1-ethyl-1-methylpyrrolidinium bromide

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: conducted under GLP using OECD guidelines

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 E (Ready biodegradability: Modified OECD Screening Test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Source and lot/batch No.of test material: 0691
- Expiration date of the lot/batch: not given
- Purity test date: 49.9% in water solution

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: 5 oC in the dark

OTHER SPECIFICS:
colorless liquid
pH = 6.84

Oxygen conditions:

aerobic

Inoculum or test system:

natural soil

Remarks:

soil from Sibersdorf

Details on inoculum:

Inuculum from soil (soil from Siebersdorf, 1988). The suitability of the soil was checked with two reference substances (sodium acetate/ sodium benzoate). 100 gr soil were suspended in 1000 ml drinking water. After stirring for 60 minutes the suspension was allowded to settle for stirring for 30 minutes. the supernatant was filtered, and the first 200 ml being discarded. The filtrate then was aerated immediately with filtered compressed air for 15 minutes.

Duration of test (contact time):

ca. 42 d

Initial conc.:

21.7 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

Principle
A predetermined amount of the test substance is dissolved in a mainly anorganic medium. The solution is inoculated with a small number of polyvalent microorganisms and aerated at 20-25 °C in the dark or at least in diffuse light only. The degradation is checked bu DOC analysis.

Procedure
One reaction vessel per group was each filled with 900 ml of the nutrient solution, containing the test substance in groups A and B respectively the reference substance in group PK and inoculated with 0.5 ml of the inoculum. The opening of the vessels was covered with aluminum foil in such a way that the exchange of air was guaranteed. Then the vessels were inserted in a shaking machune and kept there until the end of the study. Vessels were shielded from light. The room temperature was 21 °C, humidity in the range of 55-60%.

Drawing of samples
At the schedualed terms 30 ml were taken from each reaction vessel and centrifuged (4000 g, 15 min, at room temperature). The supernatants were preserved with 0.05 ml HgCl2 (1%) per 10 ml sample and were stored at 4 °C until DOC measurement.

Reference substance:

benzoic acid, sodium salt

Remarks:

34.28 mg sodium benzoate (i.e 20 mg DOC were dissolved in 1000 ml nutrient solution immediately before use.

Key result

Parameter:

% degradation (DOC removal)

Value:

ca. 9

Sampling time:

42 d

Remarks on result:

other: see remarks below

Details on results:

The DOC removal markedly less than 5% was calculated for test substance group a over the whole examination period of 42 days. In test substance group B almost the same results were gained, except for days 28 and 42 where DOC removal of 13% and 15% were calculated, degradation was also below 5% at all terms. Calculating DOC removal for both substance groups combined, biodegradation did not exceed 9%.

The degradation of the reference substance was 93% on day 7. A DOC removal of 100% was reached on day 21.

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The substance is not biodegradable under test conditions.

Executive summary:

The test substance at a concentration of about 20 mg DOC per liter was incubated in an aerobic aqueous medium according to OECD guideline 301E.

The DOC removal markedly less than 5% was calculated for test substance group a over the whole examination period of 42 days. In test substance group B almost the same results were gained, except for days 28 and 42 where DOC removal of 13% and 15% were calculated, degradation was also below 5% at all terms. Calculating DOC removal for both substance groups combined, biodegradation did not exceed 9%.

The substance is not biodegradable under test conditions.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March-July 2013

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conductd under GLP under OECD and Chinese guidelines

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)

Version / remarks:

and Chinese guidelines

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
N/A

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Activated sludge
Source: LiedeSewage Treatment Plant og Guangzhou
Treatment: the sludge was washed by centrifugation, the supernatant liquid phase was decanted and the solid material resuspended in a mineral medium. A homogenized aliquot of the final sludge suspension was weighted. Based on the ration, calculated amount of wet sludge was suspended in mineral medium to obtain a concentration equivalent to 2.4 g dry material per liter. Appropriate volume of inoculum was inculated in each bottle to give a final concentration of 28 mg dry material per liter.

Duration of test (contact time):

ca. 28 d

Initial conc.:

27 mg/L

Based on:

test mat.

Initial conc.:

32 mg/L

Based on:

test mat.

Remarks:

toxicity control

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Inoculum: Activated sludge
The sludge was washed by centrifugation, the supernatant liquid phase was decanted and the solid material resuspended in a mineral medium. A homogenized aliquot of the inal sludge suspenstion was weighted. Based on this ration, calculated amount of wet sludge was suspended in a mineral medium to obtain a concentration equivalent to 2.4 g dry material per liter. Appropriate volume of inoculum was inoculated in each bottle to give a final concentration of 28 mg dry material per liter.
20 L of test medium was prepared including inorganic salts.
Equipment: the test equipment include: erlenmeyer flasks, beakers, glass rod, pipetting tube, volumetric flasks ect.
Selection of test method: Manometric respirometry test was adopted.
Preparation of test substance stock solution: 52 mg test substance was dissolved in some test medium and then transferred to 500 ml volumetric flask. The stock solution was diluted with test medium to 500 ml and the stock solution with the effective nominal concentration of 100 mg/l was obtained (purity of the test substance is 96.5%)
Preparation of reference substance: 120 mg sodium benzoate was dissolved in test medium and then transferred to 100 ml volumetric flask. The stock solution was diluted with test medium to 100 ml and the stock solution with the concentration of 1200 mg/l was obtained.
Test groups include:
Bottles 1, 2 & A (test suspension): test substance, inoculum and test medium
Bottles 3, 4 & B (inoculum blank): inoculum and test medium
Bottle 5 (procedure control): reference substance, inoculum and test medium
Bottle 6: test substance, reference substance, inoculum and test medium
Preparation of test system:
addition of test substance: 100 ml test substance stock solution were added directly to the bottles 1, 2 and A respectively, 80 ml test substance stock solution was added to bottle 6.
addition of reference substance stock solution: 10 ml and 6.9 ml reference substance stock solution were added to bottles5 and 6 respectively.
addition of inoculum: 4.3 ml inoculum were added to bottles 1,2, A and 5; 5.0 ml inuculum were added to bottles 3,4 and B, 2.9 ml inoculum was added to bottle 6.
Addition of test medium: Test medium were added to bottles 1,2 A and 5 to make the final volume reached 365 ml; test medium were added to bottles 3, 4 and B to make the final volume reached 432 ml; Test medium was added to bottle 6 to make the final volume reached 250 ml.
Test substance effective concentration: 27 mg/l (bottles 1.2,A); 32 mg/l (bottle 6)
Reference substance concentration: 33 mg/l
inoculum concentration 28 mg/l

Test conditions: The test bottles were agitated and aerated on OxiTop control BOD at 22°C ± 1°C in the dark over 28 days period.

Reference substance:

benzoic acid, sodium salt

Key result

Parameter:

% degradation (O2 consumption)

Value:

>= 36.9 - <= 47.1

Sampling time:

28 d

Remarks on result:

other: less than 60%

Details on results:

Temperature: 21.0°C -22.8°C
Percent biodegradation:
Reference substance (14d) 71.9%
Toxicity control (14 d): 51.3%
Test substance (14d): 0% (2 replicates)
Test substance (28 d): 36.9% 47.1% (2 replicates) 42% (average)
At the end of the test, the oxygen uptake of 2 inoculum blanks were 38.2 mg/l and 33.7 mg/l
At the end of the test, pH values of test suspensions were 7.24 and 7.21, those of inoculum blanks were 7.26 and 7.27 and pH values of the procedure control and toxicity control were 7.59 and 7.58.
At the start of the test NO2- N concentrations of the test suspension and inoculum blank were <0.10 mg/l and NO3-N concentrations were 2.27 mg/l and 2.25 mg/l respectively.
At the end of the test, NO2-N concentrations of the test suspension and inoculum blank were < 0.10 mg/l NO3-N concentrations of the test suspensions were 2.92 mg/l and 2.98 mg/l NO3-N concentrations of the inoculum blank were 1.65 mg/l and 1.87 mg/l. The results revealed that there was nitrification occured.
On the 14d of the test, the percentage biodegradation of the reference substance and toxicity control were 71.9 and 51.3, which showed that the inoculum activity could meet the requirement of the test and the test material was considered not to have a toxic effect on the sewage sludge micro-organisms used in the study.
Under the test conditions, the percantage biodegradation of the test substance were 0% and 0% on the 14d. The percentage biodegradation of the test substance were 36.9% and 47.1% on the 28 d, the average was 42%.

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

Under the test conditions, the percantage biodegradation of the test substance were 0% and 0% on the 14d. The percentage biodegradation of the test substance were 36.9% and 47.1% on the 28 d, the average was 42%.
The percentage biodegradation of the test substance did not reach 60% at the 10d window of the test.

Executive summary:

Ready biodegradation was tested on MEP using Manometric Respirometry test.

Test suspension, inoculum blank, procedure control, toxicity control were test groups used in the test. Inoculum was activated sludge.

The composition of oxygen of each test bottle was determined and recorded automatically by OxiTop contol BOD test system

Results:

Reference substance (14 d) 71.9%

Toxicity control (14d) 51.3%

Test substance (14d) 0% (two replicates)

Test substance (28d) 36.9% 47.1% (two replicates)

42% average

The percentage biodegradation of the test substance did not reach 60% at the 10d-window of the test under the conditions of the study presently performed.

Description of key information

1. The test substance at a concentration of about 20 mg DOC per liter was incubated in an aerobic aqueous medium according to OECD guideline 301E.

The DOC removal markedly less than 5% was calculated for test substance group a over the whole examination period of 42 days. Calculating DOC removal for both substance groups combined, biodegradation did not exceed 9%. The substance is not biodegradable under test conditions. (1993 study)

2. Ready biodegradation was tested on MEP using Manometric Respirometry test.

Test suspension, inoculum blank, procedure control, toxicity control were test groups used in the test. Inoculum was activated sludge.

The composition of oxygen of each test bottle was determined and recorded automatically by OxiTop contol BOD test system

Results:

Reference substance (14 d) 71.9%

Toxicity control (14d) 51.3%

Test substance (14d) 0% (two replicates)

Test substance (28d) 36.9% 47.1% (two replicates)

42% average

The percentage biodegradation of the test substance did not reach 60% at the 10d-window of the test under the conditions of the study presently performed (2013 study).

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed