Tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonato-1-naphthyl]azo]-7-sulphonato-1-naphthyl]azo]benzene-1,4-disulphonate

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on three toxicity study of aquatic algae and cyanobacteria for the test chemical :1.Estimation of 96 hr EC50 value of test chemical by modelling Database - ECOSAR v.1.11. The ECOSAR Class Program is a computerized version of the ECOSAR analysis procedures as currently practiced by the Office of Pollution Prevention and Toxics (OPPT).  It has been developed within the regulatory constraints of the Toxic Substances Control Act (TSCA).  It is a pragmatic approach to SAR. The structure-activity relationships (SARs) presented in this program are used to predict the aquatic toxicity of chemicals based on their similarity of structure to chemicals for which the aquatic toxicity has been previously measured.  Most SAR calculations in the ECOSAR Class Program are based upon the octanol/water partition coefficient (Kow).  Various surfactant SAR calculations are based upon the average length of carbon chains or the number of ethoxylate units.2.To evaluate toxicity to algae by test chemical3.To evaluate toxicity of test material to algae Raphidocelis subcapitata

Specific details on test material used for the study:

- Name of test material (IUPAC name): tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonato-1-naphthyl]azo]-7-sulphonato-1-naphthyl]azo]benzene-1,4-disulphonate- Molecular formula: C29H20ClN9O12S4.4Na- Molecular weight:938.173 g/mol- Smiles notation: [Na+].[Na+].[Na+].[Na+].c1(c(cc(cc1)S(=O)(=O)[O-])\N=N\c1ccc(c2ccc(cc12)S(=O)(=O)[O-])\N=N\c1ccc(c2c(cccc12)S(=O)(=O)[O-])Nc1nc(nc(n1)N)Cl)S(=O)(=O)[O-]- InChl: 1S/C29H20ClN9O12S4.4Na/c30-27-33-28(31)35-29(34-27)32-22-10-9-20(17-2-1-3-25(26(17)22)55(49,50)51)37-36-19-7-8-21(18-12-14(52(40,41)42)4-6-16(18)19)38-39-23-13-15(53(43,44)45)5-11-24(23)54(46,47)48;;;;/h1-13H,(H,40,41,42)(H,43,44,45)(H,46,47,48)(H,49,50,51)(H3,31,32,33,34,35);;;;/q;4*+1/p-4/b37-36+,39-38+;;;;- Substance type: Organic- Physical State: Solid

Details on test solutions:

2.The test solution was prepared in aseptic condition. The test item was prepared by adding 21 mg of test item in 200ml of BBM to get the final concentration of 105mg/L. This stock solution was kept for stirring 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 remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment.

Test organisms (species):

other: 2.Chlorella vulgaris 3.Pseudokirchneriella subcapitata

Details on test organisms:

2.The fresh water green alga Chlorella vulgaris, was used as the test system (organism). Sterile, unicellular, suspension cultures of algae were obtained from National Environmental Engineering Research Institute (NEERI), and maintained at UERL. The culture was examined under the microscope to confirm that it was unicellular, healthy and not contaminated. 3.TEST ORGANISM- Common name: Raphidocelis subcapitata- Method of cultivation: Algae were routinely cultured at25±1 °C with light in Cyanobacteria BG-11 Freshwater Solution

Test type:

static

Water media type:

freshwater

Total exposure duration:

96 h

Test temperature:

2) 24 °C ±2°C3) 25±1 °C

Nominal and measured concentrations:

2) 0.3 mg/L, 0.96 mg/L, 3.072 mg/L, 9.830mg/L, 31.457 mg/L and 100.663mg/L (Nominal)

Details on test conditions:

2.Study TypeStatic Exposure Duration72 HoursTemperature24 °C ±2°C.Light QualityWhite Fluorescent LightLight Intensity1500LuxPhotoperiod16 Hour Light Period : 8 Hour Dark PeriodRPM Speed120 Revolutions per minuteWater Deionized WaterTest vessel size:100mlTest volume60mlDilution water:Bold’s Basal MediumReplicates per concentration-Three replicates for Control Two replicates for each test concentration3.TEST ORGANISM- Common name: Raphidocelis subcapitata- Method of cultivation: Algae were routinely cultured at25±1 °C with light in Cyanobacteria BG-11 Freshwater Solution

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

49.06

Nominal / measured:

estimated

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: Neutral Organics SAR (Baseline Toxicity)

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

24.34 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

cell number

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

67.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Validity criteria fulfilled:

not specified

Conclusions:

The test chemical tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonato-1-naphthyl]azo]-7-sulphonato-1-naphthyl]azo]benzene-1,4-disulphonate is likely to be toxic to aquatic algae and cyanobacteria in the concentration range of 49 - 67 mg/l

Executive summary:

Data available for the structurally similar read across chemicals and predicted study has been reviewed to determine the toxicity of aquatic algae and cyanobacteria of the test chemical tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino] -5-sulphonato-1-naphthyl]azo] -7-sulphonato-1-naphthyl]azo] benzene-1,4-disulphonate (70161 -16 -9).The studies are as mentioned below:

1.The short-term toxicity of the test substancetetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl) amino]-5-sulphonato-1-naphthyl]azo]-7- sulphonato -1-naphthyl]azo]benzene-1,4-disulphonate (CAS no.70161 -16 -9) to green algae is predicted using EPI Suite ECOSAR version 1.10. On the basis of effects observed in a static freshwater system during a 96 hr exposure, the effect concentration (EC50) for the substance is estimated to be 49.0 mg/L. Based on this value, it can be concluded that the test chemical  can be considered as toxicto green algae at environmentally relevant concentrations and can be considered to be classified in aquatic acute category 3 as per the CLP classification criteria.

