Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Ecotoxicological information

Toxicity to microorganisms

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
12-13 April 2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Effects on Biotic Systems, Activated Sludge, Respiration Inhibition Test [M]. Second Edition. Beijing: China Environmental Science Press 2013: 73-85
Qualifier:
according to guideline
Guideline:
other: The national hazardous chemical management standardizaton technical comittee. GB/T21796-2008 Chemicals-Activated Sludge, Respiration Inhibition Test[S]. Beijing: China Standards Press. 2008.
Qualifier:
according to guideline
Guideline:
other: State Environmental Protection Administration of China. HJ/T 153-2004 The Guidelines for the testing of chemical [S]. Beijing: China Environmental Science Press. 2004.
Qualifier:
according to guideline
Guideline:
EPA OPPTS 850.6800 (Modified Activated Sludge, Respiration Inhibition Test for Sparingly Soluble Chemicals)
Qualifier:
according to guideline
Guideline:
OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation))
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
Test groups
Test mixtures : FT: Containing test substance, inoculum, synthetic sewage feed and deionized water
F1-3: Containing test substance, synthetic sewage feed and deionized water.
Blank control: FTB: Containing inoculum, synthetic sewage feed and deionized water.
FHB: Containing inoculum, synthetic sewage feed, ATU and deionized water
Reference mixtures: FRL : containing reference substance, inoculum, synthetic sewage feed and deionized water.
FHR: containing reference substance, inoculum, synthetic sewage feed, ATU and deionized water.
Abiotic control: FA: containing test substance, synthetic sewage feed and deionized water.
(Note: F1-3 were prepared for the determination of pH values in the test mixtures at the start of the test, and the pH values measured as 7.19-7.24

Addition of the test substance
Since the test substance cannot be dissolved in water, direct addition was adopted for the study.
Each 5mg of the test substance was respectively weighted onto a clean and dry glass cover, and then put directed into flasks FT1 and F1.
Each 50mg of the test substance was respectively weighted onto a clean and dry glass cover, and then put directed into flasks FT2 and F2.
Each 500mg of the test substance was respectively weighted onto a clean and dry glass cover, and then put directed into flasks FT3-5 and FA1-3 and F3.
Respectively a clean and dry glass cover was added into FTB3-4
The purity of the test substance is 100% (UVCB)

Preparation of the reference substance stock solution
204 mg of the 3,5-dichlorophenol was weighed into a beaker and added with an appropriate amount of deionized water, then this was mixed with ultrasonic treatment for 10 minutes. After that, this was then put in the warm water bath (about 40˚C) to stir to make it dissolved. When it dissolved completely, this solution was transferred to 200 mL volumetric flask and further diluted to the mark with deionized water, and a stock solution with the concentration of 1000 mg/L was obtained (as the purity of 98%). The pH value of this stock solution was determined and adjusted with 40g/L NaOH to 7.27.

Preparation of the N-allylthiourea stock solution
592 mg of Allyl thiourea (ATU) was weighted into a beaker and added with some deionized water, and then this was put in the warm water bath to stir until ATU dissolved completely . The solution was then transferred to 250 mL volumetric flask and further diluted to the mark with deionized water, and the stock solution with the concentration of 2.32 g/L was obtained (as the purity of 98%).

