4-[[5-[[4-chloro-6-[(3-sulphophenyl)amino]-1,3,5-triazin-2-yl]amino]-2-sulphophenyl]azo]-4,5-dihydro-5-oxo-1-(4-sulphophenyl)-1H-pyrazole-3-carboxylic acid

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification

Justification for type of information:

QMRF report has been attached.

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Principles of method if other than guideline:

Prediction is done using QSAR Toolbox version 3.3

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (IUPAC name): 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2-sulfophenyl)diazen-1-yl]-5-oxo-1-(4-sulfophenyl)- 4,5-dihydro -1H-pyrazole-3-carboxylic acid
- Molecular formula: C25H18ClN9O12S3
- Molecular weight: 768.1192 g/mol
- Smiles notation: C1=CC(=CC(=C1)S(=O)(=O)O)NC2=NC(=NC(=N2)NC3=CC(=C(C=C3)S(=O)(=O)O)N=NC4C(=NN(C4=O)C5=CC=C(C=C5)S(=O)(=O)O)C(=O)O)Cl
- InChl: 1S/C25H18ClN9O12S3/c26-23-29-24(27-12-2-1-3-16(10-12)49(42,43)44)31-25(30-23)28-13-4-9-18(50(45,46)47)17(11-13)32-33-19-20(22(37)38)34-35(21(19)36)14-5-7-15(8-6-14)48(39,40)41/h1-11,19H,(H,37,38)(H,39,40,41)(H,42,43,44)(H,45,46,47)(H2,27,28,29,30,31)/b33-32+
- Substance type: Organic
- Physical state: Solid

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Test type:

static

Water media type:

freshwater

Total exposure duration:

72 h

Test temperature:

20.5 - 21°C

pH:

7.6

Nominal and measured concentrations:

Estimated data

Reference substance (positive control):

not specified

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

192.794 mg/L

Nominal / measured:

estimated

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other: Nontoxic

Details on results:

The EC50 was estimated to be 192.79 mg/l

The prediction was based on dataset comprised from the following descriptors: EC50
Estimation method: Takes average value from the 5 nearest neighbours
Domain  logical expression:Result: In Domain

((((((((("a" or "b" or "c" or "d" or "e" )  and "f" )  and "g" )  and "h" )  and "i" )  and "j" )  and "k" )  and "l" )  and ("m" and "n" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Substituted Triazines (Acute toxicity) by US-EPA New Chemical Categories

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Acid moiety OR Amides OR Hydrazines OR Triazines, Aromatic by Aquatic toxicity classification by ECOSAR ONLY

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as Acylation OR Acylation >> Direct Acylation Involving a Leaving group OR Acylation >> Direct Acylation Involving a Leaving group >> Acetates OR SN2 OR SN2 >> SN2 reaction at sp3 carbon atom OR SN2 >> SN2 reaction at sp3 carbon atom >> Alkyl diazo OR SNAr OR SNAr >> Nucleophilic aromatic substitution OR SNAr >> Nucleophilic aromatic substitution >> Halo-triazines by Protein binding by OECD ONLY

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as Schiff base formation OR Schiff base formation >> Pyrazolones and Pyrazolidinones derivatives OR Schiff base formation >> Pyrazolones and Pyrazolidinones derivatives >> Pyrazolones and Pyrazolidinones  OR SNAr OR SNAr >> Nucleophilic aromatic substitution on activated aryl and heteroaryl compounds OR SNAr >> Nucleophilic aromatic substitution on activated aryl and heteroaryl compounds >> Activated aryl and heteroaryl compounds by Protein binding by OASIS v1.3 ONLY

Domain logical expression index: "e"

Referential boundary: The target chemical should be classified as SN1 OR SN1 >> Nitrenium Ion formation OR SN1 >> Nitrenium Ion formation >> Unsaturated heterocyclic azo by DNA binding by OECD ONLY

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as No alert found by DNA binding by OASIS v.1.3 ONLY

Domain logical expression index: "g"

Referential boundary: The target chemical should be classified as Class 5 (Not possible to classify according to these rules) by Acute aquatic toxicity classification by Verhaar (Modified) ONLY

Domain logical expression index: "h"

