3',6'-bis(diethylamino)spiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one

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:

data from handbook or collection of data

Justification for type of information:

Data is from authoritative database J-CHECK.

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

Biodegradation study was conducted for 28 days for evaluating the percentage biodegradability of test substance C.I. Solvent Red 49.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in study report): C.I. Solvent Red 49
- Molecular formula : C28H30N2O3
- Molecular weight : 442.556 g/mol
- Smiles notation : C12(c3c(Oc4c1ccc(c4)N(CC)CC)cc(N(CC)CC)cc3)c1c(cccc1)C(O2)=O
- InChI: 1S/C28H30N2O3/c1-5-29(6-2)19-13-15-23-25(17-19)32-26-18-20(30(7-3)8-4)14-16-24(26)28(23)22-12-10-9-11-21(22)27(31)33-28/h9-18H,5-8H2,1-4H3
- Substance type: Organic
- Physical state: Solid

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- Concentration of sludge: 30mg/L

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

TOC removal

Parameter followed for biodegradation estimation:

other: BOD and HPLC

Details on study design:

Standard type

Key result

Parameter:

% degradation (O2 consumption)

Remarks:

BOD

Value:

0

Sampling time:

28 d

Remarks on result:

other: Other details not known

Key result

Parameter:

% degradation (TOC removal)

Value:

1

Sampling time:

28 d

Remarks on result:

other: Other details not known

Key result

Parameter:

% degradation (test mat. analysis)

Remarks:

HPLC

Value:

1

Sampling time:

28 d

Remarks on result:

other: Other details not known

Details on results:

Test substance undergoes 0 and 1% degradation by BOD, TOC removal and HPLC in 28 days.

Validity criteria fulfilled:

not specified

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The percentage degradation of test substance was determined to be 0 and 1% degradation by BOD, TOC removal and HPLC in 28 days. Thus, based on percentage degradation, C.I. Solvent Red 49 was considered to be not readily biodegradable in water.

Executive summary:

Biodegradation study was conducted for 28 days for evaluating the percentage biodegradability of test substance C.I. Solvent Red 49 (CAS no. 509 -34 -2). Concentration of inoculum i.e, sludge used was 30 mg/l and initial test substance conc. used in the study was 100 mg/l. The percentage degradation of test substance was determined to be 0 and 1% degradation by BOD, TOC removal and HPLC in 28 days.  Thus, based on percentage degradation, C.I. Solvent Red 49 was considered to be not readily biodegradable in nature.

Description of key information

Biodegradation study was conducted for 28 days for evaluating the percentage biodegradability of test substance C.I. Solvent Red 49 (CAS no. 509 -34 -2) (J-CHECK, 20016). Concentration of inoculum i.e, sludge used was 30 mg/l and initial test substance conc. used in the study was 100 mg/l. The percentage degradation of test substance was determined to be 0 and 1% degradation by BOD, TOC removal and HPLC in 28 days.  Thus, based on percentage degradation, C.I. Solvent Red 49 was considered to be not readily biodegradable in nature.

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed

Additional information

1 experimental study and 2 predicted data for the test compound C.I. Solvent Red 49 (CAS No. 509-34-2) and the 4 different studies for its closest read across substances using log kow as the primary descriptor were reviewed for the biodegradation endpoint, which are summarized as below:

 

Study 1 - Biodegradation study was conducted for 28 days for evaluating the percentage biodegradability of test substance C.I. Solvent Red 49 (CAS no. 509 -34 -2) (J-CHECK, 20016). Concentration of inoculum i.e, sludge used was 30 mg/l and initial test substance conc. used in the study was 100 mg/l. The percentage degradation of test substance was determined to be 0 and 1% degradation by BOD, TOC removal and HPLC in 28 days.  Thus, based on percentage degradation, C.I. Solvent Red 49 was considered to be not readily biodegradable in nature.

 

Study 2 - Estimation Programs Interface Suite (EPI suite, 2017) was run to predict the biodegradation potential of the test compound C.I. Solvent Red 49 (CAS no. 509 -34 -2) 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 C.I. Solvent Red 49 is expected to be not readily biodegradable.

 

Study 3- Biodegradability of C.I. Solvent Red 49 (CAS no. 509 -34 -2) is predicted using QSAR toolbox version 3.4 (2017). Test substance undergoes 15.77% degradation by CO2 evolution in 28 days. Thus, based on percentage degradation, the test chemical C.I. Solvent Red 49 was estimated to be not readily biodegradable in water.

