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Reference
Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
DIN 38412-8 (Pseudomonas Zellvermehrungshemmtest)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
activated sludge
Details on inoculum:
- Laboratory culture: originated from the sewage treatment plant from Hildesheim
- Preparation of inoculum for exposure: after sedimentation washed twice with declorinated and sterilized tap water
- Initial biomass concentration: the biomass was 3.32 g/L
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
3 h
Test temperature:
18-22-°C
Reference substance (positive control):
yes
Remarks:
Copper (II)sulfate-pentahydrate
Duration:
3 h
Dose descriptor:
other: EC20
Effect conc.:
84.5 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Duration:
3 h
Dose descriptor:
other: EC20
Effect conc.:
130 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Key result
Duration:
16 h
Dose descriptor:
EC50
Effect conc.:
> 650 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
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
Remarks:
respiration rate
Details on results:
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: not reported
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: EC50 = 120 mg/L, need to be within 96.9 to 135.3 mg/L., hence the refrence substance indicated that the test system is valid
Reported statistics and error estimates:
Statistics performed according to "Ausgewählte Methoden der Wasseruntersuchung", Band II, VEB Fischer Verlag

Table 1. Repiration rates and inhibition in the test and reference substance

 

Substance

Concentration

[mg test substance/L]

Respiration rate

[mg O2/h/h]

Inhibition

[%]

Control 1

-

34.8

-

Control 2

-

36.4

-

Test substance

32

32.0

10

 

58

31.6

11

 

100

28.8

19

 

180

25.6

28

 

320

23.6

34

 

580

24.0

33

 

1000

20.4

43

Reference substance

58

31.6

11

 

100

20.8

42

 

180

6.4

82

 

Validity criteria fulfilled:
yes
Executive summary:

In the Klimisch 1 study from Noack (1991) the respiration inhibition of Netzer SB 10 (which contains 65% a.i.) to activated sludge was determined in a 3 h static GLP respiration inhibition test according to OECD 209. The nominal test concentrations were 0 (control), 32, 100, 180, 320, 580 and 1000mg Netzer SB 10/L, corresponding to 0 (control), 20.8, 65, 117, 208, 377 and 650 mg a.i./L. Two control replicates were set up. Dose verification analysis was not performed. The respiration was measured using an oxygen probe. The EC20, EC50 and EC80 was calculate by regression of the log transformed concentration vs. probit transformed percent inhibition relative to the average control respiration. The EC20 and EC50 were 130 and > 1000 mg Netzer SB 10 /L, respectively. These values correspond to EC20 and EC50 values of 84.5 and > 650 mg a.i./L.

This study is considered to be acceptable for the risk assessment.

Description of key information

OECD 209: 3 h EC50: > 650 mg/L

Key value for chemical safety assessment

EC50 for microorganisms:
650 mg/L

Additional information

In the Klimisch 2 study from Noack (1991) the respiration inhibition of Netzer SB 10 (which contains 65% a.i.) to activated sludge was determined in a 3 h static GLP respiration inhibition test according to OECD 209. The nominal test concentrations were 0 (control), 32, 100, 180, 320, 580 and 1000mg Netzer SB 10/L, corresponding to 0 (control), 20.8, 65, 117, 208, 377 and 650 mg a.i./L. Two control replicates were set up. Dose verification analysis was not performed. The respiration was measured using an oxygen probe. The EC20, EC50 and EC80 was calculate by regression of the log transformed concentration vs. probit transformed percent inhibition relative to the average control respiration. The EC20 and EC50 were 130 and > 1000 mg Netzer SB 10 /L, respectively. These values correspond to EC20 and EC50 values of 84.5 and > 650 mg a.i./L.

This study is considered to be acceptable for the risk assessment.