reaction product of: saturated, monounsaturated and multiple unsaturated long-chained partly estrified alcohols of vegetable origin (Brassica napus L., Brassica rapa L., Helianthus annuus L., Glycine hispida, Gossypium hirsutum L., Cocos nucifera L., Elaeis guineensis) with O,O-diisobutyldithiophosphate and 2-ethylhexylamine and hydrogen peroxide

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:

01-15-98 to 02-14-98

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

428-630-5

EC Name:

-

Molecular formula:

Not applicable

IUPAC Name:

reaction product of Z-9-octadecen-1-ol and O,O-diisobutyl hydrogen dithiophosphate

Details on test material:

- Name of test material (as cited in study report): BECROSAN 6920
- Physical state: clear, yellowish liquid
- Analytical purity: 100%
- Storage condition of test material: Room temperature

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge: a sample of activated sludge from ,,e sewage plant at Taunusstein-Bleidenstadt

- Laboratory culture: No

- Method of cultivation: None

- Preparation of inoculum for exposure: was washed twice with the mineral nutrient solution used in this test to eliminate organic components and carbonates from the sludge. After resuspension in the mineral nutrient medium the sludge was aerated by means of compressed humidified air for about four hours. Before use as an inoculum for the COz-Evolution-Test, the sludge was homogenized in a "Waring Blender" at low speed for 2 minutes and then filtered through a cotton filter which had been carefully rinsed with deionized water. The first 200 mL of filtrate were discarded. The filtrate was used as inoculum (1 % of the final volume of the test solution) on the same day of preparation.

Duration of test (contact time):

28 d

Initial test substance concentration

Initial test substance concentration 1

Initial conc.:

ca. 25.1 mg/L

Based on:

test mat.

Initial test substance concentration 2

Initial conc.:

ca. 24 mg/L

Based on:

test mat.

Initial test substance concentration 3

Initial conc.:

16 mg/L

Based on:

other: TOC

Initial test substance concentration 4

Initial conc.:

15 mg/L

Based on:

other: TOC

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium:
Stock 1:
8.50g KH2PO4
21.75 g K2HPO4
33.40 g Na2HPO4.2H2O
0.50 g NH4Cl
were diluted in 1L deionized water; the pH should be 7.4

Stock 2:
36.40g CaCl2.2H2O were diluted in 1L deionized water

Stock 3:

22.50g MgSO4.7H2O were diluted in 1L deionized water

Stock 4:
0.25 g FeCl3.6H2O were diluted in 1L deionized water (this solution has to be prepared freshly before use in the test series).

The test solutions contained a total volume of 3500 mL: 35 mL of the phosphate mixture solution, 3.5 mL of the calcium chloride solution, 3.5 mL of the magnesium sulfate solution, 3.5 mL, of the fenichloride solution, and 35 mL of the inoculum. The test substance was added directly to the test solutions.

- Additional substrate: None
- Test temperature: 19.5 to 21.3°C
- pH: not specified
- pH adjusted: no
- Aeration of dilution water: yes
- Continuous darkness: No

TEST SYSTEM
- Culturing apparatus: 5-litre amber carboys served as test vessels.
- Number of culture flasks/concentration: 1, two concentrations were tested.
- Method used to create aerobic conditions: aerated test medium
- Test performed in closed vessels: yes

SAMPLING
- Sampling frequency: on day 3, 7, 12, 19, 24, 28 and 29
- Sample storage before analysis: No

CONTROL AND BLANK SYSTEM
- Inoculum blank: Yes
- Abiotic sterile control: No
- Toxicity control: Yes

Reference substance

Reference substance:

benzoic acid, sodium salt

Results and discussion

Preliminary study:

Not applicable

Test performance:

The viability of the inoculum and validity of the test were supported by the results of the reference substance Na-Benzaate. The control substance was degraded 86 % within 28 days, thereby fulfilling the criteria for a valid test by reaching the pass level by Day 28.

% Degradation

Parameter:

% degradation (CO2 evolution)

Value:

ca. 6

Sampling time:

28 d

Any other information on results incl. tables

See attached document

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The test substance is not ready biodegradable under the conditions of the study.

Executive summary:

Test Guideline

OECD Guideline 301B

Method and materials

The test substance and the inoculum (sewage microorganisms from a sewage plant working with predominantly domestic sewage) were incubated at 19.4 to 22.0^oC. CO2 released from the test substance was trapped as BaCO₃using a trap system described in the indicated guideline and the protocol. The degradation was followed by CO2 analysis. After reference to the blank control the total amount of CO2 produced by the test substance was determined for the test period and calculated as a percentage of the total CO2 that the test substance, based on its carbon content, could have theoretically produced (%TCO2).

One vessel with Na-Benzoate (20 mg TOC/L, final concentration) was run in parallel with the test series in order to check the activity of the inoculum. For this test solution test performance and evaluation of results were the same as for the substance to be tested.

The test solutions contained a total volume of 3500 mL: 35 mL of the phosphate mixture solution, 3.5 mL of the calcium chloride solution, 3.5 mL of the magnesium sulfate solution, 3.5 mL, of the fenichloride solution, and 35 mL of the inoculum. The test substance was added directly to the test solutions. At time to 88 mg of the test substance were given into the first test solution, and 84 mg into the second test solution. The final (calculated) concentration in the first test solution was -16 mg TOC/L, and - 15 mg TOCL in the second test solution.

For the Blank correction two test solutions were prepared in the same way as the test solutions with the test substance, but without the addition of any test- or control substance. At the same time one test solution containing 120 mg Na- Benzoate/3.5L (I 20 mg TOCL) of test solution was run in parallel. In the same way one test solution "Toxicity control" containing 120 mg Na-Benzoate plus 85 mg test substance/ 3.5L of test solution was tested in the same way.

Before starting the test the mineral nutrient solutions with inoculum but without test substance were areated with CO2 free air for 24 h in order to purge the system of CO2. The test solutions were stirred by means of magnetic stirrers in order to distribute the test substance (or control substance) and oxygen in the test solutions at the maximum solubility.

Results

The final degradation value after the test period of 28days (+ 1 day) was 6 % (mean value of two parallel test solutions). From the data the test substance may be regarded as "not readily biodegradable". The "toxicity control" was degraded 50% within 28 days and shows that no toxicity of the test substance has reduced the activity of the inoculum.

The control substance was degraded 86 % within 28 days (values not shown in a table). The threshold for classification as "readily biodegradable" of ≥ 60 % was met within 7d. Thus, the "10-days-window" as required by the OECD Guideline 301B, was met. The CO2 evolution measured in the Blank was in the required range. Thus the test is valid.

 

Conclusion

The test substance is not ready biodegradable under the conditions of the study.