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

Hydrolysis:

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the hydrolysis half-life value of the test chemical  can be expected to be 48 yrs and 5 yrs at pH 7 and 8, respectively with a second order hydrolysis rate constant of 0.0046 L/mol-sec, respectively or > 20 days at pH range 7.4 to 7.8 and a temperature of 20.1 to 20.5⁰C, respectively. Thus, based on this half-life value, it can be concluded that the test chemical  is hydrolytically stable.

Biodegradation in water:

28-days biodegradation study was conducted following the EEC, L 251 Vol. 27 (19.09.84) 84/449/EEC,C.5 guideline to determine the ready biodegradability of the test chemical. The study was performed at a temperature of 22±2°C. The test system included control, test item and reference item. Bacteria was used as a test inoculum for the study which was collected from activated sludge of 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. Test chemical conc. used for the study was 10.3 and 20.3 mg/l, respectively while that of reference substance was 20 mg/l conc. Aniline was used as a reference substance for the study. Flasks (dark brown glass) of 2 lit equipped with gas inlet and magnetic stirrer was used as a test vessel for the study.The test medium was prepared according to the method described in the guideline. Blank test system was setup during the study containing only water as specified in the guideline containing 0.5 ml of the Nonylphenol 10EO5PO solution.1200 ml of the mineral solution with the inoculum were aerated for 24 hours in the test vessel. In 300 ml mineral solution 0.5 ml Nonylphenol 10EO5PO (solution of 30 mg in 100 ml bidist. water) and 15.5 rsp. 30.4 mg of test substance were added and homogenized. This solution was given to the test vessel which was immediately connected to the CO2 traps. Determination of the initial CO2 of the 0.05 N sodium hydroxide and the CO2, absorbed in the absorbers filled with 200 ml 0.05 N sodium hydroxide on the days 3, 7, 10, 13, 17, 20, 24, 27 and 28.The biodegradation was calculated on the basis of the theoretical carbon content of the test substance and the cumulative quantities of carbon dioxide determined on the days of measurements. Accordingly, the % degradation of the test item after 28 days of incubation at 22 ± 2°C according to test was determined to be 34% and 7% after 28 days at a chemical conc. of 10.3 & 20.3 mg/l, respectively. Based on the results, the test item, under the test conditions, was considered to be primary inherently biodegradable in nature.

Biodegradation in water and sediments:

Estimation Programs Interface (2018) prediction model was run to predict the half-life in water and sediment for the test chemical. If released in to the environment, 0.702% of the chemical will partition into water according to the Mackay fugacity model level III and the half-life period of test chemical in water is estimated to be 180 days (4320 hrs). The half-life (180 days estimated by EPI suite) indicates that the chemical is very persistent in water and the exposure risk to aquatic animals is moderate to high whereas the half-life period of test chemical in sediment is estimated to be 1620.833 days (38900 hrs). Based on this half-life value, it indicates that test chemical is very persistent in sediment.

Biodegradation in soil:

The half-life period of test chemical in soil was estimated using Level III Fugacity Model by EPI Suite version 4.1 estimation database (2018). If released into the environment, 38.7% of the chemical will partition into soil according to the Mackay fugacity model level III. The half-life period of test chemical in soil is estimated to be 360 days (8640 hrs). Based on this half-life value of test chemical, it is concluded that the chemical is very persistent in the soil environment and the exposure risk to soil dwelling animals is moderate to high.

Bioaccumulation: aquatic/sediments:

Using BCFBAF Program (v3.00) model of EPI suite (2018) the estimated bio concentration factor (BCF) for test chemical is 3.162 L/kg wet-wt at 25 deg. c which does not exceed the bioconcentration threshold of 2000. Therefore it is concluded that test chemical  is nonbioaccumulative in food chain.

Adsorption/desorption:

The Adsorption Coefficient of test substance was determined as per the HPLC method (OECD Guideline-121). The Log Koc value was determined to be 1.288 ± 0.001 at 25°C. Thus based on the result it is concluded that the test substance has a  negligible sorption to soil and sediment and therefore has rapid migration potential to ground water.

Additional information

Hydrolysis:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the half-life of the test chemical . The studies are as mentioned below:

The half-life and base catalyzed second order hydrolysis rate constant was determined using a structure estimation method of the test chemical. The second order hydrolysis rate constant of test chemical was determined to be 0.0046L/mol-sec with a corresponding half-lives of 48 yrs and 5 yrs at pH 7 and 8, respectively. Based on the half-life value, it is concluded that the test chemical was considered to be hydrolytically stable.

