Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-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

Remarks:

of read across substance

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

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on two short term toxicity study of fish for the test chemical :
1.This study was designed to assess the effect of the test item on the growth of green alga Chlorella vulgaris. The study was conducted in accordance with “OECD guideline for testing of chemicals No. 201 – Alga, growth inhibition test”.

2.To evaluate efficiency of toxicity of test chemical on Raphidocelis subcapitata

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of the test chemical: Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate
- Molecular formula: C30H27ClN8O17S5.4Na
- Molecular weight: 1055.29 g/mol
- Substance type: Organic
- Smiles: [Na+].[Na+].[Na+].[Na+].c1c(c(c(c2c(cc(cc12)S(=O)(=O)[O-])NC(CC)=O)O)\N=N\c1c(ccc(c1)Nc1nc(nc(n1)Cl)Nc1cc(ccc1)S(=O)(=O)CCOS(=O)(=O)[O-])S(=O)(=O)[O-])S(=O)(=O)[O-]
- Inchi: 1S/C30H27ClN8O17S5.4Na/c1-2-24(40)34-21-14-19(58(44,45)46)10-15-11-23(60(50,51)52)26(27(41)25(15)21)39-38-20-13-17(6-7-22(20)59(47,48)49)33-30-36-28(31)35-29(37-30)32-16-4-3-5-18(12-16)57(42,43)9-8-56-61(53,54)55;;;;/h3-7,10-14,41H,2,8-9H2,1H3,(H,34,40)(H,44,45,46)(H,47,48,49)(H,50,51,52)(H,53,54,55)(H2,32,33,35,36,37);;;;/q;4*+1/p-4/b39-38+;;;;

Analytical monitoring:

yes

Vehicle:

not specified

Test organisms (species):

other: 1.Chlorella vulgaris , 2.Raphidocelis subcapitata

Details on test organisms:

1.TEST ORGANISM
- Common name:green alga
- Strain:Chlorella vulgaris
- Source (laboratory, culture collection):laboratory
- Method of cultivation:BBM
- Culturing media and conditions (same as test or not):The medium to be used for the growth of algae was Bold’s Basal Medium (BBM). It is a medium composed of macronutrients, micronutrients, alkaline EDTA solution and Iron solution. Stock solution of each of these was prepared separately and then a complete medium was prepared and sterilized. De-ionized water was used to prepare the BBM
- Any deformed or abnormal cells observed:no

2.- Common name: Raphidocelis subcapitata
- Method of cultivation: Algae were routinely cultured in Cyanobacteria BG-11 Freshwater Solution

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Test temperature:

1) 24 °C ±2°C
2)25±1 °C

pH:

1) 6.8 ±0.3

Nominal and measured concentrations:

1)Six test concentration were:
6.25mg/l, 12.5mg/l, 25mg/l, 50mg/l, 100mg/l and 200mg/l

2) 100 mg/l

Details on test conditions:

1.- No. of vessels per concentration (replicates):Two replicates for each test concentration
- No. of vessels per control (replicates):Three replicates for Control
-Test solution: The test solution was prepared in aseptic condition. The test item was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200mg/L. This stock solution was kept for stirring for 24 hours 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.
6.2. Test vessels: All the tests were carried out in 100mL conical flasks which were carefully autoclaved and sterilized. The test solution in each of these test vessels was kept constant which is 60ml so that a sufficient amount of head space was left.
6.3. Replicates: For the assessment of algal growth, the study was conducted in replicates. The control vessel was maintained in triplicates as recommended in the OECD guideline and the test concentrations were selected in geometric series which were maintained in duplicates.
Incubation:
i) The temperature of the orbital shaking incubator was kept constant throughout the period of exposure of the experiment. The temperature was maintained at 24 ° C±2°C.
ii) The test vessels were incubated with a continuous, uniform (1500Lux).
iii) The pH of the control cultures needs to be noted during the study and the pH of the control medium should not increase by more than 1.5 units during the test.
iv) The orbital shaking incubator was set at a speed of 120 revolutions per minute throughout the study period. This is to provide constant shaking to the algal cells to keep them in suspension and to ensure that they do not settle down on the bottom of the test vessel.

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Photoperiod:16 Hour Light Period : 8 Hour Dark Period
- Light intensity and quality:White Fluorescent Light

2.-Algae were exposed to the treatment in sterile 24-well plates with 2 mL of treatment solution to the microplate well, for three days.

- Algal stock solution containing 1.01E6 cells/mL was added (20 μL) to each well to obtain a starting algal density of 1E4 cells/mL.
Plates were incubated for 72 h at 25±1 °C.
on an orbital shaker (90 rpm) in a thermostatic chamber.

- Light intensity and quality: provided by a 2 W LED Unit for each plate.

