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Biodegradation in water: screening tests

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Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
13 November 1995 - 18 December 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
1d: Test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Qualifier:
according to
Guideline:
OECD Guideline 301 A (Ready Biodegradability: DOC Die Away Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.4-A (Determination of the "Ready" Biodegradability - Dissolved Organic Carbon (DOC) Die-Away Test)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
- Name of test material: methoxy-3-propylamine
- Physical state: colourless liquid
- Analytical purity: 99.5 %
- Very soluble in water
- Lot/batch No.: 8028/95
- Carbon content: 53.9 %
Oxygen conditions:
aerobic
Inoculum or test system:
other: secondary effluent
Details on inoculum:
- Source of inoculum: purification station of Versailles
- 28000 bact/mL in the medium
Duration of test (contact time):
28 d
Initial conc.:
16.9 mg/L
Based on:
COD
Parameter followed for biodegradation estimation:
DOC removal
Details on study design:
TEST CONDITIONS
- Test temperature: 22 °C (+ or - 2 °C)
- Continuous darkness: yes

TEST SYSTEM
- 500 mL glass erlenmeyer flasks
- stoppered with cotton and gauze sterilized at 170 °C during 1 hour
Reference substance:
benzoic acid, sodium salt
Remarks:
analytical grade; initial substance concentration = 14.4 mg COD/L
Parameter:
% degradation (DOC removal)
Value:
< 10
Sampling time:
28 d
Results with reference substance:
> 70 % in 14 days
Validity criteria fulfilled:
yes
Interpretation of results:
under test conditions no biodegradation observed
Conclusions:
- Difference between the extreme values of the test flasks at the end of the test < 20 %.
- Percentage of elimination of DOC in the reference substance at day 14 is > 70 %.
- The substance is not inhibitory.
- No observed artefacts (adsorption, abiotic degradation).
Executive summary:

The aerobic biodegradation of methoxy-3-propylamine has been examinated through OECD Guideline 301 A and EU Method C.4-A.

DOC removal was examinated during 28 days from an initial concentration of 16.9 mg/l.

The results have shown that less than 10 % of the substance is degraded after 28 days.

Under test conditions, no biodegradation was observed.

The substance is not inhibitory.

Endpoint:
biodegradation in water: inherent biodegradability
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 302 B (Inherent biodegradability: Zahn-Wellens/EMPA Test)
GLP compliance:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): Methoxypropylamin
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, industrial (adaptation not specified)
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): activated sludge from BASF waste water treatment plant
Duration of test (contact time):
13 d
Initial conc.:
400 mg/L
Based on:
DOC
Details on study design:
TEST SYSTEM
- Culturing apparatus: 5 L glass bottle
- Number of culture flasks/concentration: 1
- Test performed in open system: According to the original report (81/009, 1981) the test was performed in "closed vessels". This term is misleading because all flasks were aerated with air (sparging)
- Other:
- preparation of the test assay: 423 mL stock solution of test substance (5200 mg/L); 14 mL inorganic medium; 100 mL stock solution of inoculum (30 g/L dry weight); 2462 mL tap water (=3000 mL liquid volume)
- inoculum: 1 g/L dry weight
- incubation: at room temperature (20-25 °C) on a magnetic stirrer, aerated with air (sparging)
- measurements: pH, DOC (dissolved organic carbon)

CONTROL AND BLANK SYSTEM
- Inoculum blank: no blank flask was tested in parallel but a statistically obtained mean value of 17 mg/L DOC was used to calculate the elimination
- Other:
Reference: no reference substance was tested in parallel
Parameter:
% degradation (DOC removal)
Value:
14
Sampling time:
3 h
Parameter:
% degradation (DOC removal)
Value:
5
Sampling time:
1 d
Parameter:
% degradation (DOC removal)
Value:
22
Sampling time:
4 d
Parameter:
% degradation (DOC removal)
Value:
40
Sampling time:
7 d
Parameter:
% degradation (DOC removal)
Value:
90 - 100
Sampling time:
13 d

Kinetic of elimination and pH:

Time

(days)

Test substance

(mg/L DOC)

Elimination (%)

pH

 Oxygen (mg/L)

