Dialuminium tris[2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)-3,4,5,6-tetrachlorobenzoate]

Endpoint summary

Description of key information

Short term toxicity to fish:

A fish acute toxicity test according to OECD Guideline 203 was conducted for dialuminiumtris [2-(2, 4, 5, 7-tetrabromo-6 -oxido- 3-oxo xanthen-9-yl)-3, 4, 5, 6-tetrachlorobenzoate]. The nominal exposure concentration was 0.778 mg/L i.e. the solubility limit of the tested chemical. The test organism was zebrafish (Danio rerio). One had two tanks per concentration and 4 fish per tank (8 fish in total per concentration). The water control contained the same number of fish. One observed no mortalities or abnormal behaviour in these fish tanks. The exposure time was 96 hours. The exposed fish were swimming a bit slower after 96 hours. However, no mortalities were observed in the exposure tanks. The LC0 was therefore predicted to be >0.778 mg/L. The LC0 value was considered to be above the solubility limit for the tested chemical.

Short term toxicity to aquatic invertebrates:

An acute immobilisation test was used to test how a range of concentrations of CAS 15876-58-1 exerts different degrees of toxic effects on the swimming capability ofDaphnia magnaunder otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 202 by ABITEC in Prague. The standardized protocol used to obtain the herein described results was by the testing laboratory referred to as “CSN EN ISO 6341”. The testing aim was to determine a EC50 after 48 hours of exposure toD. magna. The 48 hours EC50 was by the testing laboratory defined as the concentration estimated to immobilise 50 per cent of theDaphniaafter 48 hours of exposure. The stock solution used for the exposure assessment was prepared by dissolving dark red powder in reconstituted water. The stock solution was kept at 5 min in an ultrasonic bath. The test exposure concentrations were prepared by mixing the stock solution of the test sample with reconstituted water.Daphnidswere exposed to CAS 15876-58-1 in 50 ml beakers in a volume of 25 ml of liquid solution containing both the chemical and media as specified in OECD 202. The beakers were placed in a temperature controlled room at 20±1 degrees Celsius. TheD. magna(age ≤24) used for the test had been breed at ABITEC. The breeding stock ofD. magnaoriginated from University of Technology in Prague. The animals were exposed to medium (i.e.a beaker containing only medium) and/or the tested chemical during 48 hours (±1 hour). None of the exposed animal’s immobilization were affected by exposure to only medium. The nominal concentrations used were: 5, 10, 25, 50, 100 and 200 mg/L. There were 5Daphniaper test vessels and 4 replicates per concentration. The pH in test vessels were 7.5-7.8 mg/L. The positive control/reference substance used in the tested showed an expected result and gave a EC50 that corresponded to previous exposures with this chemical inD. magna. Immobilisation effects inD. mangaby chemical exposure was evident after 48 hours in exposure concentrations above 10 mg/L. The EC50 was defined as a concentration that immobilizes 50% of the exposedD. magna. The % of immobilization inD. magnaafter 48 hours of exposure to CAS 15876-39-8 were used in a nonlinear regression by Graphpad Software Prism 4.0 (San Diego, US). The EC50 was estimated based on the nonlinear regression. In conclusion, the EC50 for CAS 15876-58-1 was 152.3 mg/L (95% C.I: 108.7-213.4 mg/L) after 48 hours of exposure inD. magna.

Toxicity to aquatic algae and cyanobacteria:

A freshwater algal growth inhibition exposure assessment was used to test how a range of concentrations of CAS 15876-58-1 exerts different degrees of toxic effects on the growth ofDesmodesmus subspicatusunder otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 201 by ABITEC in Prague, Crez Republic. The standardized protocol used to obtain the herein described results was by the testing laboratory referred to as “CSN EN ISO 8692”. The 72 hours EC50 was by the testing laboratory defined as the concentration estimated to inhibit 50 per cent of the algae cell growth compared to unexposed cells after 72 hours of exposure. The nominal concentrations used for the exposure assessment were as follows: 5, 10, 20, 40 and 80 mg/L. The positive control/reference substance used in the tested showed an expected result and gave a EC50 that corresponded to previous exposures with this chemical inDesmodesmus subspicatus. After exposure was the EC50 calculated using nonlinear regression by the software Prism 4.0 (Graphpad Software, Inc., San Diego CA). In conclusion, the EC50 for CAS 15876-58-1 was 25.0 mg/L (95% C.I.: 19.5-32.1 mg/L) after 72 hours of exposure. Based on the EC50, Di aluminium tris [2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)benzoate] was consider as toxic and can be consider as classified in aquatic chronic 3 as per CLP classification criteria.

