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Environmental fate & pathways

Biodegradation in water: screening tests

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Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Test performed under GLP according guidelines, meeting quality criteria, sufficient info on test substance CAS 28777-98-2. Test substance was first hydrolysed and the product of the hydrolisation was tested in the closed bottle test.
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
Deviations:
yes
Remarks:
acceptable deviations
Principles of method if other than guideline:
Instead of an effluent/extract mixture, activated sludge was used as an inoculum.
The inoculum was taken from an activated sludge plant treating predominantly domestic waste water. The sludge was preconditioned to reduce endogenous respiration rates. To this end, the sludge (400 mg dry wt/liter) was aerated for a period of 7 days. The sludge was diluted to a concentration in the biochemical oxygen demand (BOD) bottles of 2 mg dry wt/liter.

Ammonium chloride was omitted from the medium to prevent nitrification
GLP compliance:
yes
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, domestic, non-adapted
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Secondary activated sludge was obtained from the WWTP Nieuwgraaf in Duiven, the Netherlands (30-03-2006). The WWTP Nieuwgraaf is an activated sludge plant treating predominantly domestic wasewater.
-The sludge was preconditioned to reduce endogenous respiration rates. To this end, the sludge (400 mg dry wt/liter) was aerated for a period of 7 days. The sludge was diluted to a concentration in the biochemical oxygen demand (BOD) bottles of 2 mg dry wt/liter.
Duration of test (contact time):
28 d
Initial conc.:
2 mg/L
Based on:
other: hydrolysed test material
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Composition of medium: The nutrient medium of the closed bottle test contained per litre of deionized water: 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.4 mg Na2HPO4.2H2O, 22.5 mg MgSO4.7H2O, 27.5 mg CaCl2, 0.25 mg FeCl3.6H2O.
- Test temperature: 19-21°C
- pH: 7.0
- pH adjusted: no
- Aeration of mineral medium: yes
- Suspended solids concentration: sludge DW in BOD bottles = 2 mg/L
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: incubator
- Number of culture flasks/concentration:
10 bottles containing only inoculum = inoculum blank
10 bottles containing test substance and inoculum =test
6 bottles containing sodium acetate and inoculum = reference control
- Method used to create aerobic conditions: aeration with pressured air
- Measuring equipment: oxygen meter

SAMPLING
- Sampling frequency: day 0, 7, 14, 21 ,28 (duplicate measurements)
- Sampling method: the bottles were discarded after the oxygen measurement

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: no
- Toxicity control: no
- Reference control: yes


CALCULATIONS:
THOD EKA SA 210 = 2.7 mg/mg
THOD sodiumacetate = 0.8 mg/mg

Oxygen consumption (mg/l) (BOD) = mean oxygen concentration (mg/L) inoculum blank - mean oxygen concentration (mg/l) test (or reference)
Biodegradation (%) = BOD/THOD *100



Reference substance:
acetic acid, sodium salt
Key result
Parameter:
% degradation (O2 consumption)
Value:
60
Sampling time:
28 d
Results with reference substance:
The reference substance sodium acetate showed 83% degradation after 14 days.

Oxygen consumption (mg/L) and the percentages biodegradation of Eka SA 210, hydrolysed (BOD/Thod) and sodium acetate (BOD/Thod) in the closed bottle test:

 Time (days) Oxygen consumption (mg/l)     Biodegradation (%)    
  Test substance Acetate  Test substance Acetate 
0.0  0.0 
1.3  4.1  24  76 
14  2.1  4.5  39  83 
21  2.8    52   
28  3.2    60   
Validity criteria fulfilled:
yes
Remarks:
endogenous respiration 0.9 mg/L, differences of replicates at day 28 < 20%; biodegradation of reference substance at day 14 = 83% and >0.5 mg/L of oxygen in the closed bottles during the test.
Interpretation of results:
readily biodegradable
Conclusions:
Eka SA 210, hydrolysed is biodegraded 60% at day 28 in the closed bottle test and should therefore be classified as readily biodegradable
Test performed under GLP according guidelines with acceptable deviations, meeting quality criteria.
Executive summary:

In order to assess the biotic degradation, a ready biodegradability test was performed which allows the biodegradability to be measured in an aerobic aqueous medium. The ready biodegradability was determined in the closed bottle test performed according to slightly modified OECD, EU and ISO Test Guidelines, and in compliance with the OECD principles of Good Laboratory Practice.

Eka SA 210, hydrolysed did not cause a reduction in the endogenous respiration. The test substance is therefore considered to be non-inhibitory to the inoculum.

Eka SA 210, hydrolysed was biodegraded 60% at day 28 in the closed bottle test. Hence this compound should be classified as readily biodegradable.

The test is valid as shown by an endogenous respiration of 0.9 mg/L and by the total mineralisation of the reference compound, sodium acetate. Sodium acetate was degraded 83% of its theoretical oxygen demand after 14 days. Finally, the most important criterion was met by oxygen concentrations >0.5 mg/L in all bottles during the test period.

