Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt

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 Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt can be expected to be > 5 days at pH 4, 7 and 9 & at a temperature of 50⁰C, respectively or > 1 years at pH 7 & at a temperature of 20⁰C, respectively. Thus, based on this half-life value, it can be concluded that the test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt is not hydrolysable.

Biodegradation 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 percentage degradation of the test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt can be expected to be range from 0 to 1.2% by using CO2 evolution & DOC removal parameter in 28 to 35 days, respectively and BOD5 can be expected to be 0 g O2/g test material. Thus, based on this, test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11 -disulfo-3,10 -triphenodioxazinediyl)bis[imino-2,1 -ethanediylimino[6 -[(2,5 -disulfophenyl)amino]-1,3,5 -triazine-4,2 -diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt is considered to be not readily biodegradable in nature.

Bioaccumulation: aquatic / sediment

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the bioaccumulation factor (BCF value) of the test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt can be expected to be range from 0.6 – 1.8 L/Kg at a conc. of 2.0 mg/l and ≤ 5.5 L/Kg at a conc. of 0.2 mg/l on the basis of total lipid content, respectively or 1.4 – 4.7 L/Kg at a conc. of 0.2 mg/l and ≤ 6.4 – 28 L/Kg at a conc. of 0.02 mg/l on the basis of whole body wieght of test organism, respectively. Thus, based on this, test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxa zinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2- diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt is considered to be non-bioaccumulative in aquatic organisms.

Adsorption / desorption

The adsorption coefficient Koc in soil and in sewage sludge of test chemical Pyridinium, 1,1'-[(6,13 -dichloro-4,11 -disulfo-3,10 -triphenodioxazinediyl)bis[imino-2,1 -ethanediylimino[6 -[(2,5 -disulfophenyl)amino]-1,3,5 -triazine-4,2 -diyl]]]bis[3 -carboxy-,dihydroxide, bis(inner salt), hexasodium salt (CAS No. 79771-28-1) was determined by the Reverse Phase High Performance Liquid Chromatographic method according to OECD Guideline No. 121 for testing of Chemicals (Experimental study report, 2018). 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 5 ml water and make with Acetonitrile up to 10 ml. Thus, the test solution concentration was 400 mg/l. The pH of test substance was 5.2. 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(Annex - 2).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 were chosen according to estimated Koc range of the test substance and generalized calibration graph was prepared. The reference substances were Acetanilide, 4 -chloroaniline, 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, Direct Red 81, Benzoic acid methylester, Carbendazim, Benzoic acid phenylester, Xylene, Ethylbenzene, Toluene, Naphthalene, 1,2,3-trichlorobenzene, Pentachlorophenol, N,N-dimethylbenzamide, 3,5 -dinitrobenzamide, Benzamide, phenanthrene having Koc value ranging from 1.25 to 4.09. The Log Koc value of test chemical Pyridinium, 1,1'-[(6,13 -dichloro-4,11 -disulfo-3,10 -triphenodioxazinediyl)bis[imino-2,1 -ethanediylimino[6 -[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt was determined to be 1.326 ± 0.003 at 25°C. This log Koc value indicates that the substance Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt has a negligible sorption to soil and sediment and therefore have 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 Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt (CAS no. 79771 -28 -1).The studies are as mentioned below:

 

The half-life of the test chemical was determined at different pH range. The study was performed according to OECD Guideline 111 (Hydrolysis as a Function of pH) at a temperature of 50°C. The average percentage recovery of the test chemical after 5 days was determined to be 99.0, 99.5 and 98.7% at pH 4, 7 and 9, respectively. As no hydrolysis of test item was observed for a period of 5 days, the half-lives was determined to be > 5 days at pH 4, 7 and 9 & at a temperature of 50⁰C, respectively. Based on the half-life values, it is concluded that the test substance is not hydrolysable.

