2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]-6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

(Q)SAR

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 was done using the OECD QSAR toolbox version 3.3.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name: 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]-6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt
- Molecular Formula: C42H29N7O13S4.xMg.xNa
- Molecular Weight: 1012.2624 g/mole
- SMILES:[Mg]_[Na]_Oc1c2ccc(Nc3ccccc3)cc2cc(S(O)(=O)=O)c1N=Nc1ccc(N=Nc2ccc(N=Nc3cc(S(O)(=O)=O)ccc3S(O)(=O)=O)c3ccc(S(O)(=O)=O)cc23)c2ccccc12
- InChI: 1S/C42H29N7O13S4.Mg.Na/c50-42-29-13-10-26(43-25-6-2-1-3-7-25)20-24(29)21-40(66(60,61)62)41(42)49-47-35-16-15-34(30-8-4-5-9-31(30)35)44-46-37-18-17-36(32-14-11-27(22-33(32)37)63(51,52) 53)45-48-38-23-28(64(54,55)56)12-19-39(38)65(57,58)59;;/h1-23,43,50H,(H,51,52,53)(H,54,55,56)(H,57,58,59)(H,60,61,62);;/q;+2;+1/p-3/b46-44+,48-45+,49-47+;;

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Test type:

static

Water media type:

freshwater

Total exposure duration:

72 h

Test temperature:

22 - 23 C

pH:

8.0 - 8.1

Nominal and measured concentrations:

Estimated data

Reference substance (positive control):

not specified

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

863.556 mg/L

Nominal / measured:

estimated

Conc. based on:

not specified

Basis for effect:

growth rate

Remarks on result:

other: Other details not known

Details on results:

The EC50 was 863.5 mg/l for 72 hrs of exposure.

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" )  and ("e" and ( not "f") )  )  and ("g" and ( not "h") )  )  and ("i" and ( not "j") )  )  and "k" )  and "l" )  and ("m" and ( not "n") )  )  and ("o" and ( not "p") )  )  and ("q" and ( not "r") )  )  and ("s" and ( not "t") )  )  and ("u" and ( not "v") )  )  and ("w" and ( not "x") )  )  and ("y" and ( not "z") )  )  and ("aa" and ( not "ab") )  )  and "ac" )  and "ad" )  and ("ae" and "af" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Amine OR Aromatic compound OR Azo compound OR Hydroxy compound OR Phenol OR Secondary amine OR Secondary aromatic amine OR Sulfonic acid OR Sulfonic acid derivative by Organic functional groups, Norbert Haider (checkmol) ONLY

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Alcohol, olefinic attach [-OH] OR Aliphatic Nitrogen, one aromatic attach [-N] OR Aliphatic Nitrogen, two aromatic attach [-N-] OR Aromatic Carbon [C] OR Azo [-N=N-] OR Hydroxy, aromatic attach [-OH] OR Hydroxy, sulfur attach [-OH] OR Magnesium [Mg] OR Miscellaneous sulfide (=S) or oxide (=O) OR Nitrogen, two or tree olefinic attach [>N-] OR No functional group found OR Olefinic carbon [=CH- or =C<] OR Oxygen, one aromatic attach [-O-] OR Suflur {v+4} or {v+6} OR Sulfinic acid [-S(=O)OH] OR Sulfonate, aromatic attach [-SO2-O] by Organic functional groups (US EPA) ONLY

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as Aromatic amine OR Aryl OR Azo OR Fused carbocyclic aromatic OR No functional group found OR Overlapping groups OR Phenol OR Sulfonic acid by Organic Functional groups (nested) ONLY

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as Aromatic amine OR Aryl OR Azo OR Fused carbocyclic aromatic OR Naphtalene OR Phenol OR Sulfonic acid by Organic Functional groups ONLY

Domain logical expression index: "e"