Endpoint:
biodegradation in water: screening tests
Type of information:
(Q)SAR
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Biodegradation prediction using BIOWIN, version 4.10, a validated QSAR model included as part of the United States Environmental Protection Agency's EPI Suite modeling program, a publicly-available modeling software contained within the OECD QSAR Toolbox; the modelling was conducted on the hydrolysis product of the substance, by read-across from an analogue substance.
Justification for type of information:
QSAR prediction: migrated from IUCLID 5.6
Principles of method if other than guideline:
Biodegradation prediction using BIOWIN, version 4.10, a validated QSAR model included as part of the United States Environmental Protection Agency's EPI Suite modeling program, a publicly-available modeling software contained within the OECD QSAR Toolbox.
GLP compliance:
no

BIOWIN (v4.10) Program Results:

==============================

SMILES : OC(=O)C(CCCCCCCCC=CCCCCCCCC)CC(=O)O

CHEM  : Octadecenylsuccinic acid

MOL FOR: C22 H40 O4

MOL WT : 368.56

--------------------------- BIOWIN v4.10 Results ----------------------------

 

  Biowin1 (Linear Model Prediction)   : Biodegrades Fast

  Biowin2 (Non-Linear Model Prediction): Biodegrades Fast

  Biowin3 (Ultimate Biodegradation Timeframe): Days-Weeks

  Biowin4 (Primary Biodegradation Timeframe): Hours-Days

  Biowin5 (MITI Linear Model Prediction)   : Biodegrades Fast

  Biowin6 (MITI Non-Linear Model Prediction): Biodegrades Fast

  Biowin7 (Anaerobic Model Prediction): Biodegrades Fast

  Ready Biodegradability Prediction: YES

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin1 FRAGMENT DESCRIPTION        COEFF  | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]    |0.1084 | 0.1084

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]            |0.0727 | 0.1454

 MolWt| * | Molecular Weight Parameter             |          |-0.1755

 Const| * | Equation Constant                     |          |0.7475

============+============================================+=========+=========

  RESULT  |   Biowin1 (Linear Biodeg Probability  |         |0.8259

============+============================================+=========+=========

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin2 FRAGMENT DESCRIPTION      | COEFF  | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]    |1.8437 | 1.8437

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]            |0.6431 | 1.2862

 MolWt| * | Molecular Weight Parameter              |          |-5.2336

============+============================================+=========+=========

  RESULT  | Biowin2 (Non-Linear Biodeg Probability)   |         |0.7120

============+============================================+=========+=========

 

 A Probability Greater Than or Equal to 0.5 indicates --> Biodegrades Fast

 A Probability Less Than 0.5 indicates --> Does NOT Biodegrade Fast

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin3 FRAGMENT DESCRIPTION      | COEFF | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]    | 0.2983 | 0.2983

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           | 0.3646 | 0.7292

 MolWt| * | Molecular Weight Parameter             |         |-0.8145

 Const| * | Equation Constant                     |         |3.1992

============+============================================+=========+=========

  RESULT  | Biowin3 (Survey Model - Ultimate Biodeg)  |      |3.4122

============+============================================+=========+=========

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin4 FRAGMENT DESCRIPTION      | COEFF  | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]     0.2691 | 0.2691

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           |0.3856 | 0.7711

 MolWt| * | Molecular Weight Parameter              |         |-0.5317

 Const| * | Equation Constant                       |        |3.8477

============+============================================+=========+=========

  RESULT  |  Biowin4 (Survey Model - Primary Biodeg |         |4.3562

============+============================================+=========+=========

 

 Result Classification:  5.00 -> hours    4.00 -> days   3.00 -> weeks

 (Primary & Ultimate)   2.00 -> months   1.00 -> longer

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin5 FRAGMENT DESCRIPTION      | COEFF  | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           |0.1812 | 0.3623

 Frag | 1 | Methyl [-CH3]                         |0.0004 | 0.0004

 Frag | 16 | -CH2- [linear]                         |0.0494 | 0.7907

 Frag | 1 | -CH-  [linear]                        |-0.0507 |-0.0507

 Frag | 2 | -C=CH [alkenyl hydrogen

0.0062 | 0.0124

 MolWt| * | Molecular Weight Parameter             |         |-1.0965

 Const| * | Equation Constant                      |          | 0.7121

============+============================================+=========+=========

  RESULT  | Biowin5 (MITI Linear Biodeg Probability |         | 0.7308

============+============================================+=========+=========

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin6 FRAGMENT DESCRIPTION      | COEFF | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           | 1.1346 | 2.2692

 Frag | 1 | Methyl [-CH3]                          | 0.0194 | 0.0194

 Frag | 16 | -CH2- [linear]                        | 0.4295 | 6.8719

 Frag | 1 | -CH-  [linear]                         |-0.0998 | -0.0998

 Frag | 2 | -C=CH [alkenyl hydrogen]              | 0.0285 | 0.0570

 MolWt| * | Molecular Weight Parameter             |         |-10.6399

============+============================================+=========+=========

  RESULT  |Biowin6 (MITI Non-Linear Biodeg Probability) |      | 0.7318

============+============================================+=========+=========

 

 A Probability Greater Than or Equal to 0.5 indicates --> Readily Degradable

 A Probability Less Than 0.5 indicates --> NOT Readily Degradable

 

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin7 FRAGMENT DESCRIPTION      | COEFF | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]    |-0.3177 | -0.3177

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           | 0.1868 | 0.3735