0

400

-

7.0

9.2

3h

360

14

-

7.7 

1

391

5

8.2

 8.2

4

319

22

7.5

 8.3

7

245

40

-

 8.4

11

12

100

-

 8.4

12

-

-

7.5

 8.8

13

14

100

-

 8.6

- Comment on the calculation of the elimination (%): samples were taken at regular intervals and the sample volume (approx. 50 mL) was always

replaced with tap water. Thus, the starting volume remained constant during the whole test (approx. 3000 mL), but the reference value to calculate

the elimination degree had to be corrected after each sampling (factor: 0,9833)

- The sigmoidal shape of the elimination curve is an indication of biodegradation

Validity criteria fulfilled:
not specified
Interpretation of results:
other: easily eliminated from water (indication of biodegradation)
Conclusions:
Indication of biodegradation was observed.
Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 August 2011 - 14 September 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Well performed study under GLP conditions, according to OECD 301F guideline, but with extended exposure time. Therefore, this test should be considered an enhanced ready biodegradability test
Qualifier:
according to
Guideline:
OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
Deviations:
yes
Remarks:
; exposure time extended to 56 days
Qualifier:
according to
Guideline:
EU Method C.4-D (Determination of the "Ready" Biodegradability - Manometric Respirometry Test)
GLP compliance:
yes
Specific details on test material used for the study:
- Name of test material (as cited in study report): 3-methoxypropylamine
- Substance type: colourless
- Physical state: liquid
- Analytical purity: 99.4%
- Purity test date: 12-04-2010
- Lot/batch No.: 9M603
- Stability under test conditions: not reported
- Storage condition of test material: lab
Oxygen conditions:
anaerobic
Inoculum or test system:
activated sludge, domestic (adaptation not specified)
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Totnes Sewage Treatment Works, Totnes, Devon, UK, collected on 9 August 2011
- Storage conditions: aerated at room temperature and pH maintained at 7.0 ± 1.0
- Storage length: not specified
- Preparation of inoculum for exposure: 7 days prior to start of exposure the activated sludge was centrifuged, washed and resuspended in the mineral medium and the solids concentration determined
- Pretreatment: none
- Concentration of sludge: 30 mg/L
- Water filtered: no
Duration of test (contact time):
54 d
Initial conc.:
100 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Composition of medium: OECD standard medium
- Test temperature: 22 ± 1 °C
- pH: 7.2-7.5 at day 0 and 6.1-6.6 at day 56
- pH adjusted: yes, to 7.4 ± 0.2
- Suspended solids concentration: 30 mg/L
- Continuous darkness: yes, test performed in dark glass bottles

TEST SYSTEM
- Culturing apparatus: dark glass 500 mL bottle with Oxitop™ bottle top, on a magnetic stirrer in a constant temperature incubator
- Number of culture flasks/concentration: 3 blank, 3 reference substance, 3 test substance
- Measuring equipment: Oxitop™ respirometer (Wissenschaftlich-Technische Werkstatten, GmbH, Germany) and piezoresistive electronic pressure sensor in bottle top
- Details of trap for CO2 and volatile organics if used: potassium hydroxide (KOH) as trap located in seal cup in the neck of the bottles

STOCK SOLUTIONS
- a primary stock solution containing 1000 mg/L 3-methoxypropylamine or 1000 mg/L sodium benzoate was prepared in RO water
- stock solutions were clear and colourless
- stock solutions were prepared on day of exposure start

SAMPLING
- Sampling frequency: oxygen uptake every 224 minutes during 56 days, pH at day 0 in single bottle of inoculum blank and positive control
- Sampling method: oxygen uptake was measured as a decrease in pressure in the exposure bottle

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: no
- Toxicity control: yes, with sodium benzoate 100 mg/L

DATA ANALYSIS
- Oxygen uptake values were corrected for the inoculum blank.

