Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates

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:

26 Nov 2018 - *******

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Updated information. Study now available and provided ahead of extended submission date of 31/05/2019.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 310 (Ready Biodegradability - CO2 in Sealed Vessels (Headspace Test)

Deviations:

yes

Remarks:

Temperature and batch of 1-octanol - see 'principles of method if other than guideline' section.

Principles of method if other than guideline:

Test Procedure and Conditions
The temperature was not entirely within the range stated in the Study Plan. Evaluation: Temperature was only slightly out of range and with a relatively short duration. This deviation therefore did not impact the results or validity of the study.

Reference Item
A different batch of 1-octanol was used as stated in the Study Plan. Evaluation: All relevant details of the used 1-octanol were comparable to the batch in the Study Plan. This deviation therefore did not impact the results or validity of the study. The details of the used reference item are included in the report.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates

EC Number:

271-672-2

EC Name:

Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates

Cas Number:

68603-74-7

Molecular formula:

The substance is a UVCB so there is no molecular formula

IUPAC Name:

Amines, N-tallow alkyltrimethylenedi-, C4-18-alkyl phosphates

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: DUTEHP Lab Blend
- Expiration date of the lot/batch: 30 April 2019

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: 30 April 2019
- Stability under test conditions: Yes
- Solubility and stability of the test substance in the solvent/vehicle: Not applicable.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing and final dilution of a dissolved solid, stock liquid or gel: The test item was added to the mineral medium to give a final organic carbon concentration of 20 mg C/L. Since the test item was not sufficiently soluble to allow preparation of an aqueous solution at a concentration of 1 g/L, weighed amounts of test item were added directly to the test vessels containing medium (107 mL) with microbial organisms and mineral components (30 mg per vessel).

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

The source of test organisms was activated sludge freshly obtained from a municipal sewage treatment plant: 'Waterschap Aa en Maas', 's-Hertogenbosch, The Netherlands, receiving predominantly domestic sewage.

The freshly obtained sludge was kept under continuous aeration until further treatment. The concentration of suspended solids was determined to be 4.3 g/L in the concentrated sludge. Before use, the sludge was allowed to settle (30 minutes) and the liquid was decanted for use as inoculum at the amount of 10 mL/L of mineral medium.

Tap-water purified by reverse osmosis (Milli-RO) and subsequently passed over activated carbon and ion-exchange cartridges (Milli-Q) (Millipore Corp., Bedford, Mass., USA).

Duration of test (contact time):

28 d

Initial test substance concentration

Initial conc.:

0 other: Not applicable

Based on:

TOC

Remarks:

See 'details on study design' section.

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

Test duration: 28 days (14 days for the procedure and toxicity control).

Test vessels : Well-sealed glass serum vessels (TFE/silicone septa and Teflon seals). Vessels were filled to give a headspace to liquid ratio of 1:2 (107 mL test medium in 160 mL-capacity vessel).

Milli-RO / Milli-Q water: Tap-water purified by reverse osmosis (Milli-RO) and subsequently passed over activated carbon and ion-exchange cartridges (Milli-Q) (Millipore Corp., Bedford, Mass., USA). Milli-RO water: carbon levels < 500 ppb.

Stock solutions of mineral components:
A) 8.50 g KH2PO4; 21.75 g K2HPO4; 67.20 g Na2HPO4.12H2O; 0.50 g NH4Cl dissolved in Milli-Q water and made up to 1 litre, pH 7.4 ± 0.2
B) 22.50 g MgSO4.7H2O dissolved in Milli-Q water and made up to 1 litre.
C) 36.40 g CaCl2.2H2O dissolved in Milli-Q water and made up to 1 litre.
D) 0.25 g FeCl3.6H2O dissolved in Milli-Q water and made up to 1 litre.

Mineral medium: 1 litre mineral medium contains: 10 mL of solution (A), 1 mL of solutions (B) to (D) and Milli-RO water.