 Frag | 1 | Methyl [-CH3]                         |-0.0796 | -0.0796

 Frag | 16 | -CH2- [linear]                        | 0.0260 | 0.4158

 Frag | 1 | -CH-  [linear]                        |-0.1659 | -0.1659

 Frag | 2 | -C=CH [alkenyl hydrogen]             | -0.0735 | -0.1470

 Const| * | Equation Constant                       |         | 0.8361

============+============================================+=========+=========

  RESULT  |  Biowin7 (Anaerobic Linear Biodeg Prob) |         | 0.9153

============+============================================+=========+=========

 

 A Probability Greater Than or Equal to 0.5 indicates --> Biodegrades Fast

 A Probability Less Than 0.5 indicates --> Does NOT Biodegrade Fast

 

Ready Biodegradability Prediction: (YES or NO)

----------------------------------------------

 Criteria for the YES or NO prediction: If the Biowin3 (ultimate survey

 model) result is "weeks" or faster (i.e. "days", "days to weeks", or

 "weeks" AND the Biowin5 (MITI linear model) probability is >= 0.5, then

 the prediction is YES (readily biodegradable). If this condition is not

 satisfied, the prediction is NO (not readily biodegradable). This   method is based on application of Bayesian analysis to ready biodegradation data

 (see Help). Biowin5 and 6 also predict ready biodegradability, but for

 degradation in the OECD301C test only; using data from the Chemicals

 Evaluation and Research Institute Japan (CERIJ) database.

 

  B

Conclusions:
The determination of ready biodegradability by the BIOWIN model is based upon the results of two modules within the model, BIOWIN3 and BIOWIN5. The BIOWIN3 module predicted ultimate degradation of the hydrolysis product of the substance in the timeframe of days to weeks, and the BIOWIN5 module predicted that the hydrolysis product of the substance biodegrades fast, resulting in overall prediction of ready biodegradability. The prediction of ready biodegradability by the BIOWIN model indicates that there are no structural components of the hydrolysis product of the substance that are resistant to rapid degradation, and that ultimate degradation is likely.
Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
October 1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
Deviations:
no
GLP compliance:
yes
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, domestic, non-adapted
Details on inoculum:
- Source of inoculum/activated sludge: A mixed population of activated sewage sludge micro-organisms was obtained on 2 September 1996 from the aeration stage of the Severn Trent Water Pie sewage treatment plant at Belper, Derbyshire, UK, which treats predominantly domestic sewage.
- Method of cultivation: maintained on continuous aeration upon receipt
- Preparation of inoculum for exposure: washed three times by settlement and resuspension in culture medium
Duration of test (contact time):
28 d
Initial conc.:
20 mg/L
Based on:
DOC
Parameter followed for biodegradation estimation:
CO2 evolution
Details on study design:
TEST CONDITIONS
- Test temperature: 21 °C
- pH adjusted: no
- Aeration of dilution water: 70 mL/minute
- Suspended solids concentration:
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: glass culture vessels
- Number of culture flasks/concentration: 2
- Method used to create aerobic conditions: sealed and bubbled with CO2-free air
- Details of trap for CO2 and volatile organics if used: collected in two 500 ml Dreschel bottles containing 350 ml of 0.05 M NaOH

SAMPLING
- Sampling frequency: on days 0, 11 2, 31 6, 8, 10, 14, 16, 20, 22, 24, 27, 28 and 29


CONTROL AND BLANK SYSTEM
- Inoculum blank: yes, treated in parallel
- Toxicity control: Preliminary investigational work was carried out to assess any toxic effect of the test material on the sewage sludge micro-organisms. A range-finding study followed the method described in OECD Guideline No 209 "Activated Sludge Respiration Inhibition Test". The test material was aerated for a period of 3 hours at 21 'C in the presence of domestic sewage sludge with the addition of a synthetic sewage as a respiratory substrate. The concentrations employed were 1 0 and 20 mg carbonhitre. After 3 hours aeration the rate of respiration was measured for a 10 minute period and compared to data for the control and a reference material, 3,5-dichlorophenol.
Reference substance:
benzoic acid, sodium salt
Preliminary study:
Additional investigational work was carried out using the Activated Sludge Respiration Inhibition test method (OECD Guideline No 209), which showed that the test material did not inhibit the respiration of sewage sludge micro-organisms atthe concentration employed in the test. In the light of this the toxicity control vessel (Pentasize 68 plus sodium benzoate) was omitted from the test.
Parameter:
% degradation (CO2 evolution)
Value:
18
Sampling time:
28 d
Details on results:
The results of the inorganic carbon analysis of the first absorber vessels on day 29 showed an increase in all replicate vessels. These increases are considered to be due to CO2 present in solution being driven off by the addition of hydrochloric acid on day 28 and resulted in an increase in the percentage degradation value for the test material from 18% on day 28 to 20% on day 29.
Inorganic carbon analysis of the second absorber on day 29 confirmed that no significant carry-over of CO2 into the second absorber vessels occurred.
Results with reference substance:
Sodium benzoate attained 87% degradation after 28 days thereby confirming the suitability of the inoculum and test conditions. The increase in inorganic carbon in the first absorber vessels on day 29 resulted in an increase in the percentage degradation value for the standard material from 87% on day 28 to 92% on day 29.

