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EC number: 700-371-5 | CAS number: 178535-25-6
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Biodegradation in water: screening tests
Administrative data
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- December 8, 2008 - January 5, 2009
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study conducted under GLP.
Cross-referenceopen allclose all
- Reason / purpose for cross-reference:
- reference to same study
- Reason / purpose for cross-reference:
- reference to other study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 009
- Report date:
- 2009
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 310 (Ready Biodegradability - CO2 in Sealed Vessels (Headspace Test)
- Qualifier:
- according to guideline
- Guideline:
- other: ISO Guideline No 14593 "Water quality - Evaluation of ultimate aerobic biodegradability of organic compunds in aqueous medium-Method by analysis of inorganic carbon in sealed vessels (CO2 headspace test)
- Deviations:
- no
- Principles of method if other than guideline:
- Not relevant
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- ethylbenzene
- EC Number:
- 700-371-5
- Cas Number:
- 178535-25-6
- IUPAC Name:
- ethylbenzene
Constituent 1
- Specific details on test material used for the study:
- Details on properties of test surrogate or analogue material (migrated information):
Not applicable
Study design
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge (adaptation not specified)
- Details on inoculum:
- A mixed population of activated sludge micro-organisms was obtained from the aeration stage of the Severn Trent Water Plc sewage treatment plant at Loghborough, Leicester, UK, which treats predominantly domestic sewage.
Upon receipt in the laboratory, the sample of effluent was filtered through coarse filter paper (first approximate 200 ml discarded).
In order to reduce the inorganic carbon (IC) content of the inoculum, the filtrate was sparged with CO2-free air* for approximately 1 hour whilst maintaining its pH at 6.5 using concentrated orthophosphoric acid. After sparging, the pH was restored to its original value of 7.3 using 7 M sodium hydroxide and the inoculum allowed to settle for approximately 1 hour prior to removal of an aliquot (2 litres) of the supernatant for use in the test. The supernatant was maintained on aeration using CO2-free air until use.
- Duration of test (contact time):
- 28 d
Initial test substance concentration
- Initial conc.:
- 20 mg/L
- Based on:
- other: carbon
Parameter followed for biodegradation estimation
- Parameter followed for biodegradation estimation:
- inorg. C analysis
- Details on study design:
- TEST CONDITIONS
Culture Medium:
Solution a KH2PO4 8.50 g/L
K2HPO4 21.75 g/L
Na2HPO4.2H20 33.40 g/L
NH4Cl 0.50 g/L
Ph = 7.4
Solution b CaCl2.2H2O 36.40 g/L
Solution c MgSO4.7H2O 22.50 g/L
Solution d FeCl3.6H2O 0.25 g/L
The following volumes of the above solutions were added to 1 litre (final volume) of purified water.
10 mL of Solution a
1 mL of Solution b
1 mL of Solution c
1 mL of Solution d
The following test preparations were prepared and incubated in 125 ml glass Wheaton bottles (total volume when full 160 ml) each containing 107 ml of solution.
a) A control consisting of inoculated mineral salts medium, 33 replicate vessels.
b) The standard material (sodium benzoate) in inoculated mineral salts medium, to give a final concentration of 20 mg carbon/l, 33 replicate vessels.
c) The test material in inoculated mineral salts medium, to give a final concentration of 20 mg carbon/l, 29 replicate vessels.
d) The test material plus the standard material in inoculated mineral salts medium, to give a final concentration of 40 mg carbon/l to act as a toxicity control, 9 replicate vessels.
Test media a-d were inoculated with the prepared inoculum at a final concentration of 100 ml/l. Aliquots (107 ml) of the test media were dispensed into replicate vessels to give a headspace to liquid ratio of 1:2. Sufficient vessels were prepared to allow a single inorganic carbon determination per vessel with triplicate vessels for the control, standard material, test material and toxicity control at each sampling occasion (five replicates for analysis on Day 28). Additional control and standard material vessels were prepared to provide samples for Dissolved Organic Carbon (DOC) analyses on days 0 and 28 (duplicate vessels per sampling occasion).
TEST SYSTEM
CO2 production in the vessels was determined by measuring the increase in the concentration of Inorganic Carbon (IC) in the headspace. Triplicate control, standard material and test material vessels were sacrificed on days 0, 2, 4, 8, 10, 14, 16 and 21 for IC analysis. On day 28, five replicate vessels were sacrificed for IC analysis. Triplicate toxicity control vessels were sacrificed on days 0, 8 and 14 for IC analysis.
