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Environmental fate & pathways

Biodegradation in water: screening tests

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Reference
Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data is from secondary source
Qualifier:
according to guideline
Guideline:
other: as mentioned below
Principles of method if other than guideline:
Preliminary screening of biodegradation of test chemical was carried out for 20 days for evaluating the percentage biodegradability of test substance 2,3,5-trimethylphenol.
GLP compliance:
not specified
Specific details on test material used for the study:
- Name of test material (as cited in study report): 2, 3, 5-Trimethylphenol
- Molecular formula :C9H12O
-Molecular weight :136.193 g/mol
- Smiles notation :c1(c(cc(C)cc1O)C)C
- InChl :1S/C9H12O/c1-6-4-7(2)8(3)9(10)5-6/h4-5,10H,1-3H3
- Substance type: Organic
- Physical state: Solid
Oxygen conditions:
aerobic
Inoculum or test system:
sewage, domestic, non-adapted
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Chapel Hill sewage treatment plant

- Concentration of sludge: 1.5 mg/l
Duration of test (contact time):
20 d
Initial conc.:
5 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Additional substrate: Standard nutrients were added to the water as was 0.5 mg/l allythiourea for control of nitrification.
- Test temperature: 18.33°C (65°F)
- Continuous darkness: Yes
- Other: Dilution water was prepared from water which had been passed through activated carbon and ion exchange columns and then glass-distilled.

TEST SYSTEM
- Culturing apparatus: BOD bottles
- Number of culture flasks/concentration: Duplicates
- Measuring equipment: O2 consumption was measured at various intervals over the 20 day period by means of a Weston and Stack dissolved oxygen meter.
- Test performed in closed vessels due to significant volatility of test substance: Test was performed in 300 ml BOD bottles which was stoppered.
- Other: The BOD bottles were filled, stoppered and incubated in the dark at 18.33°C (65°F) for 20 days.
Key result
Parameter:
% degradation (O2 consumption)
Value:
80
Sampling time:
20 d
Remarks on result:
other: Other details not known
Details on results:
Test substance undergoes 80% degradation by O2 consumption parameter in 20 days.

Table: Initial screening of test chemical found in coal conversion effluents (chemical was tested at 5 mg/l, initial oxygen conc. was 7.6 mg/l).

 

Test chemical

O2 depletion (%)

Days of incubation

 

 

2,3,5-trimethylphenol

5

10

15

20

 

56

 

75

 

79

 

80

Validity criteria fulfilled:
not specified
Interpretation of results:
readily biodegradable
Conclusions:
The percentage degradation of test substance was determined to be 80% by O2 consumption parameter in 20 days. Thus, based on percentage degradation, 2,3,5-trimethylphenol is considered to be readily biodegradable in water.
Executive summary:

Preliminary screening of biodegradation of test chemical was carried out for 20 days for evaluating the percentage biodegradability of test substance 2,3,5 -trimethylphenol. Domestic sewage was used as a test inoculum obtained from Chapel Hill sewage treatment plant. Concentration of inoculum i.e, sludge used was 1.5 mg/l and initial test substance conc. used in the study was 5 mg/l, respectively. Test chemical was added to duplicate, acid washed, 300 ml BOD bottles. Dilution water was prepared from water which had been passed through activated carbon and ion exchange columns and then glass-distilled. Standard nutrients were added to the water as was 0.5 mg/l allythiourea for control of nitrification. The dilution water was seeded with 1.5 mg/l of domestic sewage. The BOD bottles were filled, stoppered and incubated in the dark at 18.33°C (65°F) for 20 days. O2 consumption was measured at various intervals over the 20 day period by means of a Weston and Stack dissolved oxygen meter. The percentage degradation of test substance was determined to be 80% by O2 consumption parameter in 20 days. Thus, based on percentage degradation, 2,3,5 -trimethylphenol is considered to be readily biodegradable in nature.

Description of key information

Preliminary screening of biodegradation of test chemical was carried out for 20 days for evaluating the percentage biodegradability of test substance 2,3,5 -trimethylphenol (PC. Singer et. al; 1978). Domestic sewage was used as a test inoculum obtained from Chapel Hill sewage treatment plant. Concentration of inoculum i.e, sludge used was 1.5 mg/l and initial test substance conc. used in the study was 5 mg/l, respectively. Test chemical was added to duplicate, acid washed, 300 ml BOD bottles. Dilution water was prepared from water which had been passed through activated carbon and ion exchange columns and then glass-distilled. Standard nutrients were added to the water as was 0.5 mg/l allythiourea for control of nitrification. The dilution water was seeded with 1.5 mg/l of domestic sewage. The BOD bottles were filled, stoppered and incubated in the dark at 18.33°C (65°F) for 20 days. O2 consumption was measured at various intervals over the 20 day period by means of a Weston and Stack dissolved oxygen meter. The percentage degradation of test substance was determined to be 80% by O2 consumption parameter in 20 days. Thus, based on percentage degradation, 2,3,5 -trimethylphenol is considered to be readily biodegradable in nature.