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

2.074 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: Read across 1

Duration:

72 h

Dose descriptor:

other: ErC50

Effect conc.:

1.12 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: Read across 2

Results with reference substance (positive control):

2.potassium dichromate 0.10– 1.8 mg/L

Validity criteria fulfilled:

not specified

Conclusions:

The test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphony l]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]- 4-hydrox y-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate is likely to be toxic to aquatic algae and cynobacteria in the concentration range of 1.120-2.074 mg/l

Executive summary:

Data available for the functionally similar read across chemicals has been reviewed to determine the toxicity of aquatic algae and cyanobacteria of the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphony l]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]- 4-hydrox y-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate (80019 -42 -7).The studies are as mentioned below:

1.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 6.25mg/l, 12.5mg/l, 25mg/l, 50mg/l, 100mg/l and 200mg/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 2.079mg/l.

2.The 72 h algal growth inhibition test with Raphidocelis subcapitata was done according to OECD 201 (2011) for the test chemical .

Algae were routinely cultured at 25±1 °C with light in Cyanobacteria BG-11 Freshwater Solution

The culture was split twice a week, adding fresh medium. Algae were exposed to the treatment in sterile 24-well plates with 2 mL of treatment solution to the respective microplate well, for three days.

The effect concentration of test material on green algae on Raphidocelis subcapitata for growth rate after 72 h was observed to be 1.20 mg/l.

Thus, based on the above summarised studies,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[ [2-(sulphonatooxy)ethyl]sulphony l]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]- 4-hydrox y-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate and its functionally similar read across substance, it can be concluded that effect concetration value is in the range 1.120 -2.074 mg/L. Thus, comparing this value with the criteria of CLP regulation, can be classified as "Aquatic chronic-2".Hence,based on the data available for the functionally similar read across, test chemical is likely to be toxic in the concentration range of 1.120 -2.074 mg/L .

Description of key information

Toxicity to aquatic algae and cyanobacteria:

Data available for the functionally similar read across chemicals has been reviewed to determine the short term toxicity to aquatic algae and cyanobacteriaof the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphony l]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]- 4-hydrox y-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate (80019 -42 -7).The studies are as mentioned below:

1.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 6.25mg/l, 12.5mg/l, 25mg/l, 50mg/l, 100mg/l and 200mg/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 2.079mg/l.

2.The 72 h algal growth inhibition test with Raphidocelis subcapitata was done according to OECD 201 (2011) for the test chemical .

Algae were routinely cultured at 25±1 °C with light in Cyanobacteria BG-11 Freshwater Solution

The culture was split twice a week, adding fresh medium. Algae were exposed to the treatment in sterile 24-well plates with 2 mL of treatment solution to the respective microplate well, for three days.

The effect concentration of test material on green algae on Raphidocelis subcapitata for growth rate after 72 h was observed to be 1.20 mg/l.

Thus, based on the above summarised studies,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[ [2-(sulphonatooxy)ethyl]sulphony l]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]- 4-hydrox y-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate and its functionally similar read across substance, it can be concluded that effect concetration value is in the range 1.120 -2.074 mg/L. Thus, comparing this value with the criteria of CLP regulation, can be classified as "Acute aquatic-2 ".Hence,based on the data available for the functionally similar read across, test chemical is likely to be toxic in the concentration range of 1.120 -2.074 mg/L .

Key value for chemical safety assessment

EC50 for freshwater algae:

2.074 mg/L

Additional information

Toxicity to aquatic algae and cyanobacteria:

Data available for the functionally similar read across chemicals has been reviewed to determine the toxicity of aquatic algae and cyanobacteria of the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphony l]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]- 4-hydrox y-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate (80019 -42 -7).The studies are as mentioned below:

1.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 6.25mg/l, 12.5mg/l, 25mg/l, 50mg/l, 100mg/l and 200mg/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 2.079mg/l.

2.The 72 h algal growth inhibition test with Raphidocelis subcapitata was done according to OECD 201 (2011) for the test chemical .

Algae were routinely cultured at 25±1 °C with light in Cyanobacteria BG-11 Freshwater Solution

The culture was split twice a week, adding fresh medium. Algae were exposed to the treatment in sterile 24-well plates with 2 mL of treatment solution to the respective microplate well, for three days.

The effect concentration of test material on green algae on Raphidocelis subcapitata for growth rate after 72 h was observed to be 1.20 mg/l.

Thus, based on the above summarised studies,Tetrasodium 3-[[5-[[4-chloro-6-[[3-[ [2-(sulphonatooxy)ethyl]sulphony l]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]- 4-hydrox y-5-[(1-oxopropyl)amino]naphthalene-2,7-disulphonate and its functionally similar read across substance, it can be concluded that effect concetration value is in the range 1.120 -2.074 mg/L. Thus, comparing this value with the criteria of CLP regulation, can be classified as "Aquatic chronic-2 ".Hence,based on the data available for the functionally similar read across, test chemical is likely to be toxic in the concentration range of 1.120 -2.074 mg/L .