  Day %degradation sodium benzoate   %degradation pentasize 68  
17 
32 
10  31 
14  67 
16  62 
20  73  15 
22  74  11 
24  77  11 
27  83  14 
28  87  18 
29  92  20 
Validity criteria fulfilled:
yes
Remarks:
reference substance > 60% biodegradation at day 14; differences between replicates at end of test <20%; Ic content at start of test <5% of the TC; CO2 evolution in inoculum blank < 40 mg/L and total CO2 concentrations < 70 mg/L
Interpretation of results:
inherently biodegradable
Conclusions:
Pentasize 68 was biodegraded 20% in the CO2 evolution test (OECD 301B) and therefore can be regarded as inherently biodegradable.
Executive summary:

A study was performed to assess the ready biodegradability of the test material in an aerobic aqueous media. The method followed that described in the OECD Guidelines for Testing of Chemicals (1992) No 301 B, "Ready Biodegradability; CO2 Evolution Test" referenced as Method C.4-C of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

The test material was exposed to activated sewage sludge micro-organ isms at a concentration of 20 mg C/L with culture medium in sealed culture vessels in the dark at 21 'C for 29 days. The degradation of the test material was assessed by the determination of carbon dioxide produced. Control solutions with inoculum and the standard material, sodium benzoate, were used for validation purposes.

The test material attained 20% degradation after 29 days and therefore cannot be considered to be readily biodegradable under the strict terms and conditions of OECD Guideline No. 301 B. However, the test substance could be regarded as inherently biodegradable.

 

Endpoint:
biodegradation in water: ready biodegradability
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
October 1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
This study was conducted on 2,5-furandione, dihydro-, mono-C15- 20-alkenyl derivatives (CAS 68784-12-3), an analogue substance used as the source of information for the assessment of the target substance through read-across. Therefore, this study is informative for evaluation of the environmental fate and toxicity of the target substance, Reaction products of furan-2,5-dione and octadec-1-ene (known here as n-ODSA EC 701-338-8; no CASRN available), and it is adequate for classification and risk assessment.

Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
As a result of increasingly rigorous criteria being applied to the analysis of commercial material used in physical property/toxicity testing, the identity of the material has been modified to reveal a more accurate and precise depiction of the commercial substance. This enhancement is reflected in changes in chemical identifiers such as EC and/or CAS numbers from those noted in earlier versions of data records or in study reports.

Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, domestic, non-adapted
Details on inoculum:
- Source of inoculum/activated sludge: A mixed population of activated sewage sludge micro-organisms was obtained on 2 September 1996 from the aeration stage of the Severn Trent Water Pie sewage treatment plant at Belper, Derbyshire, UK, which treats predominantly domestic sewage.
- Method of cultivation: maintained on continuous aeration upon receipt
- Preparation of inoculum for exposure: washed three times by settlement and resuspension in culture medium
Duration of test (contact time):
28 d
Initial conc.:
20 mg/L
Based on:
DOC
Parameter followed for biodegradation estimation:
CO2 evolution
Details on study design:
TEST CONDITIONS
- Test temperature: 21 °C
- pH adjusted: no
- Aeration of dilution water: 70 mL/minute
- Suspended solids concentration:
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: glass culture vessels
- Number of culture flasks/concentration: 2
- Method used to create aerobic conditions: sealed and bubbled with CO2-free air
- Details of trap for CO2 and volatile organics if used: collected in two 500 ml Dreschel bottles containing 350 ml of 0.05 M NaOH

SAMPLING
- Sampling frequency: on days 0, 11 2, 31 6, 8, 10, 14, 16, 20, 22, 24, 27, 28 and 29


CONTROL AND BLANK SYSTEM
- Inoculum blank: yes, treated in parallel
- Toxicity control: Preliminary investigational work was carried out to assess any toxic effect of the test material on the sewage sludge micro-organisms. A range-finding study followed the method described in OECD Guideline No 209 "Activated Sludge Respiration Inhibition Test". The test material was aerated for a period of 3 hours at 21 'C in the presence of domestic sewage sludge with the addition of a synthetic sewage as a respiratory substrate. The concentrations employed were 1 0 and 20 mg carbonhitre. After 3 hours aeration the rate of respiration was measured for a 10 minute period and compared to data for the control and a reference material, 3,5-dichlorophenol.
Reference substance:
benzoic acid, sodium salt
Preliminary study:
Additional investigational work was carried out using the Activated Sludge Respiration Inhibition test method (OECD Guideline No 209), which showed that the test material did not inhibit the respiration of sewage sludge micro-organisms atthe concentration employed in the test. In the light of this the toxicity control vessel (Pentasize 68 plus sodium benzoate) was omitted from the test.
Parameter:
% degradation (CO2 evolution)
Value:
18
Sampling time:
28 d
Details on results:
The results of the inorganic carbon analysis of the first absorber vessels on day 29 showed an increase in all replicate vessels. These increases are considered to be due to CO2 present in solution being driven off by the addition of hydrochloric acid on day 28 and resulted in an increase in the percentage degradation value for the test material from 18% on day 28 to 20% on day 29.
Inorganic carbon analysis of the second absorber on day 29 confirmed that no significant carry-over of CO2 into the second absorber vessels occurred.
Results with reference substance:
Sodium benzoate attained 87% degradation after 28 days thereby confirming the suitability of the inoculum and test conditions. The increase in inorganic carbon in the first absorber vessels on day 29 resulted in an increase in the percentage degradation value for the standard material from 87% on day 28 to 92% on day 29.