Sampling points and the number of vessels (between brackets)
Inoculum blank: day 1(3), 7 (3), 14 (5), 21 (3) and 28 (5)
Procedure control: day 1 (3), 7 (3) and 14 (5).
Test item: day 1(3), 7 (3), 14 (3), 21 (3) and 28 (5).
Toxicity control: day 1 (3), 7 (3) and 14 (5).
Abiotic control: day 1 (3) and 28 (5).

Three parallel reserve vessels for each test series were prepared. These vessels were not used and were removed at the end of the test.

Furthermore, an additional vessel of each test series was prepared for pH measurement. The vessel was prepared identically to the test vessels.

Incubation: In the dark on an orbital shaker, with a shaking rate sufficient to keep the vessel well mixed and in suspension.

7 M NaOH: 7M NaOH (Merck KGaA, Darmstadt, Germany) was prepared freshly at the start of the test.

Sample pre-treatment: 1 mL 7 M NaOH was injected through the septum of each vessel and the vessels were returned to the orbital shaker for 1 h. Thereafter, the vessels were removed from the shaker and allowed to settle. The supernatant was used for IC analysis.

Reference substance

Reference substance 1

Reference substance:

other: 1-octanol

Reference substance 2

Reference substance:

other: Sodium benzoate

Results and discussion

Preliminary study:

Not applicable.

Test performance:

The reference item was biodegraded by at least 60% (96%) within 14 days.
The mean amount of TIC present in the inoculum blanks at the end of the test was
< 3 mg C/L (1.5 mg C/L).
Since all criteria for acceptability of the test were met, this study was considered to be valid.

% Degradation

% Degradation 1

Key result

Parameter:

% degradation (inorg. C analysis)

Value:

0

Sampling time:

1 d

Remarks on result:

other: Not readily biodegradable under the conditions of the Headspace Test presently performed

% Degradation 2

Key result

Parameter:

% degradation (inorg. C analysis)

Value:

1

Sampling time:

7 d

Remarks on result:

other: Not readily biodegradable under the conditions of the Headspace Test presently performed

% Degradation 3

Key result

Parameter:

% degradation (inorg. C analysis)

Value:

1

Sampling time:

14 d

Remarks on result:

other: Not readily biodegradable under the conditions of the Headspace Test presently performed

% Degradation 4

Key result

Parameter:

% degradation (inorg. C analysis)

Value:

3

Sampling time:

21 d

Remarks on result:

other: Not readily biodegradable under the conditions of the Headspace Test presently performed

% Degradation 5

Key result

Parameter:

% degradation (inorg. C analysis)

Value:

3

Sampling time:

28 d

Remarks on result:

other: Not readily biodegradable under the conditions of the Headspace Test presently performed

Details on results:

Assuming 100% mineralization of the test item to CO2, the Theoretical maximum IC production (ThIC) in excess of that produced in the inoculum blanks equals the Total Organic Carbon (TOC) added to each test vessel at the start of the test. The TOC content of the test item was determined to be 65.35%. Thus, the ThIC of the test item was calculated to be 0.654 mg/mg.

BOD5 / COD results

Results with reference substance:

The ThIC of the reference item (1-octanol) was calculated to be 0.74 mg/mg.
The ThIC of the reference item (sodium benzoate) was calculated to be 0.58 mg/mg

Any other information on results incl. tables

              

Biodegradation

Results of inorganic carbon measurement and percentages of biodegradation are listed inTable3andTable4, respectively. An overview of the mean group % biodegradation is listed inTable1. The curves for biodegradation are shown inFigure 1.

The relative biodegradation values calculated from the measurements performed during the test period revealedno biologically relevantbiodegradation of the test item.

In the toxicity control more than 25% biodegradation occurred within 14 days (51%, based on ThIC). Therefore, the test item was assumed not to inhibit microbial activity.

Table1          
Mean Biodegradation in Percentage of theTheoretical maximum IC Production (ThIC)

Nominal day		Biodegradation in percentage of the ThIC				Degradation
		Reference Item		Test Item		Toxicity Control		Abiotic Control
1		0		0		0		0
7		66		1		31		-
14		96		1		51		-
21		-		3		-		-
28		-		3		-		0

- = Not measured

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

The test item was not readily biodegradable under the conditions of the Headspace Test presently performed.