Prior to sampling, concentrated orthophosphoric acid (1 mL) was injected through the septum of each vessel taken for analysis in order to lower the pH of the medium to < 3*. The vessels were then continued to be shaken at approximately 150 rpm (INFORS TR-225 orbital platform shaker) for 1 hour at 20±1 degC before a sample was withdrawn from the headspace and analysed for IC.
SAMPLING
On Days 0 and 28 samples (20 ml) were removed from the control and standard material vessels prepared for DOC analysis and filtered through Gelman 0.45 µm Acrocap filters (approximately 5 ml discarded) prior to DOC analysis. The samples were analysed for DOC using a Shimadzu TOC-5050A TOC analyser. See analytical methods for further details.
On day 28, 1 mL of concentrated hydrochloric acid was added to each vessel to drive off any inorganic carbonates formed. The vessels were resealed, aerated overnight and the final samples taken from both absorber vessels on Day 29.
Reference substance
- Reference substance:
- other: sodium benzoate
Results and discussion
- Preliminary study:
- Preliminary work showed that the test material was volatile and hence the conditions employed for a Modified CO2 Evolution Test (OECD Guideline No 301B) would not be suitable for the determination of the biodegradability of the test material. The test method was therefore changed to a CO2 Headspace Test, which is recommended for volatile materials.
The test material was added directly to the test vessels using a high precision volumetric syringe. Preliminary work conducted showed that a volume of 2.7 µl of test material injected into a test vessel using a gas tight micro-syringe gave a measured weight of 2.4 mg, mean of 15 separate weighings. - Test performance:
- No further information.
% Degradation
- Parameter:
- % degradation (inorg. C analysis)
- Value:
- 15
- Sampling time:
- 28 d
- Details on results:
- The biodegradation was assessed by measuring the inorganic carbon present in the headspace of the vessels. Total inorganic carbon values for the test material, standard material, toxicity control and control vessels was measured and are reported in the table below.
The mean TIC in the control vessels on Day 28 was 0.10 mg/l; equivalent to 5% of the organic carbon added initially as test material to the test vessels and therefore satisfied the validation criterion given in the Test Guideline. The test material attained 15% degradation after 28 days and hence cannot be considered to be readily biodegradable.
The toxicity control attained 41% degradation after 8 days and 44% degradation after 14 days thereby confirming that the test material was not toxic to the sewage treatment micro-organisms used in the study.
DOC analysis was also conducted and the results tabulated below. Samples taken from the standard material vessels on Days 0 and 28 (see Table 3) gave percentage degradation values of 100% and 99% respectively for Replicates R1 and R2. The degradation rates for the standard material were higher than those determined by IC analyses. This was considered to be due to incorporation of sodium benzoate into the microbial biomass prior to degradation and hence CO2 evolution occurring.
BOD5 / COD results
- Results with reference substance:
- Sodium benzoate attained 81% degradation after 14 days and 87% degradation 28 days thereby confirming the suitability of the inoculum and test conditions.
Any other information on results incl. tables
The results from the biodegradation tests are below:
Table1 Inorganic Carbon Values on Each Analysis Occasion
Day |
Control (mg TIC) |
Sodium Benzoate (mg TIC) |
Test Material (mg TIC) |
Toxicity Control (mg TIC) |
||||||||||||||||
R1 |
R2 |
R3 |
R4 |
R5 |
R1 |
R2 |
R3 |
R4 |
R5 |
R1 |
R2 |
R3 |
R4 |
R5 |
R1 |
R2 |
R3 |
R4 |
R5 |
|
0 |
0.09 |
0.13 |
0.11 |
- |
- |
0.09 |
0.09 |
0.09 |
- |
- |
0.10 |
0.09 |
0.09 |
- |
- |