  Day %degradation sodium benzoate   %degradation pentasize 68  
17 
32 
10  31 
14  67 
16  62 
20  73  15 
22  74  11 
24  77  11 
27  83  14 
28  87  18 
29  92  20 
Validity criteria fulfilled:
yes
Remarks:
reference substance > 60% biodegradation at day 14; differences between replicates at end of test <20%; Ic content at start of test <5% of the TC; CO2 evolution in inoculum blank < 40 mg/L and total CO2 concentrations < 70 mg/L
Interpretation of results:
inherently biodegradable
Conclusions:
Pentasize 68 was biodegraded 20% in the CO2 evolution test (OECD 301B) and therefore can be regarded as inherently biodegradable.
Executive summary:

A study was performed to assess the ready biodegradability of the test material in an aerobic aqueous media. The method followed that described in the OECD Guidelines for Testing of Chemicals (1992) No 301 B, "Ready Biodegradability; CO2 Evolution Test" referenced as Method C.4-C of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

The test material was exposed to activated sewage sludge micro-organ isms at a concentration of 20 mg C/L with culture medium in sealed culture vessels in the dark at 21 'C for 29 days. The degradation of the test material was assessed by the determination of carbon dioxide produced. Control solutions with inoculum and the standard material, sodium benzoate, were used for validation purposes.

The test material attained 20% degradation after 29 days and therefore cannot be considered to be readily biodegradable under the strict terms and conditions of OECD Guideline No. 301 B. However, the test substance could be regarded as inherently biodegradable.

 

Endpoint:
biodegradation in water: ready biodegradability
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Test performed under GLP according guidelines, meeting quality criteria, sufficient info on test substance CAS 28777-98-2. Test substance was first hydrolysed and the product of the hydrolisation was tested in the closed bottle test. According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (2012)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Justification for type of information:
This study was conducted on octadecenyl succinic anhydride (CAS 28777-98-2, also known as ODSA), an analogue substance used as the source of information for the assessment of the target substance through read-across. Therefore, this study is informative for evaluation of the environmental fate and toxicity of the target substance, Reaction products of furan-2,5-dione and octadec-1-ene (known here as n-ODSA EC 701-338-8; no CASRN available), and it is adequate for classification and risk assessment.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
Deviations:
yes
Remarks:
acceptable deviations
Principles of method if other than guideline:
Instead of an effluent/extract mixture, activated sludge was used as an inoculum.
The inoculum was taken from an activated sludge plant treating predominantly domestic waste water. The sludge was preconditioned to reduce endogenous respiration rates. To this end, the sludge (400 mg dry wt/liter) was aerated for a period of 7 days. The sludge was diluted to a concentration in the biochemical oxygen demand (BOD) bottles of 2 mg dry wt/liter.

Ammonium chloride was omitted from the medium to prevent nitrification
GLP compliance:
yes
Specific details on test material used for the study:
As a result of increasingly rigorous criteria being applied to the analysis of commercial material used in physical property/toxicity testing, the identity of the material has been modified to reveal a more accurate and precise depiction of the commercial substance. This enhancement is reflected in changes in chemical identifiers such as EC and/or CAS numbers from those noted in earlier versions of data records or in study reports.

Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, domestic, non-adapted
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Secondary activated sludge was obtained from the WWTP Nieuwgraaf in Duiven, the Netherlands (30-03-2006). The WWTP Nieuwgraaf is an activated sludge plant treating predominantly domestic wasewater.
-The sludge was preconditioned to reduce endogenous respiration rates. To this end, the sludge (400 mg dry wt/liter) was aerated for a period of 7 days. The sludge was diluted to a concentration in the biochemical oxygen demand (BOD) bottles of 2 mg dry wt/liter.
Duration of test (contact time):
28 d
Initial conc.:
2 mg/L
Based on:
other: hydrolysed test material
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Composition of medium: The nutrient medium of the closed bottle test contained per litre of deionized water: 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.4 mg Na2HPO4.2H2O, 22.5 mg MgSO4.7H2O, 27.5 mg CaCl2, 0.25 mg FeCl3.6H2O.
- Test temperature: 19-21°C
- pH: 7.0
- pH adjusted: no
- Aeration of mineral medium: yes
- Suspended solids concentration: sludge DW in BOD bottles = 2 mg/L
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: incubator
- Number of culture flasks/concentration:
10 bottles containing only inoculum = inoculum blank
10 bottles containing test substance and inoculum =test
6 bottles containing sodium acetate and inoculum = reference control
- Method used to create aerobic conditions: aeration with pressured air
- Measuring equipment: oxygen meter

SAMPLING
- Sampling frequency: day 0, 7, 14, 21 ,28 (duplicate measurements)
- Sampling method: the bottles were discarded after the oxygen measurement

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: no
- Toxicity control: no
- Reference control: yes


CALCULATIONS:
THOD EKA SA 210 = 2.7 mg/mg
THOD sodiumacetate = 0.8 mg/mg

Oxygen consumption (mg/l) (BOD) = mean oxygen concentration (mg/L) inoculum blank - mean oxygen concentration (mg/l) test (or reference)
Biodegradation (%) = BOD/THOD *100



Reference substance:
acetic acid, sodium salt
Key result
Parameter:
% degradation (O2 consumption)
Value:
60
Sampling time:
28 d
Results with reference substance:
The reference substance sodium acetate showed 83% degradation after 14 days.

