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Reference
Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 September 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation))
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 850.6800 (Modified Activated Sludge, Respiration Inhibition Test for Sparingly Soluble Chemicals)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Name: AMINOX®
Chemical name: 2-Propanone, reaction products with diphenylamine
CAS number: 68412-48-6
Batch/Lot Number: T7C16001
Description: Green to brown flakes
Purity: 100% (as a UVCB)
Expiry date: 14 March 2019
Storage conditions: Controlled room temperature (15-25 ºC, below 70 RH%)
Safety precautions: Routine safety precautions (lab coat, gloves, safety glasses, face mask) for unknown materials were applied to assure personnel health and safety.
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
REFERENCE ITEM
Reference item: 3,5-Dichlorophenol
Manufacturer: ACROS ORGANICS
Batch No.: A0376465
Expiry Date: 31 August 2021
Storage: In original container, at room temperature
Formulation
Because of the low solubility, the powdered test item was added by directly weighing the test item into the bottles (and using ultrasonic bath for 10 minutes). The test formulation was freshly prepared at the beginning of the experiment, in the testing laboratory.
The pH of the solutions was checked but not adjusted due to the direct application of the test substance.

CONTROLS
Abiotic Control (A)
In parallel to the Study, abiotic control flasks (A1, A2, A3, A4 and A5) were tested without inoculum, but with the highest concentration of the test item (1000 mg/L nominal concentration), under identical test conditions.

Untreated Control (B)
1-1 member of five replicates (B1, B2, B3, B4 and B5) of the untreated control group (deionised water, synthetic sewage and inoculum, without addition of the test item) was tested in parallel with all of the test series.

Reference Control (REF)
In parallel with the test item, the reference item 3,5-Dichlorophenol (REF1, REF2, REF3, REF4) was tested (at nominal test concentrations of 1.0; 3.2; 10 and 32 mg/L) under identical test conditions.
A stock solution of 3,5-Dichlorophenol was prepared according to the OECD Guideline No. 209 (1.0 g of powdered 3,5-Dichlorophenol dissolved in 1000 mL of water). Warm deionised water was used to help the dissolution of the reference substance and the volume of the solution was made up to 1.0 L with deionised water, then cooled to room temperature. The pH of the solution was checked and adjusted with NaOH solution (2 mol/L) to pH 7.49.

