Fatty acids, vegetable-oil, Me esters, sulfurized

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:

From 13 OCT 2017 to 04 JUN 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Temperature range was 19.7 – 21.3 °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)

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, vegetable-oil, Me esters, sulfurized

EC Number:

276-337-4

EC Name:

Fatty acids, vegetable-oil, Me esters, sulfurized

Cas Number:

72102-30-8

IUPAC Name:

Fatty Acids, vegetable-oil, Me-Esters, sulfurized

Test material form:

liquid

Details on test material:

- Density: 0.95 g/mL
- Batch no.: 71012767

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge: The sludge was taken from the activation basin of the ESN (Stadtentsorgung Neustadt) sewage treatment plant, Im Altenschemel, NW-Lachen-Speyerdorf
Date of collection: 17. Nov. 2017, batch no: 20171117
- Storage conditions:
- Storage length:
- Preparation of inoculum for exposure: The sludge was filtrated, washed with test medium (2x) and re-suspended in test medium. It was then aerated until use.
- Pretreatment: The carbon content of 65.85 % was determined by elemental analysis under non-GLP conditions, performed at the accredited laboratory Mikroanalytisches Labor Pascher, An der Pulvermühle 1, D-53424 Remagen, Germany. The test item was added directly, the amounts were calculated from the carbon content of the test item.
- Concentration of sludge: The dry matter was determined as 4120 mg suspended solids/L.
- Initial cell/biomass concentration:
- Water filtered: not stated

Duration of test (contact time):

28 d

Initial test substance concentration

Initial conc.:

30.4 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:
Solution a 10 mL
Solution b 1 mL
Solution c 1 mL
Solution d 1 mL
H2O demin. ad 1000 mL

Solution a
Potassium dihydrogen phosphate (KH2PO4) 8.5 g
Dipotassium hydrogen phosphate (K2HPO4) 21.75 g
Disodiumhydrogen phosphate dihydrate (Na2HPO4*2H2O) 33.4 g
Ammonium chloride (NH4Cl) 0.5 g
H2O demin. ad 1000 mL

Solution b
Calcium chloride (CaCl2) 27.5 g
H2O demin. ad 1000 mL

Solution c
Magnesium sulphate heptahydrate (MgSO4*7H2O) 22.5 g
H2O demin. ad 1000 mL

Solution d
Iron(III) chloride hexahydrate (FeCl3*6H2O) 0.25 g
Di-sodium-ethylene diaminetetraacetate dihydrate (Na2EDTA*2H2O) 0.4 g
H2O demin ad 1000 mL

- Test temperature: 19.7 – 21.3 °C
- pH: 7.1 - 7.8
- pH adjusted: no
- Aeration of dilution water: The test vessels were filled with medium and inoculum. Then, all flasks were aerated for 72 hours with purified, CO2-free, moistened air to purge the system of CO2.
- Suspended solids concentration: The dry matter was determined as 4120 mg suspended solids/L
- Continuous darkness: yes


TEST SYSTEM
- Culturing apparatus:
- Number of culture flasks/concentration: 2
- Measuring equipment: Carbon analyser TOC multi N/C 2100S, Analytik Jena
- Test performed in open system: 2000 mL-SCHOTT-flasks were used as test vessels, 100 mL scrubber flasks as absorbent vessels.
- Details of trap for CO2 and volatile organics if used: The test vessels were aerated with purified (by activated charcoal), CO2-scrubbed, moistened air. The scrubbing of carbon dioxide was achieved by bubbling the purified air through a flask containing 1.5 M NaOH. To control the absence of CO2, the air was then led through a flask containing a solution of Ba(OH)2 before reaching the test vessels.

SAMPLING
- Sampling frequency: on day 0, 2, 4, 7, 9, 11, 14, 18, 23 and 29

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

Reference substance

Reference substance:

aniline

Results and discussion

% Degradation

Key result

Parameter:

% degradation (CO2 evolution)

Value:

37.5 - 38.6

Sampling time:

29 d

Details on results:

Day Positive Positive Positive Test 1 Test 2 Test Abiotic Toxicity
Control 1 Control 2 Control Mean Mean Control Control
2 -0.5 -2.3 -1.4 4.3 0.1 2.2 0.9 1.9
4 28.1 -3.3 12.4 16.3 8.9 12.6 0.5 16.5
7 68.3 17.5 42.9 25.6 10.3 17.9 0.6 41.3
9 76.6 45.3 61.0 29.6 24.7 27.2 2.0 51.2
11 87.1 64.6 75.9 30.7 28.5 29.6 0.3 56.2
14 88.1 76.3 82.2 32.9 31.6 32.3 -0.1 58.3
18 91.3 80.6 86.0 33.9 33.7 33.8 0.1 59.3
23 88.7 85.8 87.2 35.1 34.3 34.7 0.6 59.5
29 90.7 94.9 92.8 38.6 37.5 38.1 -1.5 62.1

BOD5 / COD results

Results with reference substance:

Aniline was chosen as positive control.
Activated sludge was used as inoculum (concentration in the test 25.0 mg dry matter/L).
The test was left running for 28 days.
All validity criteria were met. Degradation of the positive control was 61 % after 9 days.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

Results for the Test Item Fatty Acids, vegetable-oil, Me-Esters, sulfurized
The test item Fatty Acids, vegetable-oil, Me-Esters, sulfurized is considered as “not
readily biodegradable“.
The degree of biodegradation reached 38 % after 28 days.
The 10-day-window began on day 4, at its end, 32 % degradation were reached, missing the pass level of 60 % given in the OECD guideline.
Because the test item is a mixture, the 10-day window has not to be taken into account.
Degradation missed 60 % within 28 days, too. Therefore, regardless of the 10-day-window, the test item is considered as “not readily biodegradable within 28 days”.
Abiotic degradation was not observed.