Toxicity to microorganisms:

Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on microorganisms was predicted for 2',4',5',7'-tetrabromo-4,5,6,7-tetrachloro-3',6' -dihydroxy -3H-spiro[2-benzofuran-1,9'-xanthen]-3-one (15876-58-1). IGC50 value was estimated to be 9.062mg/l when 2',4',5',7'-tetrabromo-4,5, 6,7 -tetrachloro-3',6'-dihydroxy-3H- spiro[2-benzofuran- 1,9'- xanthen]-3-one was exposed to Tetrahymena pyriformis for 48 hours.

Additional information

Toxicity of the chemical dialuminiumtris [2-(2, 4, 5, 7-tetrabromo-6 -oxido- 3-oxo xanthen-9-yl)-3, 4, 5, 6-tetrachlorobenzoate] (15876 -58 -1) to the aquatic fish,invertebrates, algae and microorganisms have been summarized by collecting the data from various sources:

Short term toxicity to fish:

A fish acute toxicity test according to OECD Guideline 203 was conducted for dialuminiumtris [2-(2, 4, 5, 7-tetrabromo-6 -oxido- 3-oxo xanthen-9-yl)-3, 4, 5, 6-tetrachlorobenzoate]. The nominal exposure concentration was 0.778 mg/L i.e. the solubility limit of the tested chemical. The test organism was zebrafish (Danio rerio). One had two tanks per concentration and 4 fish per tank (8 fish in total per concentration). The water control contained the same number of fish. One observed no mortalities or abnormal behaviour in these fish tanks. The exposure time was 96 hours. The exposed fish were swimming a bit slower after 96 hours. However, no mortalities were observed in the exposure tanks. The LC0 was therefore predicted to be >0.778 mg/L. The LC0 value was considered to be above the solubility limit for the tested chemical.

Short term toxicity to aquatic invertebrates:

Based on the various experimental data for the target chemical study have been reviewed to determine the toxic nature of Dialuminium tris[2-(2,4,5,7 tetrabromo- 6-oxido-3-oxoxanthen-9-yl)-3,4,5,6-tetrachlorobenzoate] (15876 -58 -1). The studies are as mentioned below:

In the first key study for 15876 -58 -1, from report, An acute immobilisation test was used to test how a range of concentrations of CAS 15876-58-1 exerts different degrees of toxic effects on the swimming capability of Daphnia magna under otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 202 by ABITEC in Prague. The standardized protocol used to obtain the herein described results was by the testing laboratory referred to as “CSN EN ISO 6341”. The testing aim was to determine a EC50 after 48 hours of exposure toD. magna. The 48 hours EC50 was by the testing laboratory defined as the concentration estimated to immobilise 50 per cent of theDaphnia after 48 hours of exposure. The stock solution used for the exposure assessment was prepared by dissolving dark red powder in reconstituted water. The stock solution was kept at 5 min in an ultrasonic bath. The test exposure concentrations were prepared by mixing the stock solution of the test sample with reconstituted water.Daphnids were exposed to CAS 15876-58-1 in 50 ml beakers in a volume of 25 ml of liquid solution containing both the chemical and media as specified in OECD 202. The beakers were placed in a temperature controlled room at 20±1 degrees Celsius. TheD. magna(age ≤24) used for the test had been breed at ABITEC. The breeding stock ofD. magna originated from University of Technology in Prague. The animals were exposed to medium (i.e.a beaker containing only medium) and/or the tested chemical during 48 hours (±1 hour). None of the exposed animal’s immobilization were affected by exposure to only medium. The nominal concentrations used were: 5, 10, 25, 50, 100 and 200 mg/L. There were 5Daphniaper test vessels and 4 replicates per concentration. The pH in test vessels were 7.5-7.8 mg/L. The positive control/reference substance used in the tested showed an expected result and gave a EC50 that corresponded to previous exposures with this chemical inD. magna. Immobilisation effects inD. mangaby chemical exposure was evident after 48 hours in exposure concentrations above 10 mg/L. The EC50 was defined as a concentration that immobilizes 50% of the exposedD. magna. The % of immobilization inD. magnaafter 48 hours of exposure to CAS 15876-39-8 were used in a nonlinear regression by Graphpad Software Prism 4.0 (San Diego, US). The EC50 was estimated based on the nonlinear regression. In conclusion, the EC50 for CAS 15876-58-1 was 152.3 mg/L (95% C.I: 108.7-213.4 mg/L) after 48 hours of exposure in D. magna.

