3,7-bis(diethylamino)phenoxazin-5-ium tetrachlorozincate (2:1)

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

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:

Data is from OECD QSAR toolbox version 3.3 and the QMRF report has been attached.

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation 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 of test material (IUPAC name): 3,7-bis(diethylamino)phenoxazin-5-ium tetrachlorozincate (2:1)
- Molecular formula: C40H52Cl4N6O2Zn
- Molecular weight: 856.093 g/mol
- Smiles notation: [o+]1c2cc(ccc2nc2ccc(N(CC)CC)cc12)N(CC)CC.[Zn+2].[ClH-].[ClH-].[ClH-].[ClH-].[o+]1c2cc(ccc2nc2c1cc(cc2)N(CC)CC)N(CC)CC
- InChl: 1S/2C20H26N3O.4ClH.Zn/c2*1-5-22(6-2)15-9-11-17-19(13-15)24-20-14-16(23(7-3)8-4)10-12-18(20)21-17;;;;;/h2*9-14H,5-8H2,1-4H3;4*1H;/q2*+1;;;;;+2/p-4
- Substance type: Organic
- Physical state:

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

- Common name: Water flea

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Hardness:

250 mg/L as CaCO3

Test temperature:

20°C to 21°C

pH:

8.38 and 8.40

Dissolved oxygen:

7.7 - 8.1 mg/l

Nominal and measured concentrations:

Estimated

Reference substance (positive control):

not specified

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.163 mg/L

Nominal / measured:

estimated

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Chemical was toxic to daphnia magna

Details on results:

The EC50 was 0.1632 mg/l after 48hrs 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" )  and ("d" and ( not "e") )  )  and ("f" and ( not "g") )  )  and "h" )  and ("i" and ( not "j") )  )  and ("k" and "l" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Zinc metal and salts by OECD HPV Chemical Categories

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Soluble complexes of Zinc by US-EPA New Chemical Categories

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as SN1 AND SN1 >> Nitrenium Ion formation AND SN1 >> Nitrenium Ion formation >> Tertiary aromatic amine by DNA binding by OECD

Domain logical expression index: "d"

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

Domain logical expression index: "e"

Referential boundary: The target chemical should be classified as Non binder, non cyclic structure OR Non binder, without OH or NH2 group OR Strong binder, OH group by Estrogen Receptor Binding

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as Halogens AND Non-Metals AND Transition Metals by Groups of elements

Domain logical expression index: "g"

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

Domain logical expression index: "h"

Similarity boundary:Target: CCN(CC)c1ccc2c(c1)o{+}(.[Zn]{2-}(Cl)(Cl)(Cl)(Cl).o{+}1c3cc(N(CC)CC)ccc3nc3ccc(N(CC)CC)cc13)c1cc(N(CC)CC)ccc1n2
Threshold=20%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as (!Undefined)Group All Lipid Solubility < 0.01 g/kg AND Group All log Kow > 9 AND Group All Melting Point > 200 C by Skin irritation/corrosion Exclusion rules by BfR

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as (!Undefined)Group CNS Surface Tension > 62 mN/m by Skin irritation/corrosion Exclusion rules by BfR

Domain logical expression index: "k"

Parametric boundary:The target chemical should have a value of log Kow which is >= 6.12

Domain logical expression index: "l"

Parametric boundary:The target chemical should have a value of log Kow which is <= 11.3

Validity criteria fulfilled:

not specified

Conclusions:

Based on the immobilisation of daphnia magna due to the exposure of chemical 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1), the EC50 was 0.1632 mg/l for 48hrs of exposure.

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 daphnia magna was predicted for 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) (63589-47-9). The EC50 value was estimated to be 0.1632 mg/l when 3,7-bis(diethylamino)phenoxazin-5- iumtetrachloro- zincate (2:1) exposed to daphnia magna for 48hrs. Thus, based on this value, it can be concluded that the test chemical 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) can be considered as toxic to aquatic organisms and thus can be classified as aquatic acute category 1 as well as chronic 1 as per the criteria mentioned in CLP.

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 daphnia magna was predicted for 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) (63589-47-9). The EC50 value was estimated to be 0.1632 mg/l when 3,7-bis(diethylamino)phenoxazin-5- iumtetrachloro- zincate (2:1) exposed to daphnia magna for 48hrs. Thus, based on this value, it can be concluded that the test chemical 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) can be considered as toxic to aquatic organisms and thus can be classified as aquatic acute category 1 as well as chronic 1 as per the criteria mentioned in CLP.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.163 mg/L

Additional information

Based on the various experimental data and prediction data for the target chemical as well as RA chemical which was selected on the basis of structure similarity study have been reviewed to determine the toxic nature of 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) (63589-47-9) on the mobility of daphnia magna. The studies are as mentioned below: 

In the first predicted weight of evidence report for the 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) (63589-47-9) from SSS (QSAR, 2017) toxicity 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 daphnia magna was predicted for 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) (63589-47-9). The EC50 value was estimated to be 0.1632 mg/l when 3,7-bis(diethylamino)phenoxazin-5- iumtetrachloro- zincate (2:1) exposed to daphnia magna for 48hrs. Thus, based on this value, it can be concluded that the test chemical 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) can be considered as toxic to aquatic organisms and thus can be classified as aquatic acute category 1 as well as chronic 1 as per the criteria mentioned in CLP.