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

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as AN2 OR AN2 >> Carbamoylation after isocyanate formation OR AN2 >> Carbamoylation after isocyanate formation >> N-Hydroxylamines OR Non-covalent interaction OR Non-covalent interaction >> DNA intercalation OR Non-covalent interaction >> DNA intercalation >> Fused-Ring Primary Aromatic Amines OR Radical OR Radical >> Radical mechanism via ROS formation (indirect) OR Radical >> Radical mechanism via ROS formation (indirect) >> Fused-Ring Primary Aromatic Amines OR Radical >> Radical mechanism via ROS formation (indirect) >> N-Hydroxylamines OR SN1 OR SN1 >> Alkylation after metabolically formed carbenium ion species OR SN1 >> Alkylation after metabolically formed carbenium ion species >> Polycyclic Aromatic Hydrocarbon Derivatives OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation >> Fused-Ring Primary Aromatic Amines OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation >> N-Hydroxylamines OR SN2 OR SN2 >> Alkylation, direct acting epoxides and related after P450-mediated metabolic activation OR SN2 >> Alkylation, direct acting epoxides and related after P450-mediated metabolic activation >> Polycyclic Aromatic Hydrocarbon Derivatives by DNA binding by OASIS v.1.3

Domain logical expression index: "g"

Referential boundary: The target chemical should be classified as No alert found by DNA binding by OECD

Domain logical expression index: "h"

Referential boundary: The target chemical should be classified as Michael addition OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals >> Alkyl phenols OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals >> Arenes OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals >> Hydroquinones OR SN1 OR SN1 >> Nitrenium Ion formation OR SN1 >> Nitrenium Ion formation >> Primary aromatic amine by DNA binding by OECD

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Non binder, MW>500 AND Non binder, non cyclic structure by Estrogen Receptor Binding

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as Moderate binder, OH grooup OR Non binder, impaired OH or NH2 group OR Non binder, without OH or NH2 group OR Very strong binder, OH group OR Weak binder, OH group by Estrogen Receptor Binding

Domain logical expression index: "k"

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: "l"

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

Domain logical expression index: "m"

Referential boundary: The target chemical should be classified as Acid moiety AND Inorganic Compound AND Phenols AND Salt AND SHOULD NOT BE PROFILED by Aquatic toxicity classification by ECOSAR

Domain logical expression index: "n"

Referential boundary: The target chemical should be classified as Not classified OR Not Related to an Existing ECOSAR Class by Aquatic toxicity classification by ECOSAR

Domain logical expression index: "o"

Referential boundary: The target chemical should be classified as Aromatic alcohol  [-OH] AND Aromatic amine   [-NH2  or  -NH-] AND Aromatic-H AND Azo group   [-N=N-] AND Benzene AND Naphthalene AND No alert found AND Number of fused 6-carbon aromatic rings AND Sulfonic acid / salt -> aromatic attach AND Unsubstituted phenyl group (C6H5-) by Bioaccumulation - metabolism alerts

Domain logical expression index: "p"

Referential boundary: The target chemical should be classified as -CH-   [linear] OR -CH2-  [linear] OR Fluorine  [-F] OR Linear C4 terminal chain  [CCC-CH3] OR Methyl  [-CH3] OR Quaternary amine OR Tin  [Sn] by Bioaccumulation - metabolism alerts

Domain logical expression index: "q"

Referential boundary: The target chemical should be classified as Alkali Earth AND Alkaline Earth AND Non-Metals by Groups of elements

Domain logical expression index: "r"

Referential boundary: The target chemical should be classified as Halogens by Groups of elements

Domain logical expression index: "s"

Referential boundary: The target chemical should be classified as Group 1 - Alkali Earth Li,Na,K,Rb,Cs,Fr AND Group 14 - Carbon C AND Group 15 - Nitrogen N AND Group 16 - Oxygen O AND Group 16 - Sulfur S AND Group 2 - Alkaline Earth Be,Mg,Ca,Sr,Ba,Ra by Chemical elements

Domain logical expression index: "t"

Referential boundary: The target chemical should be classified as Group  3 - Lanthanoids by Chemical elements

Domain logical expression index: "u"

Referential boundary: The target chemical should be classified as Group 1 - Alkali Earth Li,Na,K,Rb,Cs,Fr AND Group 14 - Carbon C AND Group 15 - Nitrogen N AND Group 16 - Oxygen O AND Group 16 - Sulfur S AND Group 2 - Alkaline Earth Be,Mg,Ca,Sr,Ba,Ra by Chemical elements

Domain logical expression index: "v"

Referential boundary: The target chemical should be classified as Group 8 - Trans.Metals Fe,Ru,Os by Chemical elements

Domain logical expression index: "w"