 

2.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 0.3 mg/L, 0.96 mg/L, 3.072 mg/L, 9.830 mg/L, 31.457 mg/L and 100.663 mg/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 24.342 mg/L.

3.The 72 h algal growth inhibition test with Raphidocelis subcapitata was done according to OECD 201. Algae were routinely cultured at 25±1 °C with light in Cyanobacteria BG-11 Freshwater Solution. Algae were exposed to the treatment in sterile 24-well plates with 2 mL of test material.

 The effect concentration (EC50) for the test material against Raphidocelis subcapitata was observed to be 67.2 mg/l.

Thus, based on the above summarised studies, and it’s structurally similar read across substance, it can be concluded that effect concetration value is in the range of 49 - 67 mg/L. Thus, comparing this value with the criteria of CLP regulation, tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonato-1-naphthyl]azo]-7-sulphonato-1-naphthyl]azo]benzene-1,4-disulphonate can be classified as aquatic acute 3 for toxicity for aquatic algae and cyanobacteria.

Description of key information

Toxicity to aquatic algae and cyanobacteria:

Data available for the structurally similar read across chemicals and predicted study has been reviewed to determine the toxicity of aquatic algae and cyanobacteria of the test chemical tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino] -5-sulphonato-1-naphthyl]azo] -7-sulphonato-1-naphthyl]azo] benzene-1,4-disulphonate (70161 -16 -9).The studies are as mentioned below:

1.The short-term toxicity of the test substancetetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl) amino]-5-sulphonato-1-naphthyl]azo]-7- sulphonato -1-naphthyl]azo]benzene-1,4-disulphonate (CAS no.70161 -16 -9) to green algae is predicted using EPI Suite ECOSAR version 1.10. On the basis of effects observed in a static freshwater system during a 96 hr exposure, the effect concentration (EC50) for the substance is estimated to be 49.0 mg/L. Based on this value, it can be concluded that the test chemical  can be considered as toxicto green algae at environmentally relevant concentrations and can be considered to be classified in aquatic acute category 3 as per the CLP classification criteria.

 

2.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 0.3 mg/L, 0.96 mg/L, 3.072 mg/L, 9.830 mg/L, 31.457 mg/L and 100.663 mg/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 24.342 mg/L.

3.The 72 h algal growth inhibition test with Raphidocelis subcapitata was done according to OECD 201. Algae were routinely cultured at 25±1 °C with light in Cyanobacteria BG-11 Freshwater Solution. Algae were exposed to the treatment in sterile 24-well plates with 2 mL of test material.

 The effect concentration (EC50) for the test material against Raphidocelis subcapitata was observed to be 67.2 mg/l.

Thus, based on the above summarised studies, and it’s structurally similar read across substance, it can be concluded that effect concetration value is in the range of 49 - 67 mg/L. Thus, comparing this value with the criteria of CLP regulation, tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonato-1-naphthyl]azo]-7-sulphonato-1-naphthyl]azo]benzene-1,4-disulphonate can be classified as aquatic acute 3 for toxicity for aquatic algae and cyanobacteria.

Key value for chemical safety assessment

EC50 for freshwater algae:

49 mg/L

Additional information

Toxicity to aquatic algae and cyanobacteria:

Data available for the structurally similar read across chemicals and predicted study has been reviewed to determine the toxicity of aquatic algae and cyanobacteria of the test chemical tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino] -5-sulphonato-1-naphthyl]azo] -7-sulphonato-1-naphthyl]azo] benzene-1,4-disulphonate (70161 -16 -9).The studies are as mentioned below:

1.The short-term toxicity of the test substancetetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl) amino]-5-sulphonato-1-naphthyl]azo]-7- sulphonato -1-naphthyl]azo]benzene-1,4-disulphonate (CAS no.70161 -16 -9) to green algae is predicted using EPI Suite ECOSAR version 1.10. On the basis of effects observed in a static freshwater system during a 96 hr exposure, the effect concentration (EC50) for the substance is estimated to be 49.0 mg/L. Based on this value, it can be concluded that the test chemical  can be considered as toxicto green algae at environmentally relevant concentrations and can be considered to be classified in aquatic acute category 3 as per the CLP classification criteria.

 

2.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 0.3 mg/L, 0.96 mg/L, 3.072 mg/L, 9.830 mg/L, 31.457 mg/L and 100.663 mg/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 24.342 mg/L.

3.The 72 h algal growth inhibition test with Raphidocelis subcapitata was done according to OECD 201. Algae were routinely cultured at 25±1 °C with light in Cyanobacteria BG-11 Freshwater Solution. Algae were exposed to the treatment in sterile 24-well plates with 2 mL of test material.

 The effect concentration (EC50) for the test material against Raphidocelis subcapitata was observed to be 67.2 mg/l.

Thus, based on the above summarised studies, and it’s structurally similar read across substance, it can be concluded that effect concetration value is in the range of 49 - 67 mg/L. Thus, comparing this value with the criteria of CLP regulation, tetrasodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonato-1-naphthyl]azo]-7-sulphonato-1-naphthyl]azo]benzene-1,4-disulphonate can be classified as aquatic acute 3 for toxicity for aquatic algae and cyanobacteria.

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