In the another support study for RA chemical rose bengal (632 -68 -8),1977. Short term toxicity study of rose bengal to Artemia Salina was carried out for 24-48 hrs. The test chemical conc. used for the study was 1049.84 mg/l, respectively. A. salina eggs (encysted dried gastrulae) were commercially obtained, and were stored at -20°C, Eggs used in experiments were washed and stored at room temperature in a desiccator over anhydrous granular CaCl, . 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 toward a light source. Food dyes of various concentrations were placed in a petri dish, and sea water containing 20 to 30 larva ewas added. After this was incubated at 30°Cfor 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. The study was performed under static conditions for 24 – 48 hrs at 30°C. 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. Based on death rate or mortality of test organism, the 48 hr LC100 value was found to be 1049.84 mg/l. According to the result the test chemical rose bengal was not classified according to the CLP regulations.

In the another supporting study for RA chemical 6441 -77 -6, Short term toxicity study of Phloxin to Artemia Salina was carried out for 24-48 hrs. The test chemical conc. used for the study was 792.96 mg/l, respectively. A. salina eggs (encysted dried gastrulae) were commercially obtained, and were stored at -20°C, Eggs used in experiments were washed and stored at room temperature in a desiccator over anhydrous granular CaCl, . 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 toward a light source. Food dyes of various concentrations were placed in a petri dish, and sea water containing 20 to 30 larva ewas 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.The study was performed under static conditions for 24 – 48 hrs at 30°C. 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 48 hours. Based on death rate or mortality of test organism, the LC 100 value was 792.96 mg/l. The chemical Phloxin was considered as non toxic and not classified according to the CLP regulation.

Based on the data for target chemical considering RA chemical and target chemical, Dialuminium tris[2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)-3,4,5,6-tetrachlorobenzoate] was consider as non toxic and can be classified as not classified as per CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria:

A freshwater algal growth inhibition exposure assessment was used to test how a range of concentrations of CAS 15876-58-1 exerts different degrees of toxic effects on the growth ofDesmodesmus subspicatusunder otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 201 by ABITEC in Prague, Crez Republic. The standardized protocol used to obtain the herein described results was by the testing laboratory referred to as “CSN EN ISO 8692”. The 72 hours EC50 was by the testing laboratory defined as the concentration estimated to inhibit 50 per cent of the algae cell growth compared to unexposed cells after 72 hours of exposure. The nominal concentrations used for the exposure assessment were as follows: 5, 10, 20, 40 and 80 mg/L. The positive control/reference substance used in the tested showed an expected result and gave a EC50 that corresponded to previous exposures with this chemical inDesmodesmus subspicatus. After exposure was the EC50 calculated using nonlinear regression by the software Prism 4.0 (Graphpad Software, Inc., San Diego CA). In conclusion, the EC50 for CAS 15876-58-1 was 25.0 mg/L (95% C.I.: 19.5-32.1 mg/L) after 72 hours of exposure. Based on the EC50, Di aluminium tris [2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)benzoate] was consider as toxic and can be consider as classified in aquatic chronic 3 as per CLP classification criteria.

Toxicity to microorganisms:

Based on the data for the terget chemical 2',4',5',7'-tetrabromo-4,5, 6,7 -tetrachloro-3',6'-dihydroxy-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one, study have been reviewed to determine the toxic nature of chemical (15876-58-1) on the growth of microorganisms.