Similarly in the second prediction study was performed based on the prediction done by EPI suite, ECOSAR version 1.1, on the basis of similarity of structure to chemicals for which the aquatic toxicity has been previously measured by structure-activity relationships (SARs) program, the LC 50 value for short term toxicity to aquatic invertebrates was predicted. On the basis of this programe, the LC 50 value for short term toxicity to aquatic invertebrates was predicted to be 0.222 mg/l for 3,7-bis(diethylamino)phenoxazin-5-ium tetrachlorozincate (2:1) in 48 hrs. Based on this value it can be concluded that the substance 3,7-bis(diethylamino)phenoxazin-5-ium tetrachlorozincate (2:1) is considered to be toxic to aquatic environment and can be classified as

aquatic acute category 1 as well as chronic 1 as per the criteria mentioned in CLP.

 

Similarly in the third weight of evidence study for the read across (548-62-9) (from Intox Pvt. Ltd. 2010) toxicity was conducted on daphnia magna to determine the toxic nature of chemical. Acute Immobilisation test in Daphnia magna was performed in compliance with the OECD Guidelines for Testing of Chemicals, Section 2, No. 202, "Daphnia Sp., Acute Immobilisation Test" . N-(4-{bis[4-(dimethylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride was tested for acute toxic effects on the swimming capabilities of Daphnia magna employing a static test procedure. In the main study, six groups of daphnids, each comprising 5 daphnids per replicate, were exposed to the test article at concentration levels of 0.025 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.4 mg and 0.8 mg/L. Observations were made for immobilisation at 24 hours and 48 hours of treatment. A concurrent blank control was also run simultaneously. Potassium Dichromate was employed as a positive control and tested at the concentration of   0.4 mg, 0.63 mg and 1 mg/L. The animals were observed for immobility after 24 hours and 48 hours of exposure. Those daphnids which were unable to swim within 15 seconds after gentle agitation of the test container were considered to be immobile. The rates of immobilisation observed at 24 hours were 0%, 0%, 0%, 10%, 25% and 75% at the test concentrations of 0.025 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.4 mg and 0.8 mg/L respectively. The rates of immobilisation observed at 48 hours were 0%, 0%, 10%, 25%, 55% and 85% at the test concentrations of 0.025 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.4 mg and 0.8 mg/L respectively. The Positive control, Potassium Dichromate induced immobilisation in 20%, 50% and 80% of the exposed daphnids at 24 hours and 25%, 65% and 100% immobilization at 48 hours at the concentrations of 0.4 mg, 0.63 mg and 1 mg/L respectively. No immobilisation was observed in blank control group. The acute immobilisation EC50value of N-(4-{bis[4-(dimethylamino) - phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride to Daphnia magna at 24 hour exposure period was determine to be 0.53 mg/L with fiducial limits between 0.41 to 0.69 mg/L at 95% confidence level. The acute immobilisation EC50 value of N-(4-{bis[4-(dimethylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride to Daphnia magna at 48 hour exposure period was determine to be 0.35 mg/L with fiducial limits between 0.24 to 0.50 mg/L at 95% confidence level. On the basis of EC50 value obtained it can be concluded that the substance N-(4-{bis[4-(dimethylamino) phenyl]methylene} cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride is toxic to the aquatic environment and can be considered as "Aquatic Acute 1" and chronic 1 as per CLP regulation.

 

Similarly in the fourth weight of evidence study for the read across (569 -64 -2) from j-check, 2017. Short term toxicity to aquatic invertebrate was performed in daphnia magna for 48 hrs. After the experiment, the EC 50 value for N-(4-{[4-(dimethylamino)phenyl](phenyl)methylene}cyclohexa -2,5-dien-1-ylidene)-N-methylmethanaminium chloride was determined to be 0.13 mg/l. Thus, based on the EC50 value, it can be concluded that the substance N-(4 -{[4 -(dimethylamino)phenyl](phenyl)methylene}cyclohexa-2,5 -dien-1 -ylidene)-N- methylmethanaminium chloride can be considered as toxic to aquatic organisms and can be consider to be classified as aquatic acute category 1 as well as chronic 1 as per the criteria mentioned in CLP.

Thus based on the above data sources for target chemical 3,7-bis(diethylamino)phenoxazin-5-iumtetrachloro- zincate (2:1) (63589-47-9) from various experimental studies and predicted studies from QSAR, EPIsuite, Intox 2010, it was summarized that the chemical was toxic to aquatic organisms and thus can be classified as aquatic acute category 1 as well as chronic 1 as per the criteria mentioned in CLP.