Oxygen consumption (mg/L) and the percentages biodegradation of Eka SA 210, hydrolysed (BOD/Thod) and sodium acetate (BOD/Thod) in the closed bottle test:

 Time (days) Oxygen consumption (mg/l)     Biodegradation (%)    
  Test substance Acetate  Test substance Acetate 
0.0  0.0 
1.3  4.1  24  76 
14  2.1  4.5  39  83 
21  2.8    52   
28  3.2    60   
Validity criteria fulfilled:
yes
Remarks:
endogenous respiration 0.9 mg/L, differences of replicates at day 28 < 20%; biodegradation of reference substance at day 14 = 83% and >0.5 mg/L of oxygen in the closed bottles during the test.
Interpretation of results:
readily biodegradable
Conclusions:
Eka SA 210, hydrolysed is biodegraded 60% at day 28 in the closed bottle test and should therefore be classified as readily biodegradable
Test performed under GLP according guidelines with acceptable deviations, meeting quality criteria.
Executive summary:

In order to assess the biotic degradation, a ready biodegradability test was performed which allows the biodegradability to be measured in an aerobic aqueous medium. The ready biodegradability was determined in the closed bottle test performed according to slightly modified OECD, EU and ISO Test Guidelines, and in compliance with the OECD principles of Good Laboratory Practice.

Eka SA 210, hydrolysed did not cause a reduction in the endogenous respiration. The test substance is therefore considered to be non-inhibitory to the inoculum.

Eka SA 210, hydrolysed was biodegraded 60% at day 28 in the closed bottle test. Hence this compound should be classified as readily biodegradable.

The test is valid as shown by an endogenous respiration of 0.9 mg/L and by the total mineralisation of the reference compound, sodium acetate. Sodium acetate was degraded 83% of its theoretical oxygen demand after 14 days. Finally, the most important criterion was met by oxygen concentrations >0.5 mg/L in all bottles during the test period.

Endpoint:
biodegradation in water: screening tests
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Biodegradation prediction using BIOWIN, version 4.10, a validated QSAR model included as part of the United States Environmental Protection Agency's EPI Suite modeling program, a publicly-available modeling software contained within the OECD QSAR Toolbox; the modelling was conducted on the hydrolysis product of the substance, by read-across from an analogue substance.
Justification for type of information:
QSAR prediction: migrated from IUCLID 5.6. This study was conducted on octadecenyl succinic acid (CAS 28299-29-8), an analogue substance used as the source of information for the assessment of the target substance through read-across. Therefore, this study is informative for evaluation of the environmental fate and toxicity of the target substance, Reaction products of furan-2,5-dione and octadec-1-ene (known here as n-ODSA EC 701-338-8; no CASRN available), and it is adequate for classification and risk assessment.
Reason / purpose for cross-reference:
read-across source
Principles of method if other than guideline:
Biodegradation prediction using BIOWIN, version 4.10, a validated QSAR model included as part of the United States Environmental Protection Agency's EPI Suite modeling program, a publicly-available modeling software contained within the OECD QSAR Toolbox.
GLP compliance:
no

BIOWIN (v4.10) Program Results:

==============================

SMILES : OC(=O)C(CCCCCCCCC=CCCCCCCCC)CC(=O)O

CHEM  : Octadecenylsuccinic acid

MOL FOR: C22 H40 O4

MOL WT : 368.56

--------------------------- BIOWIN v4.10 Results ----------------------------

 

  Biowin1 (Linear Model Prediction)   : Biodegrades Fast

  Biowin2 (Non-Linear Model Prediction): Biodegrades Fast

  Biowin3 (Ultimate Biodegradation Timeframe): Days-Weeks

  Biowin4 (Primary Biodegradation Timeframe): Hours-Days

  Biowin5 (MITI Linear Model Prediction)   : Biodegrades Fast

  Biowin6 (MITI Non-Linear Model Prediction): Biodegrades Fast

  Biowin7 (Anaerobic Model Prediction): Biodegrades Fast

  Ready Biodegradability Prediction: YES

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin1 FRAGMENT DESCRIPTION        COEFF  | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]    |0.1084 | 0.1084

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]            |0.0727 | 0.1454

 MolWt| * | Molecular Weight Parameter             |          |-0.1755

 Const| * | Equation Constant                     |          |0.7475

============+============================================+=========+=========

  RESULT  |   Biowin1 (Linear Biodeg Probability  |         |0.8259

============+============================================+=========+=========

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin2 FRAGMENT DESCRIPTION      | COEFF  | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]    |1.8437 | 1.8437

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]            |0.6431 | 1.2862

 MolWt| * | Molecular Weight Parameter              |          |-5.2336

============+============================================+=========+=========

  RESULT  | Biowin2 (Non-Linear Biodeg Probability)   |         |0.7120

============+============================================+=========+=========

 

 A Probability Greater Than or Equal to 0.5 indicates --> Biodegrades Fast

 A Probability Less Than 0.5 indicates --> Does NOT Biodegrade Fast

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin3 FRAGMENT DESCRIPTION      | COEFF | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]    | 0.2983 | 0.2983

