N1,N3-diallylpropane-1,3-diamine dihydrochloride

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020-03-06 to 2020-05-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 1992

Deviations:

no

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

Activated sludge from the municipal wastewater treatment plant Breisgauer Bucht was used as inoculum with a concentration corresponding to 30 mg dry solids per litre. The treatment plant clarifies predominantly domestic wastewater and has a capacity of 600.000 inhabitant equivaluents. Sampling date of activated sludge was 18 March 2020. The dry solids (d.s.) content of the activated sludge was 5.2 g/L. It was determined by weight measurements after drying at 105°C for about 4 hours (mean of triplicate measurements).

Duration of test (contact time):

>= 28 d

Initial conc.:

>= 44.4 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium:
Mineral medium:
A: Potassium dihydrogenphosphate KH2PO4 8.50 g
Dipotassium hydrogenphosphate K2HPO4 21.75 g
Disodium hydrogenphosphate dihydrate Na2HPO4 * 2 H2O 33.40 g
Ammonium chloride NH4Cl 0.50 g
are dissolved in demineralised water and made up to 1 litre. The pH of the solution should be 7.4.
B: Calcium chloride dihydrate CaCl2 * 2 H2O 36.4 g
is dissolved in demineralised water and made up to 1 litre.
C: Magnesium sulfate heptahydrate MgSO4 * 7 H2O 22.5 g
is dissolved in demineralised water and made up to 1 litre.
D: Iron (III) chloride hexahydrate FeCl3 * 6 H2O 0.25 g
is dissolved in demineralised water, made up to 1 litre and stabilised with one drop of concentrated HCl.

- Temperature: 21.6 - 21.9 °C

TEST SYSTEM
- Culturing apparatus: The systems OxiTop® (WTW / Xylem) and Sensomat (AQUALYTIC®) were used as test systems.
- Number of culture flasks/concentration: 3
- Method used to create aerobic conditions: magnetic stirrer

SAMPLING
- Sampling frequency: 112 min
- Sampling method: pressure measurement

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: yes
- Toxicity control: yes

Reference substance:

benzoic acid, sodium salt

Key result

Parameter:

% degradation (O2 consumption)

Value:

ca. 2.4

Sampling time:

28 d

Details on results:

Degradation was 2.4% based on ThOD (NO3) and 3.2% based on ThOD (NH4), each mean of two replicates.

Results with reference substance:

The reference compound sodium acetate reached the pass level of 60% ThOD within 8 days.

Test item:

In one of the replicate test vessels, a significantly higher oxygen consumption was measured, resulting in a final degradation value of >100%. At the end of the test, when the bottles were emptied, it was found that this test vessel was contaminated with organic material that had not been noticed when filling the vessels. Since the contamination was obviously the cause of the increased values, this test replicate was regarded as an outlier, and was not considered in the test evaluation.

N-determinations:

	NO3-N [mg/L]	NO2-N [mg/L]
Test start
Blank before addition of inoculum	0.023	<LOD
Blank after addition of inoculum	0.196	<LOD
Stock solution test item before addition of inoculum	0.061	<LOD
Test end
Blank vessel 1	2.43	<LOD
Blank vessel 2	2.35	<LOD
Blank vessel 3	2.47	<LOD
Test vessel 2	2.40	<LOD
Test vessel 3	2.35	<LOD

LODNO₂-N: 0.012 mg/L

The increase of the concentration of NO₃-N during the test duration indicates, that nitrification took place.

The N-determinations are not considered in the test evaluation, because only insignificant differences between test and blank vessels were observed.

pH:

The pH in the blank, test and abiotic control vessels was inside the range 6 – 8.5. In the reference vessels, the pH was above >8.5, but the oxygen consumption by the reference substance was greater than 60% of ThOD. To apply this criterion to the toxicity control is not practical.

N1,N3 -diallylpropane-1,3 -diamine dihydrochloride - Biodegradation in % of ThOD_NO3

	Test item**			Toxicity control***	Reference item****			Abiotic control
Test item [ml/164 ml]	164	164	164	164				164
Reference item [ml/164ml]				164	164	164	164
ThODNO3[mg/l]	99.9	99.9	99.9	199.9	100.1	100.1	100.1	99.9
Day	Degradation [%]
0	0.0*	0.0	0.0	0.0	0.0	0.0	0.0	0.0
4	5.7	0.0	0.0	31.2	59.5	59.5	59.5	0.0
8	8.5	0.0	0.0	38.3	73.5	73.5	73.5	0.0
12	15.1	6.6	3.8	43.2	83.2	80.2	83.2	0.0
16	28.3	5.7	0.0	43.8	82.4	82.4	82.4	0.0
20	67.2	4.7	-3.8	43.5	80.8	80.8	80.8	0.0
24	88.1	2.8	-5.7	44.0	78.9	78.9	81.9	0.0
28	108.9	3.8	1.0	47.4	82.7	85.7	85.7	0.0

* This replicate was an outlier with >100% degradation. At the end of the study it was discovered, that the test vessel had been contaminated with organic substances. Therefore, this replicate was not considered for analysis of biodegradation of the test item.