ADMINISTRATION OF THE TEST ITEM
Test Concentrations
The test was performed at three concentrations of the test item: 10 (TI11, TI12, TI13, TI14, TI15), 100 (TI21, TI22, TI23, TI24, TI25) and 1000 mg/L (TI31, TI32, TI33, TI34, TI35) were used.
Test organisms (species):
activated sludge of a predominantly domestic sewage
Details on inoculum:
Preparations of the test flasks
One test solution with a final volume of 400 mL (ratio of composition of each test mixture referring to 500 mL according to the guideline) was prepared per treatment in a glass flask.
A volume of 12.8 mL synthetic sewage and an adequate amount of the test item were placed into the test flask by direct addition and filled up with deionised water to 200 mL before the start of the incubation. At the start of the test, 200 mL activated sludge inoculum with a sludge concentration of 3 g per litre of suspended solids was added to all flasks, with the exception of the abiotic control flasks.
Abiotic controls in 5 parallels (A1, A2, A3, A4 and A5), and one untreated blank (B1) control was started as the first step of the test. Then at appropriate time intervals of approximately 30 minutes the further test groups were started. The second series were 4 different concentrations (without replicates) of the reference item (REF1, REF2, REF3, REF4) and to the second blank control (B2). The third series was the 5 replicates of the test item concentration of 10 mg/L nominal concentration (TI11, TI12, TI13, TI14 and TI15) and the third blank control (B3). The fourth series was the five replicates of test item concentration 100 mg/L nominal concentration (TI21, TI22, TI23, TI24 and TI25) and the fourth blank control (B4). The final series was the five replicates of test item concentration 1000 mg/L nominal concentration (TI31, TI32, TI33, TI34 and TI35) and the fifth blank control (B5).
In summary, 3 test item concentration series and one abiotic control series were prepared in 5 parallels with one blank control series by series. In parallel with the above test mixtures four reference item concentrations without replicates were also prepared with one blank control.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
3 h
Hardness:
Not specified
Test temperature:
21.5°C
pH:
7.38
Dissolved oxygen:
Not specified
Salinity:
Not specified
Conductivity:
Not specified
Nominal and measured concentrations:
The test was performed at three concentrations of the test item: 10 (TI11, TI12, TI13, TI14, TI15), 100 (TI21, TI22, TI23, TI24, TI25) and 1000 mg/L (TI31, TI32, TI33, TI34, TI35)
Details on test conditions:
TEST SYSTEM AND TEST CONDITIONS
Species: Activated sludge, microorganisms from a domestic waste water treatment plant.
Source: The activated sludge was supplied from the sewage plant for domestic sewage in Veszprém, Hungary.
Conditioning: The activated sludge was supplied by the sewage plant for domestic sewage two days before the start of the experiment. During holding prior to use the sludge was fed daily with 50 mL synthetic sewage (see 3.7.2.) per litre and kept aerated at 20 ± 2°C until use.
The activated sludge used for this study was washed and centrifuged and the supernatant liquid phase was decanted. The solid material was re-suspended in chlorine-free tap water and again centrifuged.
An aliquot of the final sludge suspension was weighed, dried and the ratio of wet sludge to dry weight determined.
Based on this ratio, calculated amounts of wet sludge were suspended in chlorine-free tap water to yield a concentration equivalent to 3 g per litre (on dry weight basis). The pH of the activated sludge inoculum was determined to be pH 7.38. The activated sludge was used directly after conditioning.

Test units
Type and size: Appropriate glass beakers for 500 mL volume and BOD bottles with 300 mL volume.
Identification: Each test flask was uniquely identified with study code, treatment and replicate codes.

Test conditions
Surrounding type: Temperature controlled laboratory
Temperature: 21.2 – 21.5 °C
Aeration: With compressed air (about 1 L/min)
Recording: Test conditions were measured with suitable instruments and documented in the raw data.

Synthetic Sewage Feed (ratio of composition of culture media referring to 1000 mL)
Peptone 16.0 g
Meat extract 11.0 g
Urea 3.0 g
NaCl 0.7 g
CaCl2 x 2H2O 0.4 g
MgSO4 x 7H2O 0.2 g
K2HPO4 2.8 g
Deionised water add to 1000.0 mL

Measurement of Respiration Rate (Dissolved Oxygen)
For the measurement of the respiration rate a well-mixed sample of each treatment was poured into a BOD flask after about 3 hours incubation time, and was not further aerated. The oxygen concentration was measured for about fifteen minutes, with an O2 electrode and was recorded in every half minute. The oxygen consumption (in mg O2 L-1 minute-1) was determined from the most linear part of the respiration curve.

Measurement of pH and temperature
The pH of the test mixtures was determined at the start and at the end of the incubation period in all test vessels. The pH data are presented in Table 2 (Section 4.). The temperature was measured in the laboratory with a min/max thermometer during the experiment.
Reference substance (positive control):
yes
Remarks:
3,5-Dichlorophenol (3,5-DCP)
Key result
Duration:
3 h
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Details on results:
In comparison to the inoculum controls the inhibition of the respiration rate in the case of the activated sludge was between -0.89 % and -4.15 % in the examined range of 10 – 1000 mg/L of test item.
The average specific respiration rate (RS) of the untreated (Blank) controls (B1-B5) was 33.16 mg O2/L/h/g solid.
The variation coefficient of oxygen uptake rate in above untreated replicates was 3.54 %.
Results with reference substance (positive control):
The following nominal concentrations of the reference control 3,5-Dichlorophenol were tested on the same activated sludge and under identical conditions as in the case of the test item: 1.0, 3.2, 10.0 and 32.0 mg/L.
In comparison to the controls the inhibition of the respiration rate of the activated sludge was 79.73 % at the highest nominal concentration of 32.0 mg/L.
At the nominal concentrations of 1.0, 3.2 and 10.0 mg/L 5.21 %, 12.04% and 21.00 % inhibition of the respiration rate was calculated respectively.
The 3-hour EC50 of 3,5-Dichlorophenol was calculated to be 15.84 mg/L with 95% confidence limits of 12.65 – 19.84 mg/L.

