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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:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Experimental result using OECD guidelines.
Qualifier:
according to
Guideline:
OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
Principles of method if other than guideline:
Biodegradation study was conducted according to OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test) for evaluating the percentage biodegradability of test substance 3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,Ntrimethylanilinium chloride (CAS No 68391-31-1) .
GLP compliance:
no
Oxygen conditions:
not specified
Inoculum or test system:
other: Mixed inoculum
Details on inoculum:
Mixed Inoculum Preparation:Polyseed were used for this study. 1 polyseed capsule was added in 500 ml D.I water and then stirred for 1 hour for proper mixing and functioning of inoculum. This gave the bacterial count as 10E7 to 10E8 CFU/ml. At the regular interval microbial plating was also performed on agar to confirm the vitality and CFU count of microorganism.

Duration of test (contact time):
28 d
Initial conc.:
4 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Composition of medium: OECD mineral medium was used for the study
- Test temperature: 20°C
- Continuous darkness: Yes
- Other: The water used in this study is deionized water.

TEST SYSTEM
- Culturing apparatus: The apparatus used in this study is BOD bottles; with glass stoppers (125 ml), BOD incubator & oxygen electrode and meter.

CONTROL AND BLANK SYSTEM
- Inoculum blank: it contains only test inoculum
- Procedure control: contains reference compound and inoculum
Reference substance:
other: Sodium Benzoate
Key result
Parameter:
% degradation (O2 consumption)
Value:
40.69
Sampling time:
28 d
Remarks on result:
other: Other details not known
Details on results:
The oxygen consumed by the test systems was corrected for oxygen consumption occurring in the blank test systems. The BOD Values (mgO2/mg) and percent biodegradation results for each test system are reported in Tables 2 and 3, respectively. The BOD28 value of 3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride (CAS No.68391-31- 1) was observed to be 0.7 mgO2/mg. ThOD was determined by calculation as 1.72 mgO2/mg. % Degradation was calculated using the values of BOD and ThOD for test item and was found to be 40.69% at 20 ± 1°C. The % degradation of procedure control (reference item) was also calculated using BOD & ThOD and was determined to be 78.31%. Degradation of Sodium benzoate exceeds 13.25% on 7 days & 61.44% on 14th day. The activity of the inoculum was thus verified and the test can be considered as valid
Results with reference substance:
The % degradation of procedure control (reference item) was also calculated using BOD & ThOD and was determined to be 78.31%. Degradation of Sodium benzoate exceeds 13.25% on 7 days & 61.44% on 14th day. The activity of the inoculum was thus verified and the test can be considered as valid

TABLE 1:D.O VALUES(mg/L)

No. of Days

Inoculum Blank
(Control)

Test Suspension

Procedure Control
(Reference Item)

0

7.3

6.9

7

7

7

5.6

5.8

14

6.8

4.1

2.4

21

6.4

3.8

1.2

28

6.2

3

0.7

 

TABLE 2:BOD VALUES (mgO2/mg)

No. of Days

Test Suspension

Procedure Control
(Reference Item)

0

0

0

7

0.25

0.22

14

0.57

1.02

21

0.55

1.22

28

0.7

1.3

 

TABLE 3:PERCENT BIODEGRDATION RESULTS

No. of Days

Test Suspension

Procedure Control
(Reference Item)

0

0%

0%

7

14.53%

13.25%

14

33.13%

61.44%

21

31.97%

73.49%

28

40.69%

78.31%

 

TABLE 4:BOD28, THOD AND % BIODEGRADATION VALUES

Method details

BOD28 (mgO2/mg)

ThOD (mgO2/mg)

% Biodegradation

Test Item

0.7

1.72

40.69

Reference Item

1.3

1.66

78.31

 

Table 5: pH OF CLOSED BOTTLE TEST

No. of Days

Inoculum Blank
(Control)

Test Suspension

Procedure Control
(Reference Item)

0

6.5

7.6

7.9

7

6.6

6.6

5.8

14

6.7

6.4

6.7

21

6.2

6.3

6.2

28

6.2

6.3

6.5

 

Validity criteria fulfilled:
yes
Interpretation of results:
other: ultimate inherently biodegradable
Conclusions:
The 3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride(CAS No.68391-31-1) undergoes 40.69% biodegradation after 28 days in the test condition. Thus, the test item was considered to be ultimate inherently biodegradable in nature.
Executive summary:

28-days Closed Bottle test following the OECD guideline 301 D to determine the ready biodegradability of 3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl- 1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride, CAS No. 68391-31-1. The test system included control, test item and reference item. The concentration of test and reference item ( Sodium benzoate) chosen for both the study was 4 mg/L, while that of inoculum was 32 ml/l. ThOD (Theoretical oxygen demand) of test and reference item was determined by calculation. % degradation was calculated using the values of BOD and ThOD for test item and reference item. The BOD28 value of 3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,Ntrimethylanilinium chloride, CAS No. 68391-31-1 was observed to be 0.7 mgO2/mg. ThOD was calculated as 1.72 mgO2/mg. Accordingly, the % degradation of the test item after 28 days of incubation at 20 ± 1°C according to Closed Bottle test was determined to be 40.69%. Based on the results, the test item, under the test conditions, was considered to be ultimate inherently biodegradable.

 

Description of key information

The 3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride(CAS No.68391-31-1) undergoes 40.69% biodegradation after 28 days in the test condition. Thus, the test item was considered to be ultimate inherently biodegradable in nature.

Key value for chemical safety assessment

Biodegradation in water:
inherently biodegradable

Additional information

28-days Closed Bottle test following the OECD guideline 301 D to determine the ready biodegradability of 3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl- 1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride, CAS No. 68391-31-1. The test system included control, test item and reference item. The concentration of test and reference item ( Sodium benzoate) chosen for both the study was 4 mg/L, while that of inoculum was 32 ml/l. ThOD (Theoretical oxygen demand) of test and reference item was determined by calculation. % degradation was calculated using the values of BOD and ThOD for test item and reference item. The BOD28 value of 3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,Ntrimethylanilinium chloride, CAS No. 68391-31-1 was observed to be 0.7 mgO2/mg. ThOD was calculated as 1.72 mgO2/mg. Accordingly, the % degradation of the test item after 28 days of incubation at 20 ± 1°C according to Closed Bottle test was determined to be 40.69%. Based on the results, the test item, under the test conditions, was considered to be ultimate inherently biodegradable.