The test solutions were aerated with 500 mL test vessels inserted into the outlet pipe. Test substance, reference substance, synthetic sewage feed, ATU and deionized water were added into the corresponding test vessels, respectively. The blank controls were prepared in the mean time.
After starting the test, 250 mL of activated sludge suspensions were respectively added into each test vessel in turn, with a final volume of 500 mL followed by aerating and agitating to stay suspending. After the exposure times of three hours, dissolved oxygen concentrations and the test temperatures from each test group were determined and recoded. The pH values of each solution at the start and end of the test were also determined at the meantime.
Test organisms (species):
activated sludge
Details on inoculum:
Name: Activated sludge
Source: Near the exit from the aeration tank of Water Purification Plant
Batch: IAs20210412-1
Selection reason: Recommended by “The Guidelines for the Testing of Chemicals”
Treatment: After collection, the sludge was removed of any coarse particles and impurities by sieving of a strainer, then washed and re-suspended in chlorine free tap water. Before the study, the solids dry weight of the activated sludge suspension was determined, and the sludge suspension initial concentration was adjusted to 3.0 g/L with the deionized water, after that this was cultivated for one day by 50ml if synthetic sewage feed per liter of activated sludge was fed and aerated under the test temperature of 20˚C±2˚C. The pH value was determined as 7.33 before used.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
3 h
Test temperature:
20°C±2°C.
pH:
7.19-7.24
Dissolved oxygen:
2.5-3.39mg/L
Nominal and measured concentrations:
Nominal: 10, 100, 1000 mg/L
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol
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:
The results of the activated sludge sensitivity test
The test temperatures were in range of 18.4 – 19.5˚C during the test and the pH values of each group at the start and end of the test were in the range of 7.67 – 7.78 and 7.92-8.20. At the end of the test, the average oxygen uptake rates of the blank controls (without ATU) was 22.3 mg/(g.h), which was greater than 20 mg/(g.h). the coefficients of variation of the blank controls were within 0.7-1.4%, which were less than 30% and met the validity of the test.
The 3h-EC50 of 3,5-DCP for total oxygen uptake was 4.5mg/L (within the 95% confidence interval limits of 3.9 mg/L – 5.2 mg/L), within the range of 2 mg/L to 25mg/L . 3h-EC50 for heterotrophic respiration was 9.0 mg/L (within the 95% confidence interval limits of 7.4 mg/L – 11mg/L), within the range of 5-40 mg/L. All of these results revealed that the sensitivity of the activated sludge used in the test could meet the requirement of the test.

The results for the Range-finding test
Test conditions
The test temperature was withing the rage of 18.1˚C during the test, and the pH values of test mixtures at the start and the end of the test were in the range of 7.61 – 7.68 and 8.03 – 8.77, the pH values of abiotic control at the start were in the range of 7.30 – 7.31 and the end of the test were in the range of 7.30-7.33.
Results
The average oxygen uptake rates of the blank controls (without ATU) was 22.4 mg/(g.h), which was greater than 20 mg/(g.h). the coefficient of variation of the blank controls was 1.9%, which was less than 30% and met the validity of the test.
Test substance with the exposure concentrations of 10, 100, 1000 mg/L (three replicates) were containing, and the abiotic control with the concentration of 100 mg/L in three replicates were included in the meantime. After an exposure time of three hours, the test results showed that the 3h-EC50 of the test substance for the total oxygen uptake was higher than 1000 mg/L. the average of oxygen uptake rats of the abiotic control was 0mg/(L.h), which revealed that there was no obvious abiotic oxygen consumption during the study period.
The absence of inhibition of oxygen consumption by the test substance in the range-finding test demonstrated that a definitive test was unnecessary.
Results with reference substance (positive control):
(1) The oxygen uptake rates of blank controls (without the test substance or reference substance) were greater than 20 mg/(g.h).
(2) The coefficients of variation of the oxygen uptake rates in the blank controls at the end of the test were less than 30%.
(3) In the sensitivity test of the activated sludge, the 3h-EC50 of 3,5-DCP for total oxygen uptake was 4.5 mg/L, which was within range (2-25 mg/L). 3h-EC50 for heterotrophic respiration was 9.0 mg/L, which was within the range 5mg/L to 40mg/L.
Validity criteria fulfilled:
yes
Conclusions:
Under the present test conditions, the 3h-EC50 of the test substance for the total oxygen uptake was higher than 1000 mg/L.
Executive summary:

A study was carried out on the test substance to determine the EC50 value of the respiration Inhibition of the test substance on activated sludge.


The pxygen uptake rates of samples of activated sludge fed with sunthtic sewage were measured in an enclosed cell containing an oxygen electrode after three hours. The sensitivity of each batch of activated sludge was checked with a suitable reference substance of 3,5-dichlorophenol on the day of exposure. Any oxygen uptake arising from abiotic processes was detected by determining the rate in mixtures of test substance, synthetic sewage medium and water, omiting activated sludge.


Under the present test conditions, the 3h-EC50 of the test substance for the total oxygen uptake was higher than 1000 mg/L.

Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17 May 2019
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))
Version / remarks:
OECD Guidelines for Testing of Chemicals, No.: 209 Adopted: 22 July 2010.
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)
Version / remarks:
Commission Regulation (EU) 2017/735 of 14 February 2017 amending, for the purpose of its adaptation to technical progress, the Annex to Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), Part C, C.11
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 850.6800 (Modified Activated Sludge, Respiration Inhibition Test for Sparingly Soluble Chemicals)
Version / remarks:
Product Properties Test Guideline OCSPP 850.3300 (Public Draft OPPTS 850.6800, April 1996) of the United States Environment Protection Agency (EPA) "Modified Activated Sludge, Respiration Inhibition Test", June 2012
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
No further details specified in the study report.
Analytical monitoring:
not specified
Vehicle:
no
Details on test solutions:
The test item was added by directly weighing the test item into the bottles and mixing using an 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.
Test organisms (species):
activated sludge of a predominantly domestic sewage
Details on inoculum:
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 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 7.70. The activated sludge was used directly after conditioning.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
3 h
Post exposure observation period:
No post exposure observation period specified int he study report.
Hardness:
Not specified
Test temperature:
19.8 – 21.6 °C
pH:
7.12 - 7.62 (start)
7.65 - 8.11 (end)
Dissolved oxygen:
Not appicable - respiration study
Salinity:
Not applicable
Conductivity:
Not specified
Nominal and measured concentrations:
Nominal concentrations: 10, 100 and 1000 mg/L
Details on test conditions:
CONTROLS
Abiotic Control (A)
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 and ultrasonic bath 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 7.42.

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: 19.8 – 21.6 °C
Aeration: With compressed air (about 1 L/min)
Recording: Test conditions were measured with suitable instruments and documented in the raw data.

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.

PERFORMANCE OF THE TEST
Preparation 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.

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 temperature was measured in the laboratory with a min/max thermometer during the experiment.
Reference substance (positive control):
yes
Remarks:
3,5-Dichlorophenol
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 -0.98 % and -7.59 % 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 29.60 mg O2/L/h/g solid.
The variation coefficient of oxygen uptake rate in above untreated replicates was 5.08 %.
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 75.65 % at the highest nominal concentration of 32.0 mg/L.
At the nominal concentrations of 1.0, 3.2 and 10.0 mg/L, the respiration rate was inhibited by 19.75 %, 29.71 % and 42.26 % respectively.
The 3-hour EC50 of 3,5-Dichlorophenol was calculated to be 9.61 mg/L with 95% confidence limits of 6.97 – 13.24 mg/L.
Reported statistics and error estimates:
Not specified

Influence if test item on oxygen consumption 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

-0.54

-

-

7.12

7.87

19.8/21.6

-

A2

1000

-1.03

-

-

7.26

7.89

-

A3

1000

-0.41

-

-

7.28

7.89

-

A4

1000

-0.74

-

-

7.31

7.88

-

A5

1000

-1.82

-

-

7.34

7.90

-

 

B1

Blank

0.00

40.80

27.20

-

7.17

7.65

19.8/21.6

5.08

B2

0.00

46.50

31.00

-

7.41

7.91

B3

0.00

43.89

29.26

-

7.58

7.96

B4

0.00

44.84

29.89

-

7.51

8.09

B5

0.00

46.00

30.67

-

7.61

8.10

 

REF1

Ref. item

1.00

34.73

23.15

19.75

7.47

7.89

19.8/21.6

-

REF2

3.20

30.30

20.20

29.71

7.43

7.90

-

REF3

10.00

24.73

16.49

42.26

7.42

7.91

-

REF4

32.00

9.90

6.60

75.65

7.41

7.93

-

 

TI1

Test item

10

43.93

29.29

-0.98

7.47

7.97

19.8/21.6

-

TI2

100

45.38

30.25

-4.24

7.51

8.10

-

TI3

1000

46.86

31.24

-7.59

7.62

8.11

-

#: Individual pH value 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)

190432598625

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

A1

A2

A3

A4

A5

B1

1

8.50

8.34

8.55

8.55

8.21

7.66

2

8.65

8.55

8.69

8.78

8.55

7.05

3

8.64

8.60

8.70

8.80

8.61

6.69

4

8.66

8.63

8.68

8.77

8.65

6.36

5

8.64

8.66

8.70

8.76

8.66

6.03

6

8.65

8.66

8.68

8.76

8.68

5.69

7

8.65

8.63

8.70

8.76

8.68

5.35

8

8.64

8.63

8.68

8.77

8.69

5.02

9

8.66

8.63

8.69

8.77

8.68

4.68

10

8.65

8.60

8.68

8.76

8.68

4.33

11

8.64

8.60

8.69

8.76

8.69

3.99

12

8.63

8.61

8.68

8.75

8.69

3.66

13

8.64

8.60

8.66

8.74

8.69

3.31

14

8.62

8.58

8.68

8.74

8.68

2.97

15

8.63

8.59

8.67

8.76

8.68

2.63

16

8.62

8.61

8.66

8.74

8.68

2.29

17

8.62

8.58

8.66

8.75

8.68

1.95

18

8.62

8.61

8.66

8.74

8.68

1.62

19

8.62

8.60

8.67

8.73

8.68

1.29

20

8.64

8.60

8.66

8.74

8.67

0.97

21

8.63

8.57

8.65

8.73

8.67

0.67

22

8.62

8.58

8.65

8.73

8.67

0.41

23

8.62

8.58

8.65

8.72

8.67

0.23

24

8.64

8.58

8.65

8.72

8.66

0.19

25

8.63

8.58

8.65

8.72

8.66

0.18

26

8.61

8.60

8.64

8.72

8.66

0.16

27

8.61

8.59

8.65

8.72

8.66

0.16

28

8.62

8.60

8.65

8.73

8.67

0.16

29

8.59

8.56

8.64

8.73

8.66

0.16

30

8.63

8.59

8.65

8.73

8.65

0.15

Q1

8.50

8.34

8.55

8.55

8.21

6.03

Q2

8.63

8.59

8.65

8.73

8.65

2.63

dt

14.5

14.5

14.5

14.5

14.5

5.0

RT(mg O2/L/minute)

-0.01

-0.02

-0.01

-0.01

-0.03

0.68

RT(mg O2/L/hour)

-0.54

-1.03

-0.41

-0.74

-1.82

40.80

RT(average)

-0.97

--

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

--

27.20

 

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)

190432598625

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

7.54

7.60

7.78

8.13

--

7.35

2

6.99

7.25

7.55

8.26

--

6.33

3

6.71

7.03

7.36

8.19

--

5.91

4

6.44

6.79

7.16

8.11

--

5.55

5

6.18

6.56

6.98

8.05

--

5.16

6

5.88

6.33

6.78

7.96

--

4.77

7

5.60

6.08

6.55

7.89

--

4.39

8

5.31

5.84

6.36

7.82

--

4.01

9

5.03

5.60

6.16

7.72

--

3.63

10

4.73

5.35

5.96

7.64

--

3.24

11

4.44

5.10

5.75

7.57

--

2.85

12

4.14

4.84

5.55

7.48

--

2.45

13

3.85

4.59

5.34

7.40

--

2.07

14

3.55

4.33

5.13

7.33

--

1.69

15

3.25

4.08

4.93

7.25

--

1.31

16

2.96

3.84

4.73

7.16

--

0.95

17

2.66

3.58

4.51

7.08

--

0.62

18

2.36

3.31

47.31

7.00

--

0.34

19

2.07

3.06

4.11

6.92

--

0.19

20

1.78

2.81

3.90

6.82

--

0.17

21

1.48

2.55

3.70

6.75

--

0.16

22

1.18

2.29

3.49

6.66

--

0.15

23

0.89

2.03

3.28

6.59

--

0.15

24

0.61

1.76

3.07

6.51

--

0.15

25

0.35

1.48

2.87

6.42

--

0.15

26

0.14

1.22

2.66

6.34

--

0.14

27

0.11

0.95

2.45

6.24

--

0.14

28

0.10

0.69

2.24

6.16

--

0.14

29

0.10

0.43

2.03

6.09

--

0.14

30

0.09

0.20

1.82

6.00

--

0.14

Q1

6.99

6.33

6.98

6.75

--

5.55

Q2

2.36

2.29

2.24

6.09

--

2.45

dt

8.0

8.0

11.5

4.0

--

4.0

RT(mg O2/L/minute)

0.58

0.51

0.41

0.17

--

0.78

RT(mg O2/L/hour)

34.73

30.30

24.73

9.90

--

46.50

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

23.15

20.20

16.49

6.60

--

31.00

IT= [1 –(RT– RTA)/RTB] x 100

IT(%)

19.75

29.71

42.26

75.65

--

--

           

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)

