Reaction mass of Reaction product of 1-chloro-3-{[1-chloro-3-(dodecyloxy)propan-2-yl]oxy}propan-2-ol with methyl diethanolamine and [3-(dodecyloxy)-2-hydroxypropyl]bis(2-hydroxyethyl)methylammonium chloride

Biodegradation in water: screening tests

Administrative data

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-12-09 to 2017-01-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

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 18.7-21.7°C instead of 20-24°C. However, as degradation of the positive control was in the normal range, this is considered as uncritical for the outcome of the study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of Reaction product of 1-chloro-3-{[1-chloro-3-(dodecyloxy)propan-2-yl]oxy}propan-2-ol with methyl diethanolamine and [3-(dodecyloxy)-2-hydroxypropyl]bis(2-hydroxyethyl)methylammonium chloride

IUPAC Name:

Reaction mass of Reaction product of 1-chloro-3-{[1-chloro-3-(dodecyloxy)propan-2-yl]oxy}propan-2-ol with methyl diethanolamine and [3-(dodecyloxy)-2-hydroxypropyl]bis(2-hydroxyethyl)methylammonium chloride

Test material form:

other: solid, very viscous yellow paste

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge: sludge taken from activation basin of the ESN (Stadtentsorgung Neustadt) sewage treatment plant, Im Altenschemel, NW-Lachen-Speyerdorf, Germany
- Sampling date: 9 Dec 2016
- Preparation of inoculum for exposure:
-- Pretreatment: The sludge was filtrated, washed with tap water (2x), then washed with and re-suspended in test medium. It was then aerated until use.
- Concentration of sludge: 4160 mg suspended solids/L

Duration of test (contact time):

28 d

Initial test substance concentration

Initial test substance concentration 1

Initial conc.:

33.4 mg/L

Based on:

test mat.

Initial test substance concentration 2

Initial conc.:

18.3 mg/L

Based on:

TOC

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: mineral medium prepared according to guideline
- Test temperature: 18.7-21.7 °C
- pH: 7.3 for test samples
- Suspended solids concentration/inoculum concentration: 25mg/L
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: test vessels 2L SCHOTT flasks
- Number of culture flasks/concentration: 2 replicate positive controls (aniline), 2 replicates test sample, 1 abiotic control, 1 toxicity control.
- Method used to create aerobic conditions: aerated with purified CO2 scrubbed moistened air. To control the absence of CO2, the air was led through flask of Ba(OH)2.
- Measuring equipment: IC measurement for CO2 determination
- Details of trap for CO2 and volatile organics if used: traps with 0.25 M NaOH. 2 scrubbers each containing 100 mL were connected in series. Only front scrubber is sampled.

SAMPLING
- Sampling frequency: 10 samples on day: 0, 2, 4, 7, 9, 11, 15, 17, 23 and 29.
- Sampling method: determination of emitted volume in 1 mL sample. The resulting change in volume is taken into account for calculations.


CONTROL AND BLANK SYSTEM
- Inoculum blank: 2 replicates
- Abiotic sterile control: 1 replicate with 18.3 mg/L organic C
- Toxicity control: 1 replicate, 26.2 mg/L aniline + 33.4 mg/L test item - total organic C: 38.3 mg/L


STATISTICAL METHODS: Not applicable

Reference substance

Reference substance:

aniline

Results and discussion

Preliminary study:

no preliminary study performed

% Degradation

Key result

Parameter:

% degradation (CO2 evolution)

Value:

-2.35

St. dev.:

0.21

Sampling time:

29 d

Details on results:

No biodegradation observed after 28 days. No 10-day window detected.
In toxicity control: degradation was 29% after 14 days, therefore the test item is considered 'not toxic towards the inoculum at concentration of 33.4 mg/L'.

Any other information on results incl. tables

Degradation values (%)

Day	positive control		Test item		Abiotic control	Toxicity control
	Rep 1	Rep 2	Rep 1	Rep 2
2	-0.4	1.3	-0.3	-2.9	1.6	0.2
4	21.5	24.2	-1.2	-4.9	1.3	-0.9
7	60.9	66	-1.2	-7.7	1.1	14.1
9	72.3	81.2	-0.6	-5.2	1	25.7
11	77.3	84.9	-1.2	-5.1	0.9	28
15	80.9	87.4	-2.3	-5.5	0.2	28.9
17	85.8	95	-1.3	-3.3	-5.8	32.4
23	86.2	90.4	-2.7	-2.5	0.4	31.2
29	87.2	92.2	-2.2	-2.5	-0.1	35.5

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

The test substance was found to be not ready biodegradable within 28 days. The 10-day window was not detected.

Executive summary:

The biodegradability of the test substance was assessed according to OECD 301B, CO2 evolution test (modified Sturm test). The test item was incubated for 28 days under aerobic conditions. As positive control, aniline was used. All validity criteria were met and the degradation of the positive control was 63% after 7 days. In contrast, the test item was found to be not ready biodegradable. The 10 -day window was not detected and at the end of the test the degradation measured was 0%.

The degradation after 14 days in the toxicity control containing test item and aniline was 29% after 14 days indicating that the test substance is not toxic to the inoculum at a concentration of 33.4 mg/L.