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           | 0.3646 | 0.7292

 MolWt| * | Molecular Weight Parameter             |         |-0.8145

 Const| * | Equation Constant                     |         |3.1992

============+============================================+=========+=========

  RESULT  | Biowin3 (Survey Model - Ultimate Biodeg)  |      |3.4122

============+============================================+=========+=========

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin4 FRAGMENT DESCRIPTION      | COEFF  | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]     0.2691 | 0.2691

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           |0.3856 | 0.7711

 MolWt| * | Molecular Weight Parameter              |         |-0.5317

 Const| * | Equation Constant                       |        |3.8477

============+============================================+=========+=========

  RESULT  |  Biowin4 (Survey Model - Primary Biodeg |         |4.3562

============+============================================+=========+=========

 

 Result Classification:  5.00 -> hours    4.00 -> days   3.00 -> weeks

 (Primary & Ultimate)   2.00 -> months   1.00 -> longer

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin5 FRAGMENT DESCRIPTION      | COEFF  | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           |0.1812 | 0.3623

 Frag | 1 | Methyl [-CH3]                         |0.0004 | 0.0004

 Frag | 16 | -CH2- [linear]                         |0.0494 | 0.7907

 Frag | 1 | -CH-  [linear]                        |-0.0507 |-0.0507

 Frag | 2 | -C=CH [alkenyl hydrogen

0.0062 | 0.0124

 MolWt| * | Molecular Weight Parameter             |         |-1.0965

 Const| * | Equation Constant                      |          | 0.7121

============+============================================+=========+=========

  RESULT  | Biowin5 (MITI Linear Biodeg Probability |         | 0.7308

============+============================================+=========+=========

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin6 FRAGMENT DESCRIPTION      | COEFF | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           | 1.1346 | 2.2692

 Frag | 1 | Methyl [-CH3]                          | 0.0194 | 0.0194

 Frag | 16 | -CH2- [linear]                        | 0.4295 | 6.8719

 Frag | 1 | -CH-  [linear]                         |-0.0998 | -0.0998

 Frag | 2 | -C=CH [alkenyl hydrogen]              | 0.0285 | 0.0570

 MolWt| * | Molecular Weight Parameter             |         |-10.6399

============+============================================+=========+=========

  RESULT  |Biowin6 (MITI Non-Linear Biodeg Probability) |      | 0.7318

============+============================================+=========+=========

 

 A Probability Greater Than or Equal to 0.5 indicates --> Readily Degradable

 A Probability Less Than 0.5 indicates --> NOT Readily Degradable

 

 

------+-----+--------------------------------------------+---------+---------

 TYPE | NUM |      Biowin7 FRAGMENT DESCRIPTION      | COEFF | VALUE 

------+-----+--------------------------------------------+---------+---------

 Frag | 1 | Linear C4 terminal chain [CCC-CH3]    |-0.3177 | -0.3177

 Frag | 2 | Aliphatic acid  [-C(=O)-OH]           | 0.1868 | 0.3735

 Frag | 1 | Methyl [-CH3]                         |-0.0796 | -0.0796

 Frag | 16 | -CH2- [linear]                        | 0.0260 | 0.4158

 Frag | 1 | -CH-  [linear]                        |-0.1659 | -0.1659

 Frag | 2 | -C=CH [alkenyl hydrogen]             | -0.0735 | -0.1470

 Const| * | Equation Constant                       |         | 0.8361

============+============================================+=========+=========

  RESULT  |  Biowin7 (Anaerobic Linear Biodeg Prob) |         | 0.9153

============+============================================+=========+=========

 

 A Probability Greater Than or Equal to 0.5 indicates --> Biodegrades Fast

 A Probability Less Than 0.5 indicates --> Does NOT Biodegrade Fast

 

Ready Biodegradability Prediction: (YES or NO)

----------------------------------------------

 Criteria for the YES or NO prediction: If the Biowin3 (ultimate survey

 model) result is "weeks" or faster (i.e. "days", "days to weeks", or

 "weeks" AND the Biowin5 (MITI linear model) probability is >= 0.5, then

 the prediction is YES (readily biodegradable). If this condition is not

 satisfied, the prediction is NO (not readily biodegradable). This   method is based on application of Bayesian analysis to ready biodegradation data

 (see Help). Biowin5 and 6 also predict ready biodegradability, but for

 degradation in the OECD301C test only; using data from the Chemicals

 Evaluation and Research Institute Japan (CERIJ) database.

 

  B

Conclusions:
The determination of ready biodegradability by the BIOWIN model is based upon the results of two modules within the model, BIOWIN3 and BIOWIN5. The BIOWIN3 module predicted ultimate degradation of the hydrolysis product of the substance in the timeframe of days to weeks, and the BIOWIN5 module predicted that the hydrolysis product of the substance biodegrades fast, resulting in overall prediction of ready biodegradability. The prediction of ready biodegradability by the BIOWIN model indicates that there are no structural components of the hydrolysis product of the substance that are resistant to rapid degradation, and that ultimate degradation is likely.

Description of key information

The key study is a closed bottle ready biodegradability test conducted in accordance with an established guideline and GLP principles, on an analogue substance based upon read-across. 

