Reaction mass of N,N'-ethylenebis[N-methyl-2-[(1-oxo-9-octadecenyl)oxy]-N-[2-[(1-oxo-9-octadecenyl)oxy]propyl]propylammonium] dimethyl disulphate and [2-[bis(2-hydroxypropyl)amino]ethyl]bis(2-hydroxypropyl)(methyl)ammonium methyl sulphate, dioleate (ester)

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:

29.10.1998-26.11.1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 98/Ba736/1 11 09 98
- Expiration date of the lot/batch: not specified
- Purity test date: not specified

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room tempearture, away from moisture
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: no solubility
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not reported

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: not specified
- Preliminary purification step (if any): not needed
- Final dilution of a dissolved solid, stock liquid or gel: not applicable
- Final preparation of a solid: not applicable

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): The source of test organisms was activated sludge freshly obtained from a municipal sewage treatment plant: Chantilly 60, France, receiving predominantly domestic sewage.
- Storage conditions: used immidiately
- Storage length: used immidiately
- Pretreatment: The freshly obtained sludge was used immediately. The concentration of suspended micro-organisms was determined to be 1.10^5 /mL in the concentrated sludge. Before use, the sludge was allowed to settle and the supernatant liquid was used as inoculum at the amount of 30 mL/L of mineral medium.
- Initial cell/biomass concentration: 1.10^5 micro-organisms/mL

Duration of test (contact time):

ca. 28 d

Initial conc.:

>= 10 - <= 20 mg/L

Based on:

TOC

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: see table 1 below
- Additional substrate: not specified
- Test temperature: ca. 20C
- pH: 7.55
- pH adjusted: not specified
- CEC (meq/100 g): not specified
- Aeration of dilution water: CO2 free compressed air
- Suspended solids concentration: not specified
- Continuous darkness: yes

TEST SYSTEM
- Number of culture flasks/concentration: 2
- Method used to create aerobic conditions:
- Method used to create anaerobic conditions:
- Measuring equipment:
- Details of trap for CO2 and volatile organics if used:

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

Reference substance:

aniline

Key result

Parameter:

% degradation (CO2 evolution)

Value:

ca. 27

St. dev.:

3

Sampling time:

28 d

Table 1: CO degradation

Day	CO2 Degradation						Cumulative CO2 production			Theoretical CO2 %
	Blanks			Assay			Assay			Assay
	FB1+FB2	FB3	FB average	FB9	FB10	FB total	FB9	FB10	FB total	FB9	FB10	FB average	FB total
1	2.09	5.28	3.66	1.65	1.98	3.74	0	0	0.05	0	0	0	0.02
4	5.83	11.44	8.83	7.37	8.89	43.67	0	0.05	35.05	0	0.02	0.01	15.92
5	13.66	15.62	14.74	17.93	22.66	81.52	3.19	7.92	70.78	2.9	3.6	3.25	34.9
6	14.74	15.95	15.34	19.47	27.5	133.96	4.12	12.15	118.83	3.75	5.52	4.84	53.92
7	16.39	16.81	16.61	21.56	31.46	153.45	4.95	14.85	136.64	4.5	6.75	5.82	62.2
8	17.93	17.49	17.71	24.2	35.66	167.88	6.49	18	150.15	5.9	8.25	7.07	68.25
11	18.15	19.03	18.59	28.6	44	186.78	10.01	15	168.18	9.1	11.56	10.32	76.45
12	18.15	19.8	18.97	30.36	42.74	193.34	11.38	25.41	174.4	10.35	13.07	11.71	79.27
14	18.37	20.02	19.19	32.56	52.14	200.75	13.38	28.78	181.55	12.15	14.97	13.56	82.52
15	18.58	20.13	19.36	34.32	55.33	205.15	14.96	32.94	185.79	13.6	16.35	14.97	84.75
18	18.58	20.57	19.56	37.16	59.07	214.83	17.6	35.97	195.25	16	18.2	17.1	88.75
20	19.25	21.01	20.13	39.27	63.14	220.33	19.14	40.04	200.2	17.4	19.55	18.47	91
22	20.66	21.01	20.84	42.48	65.47	222.42	21.61	43.01	201.52	18.85	20.32	20.01	91.42
25	21.56	21.01	21.28	45.87	66.42	222.42	24.58	44.82	201.13	22.35	21.42	21.89	91.42
27	22.66	21.01	21.83	47.65	66.63	223.96	26.01	47.13	202.12	23.85	21.72	22.69	91.87
28	22.99	21.01	22	54.34	74.25	225.3	32.34	47.79	203.5	29.4	23.75	26.57	92.5

Table 2: Theoretical CO2 %

	Stepantex X 1240
Flask	FB 9 10 mg/L	FB 10 20 mg/L
Days	-	-
4	0.0%	0.0%
7	4.5%	6.7%
14	12.1%	15.0%
22	19.6%	20.4%
28	29.4%	23.7%

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

Under the conditions and duration of the test the product Stepantex X 1240 shows a low degradation.

Executive summary:

Objective of the study

Determine under defined conditions, the biodegradability of an organic product in a mineral liquid medium inoculates a microbial flora of wastewater.

The biodegradability is experimee in% of theoretical CO2 produced.

Method of the study:

OECD Guideline for the Testing of Chemicals: Ready Biodegradability - Method 301B (17 July 1992): CO2 evolution test (modified Sturm).

Substances tested:

Agent 2076 -43D

Agent X 2197 -7 -7

Stepantex X 1040/2

Stepantex X 1040/1

Stepantex X 1240

Concentrations used:

10 and 20 mg / l organic carbon (initial, nominal)

Reference substance:

Aniline; 20 mg/l organic carbon

Inoculum:

Prepared from a sample of activated sludge from an urban wastewater treatment plant.

Duration of the study:

28 days

Results of the tests

The biodegradability of the five substances is evaluated at the end of the test (28 days):

As a percentage of theoretical CO2:

Concentration	Agent 2076-43D	Agent X 2197-7-7	Stepantex X 1040/2	Stepantex X 1040/1	Stepantex X 1240	Aniline (ref.)
10 mg/L	50%	37%	70%	74%	29%	-
20mg/L	53%	43%	69%	77%	24%	93%
Average	51%	40%	70%	75%	27%	-

As percentage of TOC removal:

Concentration	Agent 2076-43D	Agent X 2197-7-7	Stepantex X 1040/2	Stepantex X 1040/1	Stepantex X 1240	Aniline (ref.)
10 mg/L	77%	60%	84%	80%	49%	-
20mg/L	72%	66%	92%	90%	44%	98%
Average	75%	63%	88%	85%	47%	-

Conclusion:

Under the conditions and the test period, only the two ST and SP substances meet the criteria for ready biodegradability.

Description of key information

Under the conditions and duration of the test the product Stepantex X 1240 shows a low degradation.

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed

Type of water:

freshwater