In the first weight of evidence study for 2',4',5',7'-tetrabromo-4,5, 6,7 -tetrachloro-3',6'-dihydroxy-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one, QSAR 2017,Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on microorganisms was predicted for 2',4',5',7'-tetrabromo-4,5,6,7-tetrachloro-3',6' -dihydroxy-3H-spiro [2-benzofuran- 1,9'-xanthen]-3-one (15876-58-1). IGC50 value was estimated to be 9.062mg/l when 2',4',5',7'-tetrabromo-4,5,6,7-tetrachloro-3',6'-dihydroxy -3H- spiro [2-benzofuran-1,9'-xanthen]-3-one was exposed to Tetrahymena pyriformis for 48 hours.

In the second weight of evidence study for RA chemical rose bengal (632 -68 -8),The main objective of the study to evaluate the toxic effect of food dyeRose Bengal Paramecium caudatum. The effect of food dye on leucinea minopeptidase,acid phosphatise and y-glutamyl transpeptidasea ctivity in P. caudatum was also studied in order to investigate the mechanism of toxicity. In mean survival study test organism measures microscopically for 20 min. The concentration of test material in the bathing fluid was 0.1 or 1 .0%. The mean survival time of PC in 0.1% concentration varied with the chemical structure of the dye. Xanthene dye rose Bengal containing halogen atoms in their molecules showed smaller survival times,and thus these dyes were considered to be more toxic than other groups of dyes.The death rate of PC in 0.1% solution of these dyes was 100%. While in enzyme inhibition study test substance i.e Rose bengal inhibited LAP activity almost completely. The experimental results suggest that xanthene dye such as rose Bengal is most toxic to Paramecium caudatum when observed for 20 min at dose concentration of 0.1 %, the EC100 was 1000mg/l (0.1%) and MIC was 0.0095 mg/l on the basis of Mean Survival and enzyme inhibition effects. Thus overall experimental observation suggest that the test substance Rose Bengal (Cas no.632-68-8) is toxic to micro-organism i.e Paramecium caudatum.

In the another weight of evidence study by considering RA chemical (6441 -77 -6) phloxin,The main objective of the study to evaluate the toxic effect of food dye phloxin to Paramecium caudatum. The effect of food dye on leucinea minopeptidase,acid phosphatise and y-glutamyl transpeptidasea ctivity in P. caudatum was also studied in order to investigate the mechanism of toxicity. In mean survival study test organism measures microscopically for 20 min. The concentration of test material in the bathing fluid was 0.1 or 1 .0%. The mean survival time of PC in 0.1% concentration varied with the chemical structure of the dye. Xanthene dye phloxin containing halogen atoms in their molecules showed smaller survival times,and thus these dyes were considered to be more toxic than other groups of dyes.The death rate of PC in 0.1% solution of these dyes was 100%. While in enzyme inhibition study test substance i.e phloxinl inhibited LAP, AP and y-Glutamyl transpeptidase activity almost completely. The experimental results suggest that xanthene dye such as phloxin is most toxic to Paramecium caudatum when observed for 20 min at dose concentration of 0.1 %, the EC100 was 1000mg/l (0.1%) and MIC was 0.126 mg/l on the basis of Mean Survival and enzyme inhibition effects. Thus overall experimental observation suggest that the test substance phloxin is toxic to micro-organism i.e Paramecium caudatum. Thus overall experimental observation suggest that the test substance is toxic to micro-organism i.e Paramecium caudatum. Based on the data for the target chemical 2',4',5',7'-tetrabromo-4,5, 6,7 -tetrachloro-3',6'-dihydroxy-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one, it was concluded thet the chemical was toxic to the growth of microorganisms.

Based on the data from various sources for daphnia magna, aquatic algae, for the target chemical 2',4',5',7'-tetrabromo-4,5, 6,7 -tetrachloro-3',6'-dihydroxy-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one (15876 -58 -1), it was concluded that the chemical can be consider as toxic to the aquatic life and can be consider to be classified in aquatic chronic category 3 as per the CLP classification criteria.