2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl-3H-indolium chloride

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reveiwed journal

Justification for type of information:

Data is from peer reveiwed journal

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

Study was conducted to determine the degradation of chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride by the methanogenic bacteria.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (IUPAC name): 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride
- Common name: Astrazonrot 6B
- Molecular formula: C24H30ClN2Cl
- Molecular weight: 417.421 g/mol
- Smiles notation: C1(=[N+](c2ccccc2C1(C)C)C)\C=C/ c1c(cc(N(CCCl)CC)cc1)C.[ClH-]
- InChl: 1S/C24H30ClN2.ClH/c1-6-27(16-15-25)20-13-11-19(18(2)17-20)12-14-23-24(3,4)21-9-7-8-10-22(21)26(23)5;/h7-14,17H,6,15-16H2,1-5H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid
- Source of chemical: Comodepur S.p.a., Como Italy

Oxygen conditions:

anaerobic

Inoculum or test system:

other: Anaerobic granular sludge

Details on inoculum:

- Laboratory culture: UASB reactor

Duration of test (contact time):

70 d

Initial conc.:

0 mg/L

Based on:

test mat.

Initial conc.:

300 mg/L

Based on:

test mat.

Initial conc.:

600 mg/L

Based on:

test mat.

Initial conc.:

1 200 mg/L

Based on:

test mat.

Initial conc.:

2 400 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

other: % degradation

Details on study design:

- Composition of medium: The basal medium M9 was used with laboratory grade yeast extract (1 g/l), trypticase (1 g/l), sodium acetate (2 g/l); a microelements solution.
- Additional substrate: 0.5 ml of whole granules of the sludge concentrated dye solutions to obtain final dye concentrations

TEST SYSTEM
- Culturing apparatus: Serum vials (120 ml total volume) with butyl rubber Stoppers
- Number of culture flasks/ concentration: four replicate vials

Reference substance:

not specified

Preliminary study:

No data available

Test performance:

No data available

Key result

Parameter:

% degradation (test mat. analysis)

Value:

0

Sampling time:

72 h

Remarks on result:

other: No degradation observed

Details on results:

As the chemical shows high toxicity on the methanogenic bacteria (anaerobic granular sludge) and kill 100% bacterial growth at all the concentrations, thus no biodegradation was observed by the bacteria.

Results with reference substance:

No data available

Fig: Biodegradation result

Dyes initial concentrations (mg/l)	% of chemical in supernatant	Growth of organisms
300	100	0
600	100	0
1200	100	0
2400	100	0

Validity criteria fulfilled:

not specified

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

As the chemical shows high toxicity on the methanogenic bacteria (anaerobic granular sludge) and kill 100% bacterial growth at all the concentrations, thus no biodegradation was observed by the bacteria. Based on the result, 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride was not biodegradable.

Executive summary:

Study was conducted to determine the degradation of chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride by the methanogenic bacteria. Anaerobic granular sludge was used in the test.

 

Serum vials (120 ml total volume) with butyl rubber stoppers were used to test the biodegradability of the dyes. Anaerobic granular sludge sampled from a UASB reactor fed with soft-drink wastewater added as active biomass. The basal medium M9 was used with laboratory grade yeast extract (1 g/l), trypticase (1 g/l), sodium acetate (2 g/l); a microelements solution. All modification was carried out in an anaerobic chamber. The autoclaved medium was aliquoted in the vials (50 ml/each). Test conducted at various concentrations 0, 300, 600, 1200 and 2400 mg/l. The gas phase in the vials was replaced by pure O,-free sterile nitrogen. For each dye and each concentration four replicate vials were

Prepared. Methane and hydrogen production were monitored in each vial every 24 h for 3 or 4 days.

 

As the chemical shows high toxicity on the methanogenic bacteria (anaerobic granular sludge) and kill 100% bacterial growth at all the concentrations after 72hrs, thus no biodegradation was observed by the bacteria. Based on the result, 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride was not biodegradable.

Description of key information

Study was conducted to determine the degradation of chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride by the methanogenic bacteria. Anaerobic granular sludge was used in the test. Serum vials (120 ml total volume) with butyl rubber stoppers were used to test the biodegradability of the dyes. Anaerobic granular sludge sampled from a UASB reactor fed with soft-drink wastewater was used as active biomass. The basal medium M9 was used with laboratory grade yeast extract (1 g/l), trypticase (1 g/l), sodium acetate (2 g/l); a microelements solution. All modification was carried out in an anaerobic chamber. The autoclaved medium was aliquoted in the vials (50 ml/each). Test conducted at various concentrations 0, 300, 600, 1200 and 2400 mg/l. The gas phase in the vials was replaced by pure O,-free sterile nitrogen. For each dye and each concentration four replicate vials were prepared. Methane and hydrogen production were monitored in each vial every 24 h for 3 or 4 days. As the chemical shows high toxicity on the methanogenic bacteria (anaerobic granular sludge) and kill 100% bacterial growth at all the concentrations after 72hrs, thus no biodegradation was observed by the bacteria. Based on the result, 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride was not biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed

Additional information

Various predicted and experimental data for the target compound 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl-3H-indolium chloride (6441 -82 -3) and supporting weight of evidence studies for its closest read across substance with logKow as the primary descriptor were reviewed for the biodegradation end point which are summarized as below:

In a first experimental report for the 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl-3H-indolium chloride done by Bioresource Technology, 1998 biodegradation study was conducted. Study was conducted to determine the degradation of chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride by the methanogenic bacteria. Anaerobic granular sludge was used in the test. Serum vials (120 ml total volume) with butyl rubber stoppers were used to test the biodegradability of the dyes. Anaerobic granular sludge sampled from a UASB reactor fed with soft-drink wastewater was used as active biomass. The basal medium M9 was used with laboratory grade yeast extract (1 g/l), trypticase (1 g/l), sodium acetate (2 g/l); a microelements solution. All modification was carried out in an anaerobic chamber. The autoclaved medium was aliquoted in the vials (50 ml/each). Test conducted at various concentrations 0, 300, 600, 1200 and 2400 mg/l. The gas phase in the vials was replaced by pure O,-free sterile nitrogen. For each dye and each concentration four replicate vials were prepared. Methane and hydrogen production were monitored in each vial every 24 h for 3 or 4 days. As the chemical shows high toxicity on the methanogenic bacteria (anaerobic granular sludge) and kill 100% bacterial growth at all the concentrations after 72hrs, thus no biodegradation was observed by the bacteria. Based on the result, 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride was not biodegradable.

 

In the second supporting study using Estimation Programs Interface Suite (EPI suite, 2017) was run to predict the biodegradation potential of the test compound  2-[2-[4-[(2-chloroethyl) ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl- 3H-indolium chloride (CAS no.6441 -82 -3) in the presence of mixed populations of environmental microorganisms. The biodegradability of the substance was calculated using seven different models such as Linear Model, Non-Linear Model, Ultimate Biodegradation Timeframe, Primary Biodegradation Timeframe, MITI Linear Model, MITI Non-Linear Model and Anaerobic Model (called as Biowin 1-7, respectively) of the BIOWIN v4.10 software. The results indicate that the test chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl] -1,3,3-trimethyl-3H-indolium chloride is expected to be not readily biodegradable.

 

In the third supporting study for the target chemical, study was done (SSS) using Biodegradability of 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl-3H-indolium chloride (CAS no.6441 -82 -3) is predicted using OECD QSAR toolbox version 3.3 with logKow as the primary descriptor. Test substance undergoes 0.58000 % degradation by BOD in 28 days. Thus, based on percentage degradation, the test chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl-3H-indolium chloride was estimated to be not readily biodegradable in water.

 

Similarly another biodegradation study for read across chemical (127-90-2) which was similar to the target chemical (from authoritative database J-CHECK, 2017), was conducted. 14 days of biodegradation study was conducted for evaluating the percentage biodegradability of test substance 1,1,1,2-tetrachloro-3-(2,3,3,3-tetrachloropropoxy)propane. Concentration of inoculum i.e, sludge used was 30 mg/l and initial test substance conc. used in the study was 100 mg/l, respectively. The percentage degradation of test substance was determined to 0 and 6.8 % by BOD and GC parameter in 14 days. Thus, based on percentage degradation, 1,1,1,2-tetrachloro- 3-(2,3,3,3-tetrachloropropoxy)propane is considered to be not readily biodegradable in water.

 

Similarly another biodegradation study for read across chemical (2939-80-2) which was similar to the target chemical (from authoritative database J-CHECK, 2017), was studied. Biodegradation study was conducted for evaluating the percentage biodegradability of test substance (3aS,7aR)-2-(1,1,2,2-tetrachloroethylsulfanyl)-3a,4,7,7a-tetrahydroisoindole-1,3-dione. Concentration of inoculum i.e, sludge used was 30 mg/l and initial test substance conc. used in the study was 100 mg/l, respectively. The percentage degradation of test substance was determined to 0 and 36 % by BOD and GC parameter in 28 days. Thus, based on percentage degradation, (3aS,7aR)-2- (1,1,2,2-tetrachloroethylsulfanyl)-3a,4,7,7a-tetrahydroisoindole-1,3-dione is considered to be not readily biodegradable in water.

 

On the basis of above results for target chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl-3H-indolium chloride (6441 -82 -3) (from peer reviewed journal, OECD QSAR toolbox version 3.3 and EPI suite, 2017) and for its read across substance peer reviewed journal and authoritative from database J-CHECK, it can be concluded that the test substance 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl-3H-indolium chloride can be expected to be not readily biodegradable in nature.