Referential boundary: The target chemical should be classified as Group 1 - Alkali Earth Li,Na,K,Rb,Cs,Fr AND Group 14 - Carbon C AND Group 15 - Nitrogen N AND Group 16 - Oxygen O AND Group 16 - Sulfur S AND Group 2 - Alkaline Earth Be,Mg,Ca,Sr,Ba,Ra by Chemical elements

Domain logical expression index: "x"

Referential boundary: The target chemical should be classified as Group 4 - Trans.Metals Ti,Zr,Hf by Chemical elements

Domain logical expression index: "y"

Referential boundary: The target chemical should be classified as Group 1 - Alkali Earth Li,Na,K,Rb,Cs,Fr AND Group 14 - Carbon C AND Group 15 - Nitrogen N AND Group 16 - Oxygen O AND Group 16 - Sulfur S AND Group 2 - Alkaline Earth Be,Mg,Ca,Sr,Ba,Ra by Chemical elements

Domain logical expression index: "z"

Referential boundary: The target chemical should be classified as Group 15 - Phosphorus P by Chemical elements

Domain logical expression index: "aa"

Referential boundary: The target chemical should be classified as Group 1 - Alkali Earth Li,Na,K,Rb,Cs,Fr AND Group 14 - Carbon C AND Group 15 - Nitrogen N AND Group 16 - Oxygen O AND Group 16 - Sulfur S AND Group 2 - Alkaline Earth Be,Mg,Ca,Sr,Ba,Ra by Chemical elements

Domain logical expression index: "ab"

Referential boundary: The target chemical should be classified as Group 12 - Trans.Metals Zn,Cd,Hg by Chemical elements

Domain logical expression index: "ac"

Referential boundary: The target chemical should be classified as Alcohol, olefinic attach [-OH] AND Aliphatic Nitrogen, one aromatic attach [-N] AND Aliphatic Nitrogen, two aromatic attach [-N-] AND Aromatic Carbon [C] AND Azo [-N=N-] AND Hydroxy, aromatic attach [-OH] AND Hydroxy, sulfur attach [-OH] AND Magnesium [Mg] AND Miscellaneous sulfide (=S) or oxide (=O) AND Nitrogen, two or tree olefinic attach [>N-] AND No functional group found AND Olefinic carbon [=CH- or =C<] AND Oxygen, one aromatic attach [-O-] AND Suflur {v+4} or {v+6} AND Sulfinic acid [-S(=O)OH] AND Sulfonate, aromatic attach [-SO2-O] by Organic functional groups (US EPA) ONLY

Domain logical expression index: "ad"

Referential boundary: The target chemical should be classified as Alcohol, olefinic attach [-OH] AND Aliphatic Nitrogen, one aromatic attach [-N] AND Aliphatic Nitrogen, two aromatic attach [-N-] AND Aromatic Carbon [C] AND Azo [-N=N-] AND Hydroxy, aromatic attach [-OH] AND Hydroxy, sulfur attach [-OH] AND Magnesium [Mg] AND Miscellaneous sulfide (=S) or oxide (=O) AND Nitrogen, two or tree olefinic attach [>N-] AND No functional group found AND Olefinic carbon [=CH- or =C<] AND Oxygen, one aromatic attach [-O-] AND Suflur {v+4} or {v+6} AND Sulfinic acid [-S(=O)OH] AND Sulfonate, aromatic attach [-SO2-O] by Organic functional groups (US EPA) ONLY

Domain logical expression index: "ae"

Parametric boundary:The target chemical should have a value of log Kow which is >= 1.93

Domain logical expression index: "af"

Parametric boundary:The target chemical should have a value of log Kow which is <= 2.58

Validity criteria fulfilled:

not specified

Conclusions:

Based on the growth rate inhibition of test organism the EC50 value was estimated to be 863.5 mg/l when 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl] azo]-6-sulpho-1-naphthyl]azo]benzene-1,4- disulphonic acid, magnesium sodium salt exposed to Pseudokirchneriella subcapitata for 72 hrs.   

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 Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum) was predicted for 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]- 6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt. Based on the growth rate inhibition of test organism the EC50 value was estimated to be 863.5 mg/l when 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl] azo]-6-sulpho- 1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt exposed to Pseudokirchneriella subcapitata for 72 hrs.   