Reference substance:
benzoic acid, sodium salt
Preliminary study:
Not applicable.
Key result
Parameter:
other: BOD:ThOD ratio
Value:
57
Sampling time:
56 d
Details on results:
As a significant lag phase was observed (13% biodegradation after 35 days) the test was extended over a 56 day period.
Key result
Parameter:
BOD5
Value:
1.27 g O2/g test mat.
Results with reference substance:
- sodium benzoate attained a maximum mean level of biodegradation (based on the BOD:ThOD ratio) of 60% after 56 days
- > 60% degradation was achieved within the 10 day window as expected for a biodegradable substance, thus confirming that the activated ludge contained viable organisms
Validity criteria fulfilled:
yes
Interpretation of results:
not readily biodegradable
Conclusions:
This study determined the ready biodegradability of 100 mg/L 3-methoxypropylamine in a 56-day Manometric respirometry test according to OECD 301F guidelines and in the presence of activated sewage sludge with 30 mg/L suspende solids. After 56 days on average 57% biodegradation was observed.The results show that 3-methoxypropylamine is not readily biodegradable.
Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))
GLP compliance:
no
Oxygen conditions:
aerobic
Inoculum or test system:
mixture of sewage, soil and natural water
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): In March, June, September, and December, sludge was sampled at the following 10 places in Japan: 1. Fukogawa city sewage plant, 2. Fukashiba industry sewage plant, 3. Nakahama city sewage plant, 4. Ochiai city sewage plant, 5. Kitakami river, 6. Shinano river, 7. Yoshino river, 8. Lake Biwa, 9. Hiroshima bay, 10. Dookai bay; sampling: 1. City sewage: Returned sludge from sewage plants was taken. 2. Rivers, lake and sea: Surface water and surface soil which were in contact with atmosphere were collected.
- Method of cultivation: About 30 minutes after ceasing aeration to the sludge mixture, supernatant corresponding to about 1/3 of the whole volume was removed. Then the equal volume of dechlorinated water was added to the remaining portion and aerated again, followed by addition of synthetic sewage at a concentration of 0.1% (w/v). This procedure was repeated once every day. The culturing was carried out at 25 ± 2 °C. 5 L of the filtrate of the supernatant of old activated sludge was mixed with 500 mL of the filtrate of the supernatant of new sludge and cultured at pH 7.0 ± 1.0 under sufficient aeration using prefiltered open air. During the cultivation, appearance of the supernatant, precipitability, formation of flock, pH, dissolved oxygen concentration in the solution and temperature were checked and necessary adjustments were made, Microflora in the activated sludge was microscopically observed and sludge with no abnormal symptom was used for the test.
- Concentration of sludge: 30 mg/L
Duration of test (contact time):
28 d
Initial conc.:
100 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Composition of medium: 3 mL each of four stock solutions, as described in JIS K 0102-1986-21, are diluted to 1000 mL with purified water
- pH: 7.0
- pH adjusted: yes
- Suspended solids concentration: determined according to Method Japanese Industrial Standards (JIS) K 0102-1986-14.1

TEST SYSTEM
- Culturing apparatus: Closed system oxygen consumption measuring apparatus (Coulometer: Ohkura Electric Co., Ltd.); 300 mL vessel, absorbent for evolving carbon dioxide Soda lime No .l (extra pure reagent, Wako Pure Chemical Industries, Ltd.).
- Number of culture flasks/concentration: 1
- Measuring equipment: Coulometer, Okhura Electric Co., Ltd.
- Test performed in open system: no
- Details of trap for CO2 and volatile organics if used: soda lime, extra pure, Wako Pure Chemical Industries, Ltd.)

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: yes
- Toxicity control: no
Reference substance:
aniline
Parameter:
% degradation (O2 consumption)
Value:
3
Sampling time:
4 wk
Validity criteria fulfilled:
not specified
Interpretation of results:
under test conditions no biodegradation observed
Conclusions:
Under the test conditions, no biodegradation was observed.

Description of key information

In a Manometric respirometry test according to OECD 301F guidelines and performed under GLP, the test substance showed 57% degradation under the test conditions after 56 days (Commander, 2011).

Key value for chemical safety assessment

Biodegradation in water:
inherently biodegradable, not fulfilling specific criteria
Type of water:
freshwater

Additional information

The reliable key study determined the ready biodegradability of 100 mg/L 3-methoxypropylamine in a 56-day Manometric respirometry test according to OECD 301F guidelines and in the presence of activated sewage sludge with 30 mg/L suspended solids. The test period was extended to 56 days as a significant lag phase was observed (13% biodegradation after 35 days); after 56 days on average 57% biodegradation was observed. The results show that the substance is biodegradable but does not meet the criteria for ready biodegradability.

Significant biodegradation was observed in an OECD 302B test (BASF AG 2001) (ultimate biodegradability of 90-100%) concluding that the substance is inherently biodegradable (not fulfilling specific criteria).