to the test protocols defined by the manufacturer From eight serial dilutions, the percent concentration to decrease 20% of the luminescence of amodified strain ofVibrio fischeri(EC20)after 5 min incubation was calculated with the Microtox data analysis program [Microtox Omni Software(1999) Azur Environmental, Newark, Del.]. A solution of 1 g/l ZnSO4·7H2O was used as the positive control and 1 g/l glucose as the negative control. Each EC20 reported is the average of triplicate analysis. The concentration to decrease 50% of the bacterial luminescence in the Microtox acute assay (EC50) is normally reported. However, in most of these studies, the EC50 before or after decoloration was greater than 100% indicating that there was no toxicity or toxicity change. To better evaluate whether the decoloration process affected toxicity, the dilution required to decrease 20% of the bacterial luminescence relative to the control (EC20)was reported instead. The following rating was adapted from Coleman & Qureshi (1985) –

EC20:>100%=nontoxic;

>75–100%=slightly non-toxic;

 >50–75%=toxic;

>25–50%=moderately toxic;

<25% very toxic. The toxicity of 100mg/l of Amaranth determined in terms of EC20 (% dilution) was 44.6 ± 11.6.

Similarly in the third weight of evidence study for the same read across chemical from (Indian J Microbiol 2011). This investigation was aimed at identifying the effects of the Amaranth dye and its degradation products on microbial growth. Amaranth dye was purchased from Hi-media Laboratories Pvt. Ltd., Mumbai, India. Aspergillus ochraceus NCIM 1146 was obtained from National Chemical Laboratory, Pune, India. E. coli MTCC 452, B. subtilis MTCC 6910 and Penicillium ochrochloron MTCC 517 were obtained from Microbial Type Culture Collection and Gene Bank (MTCC), Institute of Microbial Technology, Chandigarh, India. It was regularly maintained and preserved at 4°C on nutrient agar slants contained in (g/l); bacteriological peptone 10.0, beef extract 10.0 and NaCl 5.0 Microbial toxicity of control dye amaranth and metabolites obtained after its decolorization (final concentration 1,000 ppm) was carried out in relation to E. coli, Bacillus substilis, Aspergillus ochraceus and Penicillium ochrochloron MTCC 517 and zone of inhibition (diameter in mm) was recorded. The diameter of the discs used was 10mm. Amaranth and its degradation products were not toxic to Aspergillus ochraceus and Penicillium ochrochloron MTCC 517 at 1,000 ppm concentration. Amaranth inhibited growth of E. coli and Bacillus substilis.

 

Similarly in the fourth study for the read across chemical (915-67-3) (from, TOXICOLOGY AND APPLIED PHARMACOLOGY, 1977). The death of Paramecium caudatum (PC), a unicellular animal, can be observed more readily and in far less time than that of small animals. Hence a bioassay was conducted to study the toxic effect of Amaranth. Paramecium Caudatum was maintained at 22°C on 0.15 % dried lettuce infusion and fed with Aerobacter aerogenes. Amaranth was tested in 0.1% and 1% concentration. The test concentrations were put in a hollow slide glass, and an equal volume of 0.04 M phosphate buffer, pH 7.0, was added. After 5 to 10 test organisms were added, their survival times were measured microscopically. Thirty to forty test organisms for each concentration were tested by the same method, and the mean survival time and the death rate were calculated. The survival time was defined as the time required until death was observed for each concentration. Death was assumed to have occurred when there was no movement. The death rate was defined as the percentage of deaths observed during 20 minutes. The mean survival time (in sec) of test organism Paramecium caudatum was determined to be 695 seconds.  The death rate of the test organism at 10000mg/l was 77.4%. Therefore the Effective concentration causing more than 50% death of Paramecium caudatum was reported as 10000 mg/l.

Based on the toxicity on fish, aquatic invertebrates and algae, it can be concluded that the chemical was nontoxic and can be consider to be not classified as toxic as per the CLP classification criteria.