Influence of test item on oxygen consumption of activated sludge

Test group

 

Conc. of test item in test mix. (mg/L)

 

Total oxygen consumption rate (RT) (mg O2/L/h)

 

Specific respiration rate (RS) (mg O2/L/h/g solid)

 

Inhib. (%)

 

pH

values#

Temperature (C°)

(measured in the Laboratory)*

Variation coefficient of O2consumption rate (%)

 

ID

 

Name

 

Start

End

A1

 

Abiotic

 

1000

 

0.79

 

-

-

7.59

7.92

21.2/21.5

-

A2

 

1000

 

0.70

-

-

7.61

7.92

-

A3

 

1000

 

0.58

-

-

7.59

7.92

-

A4

 

1000

 

0.58

-

-

7.58

7.91

-

A5

 

1000

 

0.58

-

-

7.57

7.91

-

B1

Blank

0.00

49.20

32.80

-

7.41

7.62

21.2/21.5

3.54

B2

0.00

51.20

34.13

-

7.43

8.03

B3

0.00

51.00

34.00

-

7.58

7.99

B4

0.00

50.40

33.60

-

7.72

8.12

B5

0.00

46.92

31.28

-

7.80

8.24

REF1

REF. item

1.00

47.80

31.87

5.21

7.39

7.98

21.2/21.5

-

REF2

3.20

44.40

29.60

12.04

7.39

7.99

-

REF3

10.00

39.94

26.63

21.00

7.39

8.02

-

REF4

32.00

10.73

 

7.15

79.73

7.42

8.06

-

TI1

Test item

10

52.45

34.97

-4.15

7.57

7.96

21.2/21.5

-

TI2

100

51.95

34.63

-3.13

7.72

8.10

-

TI3

1000

50.83

33.89

-0.89

7.80

8. 22

-

#: Individual pH values of the BOD flasks, except TI1-3, where the average values are reported

*: Minimum / Maximum temperature (°C) measured in the Laboratory, during the experiment

 


 

Oxygen consumption data of the abiotic test mixtures

 

Half minutes

ID of Oxygen meter (LO1, LO2, LO3)

LO1/120300068133

LO2/120300068181

LO3/120300068157

ID of Oxygen meter electrodes (L,R)

