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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
Guideline:
OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
GLP compliance:
no
Oxygen conditions:
aerobic
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:
benzoic acid, sodium salt
Key result
Parameter:
% degradation (O2 consumption)
Value:
49.4
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 arereported in Tables 2 and 3, respectively. The BOD28 value of test chemical was observed to be 1.25 mgO2/mg. ThOD was determined by calculation as 2.53 mgO2/mg. % Degradation was calculated using the values of BOD and ThOD for test item and was determined to be 49.4% at 20 ± 1°C. The % degradation of procedure control (reference item) was also calculated using BOD & ThOD and was determined to be 73.49% at 20 ± 1°C. Degradation of Sodium Benzoate exceeds 43.37% on 7th days & 58.43% 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 73.49% at 20 ± 1°C. Degradation of Sodium Benzoate exceeds 43.37% on 7th days & 58.43% 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

6.7

6.5

6.6

7

6.2

3.9

3.2

14

6

2

2

21

5.7

1

1

28

5.5

0.3

0.5

TABLE 2: BOD Values (mg O2/mg)

 

No. of Days

Test Suspension

Procedure Control

(Reference Item)

0

0

0

7

0.52

0.72

14

0.95

0.97

21

1.12

1.15

28

1.25

1.22     

 

TABLE 3: PERCENT BIODEGRDATION RESULTS

No. of Days

Test Suspension

Procedure Control

(Reference Item)

0

0%

0%

7

20.55%

43.37%

14

37.54%

58.43%

21

44.26%

69.27%

28

49.4%

73.49%

 

Table 4: BOD28, THOD AND % BIODEGRADATION VALUES

 

Method details

BOD28 (mgO2/mg)

ThOD (mgO2/mg)

%Biodegradation

Test Item

1.25

2.53

49.4

Reference Item

1.22

1.66

73.49

Validity criteria fulfilled:
yes
Interpretation of results:
inherently biodegradable
Conclusions:
The test chemical undergoes 49.4% biodegradation after 28 days in the test condition. Thus, the test chemical was considered to be inherently biodegradable in nature.
Executive summary:

Biodegradation study was conducted for 28-days Closed Bottle test following the OECD guideline 301 D to determine the ready biodegradability of the test chemical. The study was performed at a temperature of 20°C. The test system included control, test chemical and reference substance. Polyseed were used as a test inoculum for the study. 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. OECD mineral medium was used for the study. 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 chemical and reference substance. The % degradation of procedure control (reference item) was also calculated using BOD & ThOD and was determined to be 73.49% at 20 ± 1°C. Degradation of Sodium Benzoate exceeds 43.37% on 7th days & 58.43% on 14th day. The activity of the inoculum is thus verified and the test can be considered as valid. The BOD28 value of test chemical was observed to be 1.25 mgO2/mg. ThOD was calculated as 2.53 mgO2/mg. Accordingly, the % degradation of the test chemical after 28 days of incubation at 20 ± 1°C according to Closed Bottle test was determined to be 49.4%. Based on the results, the test chemical, under the test conditions, was considered to be inherently biodegradable in nature.

Description of key information

Biodegradation study was conducted for 28-days Closed Bottle test following the OECD guideline 301 D to determine the ready biodegradability of the test chemical (Experimental study report, 2017). The study was performed at a temperature of 20°C. The test system included control, test chemical and reference substance. Polyseed were used as a test inoculum for the study. 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. OECD mineral medium was used for the study. 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 chemical and reference substance. The % degradation of procedure control (reference item) was also calculated using BOD & ThOD and was determined to be 73.49% at 20 ± 1°C. Degradation of Sodium Benzoate exceeds 43.37% on 7th days & 58.43% on 14th day. The activity of the inoculum is thus verified and the test can be considered as valid. The BOD28 value of test chemical was observed to be 1.25 mgO2/mg. ThOD was calculated as 2.53 mgO2/mg. Accordingly, the % degradation of the test chemical after 28 days of incubation at 20 ± 1°C according to Closed Bottle test was determined to be 49.4%. Based on the results, the test chemical, under the test conditions, was considered to be inherently biodegradable in nature.

Key value for chemical safety assessment

Biodegradation in water:
inherently biodegradable

Additional information

Biodegradation study was conducted for 28-days Closed Bottle test following the OECD guideline 301 D to determine the ready biodegradability of the test chemical (Experimental study report, 2017). The study was performed at a temperature of 20°C. The test system included control, test chemical and reference substance. Polyseed were used as a test inoculum for the study. 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. OECD mineral medium was used for the study. 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 chemical and reference substance. The % degradation of procedure control (reference item) was also calculated using BOD & ThOD and was determined to be 73.49% at 20 ± 1°C. Degradation of Sodium Benzoate exceeds 43.37% on 7th days & 58.43% on 14th day. The activity of the inoculum is thus verified and the test can be considered as valid. The BOD28 value of test chemical was observed to be 1.25 mgO2/mg. ThOD was calculated as 2.53 mgO2/mg. Accordingly, the % degradation of the test chemical after 28 days of incubation at 20 ± 1°C according to Closed Bottle test was determined to be 49.4%. Based on the results, the test chemical, under the test conditions, was considered to be inherently biodegradable in nature.