The half-life of the test chemical was determined at different pH range. The study was performedat apH range7.4 to 7.8 and a temperature of 20.1 to 20.5⁰C, respectively.The test substance was dissolved in dechlorinated tap water. The concentration of the test substance was measured in the supernatant of the test solution centrifuged. The test substance was measured on day 0, 1, 3, 6, 8, 10, 13, 15, 17, and 20 over a 21-day period. The test concentration used was considered to be at saturation in water. Based on the measured concentration of test chemical in water, chemical was reported to be hydrolytically stable at pH range7.4 to 7.8 and a temperature of 20.1 to 20.5 ⁰C for 20 days. Test chemical appears to be in equilibrium and does not appear to be breaking down in water. The concentration of test chemical is not decreasing with time. Thus, the half-life value of test chemical was determined to be > 20 days and on the basis of this, test chemical was considered to be stable in water.

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the hydrolysis half-life value of the test chemical can be expected to be 48 yrs and 5 yrs at pH 7 and 8, respectively with a second order hydrolysis rate constant of 0.0046 L/mol-sec, respectively or > 20 days at pH range 7.4 to 7.8 and a temperature of 20.1 to 20.5⁰C, respectively. Thus, based on this half-life value, it can be concluded that the test chemical is hydrolytically stable.

Biodegradation in water:

Experimental studies for the target chemical were reviewed for the biodegradation end point and their results are summarized as below:

In an experimental key study from study report (1989), 28-days biodegradation study was conducted following the EEC, L 251 Vol. 27 (19.09.84) 84/449/EEC,C.5 guideline to determine the ready biodegradability of the test chemical. The study was performed at a temperature of 22±2°C. The test system included control, test item and reference item. Bacteria was used as a test inoculum for the study which was collected from activated sludge of the sewage treatment plant of CH-4106.The preparation was carried out according to the method described in the guideline. Test chemical conc. used for the study was 10.3 and 20.3 mg/l, respectively while that of reference substance was 20 mg/l conc. Aniline was used as a reference substance for the study. Flasks (dark brown glass) of 2 lit equipped with gas inlet and magnetic stirrer was used as a test vessel for the study. The test medium was prepared according to the method described in the guideline. Blank test system was setup during the study containing only water as specified in the guideline containing 0.5 ml of the Nonylphenol 10EO5PO solution.1200 ml of the mineral solution with the inoculum were aerated for 24 hours in the test vessel. In 300 ml mineral solution 0.5 ml Nonylphenol 10EO5PO (solution of 30 mg in 100 ml bidist. water) and 15.5 rsp and 30.4 mg of test substance were added and homogenized. This solution was given to the test vessel which was immediately connected to the CO2 traps. Determination of the initial CO2 of the 0.05 N sodium hydroxide and the CO2, absorbed in the absorbers filled with 200 ml 0.05 N sodium hydroxide on the days 3, 7, 10, 13, 17, 20, 24, 27 and 28.The biodegradation was calculated on the basis of the theoretical carbon content of the test substance and the cumulative quantities of carbon dioxide determined on the days of measurements. Accordingly, the % degradation of the test item after 28 days of incubation at 22 ± 2°C according to test was determined to be 34% and 7% after 28 days at a chemical conc. of 10.3 & 20.3 mg/l, respectively. Based on the results, the test item, under the test conditions, was considered to be primary inherently biodegradable in nature.

Another biodegradation study was conducted following the EEC L251, Vol. 27 (19.09.84), 84/449/EEC, C.9 guideline to determine the ready biodegradability of the test chemical. The study was performed at a temperature of 148 ± 3°C. For the preparation of the stock solution of test substance (TK 11484 ), 9.1 mg were dissolved and filled up to 100 ml with silver sulfate in sulfuric acid. The preparation of the stock solution of the reagents was carried out according the method described in the guideline. Potassium hydrogen phthalate was used as a reference substance for the study. Round bottom flasks of 250 ml equipped with a condenser was used as a test vessel for the study. Test system include the following test solutions i.e, Potassium dichromate solution, Mercury sulfate solution ,Silver sulfate solution and Sulfuric acid d=1.84 ,respectively. Bidistilled Water was used as a blank. Titration of the unreacted potassium dichromate with ammonium iron-II-sulfate was done on a METTLER Memotitrator. Titration solution used for the study was ammonium iron-II-sulfate 0.12 mol/l. The chemical oxygen demand was calculated on the basis of the amount of test substance and consumption of potassium dichromate. COD of the reference substance was determined to be 193.7 mg COD/l (limit: 192 – 208 mg COD/l) (n = 3, Srel.= 0.00%) and COD of the test chemical salts was determined to be 1.26 g O2/g test mat. after 2 hrs in the test condition. Based on the results, the test item, under the test conditions, was considered to be biodegradable in nature.