Key value for chemical safety assessment

Biodegradation in water:
readily biodegradable but failing 10-day window

Additional information

The closed bottle test ready biodegradability test conducted on the analogue substance (CAS 28777-98-2) in accordance with OECD 301D showed 60% degradation in 28 days. This level of degradation met the threshold of ready biodegradability (60% degradation), although the 10-day window was not achieved. 

This information is from the substance dihydro‐3‐(octadecenyl)furan‐2,5‐dione (CAS 28777-98-2, also known as octadecenyl succinic anhydride, ODSA and EKA SA 210TM), an analogue used for the assessment of several endpoints through read-across. The hypothesis for read-across between the substance being registered (Reaction products of furan-2,5-dione and octadec-1-ene; known here as n-ODSA EC 701-338-8; no CASRN available), and the analogue substance is a common functional group: a 2,5-furandione, dihydro- structure, also known as a succinic anhydride, to which is attached a long-chain monounsaturated olefin. In the environment, the anhydride moiety is quickly hydrolysed to form a dioic acid.  When the substance to be registered and the analogue substance are compared, slight changes in the purity of the starting olefin stock, or the location of the double bond within the carbon chain, are not anticipated to significantly affect the environmental fate properties or the toxicity of the substances. For each endpoint study based upon read-across, the analogue approach is substantiated by an evaluation provided in the Analogue Approach Report Format (AARF) attached to the endpoint study summary file. The AARF allows the read-across information to fulfil the information requirements of the REACH Annexes VII-X, to be the basis for classification and labelling decisions, and for risk assessment.

 

 

A supporting study on another read-across substance (CAS 67784-12-3), a CO2 evolution ready biodegradability test conducted in accordance with OECD Guideline 301B, showed 18% degradation in 28 days, thus failing the threshold of ready biodegradability (60%) in the test. 

This information is from the substance 2,5-furandione, dihydro-,mono-C15-20-alkenylderivatives (CAS 68784-12-3, a mixture of a hexadecenyl- and octadecenyl succinic anhydrides, and also known as PentasizeTM68, AS 1100TMand AS 1000TM), an analogue used for the assessment of several endpoints through read-across. The hypothesis for read-across between the substance being registered (Reaction products of furan-2,5-dione and octadec-1-ene; known here asn-ODSA EC 701-338-8; no CASRN available), and the analogue substance is a common functional group: a 2,5-furandione, dihydro- structure, also known as a succinic anhydride, to which is attached a long-chain monounsaturated olefin. In the environment, the anhydride moiety is quickly hydrolysed to form a dioic acid.  When the substance to be registered and the analogue substance are compared, changes in the purity of the starting olefin stock, or small differences in the length (between sixteen and twenty) or arrangement (linear or branched) of the carbon chain are not anticipated to significantly affect the environmental fate properties or the toxicity of the substances. For each endpoint study based upon read-across, the analogue approach is substantiated by an evaluation provided in the Analogue Approach Report Format (AARF) attached to the endpoint study summary file. The AARF allows the read-across information to fulfil the information requirements of the REACH Annexes VII-X, to be the basis for classification and labelling decisions, and for risk assessment.

A second supporting study provides QSAR modeling by read-across on octadecenyl succinic acid, the hydrolysis product of the substance. Read-across from the hydrolysis product of the substance is relevant, since hydrolysis of the substance has been shown to be rapid. The BIOWIN module (v4.10) within EPI Suite predicted that the hydrolysis product of the substance would be readily biodegradable. The determination of ready biodegradability by the BIOWIN model is based upon the results of two modules within the model, BIOWIN3 and BIOWIN5. The BIOWIN3 module predicted ultimate degradation of the hydrolysis product of the substance in the timeframe of days to weeks, and the BIOWIN5 module predicted that the hydrolysis product of the substance biodegrades fast, resulting in overall prediction of ready biodegradability. The prediction of ready biodegradability by the BIOWIN model indicates that there are no structural components of the hydrolysis product of the substance that are resistant to rapid degradation, and that ultimate degradation is likely.  

This information is from the substance octadecenyl succinic acid (CAS 28299-29-8), an analogue substance used for the assessment of several endpoints through read-across. The hypothesis for read-across between the substance being registered (Reaction products of furan-2,5-dione and octadec-1-ene; known here asn-ODSA EC 701-338-8; no CASRN available), and the analogue substance is a common functional group. The common functional group is a 2,5-furandione, dihydro- structure (also known as a succinic anhydride) or its hydrolysed form of butanedioic acid (also known a s succinic acid), to which is attached a linear, 18-carbon alkenyl group. The anhydride moiety is quickly hydrolysed to the dioic acid form in the environment; thus, the analogue substance (octadecenyl succinic acid) is the hydrolysed form of the substance being registered (n-ODSA EC 701-338-8). The dioic acid form of the moiety is expected to be representative of the environmental fate properties and aquatic toxicity of the anhydride form of the moiety. For each endpoint study based upon read-across, the analogue approach is substantiated by an evaluation provided in the Analogue Approach Report Format (AARF) attached to the endpoint study summary file. The AARF allows the read-across information to fulfil the information requirements of the REACH Annexes VII-X, to be the basis for classification and labelling decisions, and for risk assessment.