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Environmental fate & pathways

Biodegradation in water: screening tests

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Administrative data

Endpoint:
biodegradation in water: inherent biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Rationale was chosen based on high deviations between replicates and inconsistent COD/BOD values

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))
Version / remarks:
12 May 1981
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
1,1'-(methylenedi-p-phenylene)bismaleimide
EC Number:
237-163-4
EC Name:
1,1'-(methylenedi-p-phenylene)bismaleimide
Cas Number:
13676-54-5
Molecular formula:
C21H14N2O4
IUPAC Name:
1,1'-[methylenedi(4,1-phenylene)]di(1H-pyrrole-2,5-dione)

Study design

Oxygen conditions:
aerobic
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): Domestic waste-water plant (ARA Sissach/Switzerland)
- Pretreatment: The sludge used for this study was washed three times with tap water and an amount corresponding to of 4 g dry material per litre was mixed with Soerensen buffer solution (pH 7) and then aerated prior to incubation. About 30 minutes after stopping the aeration of the sludge, approximately 1/3 of the whole volume of the supernatant was removed. An equal volume of 0.1 % glucose, peptones and monopotassium phosphate was added to the remaining sludge, and aerated again. The pH of the sludge was adjusted to 7 ± 1.
- Concentration of sludge: 25 mg for the treated and the untreated flasks and 7.5 mg for the ANILINE treated flasks
Duration of test (contact time):
28 d
Initial test substance concentration
Initial conc.:
30 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimationopen allclose all
Parameter followed for biodegradation estimation:
O2 consumption
Parameter followed for biodegradation estimation:
DOC removal
Details on study design:
TEST CONDITIONS
- Composition of medium: according to OECD guideline 302C
- Test temperature: 22-31°C, was controlled daily during the study. The temperature was slightly above the temperature recommended by the guidelines. However, it can be stated that these transitory temperature differences had no effect on the integrity of the test.
- pH: Was monitored for all incubation flasks at the end of the incubation period.
- pH adjusted: yes, in the final test medium and the sludge
- Aeration of dilution water: At preparation of the flasks
- Continuous darkness: yes by means of brown glass bottles

TEST SYSTEM
- Culturing apparatus: BSB 1000 and BSB 601 apparatus (Wissenschaftliche-Technische Werkstätten GmbH, Weilheim, FRG)
- Number of culture flasks/concentration: 3 for the treated flasks and one each for the negative controls.
Flask 9: 250 ml bidistilled water and 7.5 mg test article
Flask 10+11+12: 250 ml final test medium, 7.5 mg test article (pH 7.0) and 25 mg inoculum.
Flask 13+14+15: 250 ml final test medium, 25 mg ANILINE and 7.5 mg inoculum.
Flask 16: 250 ml final test medium and 25 mg inoculum.
- Measuring equipment: carbon analyser (Technicon Autoanalyser II)
- Details of trap for CO2 and volatile organics if used: The CO2 was adsorbed by NaOH and the resulting drop in pressure was read on a manometer. The test vessel was opened and the NaOH replaced three times per week, except between day 8 and day 13.

SAMPLING
- Sampling frequency: daily

CONTROL AND BLANK SYSTEM
- Inoculum blank: final test medium + inoculum
- Abiotic sterile control: bidistilled water + test article

Reference substance
Reference substance:
aniline

Results and discussion

% Degradation
Key result
Parameter:
% degradation (O2 consumption)
Value:
37.8
Sampling time:
28 d
Details on results:
A degradation of 0 - 55.3 % was observed when estimated using the DOC-values. Furthermore, it has to be stated that a degradation in the same order of magnitude was observed in the flask without inoculum, allowing the assumption that the test article was not only degraded by microorganisms. It cannot be excluded that the insolubility of COMPIMIDE MDAB in aqueous solution may prevent the test article to be optimally degraded by aquatic microorganisms. Therefore, the test article may be degradable to a higher amount as indicated by the value in the present study.

BOD5 / COD results

Results with reference substance:
The standard compound, ANILINE, was readily degraded after 14 and 28 days by 64.0% and 64.3% (BOD/ThOD), respectively.

Any other information on results incl. tables

Table 1: Biochemical Oxygen Demand (BOD) of the test item and ANILINE

Flask No.

Test article concentration (mg/mL)

Cumulative BOD-values after x-days (mgO2/100 mg test article)

1

2

3

4

5

6

7

91

7.5

58.3

58.3

58.3

58.3

58.3

58.3

58.3

10

7.5

50

50

50

50

50

50

66.7

11

7.5

83.3

83.3

83.3

83.3

100

100

133.3

12

7.5

0

0

0

0

0

0

0

13

25 (ANILINE)

0

0

105

150

150

150

150

14

25 (ANILINE)

0

0

105

135

135

135

135

15

25 (ANILINE)

0

0

120

175

177.5

177.5

177.5

Flask No.

Test article concentration (mg/mL)

Cumulative BOD-values after x-days (mgO2/100 mg test article)

8

9

10

11

12

13

14

91

7.5

58.3

75

75

75

75

75

75

10

7.5

66.7

66.7

66.7

66.7

66.7

66.7

66.7

11

7.5

133.3

133.3

150

150

200

200

225

12

7.5

0

0

0

0

0

0

0

13

25 (ANILINE)

150

150

150

150

150

150

150

14

25 (ANILINE)

135

135

135

135

135

135

135

15

25 (ANILINE)

177.5

177.5

177.5

177.5

177.5

177.5

177.5

Flask No.