In a supporting weight of evidence study, biodegradation experiment was conducted for 35-days following the OECD guideline 301 D for determining the ready biodegradability of the test chemical. The study was performed at a temperature of 20°C under aerobic conditions. Aerobic conditions was provided by means of mineral media which is aerated for 20 hours prior to start of the experiment. The test system included control, test chemical and reference substance. Polyseed capsule (mixed culture) was used as a test inoculum for the study. Test inoculum polyseed capsule was composed of blend of specialized microbial cultures and food grade gelatin made by International Laboratory Supply (InterLab), LTD. No pretreatment / preconditioning was given to the test inoculum as the polyseed capsule requires only one hour of stirring to activate it. 1 Polyseed capsule was added in 500 mL DI water and then stirred for 1 hour for proper mixing and functioning of inoculum & settled for 15 minutes to eliminate the bran. Decanted polyseed solution was used as mixed inoculum. Thus, concentration of test inoculum used for the study was 32 ml/l which corresponds to 10E7 to 10E8 CFU/ml. The concentration of test and reference substance (Sodium Benzoate) chosen for both the study was 4 mg/L. OECD mineral medium was used for the study. ThOD (Theoretical oxygen demand) of test and reference chemical was determined by calculation. % degradation was calculated using the values of BOD and ThOD for test chemical and reference substance. The % degradation of procedure control (reference substance) was also calculated using BOD & ThOD and was determined to be 70.48%. Degradation of Sodium Benzoate exceeds 66.26% on 7 days & 72.28 % on 14th day. The activity of the inoculum is thus verified and the test can be considered as valid.The BOD42 value of test chemical was observed to be 1.15 mgO2/mg. ThOD was calculated as 4.54 mgO2/mg. Accordingly, the % degradation of the test chemical after 42 days of incubation at 20 ± 1°C according to Closed Bottle test was determined to be 25.33%. Based on the results, the test chemical under the test conditions, was considered to be inherently biodegradable in nature.

On the basis of above results for test chemical, it can be concluded that the test chemical was considered to be inherently biodegradable in nature.

Biodegradation in water and sediments:

Estimation Programs Interface (2018) prediction model was run to predict the half-life in water and sediment for the test chemical. If released in to the environment, 0.702% of the chemical will partition into water according to the Mackay fugacity model level III and the half-life period of test chemical in water is estimated to be 180 days (4320 hrs). The half-life (180 days estimated by EPI suite) indicates that the chemical is very persistent in water and the exposure risk to aquatic animals is moderate to high whereas the half-life period of test chemical in sediment is estimated to be 1620.833 days (38900 hrs). Based on this half-life value, it indicates that test chemical is very persistent in sediment.

Biodegradation in soil:

The half-life period of test chemical in soil was estimated using Level III Fugacity Model by EPI Suite version 4.1 estimation database (2018). If released into the environment, 38.7% of the chemical will partition into soil according to the Mackay fugacity model level III. The half-life period of test chemical in soil is estimated to be 360 days (8640 hrs). Based on this half-life value of test chemical, it is concluded that the chemical is very persistent in the soil environment and the exposure risk to soil dwelling animals is moderate to high.

Bioaccumulation: aquatic/sediments:

Predicted data of test chemical and various supporting weight of evidence studies of its read across chemical were reviewed for bioaccumulation endpoint which are summarized below.

In a prediction done using the using BCFBAF Program (v3.00) model of EPI suite (2018), bio concentration factor (BCF) for test chemical was estimated to be 3.162 L/kg wet-wt at 25°C which does not exceed the bioconcentration threshold of 2000. Therefore it is concluded that test chemical  is non bioaccumulative in food chain.

