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Ecotoxicological information

Toxicity to microorganisms

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Reference
Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test performed under GLP according guidelines, all quality criteria were met
Justification for type of information:
Read-across data is presented from quaternary ammonium compounds, C12-18-alkylbis(hydroxyethyl)methyl, chlorides, CAS Number 71808-53-2, EC Number 276-038-9. This substance is considered to be close enough in structural integrity to the target substance, bis(2 -hydroxyethyl)oleylmethylammonium chloride, CAS Number 18448 -65 -2, EC Number 242 -332 -0, so as to justify valid read-across.
Qualifier:
according to guideline
Guideline:
OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
67.2% active.
Analytical monitoring:
not required
Vehicle:
no
Details on test solutions:
The cocobis(2-hydroxyethyl) methylammonium chloride stock solution was prepared by stirring 0.47 g cocobis(2-hydroxyethyl) methylammonium chloride in 470 mL of deionized water. The cocobis(2-hydroxyethyl) methylammonium chloride stock solution was a clear solution.
pH stock solution = 5.6
Test organisms (species):
activated sludge of a predominantly domestic sewage
Details on inoculum:
Unadapted secondary activated sludge was obtained (08-02-2010) from the WWTP Nieuwgraaf in Duiven. The WWTP Duiven is an activated sludge plant treating predominantly domestic wastewater. Prior to use the activated sludge was homogenized with a syringe. The dry weight of the homogenized activated sludge was determined and subsequently concentrated by settlement to the required dry weight concentration. The dry weight of the activated sludge in the incubation vessels was 1.7 g/L.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
3 h
Test temperature:
Temperature varied from 20.0 to 20.7°C.
pH:
The pH of the reaction mixtures after the incubation period ranged from 7.4 to 7.6.
Dissolved oxygen:
aerobic during test
Details on test conditions:
-The test was performed in 250 to 300 mL Erlenmeyers.
-The homogenized activated sludge was incubated in a shaking water bath (100 rpm, 20°C) for 3 hours with various concentrations of the test
compound and synthetic sewage.
-Synthetic sewage was added to the activated sludge (1.6 mL on a total volume of 50 mL).
Synthetic sewage contained per liter: 16 g peptone
11 g meat extract
3 g urea
0.7 g NaCl
0.4 g CaCl2.2H2O
0.2 g MgSO4.7H2O
2.8 g K2HPO4
-Different concentrations (spaced by a factor two) of test compound and reference compound where added using stock solutions of respectively.
1.0 g/L and 0.5 g/L.
-Tap-water was used to fill up the total test volume to 50 mL.
- The dry weight concentration of the activated sludge in the test was 1.7 g/L.
-The tested concentrations of the reference compound 3,5-dichlorophenol where: 2.5 mg/L, 5.1 mg/L,10.2 mg/L, 20.3 mg/L and 40.6 mg/L
-The tested concentrations ofcocobis(2-hydroxyethyl) methylammonium chloride where: 5 mg/L, 10 mg/L, 20 mg/L, 40 mg/L, 80 mg/L, 160 (mg/L)
-Controls without test substance (or reference compound) are measured at the beginning and at the end of the test
-The inhibitory effect of the test substance/reference compound is expressed as a percentage of the mean of the two control respiration rates.
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol
Key result
Duration:
3 h
Dose descriptor:
EC10
Effect conc.:
10.9 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Key result
Duration:
3 h
Dose descriptor:
EC50
Effect conc.:
28.8 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Remarks on result:
other: (14.9 - 55.7)
Key result
Duration:
3 h
Dose descriptor:
EC10
Effect conc.:
11 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Key result
Duration:
3 h
Dose descriptor:
EC50
Effect conc.:
29 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Remarks on result:
other: with 95% confidence limits of 15 and 56 mg/L
Details on results:
Test conditions
The pH of the reaction mixtures after the incubation period ranged from 7.4 to 7.6. Temperature varied from 20.0 to 20.7°C. These conditions allowed respiration of the activated sludge used.