120132594001

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

A1

A2

A3

A4

A5

Bi

1

8.38

8.26

8.31

8.49

8.22

6.58

2

8.33

8.17

8.23

8.41

8.15

5.16

3

8.30

8.11

8.21

8.39

8.13

4.68

4

8.27

8.10

8.21

8.39

8.11

4.25

5

8.25

8.09

8.20

8.37

8.11

3.84

6

8.28

8.08

8.19

8.38

8.10

3.43

7

8.24

8.08

8.18

8.37

8.10

3.02

8

8.24

8.06

8.17

8.37

8.10

2.61

9

8.23

8.08

8.18

8.36

8.09

2.20

10

8.25

8.06

8.18

8.37

8.09

1.81

11

8.25

8.08

8.17

8.36

8.08

1.38

12

8.22

8.05

8.18

8.36

8.08

0.97

13

8.24

8.05

8.18

8.35

8.08

0.57

14

8.21

8.05

8.18

8.35

8.08

0.21

15

8.21

8.06

8.17

8.36

8.08

0.12

16

8.24

8.06

8.17

8.35

8.07

0.11

17

8.24

8.05

8.17

8.36

8.08

0.10

18

8.26

8.05

8.17

8.35

8.08

0.09

19

8.22

8.03

8.18

8.35

8.08

0.09

20

8.24

8.04

8.17

8.34

8.08

0.09

21

8.25

8.06

8.17

8.35

8.08

0.08

22

8.24

8.06

8.18

8.36

8.07

0.08

23

8.19

8.04

8.17

8.36

8.07

0.08

24

8.18

8.08

8.17

8.36

8.07

0.08

25

8.21

8.07

8.17

8.35

8.07

0.07

26

8.19

8.08

8.17

8.34

8.07

0.07

27

8.21

8.02

8.16

8.35

8.07

0.07

28

8.21

8.00

8.17

8.35

8.07

0.07

29

8.22

8.10

8.16

8.35

8.07

0.07

30

8.19

8.09

8.17

8.35

8.07

0.07

Q1

8.38

8.26

8.31

8.49

8.08

4.25

Q2

8.19

8.09

8.17

8.35

8.08

3.02

Dt

14.5

14.5

14.5

14.5

14.5

1.5

RT(mg O2/L/minute)

0.01

0.01

0.01

0.01

0.01

0.82

RT(mg O2/L/hour)

0.79

0.70

0.58

0.58

0.58

49.20

RT(average)

0.65

RTs(mg O2/L/h/g solid sludge)

-

32.80

 

Oxygen consumption data of the reference item test mixtures

Half minutes

ID of Oxygen meter (LO1, LO2, LO3)

LO1/120300068133

LO2/120300068181

LO3/120300068157

ID of Oxygen meter electrodes (L,R)

120132594001

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

Conc. (mg/L) of the Reference item

1.0

3.2

10.0

32.0

-

-

 

REF1

REF2

REF3

REF4

-

B2

1

6.84

6.92

6.52

8.01

-

6.53

2

5.65

5.69

5.63

7.64

-

5.15

3

5.30

5.36

5.27

7.51

-

4.65

4

4.89

4.99

4.29

7.42

-

4.21

5

4.49

4.62

4.59

7.32

-

3.78

6

4.07

4.23

4.26

7.23

-

3.36

7

3.68

3.90

3.93

7.13

-

2.93

8

3.28

3.50

3.59

7.03

-

2.50

9

2.89

3.15

3.26

6.95

-

2.08

10

2.50

2.76

2.93

6.85

-

1.65

11

2.10

2.40

2.59

6.77

-

1.23

12

1.71

2.02

2.26

6.68

-

0.81

13

1.30

1.65

1.92

6.58

-

0.40

14

0.90

1.26

1.58

6.49

-

0.13

15

0.51

0.88

1.25

6.39

-

0.10

16

0.16

0.52

0.91

6.31

-

0.09

17

0.10

0.16

0.57

6.23

-

0.08

18

0.10

0.10

0.25

6.13

-

0.08

19

0.09

0.09

0.12

6.04

-

0.08

20

0.08

0.08

0.10

5.95

-

0.07

21

0.08

0.09

0.10

5.86

-

0.07

22

0.08

0.08

0.09

5.78

-

0.07

23

0.08

0.06

0.09

5.69

-

0.07

24

0.07

0.07

0.09

5.60

-

0.07

25

0.07

0.06

0.09

5.50

-

0.06

26

0.07

0.06

0.08

5.42

-

0.06

27

0.06

0.06

0.08

5.33

-

0.06

28

0.06

0.07

0.08

5.25

-

0.06

29

0.06

0.06

0.08

5.16

-

0.06

30

0.07

0.07

0.08

5.06

-

0.06

Q1

4.89

5.36

4.92

6.77

-

4.21

Q2

2.50

2.40

2.59

5.25

-

2.93

Dt

3.0

4.0

3.5

8.5

-

1.5

RT(mg O2/L/minute)