 

Study 4 - For a read across chemical Uranine (CAS no. 518 -47 -8), a combination of two biodegradation tests (Closed Bottle test and Manometric Respiratory test, OECDs 301 D, F) was carried out to investigate the biodegradability of the read across substance Uranine (CAS no. 518-47-8) (Lukasz Gutowski et. al, 2015). Closed Bottle Test was performed according to the guidelines of the Organization for Economic Co-operation and Development OECD 301D. This test is characterized by low bacteria density (102–105 colony forming units (CFUs)/mL), low nutrient content, and constant temperature (20 ± 1 °C) and it was kept in the dark. The average biodegradation value after 28 days for Uranine monitored by measurement of the oxygen concentration was 7.6. Based on the results obtained, Uranine is expected to be not readily biodegradable.

 

Study 5 - For a read across chemical Uranine (CAS no. 518 -47 -8), a combination of two biodegradation tests (Closed Bottle test and Manometric Respiratory test, OECDs 301 D, F) was carried out to investigate the biodegradability of the read across substance Uranine (CAS no. 518-47-8) (Lukasz Gutowski et. al, 2015).

The Manometric Respiratory tests works with higher bacterial density (5–10×106CFUs/mL) and diversity as the Closed Bottle test thus increasing the probability for biodegradation. This test was performed according to the OECD guidelines (OECD, 1992) in the dark at room temperature (20 ± 1 °C) under gentle stirring. CO2 production as the parameter of the endpoint biodegradation was measured indirectly by the OxiTop OC110-system.

The concentration of standard solution for uranine was 16.7 mg/L, corresponding to the theoretical oxygen demand ThOD of 30 mg/L. Inoculum was derived from the municipal sewage treatment plant (Lüneburg, Germany).Aliquots (measuring) of 80ml of inoculum were added to 1 L of mineral medium. The test consisted of four different series: (i) a blank series (containing only the mineral medium and inoculum), (ii) quality control (containing readily biodegradable sodium acetate as the only relevant carbon source apart from the inoculum), (iii) a test series (containing the target compound) and (iv) toxicity control (containing target compound and sodium acetate as carbon source). The amount of sodium acetate for each series corresponded to ThOD of 5 mg/L. A compound is qualified as “ready biodegradable” when 60% of ThOD expressed as percentage of oxygen consumption is consumed within a period of 10 days after the oxygen uptake reached 10% of ThOD. Samples from the beginning (day 0) and the end of the test (day 28) were collected and stored at−20 °C until analysis with HPLC-FLD and LC–M/MS. No toxic effects on bacteria were observed in the toxicity control as well as no degradation was observed in the sterile control. The measurements with HPLC-FLD confirmed that no elimination of uranine occurred during the Manometric Respiratory Test. Only 0.1% degradation of test substance Uranine was observed. The reason for the negative values in Manometric Respiratory test might be interpreted as high degradation in the blank control and should be considered could be due to some background in the blanks and can be considered as 0% degradation of the test substance. Based on the results obtained in Manometric Respiratory test method, Uranine is expected to be not readily biodegradable.

 

Study 6 - In a weight of evidence study of read across chemical Uranine (CAS no. 518 -47 -8), biodegradation screening test was conducted according to OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I)) for 28 days (4 weeks) for evaluating the percentage biodegradability of the read across substance Uranine (CAS no. 518-47-8) (J-CHECK, 20016). The purity of test substance by HPLC was found to be 92.1% and water solubility value is ≥ 300 g/l at 20ᵒC (Flask method). Concentration of inoculum i.e, sludge is 30 mg/l and initial test substance conc. used in the study is 100 mg/l. The percentage degradation of read across substance is determined to be 0% by O2 consumption and by TOC removal whereas only 1% degradation was observed by HPLC. Thus, based on percentage degradation, substance Fluorescein sodium is considered to be not readily biodegradable in nature.

 

Study 7- In an another weight of evidence study of read across chemical Rhodamine B (CAS no. 81 -88 -9) , biodegradation screening test is conducted for 4 weeks(J-CHECK, 20016) for read-across substance Rhodamine B (CAS: 81 -88 -9). Concentration of inoculum i.e, sludge was 30 mg/l and initial substance conc. used in the study was 100 mg/l, respectively. The percentage degradation of read across substance is determined to be 0, 2 and 7% degradation by BOD, TOC removal and Uv-Vis parameter in 28 days. Thus, based on percentage degradation, read across substance Rhodamine B (CAS no. 81-88-9) is considered to be not readily biodegradable in nature.

 

On the basis of above results for target chemical (study 1, 2 and 3) and of its closest read across substance (study 4, 5, 6 and 7 from peer reviewed journal and authoritative database), it can be concluded that the test substance C.I. Solvent Red 49 can be expected to be not readily biodegradable in nature.