Validity of the test
The validity of the test is demonstrated by two criteria. First, the control respiration rates were within 15% of each other and secondly, the EC50 of the reference compound was 9 mg/L which is within the prescribed range of 5 to 30 mg/L .

Activated sludge respiration inhibition test
The inhibitory effect of cocobis(2-hydroxyethyl) methylammonium chloride at a particular concentration is expressed as a percentage of the two
controls. From the results EC values were calculated.
The EC50 of cocobis(2-hydroxyethyl) methylammonium chloride for activated sludge after 3 hours contact time is 29 mg/L with 95% confidence
limits of 15 and 56 mg/L. The EC10, EC20 and EC80 are 11, 15 and 55 mg/L, respectively.
Results with reference substance (positive control):
The EC50 of the reference compound was 9 mg/L at a contact time of 3 hours, respectively. These EC50 values is within the prescribed range of 5 to 30 mg/L.
Validity criteria fulfilled:
yes
Conclusions:
The EC50 of the test item, Quaternary ammonium compounds, coco alkylbis(hydroxyethy​l)​methyl, chlorides, CAS Number 70750-47-9, EC Number 274-846-6, for activated sludge after 3 hours contact time is 29 mg/L with 95% confidence
limits of 15 and 56 mg/L. The EC10, EC20 and EC80 are 11, 15 and 55 mg/L, respectively.
Executive summary:

The toxicity to microorganisms of the test substance, Quaternary ammonium compounds, coco alkylbis(hydroxyethyl)methyl chlorides, CAS Number 70750 -47 -9, EC Number 274-846-6, [Quaternary ammonium compounds, C12-18-alkylbis(hydroxyethyl)methyl, chlorides, CAS Number 71808 -53 -2, EC Number 276 -038 -9], was investigated in a study conducted according to OECD guideline 209 (Activated Sludge, Respiration Inhibition Test).

This substance is considered to be close enough in structural integrity to the target substance, bis(2 -hydroxyethyl)oleylmethylammonium chloride, CAS Number 18448 -65 -2, EC Number 242 -332 -0, so as to justify valid read-across.

The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

Under the conditions of the study the EC50 of the test item, Quaternary ammonium compounds, coco alkylbis(hydroxyethy​l)​methyl, chlorides, CAS Number 70750-47-9, EC Number 274-846-6, for activated sludge after 3 hours contact time is 29 mg/L with 95% confidence limits of 15 and 56 mg/L. The EC10, EC20 and EC80 are 11, 15 and 55 mg/L, respectively

Description of key information

The toxicity to microorganisms of the test substance, Quaternary ammonium compounds, coco alkylbis(hydroxyethyl)methyl chlorides, CAS Number 70750 -47 -9, EC Number 274-846-6, [Quaternary ammonium compounds, C12-18-alkylbis(hydroxyethyl)methyl, chlorides, CAS Number 71808 -53 -2, EC Number 276 -038 -9], was investigated in a study conducted according to OECD guideline 209 (Activated Sludge, Respiration Inhibition Test).

This substance is considered to be close enough in structural integrity to the target substance, bis(2 -hydroxyethyl)oleylmethylammonium chloride, CAS Number 18448 -65 -2, EC Number 242 -332 -0, so as to justify valid read-across.

The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

Under the conditions of the study the EC50 of the test item, Quaternary ammonium compounds, coco alkylbis(hydroxyethy​l)​methyl, chlorides, CAS Number 70750-47-9, EC Number 274-846-6, for activated sludge after 3 hours contact time is 29 mg/L with 95% confidence limits of 15 and 56 mg/L. The EC10, EC20 and EC80 are 11, 15 and 55 mg/L, respectively

Key value for chemical safety assessment

EC50 for microorganisms:
29 mg/L
EC10 or NOEC for microorganisms:
11 mg/L

Additional information