 

In an another study, the half-life of the test chemical was determined at pH 7.0. The study was performed at a temperature of 50°C. The only functional group present is the amide group, which hydrolyzes slowly at neutral pH. The half-life value of the test chemical was determined to be > 1 years at pH 7 & at a temperature of 20⁰C, respectively. Based on the half-life value, it is concluded that test chemical is not hydrolysable.

 

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 Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt can be expected to be  > 5 days at pH 4, 7 and 9 & at a temperature of 50⁰C, respectively or  > 1 years at pH 7 & at a temperature of 20⁰C, respectively. Thus, based on this half-life value, it can be concluded that the test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt is not hydrolysable.

Biodegradation in water

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the biodegradability of the test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11- disulfo-3,10 -triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt (CAS no. 79771 -28 -1). The studies are as mentioned below:

 

Biodegradation study was carried out using river die away test for 35 days for evaluating the percentage biodegradability of test substance. Initial test substance conc. used was 1 mg/l (1000 ppb). The percentage degradation of test substance was determined to be 0% after 35 days by using CO2 evolution parameter. Thus, based on percentage degradation, test chemical is considered to be not readily biodegradable in nature.

 

Biodegradation study was conducted for 5 days for evaluating the percentage biodegradability of test substance. Bacteria was used as a test inoculum for the study. The 5 day BOD value of test chemical was determined to be 0 g O2/g test material. Thus, based on this BOD5 value, test substance is considered to be not readily biodegradable in nature.

 

Biodegradation study was conducted for 28 days for evaluating the percentage biodegradability of test substance. The study was performed according to EPA OTS 796.3180 (Ready Biodegradability: Modified AFNOR Test) and OECD Guideline 301 A (Ready Biodegradability: DOC Die Away Test), respectively, under aerobic conditions. Domestic sewage effluentwas used as a test inoculum obtained from sewage treatment plant. Initial test substance concentration used for the study was 40 mg/l based on DOC (Dissolved Organic Carbon). Modifications of the guideline involve sterilisation instead of sterile filtration and bacterial concentration of 2.6000000 per ml, respectively.The percentage degradation of test substance was determined to be 1.2% by using DOC removal parameter in 28 days. Thus, based on percentage degradation, test chemical is considered to be not readily biodegradable in nature.

 

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the percentage degradation of the test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt can be expected to be range from 0 to 1.2% by using CO2 evolution & DOC removal parameter in 28 to 35 days, respectively and BOD5 can be expected to be 0 g O2/g test material. Thus, based on this, test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5 -triazine-4,2 -diyl]]]bis[3 -carboxy-, dihydroxide, bis(inner salt), hexasodium salt is considered to be not readily biodegradable in nature.

Bioaccumulation: aquatic / sediment

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the bioaccumulation potential in aquatic organisms of the test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11- disulfo-3,10 -triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt (CAS no. 79771 -28 -1). The studies are as mentioned below:

 

Bioaccumulation study was conducted on test organism Cyprinus carpiofor 6 weeks for evaluating the bioconcentration factor (BCF value) of test substance.The study was performed according to 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. Details on analytical methods involve the recovery ratio: Test water: 1st concentration area : 97.6%, 2nd concentration area : 94.8%, Fish: 71.9%, - Limit of quantitation: Test water: 1st concentration area : 0.14 ppm, 2nd concentration area : 0.015 ppm, Fish : 1.1 ppm. Test chemical nominal conc. used for the study were 2.0 mg/land 0.2 mg/l, respectively. Range finding study involve the TLm (48 hr) ≥ 1000 ppm (w/v) on Rice fish (Oryzias latipes). Lipid content of the test organism was determined to be 5.0%.The bioconcentration factor (BCF value) of test substance on Cyprinus carpio was determined to be in the range of 0.6 – 1.8 L/Kg at a conc. of 2.0 mg/l and ≤ 5.5 L/Kg at a conc. of 0.2 mg/l, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the test chemical is not expected to bioaccumulate in the food chain.

