Ferrate(4-), hexakis(cyano-C)-, Et 2-[6-(ethylamino)-3-(ethylimino)-2,7-dimethyl-3H-xanthen-9-yl]benzoate copper(2+) salts

Endpoint summary

Description of key information

Short term toxicity to fish:

Short term toxicity to fish for 96 h was evaluated for the test material . The test was performed as per EEC-Guideline L251.No stock solution was prepared.The test substance was mixed directly in the water of the tanks .10 fishes per concentration and control were used .Glass aquaria of 20 liters filled with 15 liters Dechlorinated tap water (carbon filter) was used as medium . The nominal test concentration used were 1.0, 1.8, 3.2, 5.8, 10 mg/l . The LC-50 values were calculated according to the maximum likelihood method, probit model. The lethal concentration (LC50) value of test material in zebra fish in a 96 hr study was observed to be 6.3 mg/l. Based on the above effect concentration it can be concluded that test substance was toxic for fish and hence, considered to be classified in aquatic chronic category 2 as per CLP criteria.

Long term toxicity to fish:

Based on the prediction done using ECOSAR version 1.1, the long term toxicity on fish was predicted for test substance . On the basis of effects observed in a flow through fresh water system, the NOEC value for the substance was estimated to be 0.00000412mg/l for fish for 28 days of exposure duration.Based on the NOEC value it can be concluded that the test chemical as toxic to fish at environmentally relevant concentrations and hence, considered to be classified in aquatic chronic category 1 as per the CLP classification criteria. 

Short term toxicity to aquatic invertebrate:

Short term toxicity of test material was evaluated on aquatic invertebrate Daphnia magna for 24 hr according to EEC-Guideline L251 .Cultures of daphnia were maintained in glass vessels containing approx. 2.5 1 of reconstituted waterat 20 ± 1"C (water is renewed partially thrice weekly). At each renewal the daphnia are fed a suspension of green algae (Scenedesmus subspicatus) supplemented by a suspension of TETRAMIN-extract in such quantities that the food is consumed after 24h. In the pretest 24h before the start of the test reproductive daphnia were separated from the young by sieving all individuals through a 800 µm seive. This operation is repeated 6h before the start of the test and the young (6-24h of age) are retained for the test. Test was performed in beaker covered with watch glass having fill volume of 100 ml. The nominal concentrations at which test was performed were 0.0010, 0.0018, 0.0032, 0.0058, 0.010 0.018, 0.032, 0.058, 0.10, 0.18, 0.32 0.58, 1.0 mg/l. The EC50-values were calculated according to SPEARMAN-KAERBER in: D.J. Finney, 524-530, London (1964). EC-values were graphically determined on gausso-logarithmic probability paper. The effective concentration (EC50) value of test material in aquatic invertebrate (Daphnia Magna) in a 24 hr study on the basis of mobility was estimated to be 0.023 mg/l. Based on the above effect concentration it can be considered that the test material as toxic to aquatic invertebrates.

Long term toxicity to aquatic invertebrate:

Based on the prediction done using ECOSAR version 1.1, the long term toxicity on aquatic invertebrate was predicted for test substance . On the basis of effects observed in a flow through freshwater system, the NOEC value for the test substance was estimated to be 0.0000179 mg/l for aquatic invertebrate for 21 days of exposure, Based on the NOEC value it can be concluded that the test chemical was considered as toxic to aquatic invertebrate at environmentally relevant concentrations and hence, considered to be classified in aquatic chronic category 1 as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria:

On the basis of the experimental studies of the read across chemical and applying the weight of evidence approach, the 72 hr EC50 value of the test chemical  can be expected to be in the range of 0.13 to 0.61 mg/l, respectively. Thus, based on the EC50 value, test chemical can be considered to be toxic to aquatic algae and hence, considered to be classified in 'aquatic acute category 1' as per the CLP classification criteria.

Toxicity to microorganism:

Toxicity studies to determine the inhibitory concentration of test material on the respiration of aerobic waste water bacteria were conducted according to EEC L 133, p. 118-122 (30. May 1988) guideline. The test was conducted on activated sludge from a sewage treatment plant for 3 hours at 20°C. The results were graphically determined . Activated sludge collected from the sewage treatment plant of CH-4106 Therwil on 16/10/89. The preparation was carried out according to the method described in the guideline. The sludge was separated from the aqueous layer by settling instead of centrifugation. 3,5-Dichlorophenol was used as referance substance. The inhibitory concentration (IC50) value was determined to be 15 mg/l.

Additional information

Short term toxicity to fish:

Short term toxicity to fish for 96 h was evaluated for the test material . The test was performed as per EEC-Guideline L251.No stock solution was prepared.The test substance was mixed directly in the water of the tanks .10 fishes per concentration and control were used .Glass aquaria of 20 liters filled with 15 liters Dechlorinated tap water (carbon filter) was used as medium . The nominal test concentration used were 1.0, 1.8, 3.2, 5.8, 10 mg/l . The LC-50 values were calculated according to the maximum likelihood method, probit model. The lethal concentration (LC50) value of test material in zebra fish in a 96 hr study was observed to be 6.3 mg/l. Based on the above effect concentration it can be concluded that test substance was toxic for fish and hence, considered to be classified in aquatic chronic category 2 as per CLP criteria.

Long term toxicity to fish:

Based on the prediction done using ECOSAR version 1.1, the long term toxicity on fish was predicted for test substance . On the basis of effects observed in a flow through fresh water system, the NOEC value for the substance was estimated to be 0.00000412mg/l for fish for 28 days of exposure duration.Based on the NOEC value it can be concluded that the test chemical as toxic to fish at environmentally relevant concentrations and hence, considered to be classified in aquatic chronic category 1 as per the CLP classification criteria. 

Short term toxicity to aquatic invertebrate:

Short term toxicity of test material was evaluated on aquatic invertebrate Daphnia magna for 24 hr according to EEC-Guideline L251 .Cultures of daphnia were maintained in glass vessels containing approx. 2.5 1 of reconstituted waterat 20 ± 1"C (water is renewed partially thrice weekly). At each renewal the daphnia are fed a suspension of green algae (Scenedesmus subspicatus) supplemented by a suspension of TETRAMIN-extract in such quantities that the food is consumed after 24h. In the pretest 24h before the start of the test reproductive daphnia were separated from the young by sieving all individuals through a 800 µm seive. This operation is repeated 6h before the start of the test and the young (6-24h of age) are retained for the test. Test was performed in beaker covered with watch glass having fill volume of 100 ml. The nominal concentrations at which test was performed were 0.0010, 0.0018, 0.0032, 0.0058, 0.010 0.018, 0.032, 0.058, 0.10, 0.18, 0.32 0.58, 1.0 mg/l. The EC50-values were calculated according to SPEARMAN-KAERBER in: D.J. Finney, 524-530, London (1964). EC-values were graphically determined on gausso-logarithmic probability paper. The effective concentration (EC50) value of test material in aquatic invertebrate (Daphnia Magna) in a 24 hr study on the basis of mobility was estimated to be 0.023 mg/l. Based on the above effect concentration it can be considered that the test material as toxic to aquatic invertebrates.

Long term toxicity to aquatic invertebrate:

Based on the prediction done using ECOSAR version 1.1, the long term toxicity on aquatic invertebrate was predicted for test substance . On the basis of effects observed in a flow through freshwater system, the NOEC value for the test substance was estimated to be 0.0000179 mg/l for aquatic invertebrate for 21 days of exposure, Based on the NOEC value it can be concluded that the test chemical was considered as toxic to aquatic invertebrate at environmentally relevant concentrations and hence, considered to be classified in aquatic chronic category 1 as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria:

Data available for the read across chemicals has been reviewed to determine the effect of the test chemical on aquatic algae . The studies are as mentioned below:

Toxicity to aquatic algae study was conducted for 72 hrs for assessing the effect of test chemical on green algae. The test was performed in accordance to OECD Guideline 201 (Alga, Growth Inhibition Test). Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) of strain 86.81 SAG obtained from Institute of botany of the ASCR with an initial biomass conc. 5000 cells /ml was used as a test organism. The stock solution (10 mg/L) was prepared by dissolving yellow powder in OECD growth medium. Test solutions of required concentration were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Test chemical concentrations were not verified analytically. Nominal test chemical conc. used for the study were 0, 0.02, 0.04, 0.08, 0.16, 0.32 and 0.64 mg/l, respectively. Study was performed in a static system for 72 hrs. Desmodesmus subspicatus were exposed to test chemical in 50 ml glass vessel in a volume of 15 ml of liquid solution containing both the chemical and media. Control solution vessel containing OECD medium without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 23±2°C with a continuous light intensity of 6000-8000 lx, respectively. Alongwith the test chemical, one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. The 24 hr EC50 value of the reference substance was determined to be 0.75 mg/l. Cell counting was carried out using microscope with counting chamber Cyrus I or electronic particle counter. ErC50 was calculated using non-linear regression by the software Prism 4.0. On the basis of the effect of test chemical on the growth rate of the test organism Desmodesmus subspicatus, the 72 hr median effect concentration (ErC50) value was determined to be 0.13 mg/l (95% CL: 0.11 to 0.15 mg/l). Thus, based on the EC50 value, test chemical can be considered to be toxic to aquatic algae and hence, considered to be classified in 'aquatic acute category 1' as per the CLP classification criteria.

First study was supported by the second study. The effect of test item was studied on the growth of fresh water green algaChlorella vulgaris.The study was conducted following OECD guideline 201- Alga, growth inhibition test. The test concentration chosen for the study were 0.5mg/l, 1.25mg/l, 3.125mg/l, 7.813mg/l, 19.531mg/l and 48.828mg/l. The test concentrations were prepared using stock solution of the test item 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 the test item. EC50 calculated graphically through probit analysis was observed to be 0.610 mg/L.

On the basis of the experimental studies of the read across chemical and applying the weight of evidence approach, the 72 hr EC50 value of the test chemical  can be expected to be in the range of 0.13 to 0.61 mg/l, respectively. Thus, based on the EC50 value, test chemical can be considered to be toxic to aquatic algae and hence, considered to be classified in 'aquatic acute category 1' as per the CLP classification criteria.

Toxicity to microorganism:

Toxicity of test material on microorganism was evaluated using two experimental reports described as follows:

In the first toxicity studies to determine the inhibitory concentration of test material on the respiration of aerobic waste water bacteria were conducted according to EEC L 133, p. 118-122 (30. May 1988) guideline. The test was conducted on activated sludge from a sewage treatment plant for 3 hours at 20°C. The results were graphically determined . Activated sludge collected from the sewage treatment plant of CH-4106 Therwil on 16/10/89. The preparation was carried out according to the method described in the guideline. The sludge was separated from the aqueous layer by settling instead of centrifugation. 3,5-Dichlorophenol was used as referance substance.The inhibitory concentration (IC50) value was determined to be 15 mg/l.

Similarly in the second study, respiratory inhibition study ofactivated sludge by test material was conducted according to EC Directive 88/302/EEC - activated sludge: verificationof the inhibition of respiration;corresponds to the OECD Guidelines for Testing of Chemicals 209 and ISO Standard 8192.In the short-term respiration test, the effect of test substance on the respiration of aerobicMicroorganisms (activated sludge) after a short exposure time (30 to 180 min) were investigated.As the test result, the effective concentrations are determined, in which the breathinginhibited relative to a reference value without Priifsubstanz by 20, 50 and 80% (EC20,EC50, EC80).  Effect of the test substance on the respiration of municipal activated sludge were found asEC20: after 30 min about 40 mg / l ,EC50: - After 30 min> 1000 mg /l, respectively.