0.09 |
0.08 |
0.09 |
- |
- |
2 |
0.09 |
0.12 |
0.09 |
- |
- |
1.36 |
1.32 |
1.08 |
- |
- |
0.09 |
0.09 |
0.09 |
- |
- |
- |
- |
- |
- |
- |
4 |
0.09 |
0.14 |
0.13 |
- |
- |
1.70 |
1.73 |
1.74 |
- |
- |
0.13 |
0.10 |
0.11 |
- |
- |
- |
- |
- |
- |
- |
8 |
0.18 |
0.13 |
0.13 |
- |
- |
2.06 |
2.00 |
1.94 |
- |
- |
0.25 |
0.18 |
0.17 |
- |
- |
1.90 |
1.84 |
2.00 |
- |
- |
10 |
0.08 |
0.12 |
0.13 |
- |
- |
1.88 |
1.93 |
1.82 |
- |
- |
0.23 |
0.20 |
0.21 |
- |
- |
- |
- |
- |
- |
- |
14 |
0.14 |
0.14 |
0.14 |
- |
- |
1.87 |
1.80 |
1.95 |
- |
- |
0.49 |
0.55 |
0.48 |
- |
- |
1.87 |
2.09 |
2.13 |
- |
- |
16 |
0.15 |
0.17 |
0.13 |
- |
- |
1.76 |
1.86 |
1.46 |
- |
- |
0.48 |
0.41 |
0.47 |
- |
- |
- |
- |
- |
- |
- |
21 |
0.07 |
0.18 |
0.16 |
- |
- |
1.78 |
1.74 |
1.84 |
- |
- |
0.48 |
0.59 |
0.51 |
- |
- |
- |
- |
- |
- |
- |
28 |
0.07 |
0.10 |
0.11 |
0.12 |
0.11 |
1.94 |
2.06 |
1.86 |
2.06 |
1.80 |
0.48 |
0.26 |
0.52 |
0.52 |
0.43 |
- |
- |
- |
- |
- |
Table2 Percentage Biodegradation Values
Day |
% Degradation |
||
Sodium Benzoate |
Test Material |
Toxicity Control |
|
0 |
0 |
0 |
0 |
2 |
54 |
0 |
- |
4 |
75 |
0 |
- |
8 |
86 |
2 |
41 |
10 |
83 |
5 |
- |
14 |
81 |
17 |
44 |
16 |
72 |
14 |
- |
21 |
77 |
18 |
- |
28 |
87 |
15 |
- |
Table 3 Dissolved Organic Carbon (DOC) Values in the Control and Standard Material Vessels on Days 0 and 28
DOC Concentration |
|||||||
Test Vessel |
Day 0 |
Day 28 |
|||||
mg C/l |
mg C/l Corrected for Mean Control Value |
% of Nominal Carbon Content |
mg C/l |
mg C/l Corrected for Mean Control Value |
% Degradation |
||
Control |
R1 |
2.18 |
- |
- |
<LOQ |
- |
- |
|
R2 |
2.11 |
- |
- |
1.43 |
- |
- |
Standard Material |
R1 |
23.05 |
20.90 |
105 |
1.25 |
<control |
100 |
|
R2 |
23.17 |
21.02 |
105 |
1.65 |
0.22 |
99 |
R1– R2= Replicates 1 and 2
LOQ = Limit of quantitation
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- under test conditions no biodegradation observed
- Conclusions:
- The test material attained 15% degradation after 28 days and therefore cannot be considered to be readily biodegradable.
- Executive summary:
Preliminary work showed that the test material was volatile and hence the conditions employed for a Modified CO2 Evolution Test (OECD Guideline No. 301B) would not be suitable for the determination of the biodegradability of the test material. The test method was therefore changed to a CO2 Headspace test, which is recommended for volatile materials. This test was conducted in sealed vessels and assessed the biodegradation of a test material by measuring the inorganic carbon present in the headspace of the vessels.
The test material, at a concentration of 20 mg C/l, was exposed to activated sewage sludge micro-organisms with mineral salts medium in sealed culture vessels in the dark at 20±1.0°C for 28 days.
The degeneration of the test material was assessed by the determination of carbon dioxide produced on Days 0, 2, 4, 8, 10, 14, 16, 21 and 28. Control solutions with inoculum and the standard material, sodium benzoate, together with a toxicity control were used for validation purposes.
The mean TIC in the control vessels on Day 28 was 0.10 mg/l; equivalent to 5% of the organic carbon added initially as test material to the test vessels and therefore satisfied the validation criterion given in the Test Guideline. The test material attained 15% degradation after 28 days and hence cannot be considered to be readily biodegradable.
The toxicity control attained 41% degradation after 8 days and 44% degradation after 14 days thereby confirming that the test material was not toxic to the sewage treatment micro-organisms used in the study. Sodium benzoate attained 81% degradation after 14 days and 87% degradation 28 days thereby confirming the suitability of the inoculum and test conditions. All validity criteria was therefore met.
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