Key value for chemical safety assessment

Biodegradation in water:
readily biodegradable

Additional information

Various experimental key study and supporting data for the target compound 2,3,5-trimethylphenol (CAS no. 697-82-5) were reviewed for the biodegradation end point which are summarized as below:

 

In an experimental key study, preliminary screening of biodegradation of test chemical was carried out for 20 days for evaluating the percentage biodegradability of test substance 2,3,5 –trimethylphenol (CAS no. 697-82-5) (PC. Singer et. al; 1978). Domestic sewage was used as a test inoculum obtained from Chapel Hill sewage treatment plant. Concentration of inoculum i.e, sludge used was 1.5 mg/l and initial test substance conc. used in the study was 5 mg/l, respectively. Test chemical was added to duplicate, acid washed, 300 ml BOD bottles. Dilution water was prepared from water which had been passed through activated carbon and ion exchange columns and then glass-distilled. Standard nutrients were added to the water as was 0.5 mg/l allythiourea for control of nitrification. The dilution water was seeded with 1.5 mg/l of domestic sewage. The BOD bottles were filled, stoppered and incubated in the dark at 18.33°C (65°F) for 20 days. O2 consumption was measured at various intervals over the 20 day period by means of a Weston and Stack dissolved oxygen meter. The percentage degradation of test substance was determined to be 80% by O2 consumption parameter in 20 days. Thus, based on percentage degradation, 2,3,5 -trimethylphenol is considered to be readily biodegradable in nature.

 

Another study of biodegradation was carried out for 14 days for evaluating the percentage biodegradability of test substance 2,3,5-trimethylphenol (CAS no. 697-82-5) (JAMES G. MUELLER et. al; 1991). Micro-organism was used as a test inoculum isolated from surface soil which was freshly obtained from theAmerican Creosote Works site. Microbial inoculum was prepared by mixing 25 g of creosote-contaminated surface soil (17) freshly obtained from the American Creosote Works site with 100 ml of 2.5 mM phosphate buffer (pH 7). After being mixed well, the suspension was centrifuged (2,500 rpm, 10 min) to remove larger soil particles. The resultant supernatant was decanted and used as a source of indigenous, "creosote-adapted" microorganisms for the ground water medium (GWM).Ground water medium (GWM) was used for the study.For the biodegradation study, approximately 400 liters of groundwater contaminated with creosote and PCP was recovered from an on-site sampling well. Groundwater was removed from a depth of 7 m through Teflon-coated Bev-a-line tubing (15 mm inner diameter) by means of an electric pump. Groundwater was delivered directly into two freshly rinsed, 208-liter steel drums (DOT-17E) and stored on site for ancillary testing. Five subsamples (1.0 liter) were collected in clean, sterile, 1.0-liter Wheaton bottles fitted with Teflon-lined screw caps and stored on ice for transport to the laboratory. Upon arrival at the laboratory, subsamples were stored in darkness at 2°C for subsequent biodegradation studies, toxicity and teratogenicity testing, and chemical analyses.

It contains2.5 ml of filtered groundwater (passed through a plug of silanized glass wool to remove undissolved solids) plus 12.5 ml of modified Bushnell-Haas medium.Additionally, two clean, sterile, 1.0-liter Wheaton bottles fitted with Teflon-lined screw caps received 200 ml of the same medium.Ground water contains the test chemical2,3,5-trimethylphenol. Wheaton bottles consist of 200 ml of ground water medium (which contain ground water and modified Bushnell-Haas medium. Duplicate 25 ml samples wereimmediately extracted for time zero analysis. Flasks were incubated at 30°C with shaking (200 rpm) in the dark for 14 days. Killed-cell controls were prepared for each sampling time point by adding 2.5 ml of a 37% formaldehyde solution to five of the shake flasks containing 25 ml of GWM. After 1, 3, 5, 8, and 14 days of incubation, the entire contents of two active flasks and one killed-cell control flask were separately extracted and analyzed by GC for the presence of creosote constituents. These data were compared with those obtained from untreated (non-inoculated) GWM that had been stored at 2°C during the 14-day incubation period.As the limit of detection of test chemical by GC is very low during a period 14 days, test chemical2,3,5-trimethylphenol can be considered to be readily biodegradable in nature. 

 

In a supporting study from peer reviewed journal (HIDESHI YANASE et. al; 1992), biodegradation experimented was conducted for 30 mins for evaluating the percentage biodegradability of test substance 2,3,5 –trimethylphenol (697-82-5). Phenol degrading bacteria (Agrobacterium spp.) was used as a test organism isolated from brine that had just emerged from a natural gas well under the sea near Niigata Prefecture, Japan. The identification of bacteria was carried out by the National Collection of Industrial and Marine Bacteria, Ltd., Scotland. Bacterial cells suspended in sterile artificial seawater containing 10% (vol/vol) glycerol were stored frozen at -80°C in plastic vials. Under the storage conditions, the isolates were viable and did not lose phenol degradation activity for at least 2 years. The reaction mixture contained 100 mM MOPS buffer (pH 7.0), 0.1 mM (13.61 mg/l) phenolic compound, and 4.5 mg/mi cells in a total volume of 2.2 ml in a test tube (18 x 180 ram). The reaction occurred at 50°C for 30 mins with reciprocal shaking (230 strokes/rnin).The percentage degradation of test substance was determined to be 21.4 and 20.1% degradation in 30 mins using 10% and 100% seawater, respectively. Thus, based on percentage degradation, 2,3,5 -trimethylphenol is considered to be inherently biodegradable in nature.

 

On the basis of above results for target chemical 2,3,5 -trimethylphenol (frompeer reviewed journal), it can be concluded that the test substance 2,3,5-trimethylphenol can be expected to be readily biodegradable in nature.