In a supporting weight of evidence study from authoritative database (J-check),bioaccumulation study was conducted on test organism Cyprinus carpio for 28 days for evaluating the bioconcentration factor (BCF value) of test chemical.The study was performed according to OECD Guideline 305 (Bioconcentration: Flow-through Fish Test) and other guideline"Bioaccumulation test of a chemical substance in fish or shellfish" provided in "the Notice on the Test Method Concerning New Chemical Substances", respectively. Cyprinus carpio was used as a test organism for the study. Test chemical nominal conc. used for the study were 0.46 mg/l and 0.046 mg/l, respectively. Analytical method involve the recovery ratio: Test water : 100 % (The test solution were input into analytical equipment directly.) Fish : 93.8 % - Limit of quantitation : Test water : 1st concentration area : 11 microg/L 2nd concentration area : 1.1 microg/L Fish: 120 ng/g. Range finding study involve the LC50(96h) > 200 mg/Lon Rice fish (Oryzias latipes). Lipid content of the test organism Cyprinus carpio was determined to be 2.94% and 4.14% at the start and end of the exposure. The bioconcentration factor (BCF value) of test chemical on Cyprinus carpio was determined to be≤ 0.27 L/Kg at a conc. of 0.46 mg/l and ≤ 2.7 L/Kg at a conc. of 0.046 mg/l, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the chemical is not expected to bioaccumulate in the food chain.

Another Bioaccumulation test was conducted for estimating the bioconcentration factor (BCF) of test chemical (from authoritative database, 2017). The bioconcentration factor was determined to be 75 dimesionless, from an estimated log Pow of -0.67 and a regression derived equation. This suggests that the potential for bioconcentration of test substance in aquatic organisms is low.

For the test chemical, bioaccumulation experiment was conducted on test organism Cyprinus carpio for 28 days for evaluating the bioconcentration factor (BCF value) of test chemical.The study was performed according to OECD Guideline 305 (Bioconcentration: Flow-through Fish Test) and other guideline"Bioaccumulation test of a chemical substance in fish or shellfish" provided in "the Notice on the Test Method Concerning New Chemical Substances", respectively. Cyprinus carpio was used as a test organism for the study. Test chemical nominal conc. used for the study were1.69 mg/land 0.169 mg/l, respectively. Analytical method involve the recovery ratio:Fish : 89.6 % - Limit of quantitation : Test water : 1st concentration area : 83 microg/L 2nd concentration area : 8.3 microg/L Fish: 940 ng/g.Range finding study involve the LC50(96h) > 200 mg/LonRice fish (Oryzias latipes). Lipid content of the test organism Cyprinus carpio was determined to be 5.09% and 6% at the start and end of the exposure. The bioconcentration factor (BCF value) of test substance on Cyprinus carpio was determined to be≤ 0.57 L/Kg at a conc. of 1.69 mg/l and ≤ 5.3 L/Kg at a conc. of 0.169 mg/l, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the chemical is not expected to bioaccumulate in the food chain.

By considering results of all the studies mentioned above, it waas concluded that test chemical is non-bioaccumulative in aquatic organisms

Adsorption/desorption:

The adsorption coefficient Koc in soil and in sewage sludge of test chemical was determined by the Reverse Phase High Performance Liquid Chromatographic method according to OECD Guideline No. 121 for testing of Chemicals. The solutions of the test substance and reference substances were prepared in appropriate solvents. A test item solution was prepared by accurately weighing 4 mg of test item and diluted with methanol up to 10 ml. Thus, the test solution concentration was 400 mg/l. The pH of test substance was 6.7. Each of the reference substance and test substance were analysed by HPLC at 210 nm. After equilibration of the HPLC system, Urea was injected first, the reference substances were injected in duplicate, followed by the test chemical solution in duplicate. Reference substances were injected again after test sample, no change in retention time of reference substances was observed. Retention time tR were measured, averaged and the decimal logarithms of the capacity factors k were calculated. The graph was plotted between log Koc versus log k. The linear regression parameter of the relationship log Koc vs log k were also calculated from the data obtained with calibration samples and therewith, log Koc of the test substance was determined from its measured capacity factor. The reference substances Acetanilide, 4-methylaniline(p-Tolouidine), N-methylaniline, p-toluamide, Aniline, 2,5- Dichloroaniline, 4-nitrophenol, 2 - nitrophenol, 2-nitrobenzamide, 3-nitrobenzamide, Nitrobenzene, 4- Nitrobenzamide, 1-naphtol, Benzoic acid methylester, Carbendazim, Benzoic acid phenylester, Xylene, Ethylbenzene, Toluene, Naphthalene, 1,2,3-trichlorobenzene, Phenol, N,N-dimethylbenzamide, 3,5- dinitrobenzamide, N-methylbenzamide, Benzamide, 1-naphthylamine, were chosen having Koc value range from 1.25 to 3.51. The Log Koc value of test chemical was determined to be 1.288 ± 0.001 dimensionless at 25°C.This log Koc value indicates that the substance has a negligible sorption to soil and sediment and therefore have rapid migration potential to ground water.