    

Based on this value it can be concluded that the substance 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]-6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt is considered to be nontoxic to aquatic environment and cannot 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 Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum) was predicted for 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]- 6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt. Based on the growth rate inhibition of test organism the EC50 value was estimated to be 863.5 mg/l when 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl] azo]-6-sulpho- 1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt exposed to Pseudokirchneriella subcapitata for 72 hrs. Based on this value it can be concluded that the substance 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]-6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt is considered to be nontoxic to aquatic environment and cannot classified as per the criteria mentioned in CLP regulation. 

Key value for chemical safety assessment

EC50 for freshwater algae:

863.5 mg/L

Additional information

Based on the various prediction data for the target chemical and experimental data for the structurally and functionally similar read across chemical study have been reviewed to determine the toxic nature of 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]-6-sulpho-1-naphthyl]azo] benzene-1,4-disulphonic acid, magnesium sodium salt (93941-74-3) on the growth of aquatic algae and cyanobacteria. The studies are as mentioned below:  

In the first predicted study for the target chemical 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]-6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt (93941-74-3) from QSAR toolbox 2017, study was carried out. 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 Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum) was predicted for 2-[[4-[[4- [[6-anilino- 1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]- 6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt. Based on the growth rate inhibition of test organism the EC50 value was estimated to be 863.5 mg/l when 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1 -naphthyl] azo]-6-sulpho- 1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt exposed to Pseudokirchneriella subcapitata for 72 hrs. Based on this value it can be concluded that the substance 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1 -naphthyl]azo]- 6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt is considered to be nontoxic to aquatic environment and cannot classified as per the criteria mentioned in CLP regulation. 

 

First predicted study was supported by the other experimental study for the read across chemical Brilliant black 1 (2519-30-4) from ABITEC lab report 2016. Short term toxicity study of Tetrasodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate (Brilliant black 1) to aquatic algae Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) was conducted for 72 hrs. Test was performed according to the 201 OECD guideline in a static system. The stock solution 100 mg/l was prepared by dissolving black powder in OECD growth medium. Test solutions of required concentration were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture and tested at the 0, 12, 20, 35, 60, 100 mg/l nominal concentrations. Effects on the growth rate of the organism were studied. Potassium dichromate (K2Cr2O7) were used as a reference postive control. Effects on growth rate were observed for 72 hours by using nonlinear regression by the software Prism 4.0. Based on the growth rate inhibition of test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) due to the chemical Tetrasodium 1-acetamido-2-hydroxy- 3-(4-((4-sulphonatophenylazo)- 7-sulphonato -1-naphthylazo))naphthalene -4,6-disulphonate (Brilliant black 1), the effect concentration ErC50 was 76.8 mg/l. This value indicates that the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 3 as per the CLP criteria. But as the chemical was readily biodegradable in water so on that basis chemical was consider as nontoxic and can be consider to be not classified as per the CLP classification criteria.

 

Similarly in the third weight of evidence study for the read across chemical disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalenesulphonate (3567-69-9) from ABITEC report 2016, toxicity study was carried out. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance Disodium 4-hydroxy- 3-[(4-sulphonatonaphthyl)azo] naphthalenesulphonate according to OECD Guideline 201. The test substance was dissolved in OECD growth medium and tested at the concentrations 0, 2.6, 5.6, 12.0, 27.0, 59.0 and 130 mg/L. Effects on the growth rate of the organism were studied. The median effective concentration (ErC50) for the test substance, Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalene sulphonate, in Desmodesmus subspicatus was determined to be 265.1 mg/L. Based on this ErC50 value and after comparing with CLP criteria for aquatic classification of the substance it is concluded that the substance Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl) azo] naphthalenesulphonate does not exhibit toxicity to aquatic algae (Desmodesmus subspicatus).

 

By considering the another read across chemical (15790-07-5) from UERL lab 2016 which was similar to the target chemical as supports the non-toxic classification of chemical study are as follows. 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-naphthalenesulfonic 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.

 

Thus based on the predicted data for the target chemical and experimental studies for the read across chemical obtain from various ABITEC lab reports 2016 and UERL report, it was concluded that the chemical 2-[[4-[[4-[[6-anilino-1-hydroxy-3-sulpho-2-naphthyl]azo]-1-naphthyl]azo]-6-sulpho-1-naphthyl]azo]benzene-1,4-disulphonic acid, magnesium sodium salt (93941-74-3) was consider as nontoxic and can be consider to be not classified as toxic to aquatic invertebrates as per CLP classification criteria.