N,N'-[1,4-phenylenebis(methylene)]bis{N,N-dimethyl-3-[(prop-2-enoyl)amino]propan-1-aminium} dichloride

Biodegradation in water: screening tests

Administrative data

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)

Version / remarks:

1992

Deviations:

yes

Remarks:

Temperature range was 19.5 – 21.2 °C instead of 20.0 – 24.0 °C. As degradation of the positive control was in the normal range, this is considered as uncritical concerning the outcome of the study.

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Rheinland-Pfalz, Mainz, Germany

Test material

Test material information

Constituent 1

Reference substance name:

{[4-({dimethyl[3-(prop-2-enamido)propyl]azaniumyl}methyl)phenyl]methyl}dimethyl[3-(prop-2-enamido)propyl]azanium dichloride

EC Number:

812-548-5

Cas Number:

1621424-03-0

Molecular formula:

C24H40Cl2N4O2

IUPAC Name:

{[4-({dimethyl[3-(prop-2-enamido)propyl]azaniumyl}methyl)phenyl]methyl}dimethyl[3-(prop-2-enamido)propyl]azanium dichloride

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

- Source of inoculum/activated sludge: activation basin of the ESN (Stadtentsorgung Neustadt) sewage treatment plant, NW-Lachen-Speyerdorf, Germany (12. Jan. 2018)
- Preparation of inoculum for exposure: filtrated, washed with test medium (2x) and re-suspended in test medium, aerated until use
- Concentration of sludge: 25.0 mg dry matter/L (4900 mg of suspended solids/L)
- Initial cell/biomass concentration: approx. 10^7 - 10^8 cells/L

Duration of test (contact time):

28 d

Initial test substance concentration

Initial conc.:

20 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: according to OECD 301
- Test temperature: 19.5 - 21.2°C
- pH: 7.1 - 7.7
- pH adjusted: no
- Suspended solids concentration: 25 mg/L

TEST SYSTEM
- Culturing apparatus: 2000 mL-SCHOTT-flasks as test vessels, 100 mL scrubber flasks as absorbent vessels
- Number of culture flasks/concentration: 2
- Method used to create aerobic conditions: all flasks were aerated with purified, CO2-free, moistened air to purge the system of CO2
- Measuring equipment: IC measurement using the carbon analyser TOC multi N/C 2100S, Analytik Jena, Germany
- Details of trap for CO2 and volatile organics if used: CO2 was trapped in 0.25 M NaOH. Two scrubbers containing 100 mL each were connected in series to the test vessels. The initial IC value of the 0.25 M NaOH was separately determined in each flask.

SAMPLING
- Sampling frequency: day 0, 2, 4, 7, 9, 11, 14, 18, 23 and 29
- Sampling method: From each front scrubber flask, 10 samples were taken in order to determine the emitted CO2. The sample volume was 1 mL. The resulting change in the volume of the front flask was considered in the calculation of emitted CO2

CONTROL AND BLANK SYSTEM
- Inoculum blank: 2
- Abiotic sterile control: 1
- Toxicity control: 1
- Other: Positive control: 2; Apparatus blank (mineral medium only): 2

Reference substance

Reference substance:

aniline

Results and discussion

% Degradation

Parameter:

% degradation (CO2 evolution)

Value:

9

Sampling time:

29 d

BOD5 / COD results

Results with reference substance:

Degradation of the positive control was 80 % after 9 days.

Any other information on results incl. tables

Table 1: Net IC-values in mg/L of front scrubber flasks

 

Day	Blank Control 1	Blank Control 2	Positive Control 1	Positive Control 2	Test 1	Test 2	Abiotic Control	Toxicity Control
0	2.3	2.6	2.4	2.6	2.0	2.0	2.1	2.1
2	2.7	17.1	8.7	10.0	14.6	14.5	2.5	4.6
4	4.0	28.2	25.8	84.3	24.1	24.5	4.1	82.1
7	6.9	38.2	199.1	201.3	34.4	36.4	3.7	159.5
9	11.4	45.2	279.9	278.7	42.6	45.4	3.2	263.6
11	13.9	49.2	307.6	324.3	47.0	50.3	6.1	317.0
14	18.2	52.2	339.1	342.9	49.7	56.7	3.2	333.4
18	13.5	49.4	339.4	345.5	56.5	64.7	2.2	351.2
23	22.6	57.4	374.7	369.4	61.4	69.2	2.0	356.5
29	23.8	60.9	374.8	375.9	63.9	77.5	1.1	367.9

Table 2: Net IC-values in mg/L of back scrubber flasks

 

Day	Blank Control 1	Blank Control 2	Positive Control 1	Positive Control 2	Test 1	Test 2	Abiotic Control	Toxicity Control
0	2.3	2.3	2.2	2.0	2.0	2.2	2.0	2.2
29	2.2	2.6	2.4	2.8	4.1	2.1	2.1	2.5

Table 3: Degradation values in %

 

Day	Positive Control 1	Positive Control 2	Positive Control Mean	Test 1	Test 2	Test Mean	Abiotic Control	Toxicity Control
2	-0.4	0.0	-0.2	1.7	1.7	1.7	0.1	-0.8
4	3.2	22.2	12.7	2.8	2.9	2.8	0.6	10.8
7	57.1	57.7	57.4	3.9	4.6	4.3	0.5	22.2
9	80.5	80.1	80.3	4.7	5.6	5.1	0.3	37.7
11	87.4	92.6	90.0	5.0	6.1	5.5	1.2	45.2
14	95.2	96.3	95.8	4.7	6.8	5.7	0.3	46.7
18	95.4	97.3	96.4	7.8	10.4	9.1	0.0	49.6
23	102.6	101.0	101.8	6.6	9.0	7.8	0.0	48.5
29	100.8	101.3	101.1	7.3	10.7	9.0	-0.3	49.4

Table 4: Validity criteria for OECD 301B

Criterion from the guideline	Outcome	Validity criterion fulfilled
Difference of extremes of replicate values of the removal of the test chemical at the plateau, at the end of the test or at the end of the 10-d window, as appropriate, is less than 20%.	3.4%	yes
Percentage degradation of the reference compound reached the pass level by day 14 (≥ 60%).	Pass level reached on day 9	yes
The toxicity control should degrade to at least 35% (based on DOC) or at least 25% (based on ThOD or ThCO2) within 14 d.	46.7%	yes
The IC content of the test substance suspension in the mineral medium at the beginning of the test must be less than 5% of the TC.	0%	yes
The total CO2 evolution in the inoculum blank at the end of the test should not normally exceed 40 mg/L medium.	8.9 mg/L	yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Interpretation of results:

under test conditions no biodegradation observed