0.80

0.74

0.67

0.18

-

0.85

RT(mg O2/L/hour)

47.80

44.40

39.94

10.73

-

51.20

RTs(mg O2/L/h/g solid sludge)

31.87

29.60

26.63

7.15

-

34.13

IT(%)

5.21

12.04

21.00

79.73

-

-

  Oxygen consumption data of the test mixture series with test item concentration of 10 mg/L

Half minutes

ID of Oxygen meter (LO1, LO2, LO3)

LO1/120300068133

LO2/120300068181

LO3/120300068157

ID of Oxygen meter electrodes (L,R)

120132594001

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

 

TI11

TI12

TI13

TI14

TI15

B3

1

6.50

6.73

6.47

6.67

6.33

6.66

2

5.32

5.25

5.32

5.38

5.05

5.29

3

4.95

4.87

4.84

4.89

4.67

4.91

4

4.50

4.44

4.36

4.43

4.23

4.47

5

4..08

3.97

3.95

3.98

3.81

4.05

6

3.64

3.53

3.50

3.53

3.38

3.62

7

3.23

3.11

3.05

3.08

2.96

3.20

8

2.81

2.66

2.61

2.63

2.53

2.77

9

2.38

2.23

2.16

2.18

2.11

2.35

10

1.95

1.79

1.71

1.73

1.68

1.92

11

1.52

1.33

1.27

1.28

1.26

1.50

12

1.10

0.90

0.82

0.84

0.84

1.08

13

0.68

0.47

0.39

0.41

0.44

0.66

14

0.29

0.13

0.121

0.14

0.17

0.27

15

0.10

0.09

0.10

0.12

0.15

0.10

16

0.09

0.08

0.09

0.11

0.14

0.09

17

0.08

0.07

0.09

0.11

0.13

0.08

18

0.08

0.07

0.09

0.10

0.13

0.07

19

0.07

0.07

0.08

0.10

0.12

0.07

20

0.07

0.07

0.08

0.10

0.12

0.07

21

0.07

0.07

0.08

0.10

0.12

0.06

22

0.06

0.06

0.08

0.09

0.11

0.06

23

0.07

0.06

0.07

0.09

0.11

0.06

24

0.06

0.06

0.07

0.09

0.11

0.06

25

0.07

0.05

0.07

0.09

0.11

0.06

26

0.06

0.05

0.07

0.08

0.10

0.06

27

0.06

0.05

0.07

0.08

0.10

0.05

28

0.05

0.06

0.07

0.08

0.10

0.05

29

0.05

0.05

0.06

0.08

0.10

0.05

30

0.06

0.06

0.06

0.08

0.10

0.05

Q1

4.50

4.87

4.84

4.43

4.23

4.47

Q2

2.81

2.66

2.61

2.63

2.53

2.77

Dt

2.0

2.5

2.5

2.0

2.0

2.0

RT(mg O2/L/minute)

0.85

0.88

0.89

0.90

0.85

0.85

RT(mg O2/L/hour)

50.70

53.04

53.52

 

 

 

Average RT(mg O2/L/hour)

52.45

-

RTs(mg O2/L/h/g solid sludge)

33.80

35.36

35.68

36.00

34.00

34.00

IT(%)

-4.15

Oxygen uptake of control (blank) replicates (RT(mg O2/L/hour))

B1

B2

B3

B4

B5

49.20

51.20

51.00

50.40

46.52

Variation coefficient of oxygen uptake of the blanks (B1-B5) (%)

3.54

Average Oxygen uptake of control (blank) replicates (RT(average)(mg O2/L/hour))

49.74

Average Specific Oxygen uptake of control (blank) replicates (RTs(average)(mg O2/L/h/g solid sludge))

33.16

Oxygen consumption data of the test mixture series with test item concentration of 100 mg/L

Half minutes

ID of Oxygen meter (LO1, LO2, LO3)

LO1/120300068133

LO2/120300068181

LO3/120300068157

ID of Oxygen meter electrodes (L,R)

120132594001

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

 

TI21

TI22

TI23

TI24

TI25

B4

1

6.77

6.21

6.57

7.00

6.85

7.23

2

6.02

5.22

5.45

6.02

5.66

6.10

3

5.59

4.81

4.98

5.57

5.30

5.74

4

5.16

4.36

4.53

5.13

4.87

5.32

5

4.74

3.91

4.09

4.70

4.45

4.90

6

4.31

3.46

3.65

4.26

4.02

4.48

7

3.90

3.04

3.21

3.84

3.60

4.06

8

3.47

2.62

2.77

3.40

3.17

3.65

9

3.05

2.17

2.32

2.96

2.74

3.23

10

2.61

1.74

1.88

2.53

2.32

2.80

11

2.19

1.30

1.43

2.09

1.89

2.38

12

1.76

0.87

0.98

1.65

1.46

1.96

13

1.32

0.44

0.54

1.22

1.03

1.53

14

0.90

0.12

0.16

0.78

0.61

1.11

15

0.48

0.08

0.10

0.37

0.23

0.69

16

0.13

0.08

0.09

0.13

0.13

0.30

17

0.09

0.07

0.09

0.11

0.12

0.10

18

0.08

0.07

0.08

0.10

0.11

0.08

19

0.08

0.07

0.08

0.10

0.11

0.07

20

0.07

0.06

0.08

0.09

0.10

0.07

21

0.07

0.06

0.07

0.09

0.10

0.07

22

0.06

0.06

0.07

0.09

0.10

0.06

23

0.06

0.06

0.07

0.09

0.09

0.06

24

0.06

0.05

0.07

0.08

0.09

0.06

25

0.06

0.04

0.07

0.08

0.09

0.05

26

0.05

0.04

0.07

0.08

0.09

0.05

27

0.05

0.05

0.06

0.08

0.09

0.05

28

0.06

0.04

0.06

0.08

0.09

0.05

29

0.05

0.04

0.06

0.08

0.09

0.05

30

0.05

0.05

0.06

0.08

0.08

0.05

Q1

6.02

4.81

4.53

5.57

5.30

5.74

Q2

2.61

2.62

3.21

2.53

3.17

2.80

Dt

4.0

2.5

1.5

3.5

2.5

3.5

RT(mg O2/L/minute)

0.58

0.88

0.88

0.87

0.85

0.84

RT(mg O2/L/hour)

51.15

52.56

52.80

52.11

51.12

50.40

Average RT(mg O2/L/hour)

51.95

-

RTs(mg O2/L/h/g solid sludge)

34.10

35.04

35.20

34.74

34.08

33.60

IT(%)

-3.13

  Oxygen consumption data of the test mixture series with test item concentration of 1000 mg/L

Half minutes

ID of Oxygen meter (LO1, LO2, LO3)

LO1/120300068133

LO2/120300068181

LO3/120300068157

ID of Oxygen meter electrodes (L,R)

120132594001

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

 