 

In an another study, the bioaccumulation experiment was conducted on test organism Cyprinus carpiofor 6 weeks for evaluating the bioconcentration factor (BCF value) of test substance. The study was performed according to other guideline "Bioaccumulation test of a chemical substance in fish or shellfish" provided in "the Notice on the Test Method Concerning New Chemical Substances”. Cyprinus carpio was used as a test organism for the study. Details on analytical methods involve the recovery ratio: Test water: 1st concentration area : 94.6%, 2nd concentration area : 91.3%, Fish: 85.9%, - Limit of quantitation: Fish : 0.093 ppm. Test chemical nominal conc. used for the study were 0.2 mg/land 0.02 mg/l, respectively. Dimethyl sulfoxide was used as a vehicle in the study. Range finding study involve the TLm (48 hr) ≥ 50 ppm (w/v) on Rice fish (Oryzias latipes).The bioconcentration factor (BCF value) of test substance on Cyprinus carpio was determined to be in the range of 1.4 – 4.7 L/Kg at a conc. of 0.2 mg/l and ≤ 6.4 – 28 L/Kg at a conc. of 0.02 mg/l, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that test chemical is not expected to bioaccumulate in the food chain.

 

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the bioaccumulation factor (BCF value) of the test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt can be expected to be range from 0.6 – 1.8 L/Kg at a conc. of 2.0 mg/l and ≤ 5.5 L/Kg at a conc. of 0.2 mg/l on the basis of total lipid content, respectively or  1.4 – 4.7 L/Kg at a conc. of 0.2 mg/l and ≤ 6.4 – 28 L/Kg at a conc. of 0.02 mg/l on the basis of whole body wieght of test organism, respectively. Thus, based on this, test chemical Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxa zinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2- diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt is considered to be non-bioaccumulative in aquatic organisms.

Adsorption / desorption

The adsorption coefficient Koc in soil and in sewage sludge of test chemical Pyridinium, 1,1'-[(6,13 -dichloro-4,11 -disulfo-3,10 -triphenodioxazinediyl)bis[imino-2,1 -ethanediylimino[6 -[(2,5 -disulfophenyl)amino]-1,3,5 -triazine-4,2 -diyl]]]bis[3 -carboxy-,dihydroxide, bis(inner salt), hexasodium salt (CAS No. 79771-28-1) was determined by the Reverse Phase High Performance Liquid Chromatographic method according to OECD Guideline No. 121 for testing of Chemicals (Experimental study report, 2018). 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 5 ml water and make with Acetonitrile up to 10 ml. Thus, the test solution concentration was 400 mg/l. The pH of test substance was 5.2. 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(Annex - 2).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 were chosen according to estimated Koc range of the test substance and generalized calibration graph was prepared. The reference substances were Acetanilide, 4 -chloroaniline, 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, Direct Red 81, Benzoic acid methylester, Carbendazim, Benzoic acid phenylester, Xylene, Ethylbenzene, Toluene, Naphthalene, 1,2,3-trichlorobenzene, Pentachlorophenol, N,N-dimethylbenzamide, 3,5 -dinitrobenzamide, Benzamide, phenanthrene having Koc value ranging from 1.25 to 4.09. The Log Koc value of test chemical Pyridinium, 1,1'-[(6,13 -dichloro-4,11 -disulfo-3,10 -triphenodioxazinediyl)bis[imino-2,1 -ethanediylimino[6 -[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt was determined to be 1.326 ± 0.003 at 25°C. This log Koc value indicates that the substance Pyridinium, 1,1'-[(6,13-dichloro-4,11-disulfo-3,10-triphenodioxazinediyl)bis[imino-2,1-ethanediylimino[6-[(2,5-disulfophenyl)amino]-1,3,5-triazine-4,2-diyl]]]bis[3-carboxy-, dihydroxide, bis(inner salt), hexasodium salt has a negligible sorption to soil and sediment and therefore have rapid migration potential to ground water.