Referential boundary: The target chemical should be classified as Reactive unspecified by Acute aquatic toxicity MOA by OASIS ONLY

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Not bioavailable by Lipinski Rule Oasis ONLY

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as Aromatic amine AND Aromatic heterocyclic halide AND Aryl AND Aryl halide AND Azo AND Carboxylic acid AND Pyrazolone AND Sulfonic acid AND Triazine AND Unsaturated heterocyclic amine AND Unsaturated heterocyclic fragment by Organic Functional groups ONLY

Domain logical expression index: "k"

Referential boundary: The target chemical should be classified as Aromatic amine AND Aromatic heterocyclic halide AND Aryl AND Aryl halide AND Azo AND Carboxylic acid AND Overlapping groups AND Pyrazolone AND Sulfonic acid by Organic Functional groups (nested) ONLY

Domain logical expression index: "l"

Referential boundary: The target chemical should be classified as Alcohol, olefinic attach [-OH] AND Aliphatic Carbon [CH] AND Aliphatic Nitrogen, one aromatic attach [-N] AND Aliphatic Nitrogen, two aromatic attach [-N-] AND Amide, aliphatic attach [-C(=O)N] AND Amino Triazine/Pyrazine/Pyrimidine  AND Amino, aliphatic attach [-N<] AND Amino-carbonyl compound [NCC(=O)-C] AND Aromatic Carbon [C] AND Aromatic Nitrogen AND Azo [-N=N-] AND Azomethine, aliphatic attach [-N=C] AND Carbonyl, aliphatic attach [-C(=O)-] AND Carbonyl, olefinic attach [-C(=O)-] AND Chlorine, aromatic attach [-Cl] AND Chlorine, olefinic attach [-Cl] AND Hydrazine [>N-N<] AND Hydroxy, sulfur attach [-OH] AND Miscellaneous sulfide (=S) or oxide (=O) AND Nitrogen, two or tree olefinic attach [>N-] AND Olefinic carbon [=CH- or =C<] AND Suflur {v+4} or {v+6} AND Sulfinic acid [-S(=O)OH] AND Sulfonate, aromatic attach [-SO2-O] AND Sym-Triazine ring  by Organic functional groups (US EPA) ONLY

Domain logical expression index: "m"

Parametric boundary:The target chemical should have a value of log Kow which is >= 0.646

Domain logical expression index: "n"

Parametric boundary:The target chemical should have a value of log Kow which is <= 4.33

Validity criteria fulfilled:

not specified

Conclusions:

Based on the growth rate inhibition of Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) due to tested chemical, the EC50 was estimated to be 192.79 mg/l

Executive summary:

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) predicted for 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2- ylidene] amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858-24-5).The EC50 value was estimated to be192.7 mg/l when 4-[(E)-2-(5-{[(2Z,4E) -6-chloro-4-[(3-sulfophenyl) imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acidexposed toDesmodesmus subspicatus for 72 hrs. Based on this value it can be concluded that the substance 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2 -ylidene]amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858-24-5) is considered to be not toxic to aquatic environment and not classified as per the criteria mentioned in CLP regulation.     

Description of key information

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) predicted for 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2- ylidene] amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858-24-5).The EC50 value was estimated to be192.7 mg/l when 4-[(E)-2-(5-{[(2Z,4E) -6-chloro-4-[(3-sulfophenyl) imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acidexposed toDesmodesmus subspicatus for 72 hrs. Based on this value it can be concluded that the substance 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2 -ylidene]amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858-24-5) is considered to be not toxic to aquatic environment and not classified as per the criteria mentioned in CLP regulation.     

Key value for chemical safety assessment

EC50 for freshwater algae:

1 927 mg/L

Additional information

Various predicted data for the target compound 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2-sulfophenyl)diazen-1-yl]-5-oxo-1- (4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858-24-5) and supporting weight of evidence studies for its closest read across substance with log Kow as the primary descriptor were reviewed for the toxicity on the algae end point which are summarized as below:  

 

In a prediction done by SSS (2018), using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) predicted for 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2- ylidene]amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858-24-5).The EC50 value was estimated to be 192.7 mg/l when 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl) imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3- carboxylic acid exposed to Desmodesmus subspicatus for 72 hrs. Based on this value it can be concluded that the substance 4-[(E)- 2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]- 1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2-sulfophenyl)diazen -1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858-24-5) is considered to be not toxic to aquatic environment and not classified as per the criteria mentioned in CLP regulation.     