TI31

TI32

TI33

TI34

TI35

B5

1

7.20

6.99

6.89

7.43

7.08

7.17

2

6.62

6.30

6.42

6.60

6.40

6.70

3

6.31

5.98

6.02

6.21

6.01

6.30

4

5.90

5.59

5.63

5.81

5.63

5.92

5

5.47

5.20

5.23

5.41

5.24

5.53

6

5.10

4.77

4.83

5.01

4.84

5.15

7

4.68

4.38

4.43

4.60

4.45

4.75

8

4.25

3.99

4.02

4.21

4.05

4.37

9

3.88

3.57

6.31

3.80

3.65

3.97

10

3.45

3.14

3.21

3.39

3.26

3.58

11

3.05

2.75

2.79

2.99

2.85

3.19

12

2.63

2.29

2.38

2.58

2.45

2.79

13

2.22

1.89

1.97

2.17

2.05

2.39

14

1.80

1.46

1.55

1.75

1.64

1.99

15

1.39

1.06

1.13

1.34

1.24

1.59

16

0.98

0.64

0.72

0.93

0.84

1.19

17

0.57

0.24

0.33

0.53

0.45

0.79

18

0.20

0.09

0.11

0.18

0.15

0.41

19

0.09

0.08

0.09

0.11

0.12

0.12

20

0.08

0.07

0.09

0.10

0.11

0.08

21

0.07

0.06

0.08

0.10

0.10

0.07

22

0.07

0.07

0.08

0.09

0.10

0.07

23

0.06

0.06

0.08

0.09

0.10

0.06

24

0.06

0.06

0.07

0.09

0.09

0.06

25

0.06

0.06

0.07

0.09

0.09

0.06

26

0.06

0.06

0.07

0.08

0.09

0.06

27

0.06

0.06

0.07

0.08

0.09

0.05

28

0.06

0.05

0.07

0.08

0.09

0.05

29

0.05

0.05

0.06

0.08

0.08

0.05

30

0.06

0.04

0.06

0.08

0.08

0.05

Q1

6.31

5.98

6.42

6.62

6.40

6.70

Q2

2.63

3.14

2.79

2.58

2.45

2.79

Dt

4.5

3.5

4.5

4.0

5.0

5.0

RT(mg O2/L/minute)

0.82

0.81

0.81

1.01

0.79

0.78

RT(mg O2/L/hour)

49.07

48.69

48.40

60.60

47.40

46.92

Average RT(mg O2/L/hour)

50.83

-

RTs(mg O2/L/h/g solid sludge)

32.71

32.46

32.27

40.40

31.60

31.28

IT(%)

-0.89

 

Validity criteria fulfilled:
yes
Conclusions:
Activated sludge respiration inhibition test (carbon and ammonium oxidation) was carried out with AMINOX® to evaluate the influence of the test item on the activity of activated sludge microorganisms by measuring the respiration rate under defined conditions.
Under the conditions of this Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation) the observed endpoint for the effect of AMINOX® was the following:
The 3-hour EC50: > 1000 mg/L
Based on the results of this study, the test item AMINOX® had no inhibition effect on the total respiration rate of the microorganisms of the activated sludge at 1000 mg/L. No further testing is necessary.
Executive summary:

A laboratory test was carried out with AMINOX® to evaluate the effect of the test item on microorganisms by measuring the respiration rate.

The test concentrations (10, 100 and 1000 mg/L) were chosen to permit the determination of the approximate EC50.

The reference control results showed that in this study, the inoculum and methodology used provided a good measure of inhibition of activated sludge respiration rate.

Under the conditions of this Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation) the observed endpoint for the effect of AMINOX® was the following:

The 3-hour EC50: > 1000 mg/L

Based on the results of this study, the test item AMINOX® had no inhibition effect on the total respiration rate of the microorganisms of the activated sludge at 1000 mg/L. No further testing is necessary.

Description of key information

The 3-hour EC50: > 1000 mg/L

Key value for chemical safety assessment

EC50 for microorganisms:
1 000 mg/L

Additional information

A laboratory test was carried out with AMINOX® to evaluate the effect of the test item on microorganisms by measuring the respiration rate.

The test concentrations (10, 100 and 1000 mg/L) were chosen to permit the determination of the approximate EC50.

The reference control results showed that in this study, the inoculum and methodology used provided a good measure of inhibition of activated sludge respiration rate.

Under the conditions of this Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation) the observed endpoint for the effect of AMINOX® was the following:

The 3-hour EC50: > 1000 mg/L

Based on the results of this study, the test item AMINOX® had no inhibition effect on the total respiration rate of the microorganisms of the activated sludge at 1000 mg/L. No further testing is necessary.