190432598625

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

TI11

TI12

TI13

TI14

TI15

B3

1

6.93

6.96

7.47

7.44

6.99

7.27

2

6.13

6.13

6.66

6.79

6.08

6.69

3

5.76

5.75

6.30

6.42

5.77

6.60

4

5.39

5.39

5.97

6.10

5.42

6.33

5

5.02

5.04

5.62

5.75

5.07

5.73

6

4.65

4.68

5.27

5.40

4.70

5.44

7

4.26

4.30

4.93

5.05

4.33

4.95

8

3.88

3.92

4.56

4.69

3.97

4.58

9

3.50

3.54

4.21

4.34

3.60

4.21

10

3.11

3.17

3.84

3.98

3.23

3.84

11

2.72

2.79

3.48

3.63

2.86

3.48

12

2.35

2.40

3.12

3.27

2.49

3.12

13

1.97

2.03

2.76

2.91

2.12

2.75

14

1.59

1.65

2.39

2.56

1.75

2.39

15

1.23

1.28

2.03

2.21

1.39

2.02

16

0.87

0.92

1.67

1.85

1.03

1.67

17

0.54

0.58

1.31

1.50

0.69

1.32

18

0.26

0.29

0.96

1.15

0.40

0.99

19

0.12

0.15

0.63

0.82

0.20

0.69

20

0.10

0.12

0.34

0.52

0.17

0.41

21

0.09

0.12

0.18

0.27

0.16

0.22

22

0.09

0.12

0.15

0.18

0.16

0.16

23

0.09

0.11

0.15

0.17

0.16

0.15

24

0.09

0.11

0.14

0.16

0.15

0.15

25

0.09

0.11

0.14

0.16

0.15

0.14

26

0.09

0.10

0.13

0.16

0.15

0.14

27

0.08

0.10

0.13

0.15

0.15

0.14

28

0.08

0.10

0.13

0.15

0.14

0.14

29

0.08

0.10

0.13

0.15

0.14

0.14

30

0.08

0.10

0.13

0.15

0.14

0.13

Q1

6.13

6.13

6.66

6.10

5.07

4.95

Q2

2.72

2.40

2.39

2.56

2.49

2.39

dt

4.5

5.0

6.0

5.0

3.5

3.5

RT(mg O2/L/minute)

0.76

0.75

0.71

0.71

0.74

0.73

RT(mg O2/L/hour)

45.47

44.76

42.70

42.48

44.23

43.89

Average RT(mg O2/L/hour)

43.93

--

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

30.31

29.84

28.48

28.32

29.49

29.26

IT= [1 – (RT– RTA)/RTB] x 100

IT(%)

-0.98

--

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

B1

B2

B3

B4

B5

40.80

46.50

43.89

44.84

46.00

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

5.08

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

44.40

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

29.60

 

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)

190432598625

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

TI21

TI22

TI23

TI24

TI25

B4

1

7.13

7.25

7.48

7.30

7.26

7.48

2

6.34

6.48

6.87

6.33

6.63

6.86

3

6.03

6.17

6.34

5.90

6.29

6.48

4

5.64

5.81

5.97

5.52

5.94

6.11

5

5.28

5.46

5.60

5.13

5.58

5.74

6

4.89

5.09

5.22

4.74

5.22

5.37

7

4.50

4.73

4.86

4.35

4.86

5.00

8

4.12

4.36

4.49

3.96

4.49

4.63

9

3.74

3.99

4.12

3.56

4.12

4.25

10

3.34

3.62

3.75

3.17

3.75

3.87

11

2.95

3.23

3.37

2.78

3.38

3.51

12

2.56

2.85

3.00

2.39

3.02

3.13

13

2.18

2.48

2.62

1.99

2.63

2.75

14

1.80

2.10

2.25

1.61

2.26

2.37

15

1.41

1.72

1.87

1.23

1.88

2.00

16

1.04

1.34

1.51

0.87

1.51

1.62

17

0.70

0.98

1.15

0.53

1.15

1.26

18

0.39

0.63

0.80

0.26

0.80

0.90

19

0.16

0.33

0.48

0.16

0.48

0.57

20

0.10

0.14

0.24

0.15

0.24

0.30

21

0.09

0.12

0.145

0.15

0.16

0.16

22

0.09

0.11

0.14

0.15

0.15

0.15

23

0.08

0.11

0.13

0.147

0.14

0.14

24

0.08

0.10

0.13

0.14

0.14

0.14

25

0.08

0.10

0.13

0.14

0.14

0.14

26

0.08

0.10

0.12

0.14

0.14

0.13

27

0.08

0.09

0.12

0.14

0.13

0.13

28

0.07

0.10

0.12

0.13

0.13

0.13

29

0.07

0.09

0.12

0.13

0.13

0.13

30

0.07

0.09

0.12

0.13

0.13

0.13

Q1

6.03

6.17

6.34

5.52

4.86

6.86

Q2

2.56

2.47

2.62

2.78

2.63

2.75

dt

4.5

5.0

5.0

3.5

3.0

5.5

RT(mg O2/L/minute)