 

In second prediction using the prediction done by EPI suite ECOSAR version 1.10, the short term toxicity on green algae was predicted for test substance  4-[(E)-2-(5-{[(2Z,4E)-6-chloro- 4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2-sulfophenyl)diazen-1-yl]-5-oxo-1 -(4-sulfophenyl)- 4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858 -25 -4) . On the basis of effects observed in a static freshwater system, the effect concentration EC50 value for the substance is estimated to be 1269.3 mg/l for green algae for 96 duration. Based on this value, it can be concluded that the test chemical 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl) imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2- sulfophenyl)diazen -1-yl]-5-oxo-1 -(4-sulfophenyl)- 4,5-dihydro-1H-pyrazole -3-carboxylic acid can be considered as non-toxic to green algae at environmentally relevant concentrations and can be considered not-classified as per the CLP classification criteria.

 

Similarly in the third weight of evidence study for the read across chemical (70616-89-6) from ABITEC report was carried out. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance trisodium 2-({6-[(4-amino-6-chloro-1,3,5-triazin-2-yl)(methyl) amino]-1-hydroxy- 3-sulfonato-2-naphthyl}diazenyl)naphthalene -1,5-disulfonate according to OECD Guideline 201. The stock solution 100 mg/l was prepared by dissolving redbrown powder in OECD growth medium and tested at the limit concentration i.e 100 mg/l. Effects on the growth rate of the organism were studied after the exposure period of 72hrs with the positive reference control Potassium dichromate (K2Cr2O7). Test performed in the six replicates. The inhibitory concentration for the test substance, trisodium 2-({6-[(4-amino-6-chloro-1,3,5-triazin-2-yl)(methyl) amino]-1-hydroxy-3- sulfonato-2-naphthyl}diazenyl) naphthalene-1,5- disulfonate, in Desmodesmus subspicatus was determined. At 100 mg/l inhibition was not observed in any replicates (0% inhibition was observed). This value indicates that the substance is likely to be non-hazardous to aquatic algae and can be consider as not classified as per the CLP criteria.

 

 

Similarly in the third weight of evidence study for the read across chemical (15790-07-5) from ABITEC study was carried out. The effect of test item aluminium, 6-hydroxy-5- [(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex (15790-07-5) was studied on the growth of fresh water green alga Chlorella 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, 200mg/L. The test solution was prepared in aseptic condition. The test item aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalenesulfonic acid complex was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200 mg/L. This stock solution was kept for stirring/sonication 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 concentrations chosen were set up to the water solubility limit. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment. Test was performed in static manner at proper requirement of pH and temperature. The microscopic observations were noted down in each of the control vessel. All the cells appeared healthy, round and green throughout the study duration in the control. Also, the drift in pH in the control vessels did not increase by >1.5 units when observed on 72 hours as compared to 0 hours. The average pH drift observed in the control vessels was 0 units. After 72 hours of exposure to aluminium, 6-hydroxy-5-[(4-sulfophenyl)azo]-2-naphthalene sulfonic acid complex (15790-07-5) to various nominal test concentration, EC50 calculated from equation through probit analysis was determine to be > 200 mg/l. Based on the EC50, it can be concluded that the chemical was nontoxic and can be consider to be not classified as toxic as per the CLP classification criteria.

 

On the basis of above results for target chemical 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino}-2- sulfophenyl)diazen-1-yl]-5- oxo-1- (4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid (93858-24-5) (from OECD QSAR toolbox version 3.3, and EPIsuite, 2017) and for its read across substance from ABITEC report, it can be concluded that the test substance 4-[(E)-2-(5-{[(2Z,4E)-6-chloro-4-[(3-sulfophenyl)imino]-1,2,3,4-tetrahydro-1,3,5-triazin-2-ylidene]amino} -2-sulfophenyl)diazen- 1-yl]-5-oxo-1- (4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid is not toxic and can be consider to be not classified as toxic as per the CLP classification criteria.