Test article concentration (mg/mL)

Cumulative BOD-values after x-days (mgO2/100 mg test article)

15

16

17

18

19

20

21

91

7.5

75

83.3

83 .3

83.3

83.3

83.3

100

10

7.5

66.7

66.7

66.7

66.7

83.3

83.3

83.3

11

7.5

225

225

258.3

258.3

300

300

325

12

7.5

0

0

8.3

8.3

41.7

41.7

41.7

13

25 (ANILINE)

150

150

150

150

150

150

150

14

25 (ANILINE)

135

135

137.5

137.5

137.5

137.5

137.5

15

25 (ANILINE)

177.5

177.5

177.5

177.5

177.5

177.5

177.5

Flask No.

Test article concentration (mg/mL)

Cumulative BOD-values after x-days (mgO2/100 mg test article)

22

23

24

25

26

27

28

91

7.5

100

100

100

100

100

100

100

10

7.5

83.3

83.3

83.3

83.3

83.3

83.3

83.3

11

7.5

325

375

375

375

525

533.3

533.3

12

7.5

41.7

41.7

41.7

41.7

58.3

58.3

58.3

13

25 (ANILINE)

150

150

150

150

150

150

150

14

25 (ANILINE)

137.5

137.5

137.5

137.5

137.5

137.5

137.5

15

25 (ANILINE)

177.5

177.5

177.5

177.5

177.5

177.5

177.5

1: Test article without inoculum

Table 2: Degradation of the test item and ANILINE (BOD/ThOD)

Flask No.

Test article concentration (mg/ 250mL)

Percentage BD after x days

1

2

3

4

5

6

7

91

7.5

31.1

31.1

31.1

31.1

31.1

31.1

31.1

10

7.5

26.7

26.7

26.7

26.7

26.7

26.7

35.6

112

7.5

44.4

44.4

44.4

44.4

53.3

53.3

71.1

12

7.5

0

0

0

0

0

0

0

MEAN3

 

13.4

13.4

13.4

13.4

13.4

13.4

17.8

13

25 (ANILINE)

0

0

43.6

62.2

62.2

62.2

62.2

14

25 (ANILINE)

0

0

43.6

56

56

56

56

15

25 (ANILINE)

0

0

49.8

72.6

73.7

73.7

73.7

MEAN

 

0

0

45.7

63.6

64

64

64

Flask No.

Test article concentration (mg/mL)

Percentage BD after x days

8

9

10

11

12

13

14

91

7.5

31.3

40

40

40

40

40

40

10

7.5

35.6

35.6

35.6

35.6

35.6

35.6

35.6

112

7.5

71.1

71.1

80

80

106.7

106.7

120

12

7.5

0

0

0

0

0

0

0

MEAN3

 

17.8

17.8

17.8

17.8

17.8

17.8

17.8

13

25 (ANILINE)

62.2

62.2

62.2

62.2

62.2

62.2

62.2

14

25 (ANILINE)

56

56

56

56

56

56

56

15

25 (ANILINE)

73.7

73.7

73.7

73.7

73.7

73.7

73.7

MEAN

 

64

64

64

64

64

64

64

Flask No.

Test article concentration (mg/mL)

Percentage BD after x days

15

16

17

18

19

20

21

91

7.5

40

44.4

44.4

44.4

4.4

44.4

53.3

10

7.5

35.6

35.6

35.6

35.6

44.4

44.4

44.4

112

7.5

120

120

137.8

1378

160

160

173.3

12

7.5

0

0

4.4

4.4

22.2

22.2

22.2

MEAN3

 

17.8

17.8

20

20

33.3

33.3

33.3

13

25 (ANILINE)

62.2

62.2

62.2

62.2

62.2

62.2

62.2

14

25 (ANILINE)

56

56

57.1

57.1

57.1

57.1

57.1

15

25 (ANILINE)

73.7

73.7

73.7

73.7

73.7

73.7

73.7

MEAN

 

64

64

64.3

64.3

64.3

64.3

64.3

Flask No.

Test article concentration (mg/mL)

Percentage BD after x days

22

23

24

25

26

27

28

91

7.5

53.3

53.3

53.3

53.3

53.3

53.3

53.3

10

7.5

44.4

44.4

44.4

44.4

44.4

44.4

44.4

112

7.5

173.3

200

200

200

280

284.4

284.4

12

7.5

22.2

22.2

22.2

22.2

31.1

31.1

31.1

MEAN3

 

33.3

33.3

33.3

33.3

37.8

37.8

37.8

13

25 (ANILINE)

62.2

62.2

62.2

62.2

62.2

62.2

62.2

14

25 (ANILINE)

57.1

57.1

57.1

57.1

57.1

57.1

57.1

15

25 (ANILINE)

73.7

73.7

73.7

73.7

73.7

73.7

73.7

MEAN

 

64.3

64.3

64.3

64.3

64.3

64.3

64.3

1: Test article without inoculum

2: Qutlier value

3: Mean of flask 10 and 12

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Interpretation of results:
not readily biodegradable
Conclusions:
In the present study conducted according to OECD guideline 302C (adopted 12 May 1981) activated sewage sludge was incubated with 30 mg of the test item for 28 days. Measured BOD/ThOD values relvealed a biodegradation of 37.8% within 28 days, whereas the claculated DOC-values were between 0 and 53% degradation.