0.77

0.74

0.74

0.78

0.74

0.75

RT(mg O2/L/hour)

46.27

4.40

44.64

46.97

44.60

44.84

Average RT(mg O2/L/hour)

45.38

--

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

30.84

29.60

29.76

31.31

29.73

29.89

IT= [1 –(RT– RTA)/RTB] x 100

IT(%)

-4.27

--

 

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)

190432598625

120182592012

120182592018

120162592009

120182592014

120182592021

L

R

L

R

L

R

Test mixtures

TI31

TI32

TI33

TI34

TI35

B5

1

7.33

7.33

7.41

7.27

7.27

7.45

2

6.65

6.52

6.28

6.35

6.31

6.93

3

6.37

6.19

5.85

5.91

5.92

6.84

4

5.95

5.87

5.46

5.51

5.54

6.75

5

5.58

5.47

5.06

5.10

5.16

5.83

6

5.21

5.13

4.69

4.70

4.78

5.28

7

4.83

4.76

4.28

4.29

4.40

4.89

8

4.43

4.37

3.89

3.90

4.02

4.51

9

4.05

4.01

3.50

3.49

3.64

4.12

10

3.65

3.60

3.11

3.09

3.25

3.74

11

3.25

3.23

2.72

2.69

2.86

3.36

12

2.85

2.85

2.33

2.28

2.48

2.97

13

2.46

2.47

1.95

1.88

2.09

2.59

14

2.08

2.08

1.56

1.49

1.71

2.21

15

1.69

1.70

1.19

1.10

1.33

1.83

16

1.31

1.32

0.83

0.74

0.97

1.45

17

0.94

0.95

0.51

0.41

0.63

1.09

18

0.60

0.60

0.25

0.19

0.34

0.74

19

0.30

0.31

0.16

0.16

0.17

0.43

20

0.12

0.13

0.14

0.15

0.15

0.21

21

0.10

0.12

0.14

0.15

0.15

0.15

22

0.09

0.11

0.13

0.14

0.14

0.15

23

0.09

0.11

0.13

0.14

0.14

0.14

24

0.08

0.10

0.13

0.14

0.14

0.14

25

0.08

0.10

0.12

0.14

0.13

0.14

26

0.08

0.10

0.12

0.14

0.13

0.13

27

0.08

0.09

0.12

0.13

0.13

0.13

28

0.08

0.09

0.12

0.13

0.13

0.13

29

0.08

0.09

0.12

0.13

0.13

0.13

30

0.08

0.09

0.12

0.13

0.13

0.13

Q1

4.83

5.13

5.85

5.91

5.92

4.89

Q2

2.85

2.47

2.72

3.09

2.86

2.59

dt

2.5

3.5

4.0

3.5

4.0

3.0

RT(mg O2/L/minute)

0.79

0.76

0.78

0.81

0.77

0.77

RT(mg O2/L/hour)

47.52

45.60

46.95

48.34

45.90

46.00

Average RT(mg O2/L/hour)

46.86

--

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

31.68

30.40

31.30

32.23

30.60

30.67

IT= [1 –(RT– RTA)/RTB] x 100

IT(%)

-7.59

--

 

Validity criteria fulfilled:
yes
Conclusions:
Activated sludge respiration inhibition test (carbon and ammonium oxidation) was carried out with SynNova Base Oil 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 SynNova Base Oil was the following:
The 3-hour EC50: > 1000 mg/L
Based on the results of this study, the test item SynNova Base Oil 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 SynNova Base Oil 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 SynNova Base Oil was the following:

The 3-hour EC50: > 1000 mg/L

 

Based on the results of this study, the test item SynNova Base Oil 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 SynNova Base Oil 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 SynNova Base Oil was the following:

The 3-hour EC50: > 1000 mg/L

 

Based on the results of this study, the test item SynNova Base Oil 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.