** stock solution with 44.4 mg/L test item

***stock solution with 44.4 mg/L test item and 128.2 mg/L reference item

****stock solution with 128.2 mg/L reference item

N1,N3 -diallylpropane-1,3 -diamine dihydrochloride - Biodegradation in % of ThOD_NH4

	Test item**			Toxicity control***	Reference item****			Abiotic control
Test item [ml/164 ml]	164	164	164	164				164
Reference item [ml/164ml]				164	164	164	164
ThODNH4[mg/l]	75.0	75.0	75.0	175.0	100.1	100.1	100.1	75.0
Day	Degradation [%]
0	0.0*	0.0	0.0	0.0	0.0	0.0	0.0	0.0
4	7.6	0.0	0.0	35.6	59.5	59.5	59.5	0.0
8	11.3	0.0	0.0	43.7	73.5	73.5	73.5	0.0
12	20.1	8.8	5.0	49.3	83.2	80.2	83.2	0.0
16	37.7	7.6	0.0	50.0	82.4	82.4	82.4	0.0
20	89.4	6.3	-5.1	49.6	80.8	80.8	80.8	0.0
24	117.3	3.7	-7.6	50.3	78.9	78.9	81.9	0.0
28	145.0	5.1	1.3	54.2	82.7	85.7	85.7	0.0

* This replicate was an outlier with >100% degradation. At the end of the study it was discovered, that the test vessel had been contaminated with organic substances. Therefore, this replicate was not considered for analysis of biodegradation of the test item.

** stock solution with 44.4 mg/L test item

***stock solution with 44.4 mg/L test item and 128.2 mg/L reference item

****stock solution with 128.2 mg/L reference item

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

After 28-d incubation, the test item showed 2.4% based on ThOD (NO3) and 3.2% based on ThOD (NH4) degradation, each mean of two replicates. Therefore, the test item is not readily biodegradable.

Executive summary:

The purpose of this study was to determine the ready biodegradability of the test item N1,N3-diallylpropane-1,3-diamine dihydrochloride. The test item was exposed to aerobic activated sludge from the aeration tank of a domestic waste water treatment plant for 28 days. The biodegradation was followed by the oxygen uptake of the microorganisms during exposure. As a reference item, sodium benzoate was tested simultaneously under the same conditions as the test item, and functioned as a procedure control.

This study was designed to comply with the following methods:

-       Commission Regulation 440/2008/EC, Method C.4-D of May 30, 2008: Manometric Respirometry Test (EEC Publication No. L 142/496, May 2008)

-       OECD Guideline for Testing of Chemicals No. 301 F: "Ready Biodegradability: Manometric Respirometry Test", adopted July 17, 1992

All validity criteria were fulfilled, there were no deviations from the guidelines. The degradation rate of N1,N3-diallylpropane-1,3-diamine dihydrochloride never reached 60% within the incubation period of 28 days. The mean % biodegradation and biological oxygen demand of the test item, corrected for the inoculum blank, were 2.4% (based on ThOD_NO3) and 3.2% (based on ThOD_NH4) and 34 -31 mg/L O₂, respectively, over the incubation period of 28 days. Therefore, N1,N3 -diallylpropane-1,3 -diamine dihydrochloride is not readily biodegradable under these test conditions.

Description of key information

The purpose of this study was to determine the ready biodegradability of the test item N1,N3-diallylpropane-1,3-diamine dihydrochloride. The test item was exposed to aerobic activated sludge from the aeration tank of a domestic waste water treatment plant for 28 days. The biodegradation was followed by the oxygen uptake of the microorganisms during exposure. As a reference item, sodium benzoate was tested simultaneously under the same conditions as the test item, and functioned as a procedure control.

This study was designed to comply with the following methods:

-       Commission Regulation 440/2008/EC, Method C.4-D of May 30, 2008: Manometric Respirometry Test (EEC Publication No. L 142/496, May 2008)

-       OECD Guideline for Testing of Chemicals No. 301 F: "Ready Biodegradability: Manometric Respirometry Test", adopted July 17, 1992

All validity criteria were fulfilled, there were no deviations from the guidelines. The degradation rate of N1,N3-diallylpropane-1,3-diamine dihydrochloride never reached 60% within the incubation period of 28 days. The mean % biodegradation and biological oxygen demand of the test item, corrected for the inoculum blank, were 2.4% (based on ThOD_NO3) and 3.2% (based on ThOD_NH4) and 34 -31 mg/L O₂, respectively, over the incubation period of 28 days. Therefore, N1,N3-diallylpropane-1,3-diamine dihydrochloride is not readily biodegradable under these test conditions.

Key value for chemical safety assessment

Biodegradation in water:

not biodegradable

Type of water:

freshwater