tert-butyl peroxyneodecanoate

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:

2018-05-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)

Version / remarks:

1992

Deviations:

yes

Remarks:

Please refer to "Principles of method"

Qualifier:

according to guideline

Guideline:

EU Method C.4-E (Determination of the "Ready" Biodegradability - Closed Bottle Test)

Version / remarks:

1992

Deviations:

yes

Remarks:

Please refer to "Principles of method"

Qualifier:

according to guideline

Guideline:

ISO 10707 Water quality - Evaluation in an aqueous medium of the "ultimate" aerobic biodegradability of organic compounds - Method by analysis of biochemical oxygen demand (closed bottle test)

Version / remarks:

1994

Deviations:

yes

Remarks:

Please refer to "Principles of method"

Qualifier:

according to guideline

Guideline:

other: Revised introduction to the OECD guidelines for testing of chemicals, section 3, Part 1: Principles and strategies related to the testing of degradation of organic chemicals, Paris Cedex, France.

Version / remarks:

2006

Deviations:

no

Principles of method if other than guideline:

Slight modifications to the guidelines:

a) ammonium chloride was omitted from the medium to prevent oxygen consumption due to nitrification (omission does not result in nitrogen limitation as shown by the biodegradation of the reference compound)

b) river water instead of an effluent/extract/mixture was used as inoculum.

GLP compliance:

yes (incl. QA statement)

Oxygen conditions:

aerobic

Inoculum or test system:

natural water: freshwater

Details on inoculum:

- Source of inoculum:
River water was sampled from the Rhine near Heveadorp, The Netherlands (03-01-2018). The nearest plant (Arnhem-Zuid) treating domestic wastewater biologically was 3 km upstream.

Duration of test (contact time):

28 d

Initial conc.:

2 mg/L

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium:
10 bottles containing river water only
10 bottles containing river water and silica gel
10 bottles containing silica gel with test item
6 bottles containing silica gel and the reference substances (sodium actetate)
10 bottles containing silica gel only

- Additional substrate: The river water used in the Closed Bottle test was spiked per liter of water with 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.4 mg Na2HPO4·2H2O, 22.5 mg MgSO4·7H2O, 27.5 mg CaCl2, 0.25 mg FeCl3·6H2O. Ammonium chloride was not added to the river water to prevent nitrification.

- Solubilising agent: not applicable
Accurate administering of the test substance was accomplished by preparing a solid stock of 3.0 mg of the test substance per g of silica gel in a 50-mL serum flask. Only part of the top layer of the silica gel was brought into contact with the test substance. The serum flask was closed with a screw top and the content was mixed vigorously. Subsequently, 0.20 g of silica gel with the test substance was added to the test bottles. The resulting concentration of test substance in the bottles was 2.0 mg/L. Next the bottles were filled with nutrient medium with inoculum and closed. The reference substance (sodium acetate) was added to the bottles using a stock solution of 1.0 g/L.

- Test temperature: 22.7 - 23.0 °C
- pH: 8.0
- pH adjusted: no
- Aeration of water: Each of the prepared solutions was dispensed into the respective group of BOD bottles so that all bottles were completely filled without air bubbles.
- Suspended solids concentration: River water without particles was used as inoculum. The particles were removed by sedimentation after 1 day while moderately aerating.
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: 0.30 L BOD bottles with glass stoppers
- Number of culture flasks/concentration: 10 per group; test item concentration was 2.0 mg/L
- Method used to create aerobic conditions: The river water was aerated for 7 days before use to reduce the endogenous respiration (van Ginkel and Stroo, 1992).

SAMPLING
- Sampling: Two duplicate bottles of all series were withdrawn for analyses of the dissolved oxygen concentration at day 7, 14, 21, and 28

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: no
- Toxicity control: yes
- Other: Reference substance (sodium acetate)

Reference substance:

acetic acid, sodium salt

Key result

Parameter:

% degradation (O2 consumption)

Value:

63

Sampling time:

28 d

Results with reference substance:

85 % at day 14

Theoretical oxygen demand (ThOD)

The theoretical oxygen demand (ThOD) of the test substance is 2.6 g O2/g test substance. It is assumed that the ThODs of the unknown constituents (9.9%) are equal to ThOD of the test substance. The ThOD of sodium acetate is 0.8 g/g.

Toxicity

Inhibition of the degradation of a well-degradable compound, e.g. sodium acetate by the test substance in the Closed Bottle test was not determined because possible toxicity of tert-Butyl peroxyneodecanoate to microorganisms degrading acetate is not relevant. A slight inhibition of the endogenous respiration of the inoculum by the test substance was detected only at day 7. Therefore, no inhibition of the biodegradation due to the "high" initial test substance concentration is expected.

Test conditions

The pH of the media was 8.0 at the start of the test. The pH of the medium at day 28 was 8.0 (controls) and 7.9 (test). The temperature ranged from 22.7 to 23.0°C which is within the prescribed temperature range of 22 to 24°C.

Validity of the test

The validity of the test is demonstrated by an endogenous respiration of 1.3 mg/L at day 28. Furthermore, the differences of the replicate values at day 28 were less than 20%. The biodegradation percentage of the reference compound, sodium acetate, at day 14 was 85. Finally, the validity of the test is shown by oxygen concentrations >0.5 mg/L in all bottles during the test period.

Biodegradability

tert-Butyl peroxyneodecanoate was biodegraded by 63% at day 28 in the Closed Bottle test. The time-day window concept assumes that biodegradation of a single organic compound in a ready biodegradability test is a growth-linked process which follows an S-shaped growth curve. tert-Butyl peroxyneodecanoate is a substance consisting of two chemicals linked together by a peroxide bond. Upon hydrolysis two compounds are formed i.e. neodecanoate and tert-butanol. The biodegradation kinetics (lag period, growth rate, and yield) of the individual hydrolysis products are not necessarily same. The biodegradation of the peroxide is an addition of different biodegradation curves. It is thus possible that individual compounds meet the time window criterion whereas the biodegradability curve of the peroxide suggests that the test substance is not readily biodegradable. The time window is therefore not considered applicable to tert-Butyl peroxyneodecanoate (OECD, 2006). tert-Butyl peroxyneodecanoate should therefore be classified as readily biodegradable.

Table 1 Oxygen consumption (mg/L) and the percentages biodegradation of the test substance, tert-Butyl peroxyneodecanoate (BOD/ThOD) and sodium acetate (BOD/ThOD) in the Closed Bottle test.

Time (days)	Oxygen consumption (mg/L)		Biodegradation (%)
	Test substance	Acetate	Test substance	Acetate
0	0.0	0.0	0	0
7	-0.1	4.0	0	74
14	1.2	4.6	23	85
21	2.7	-	52	-
28	3.3	-	63	-

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

Under the conditions of this study, the test item was biotically degraded by 63 % after 28 d. Therefore, the test item is considered readily biodegradable.

Executive summary:

In order to assess the biotic degradation of the test item, a ready biodegradability test was performed which allows the biodegradability to be measured in an aerobic aqueous medium. The ready biodegradability was determined in the Closed Bottle test performed according to slightly modified OECD, EU and ISO Test Guidelines, and in compliance with the OECD principles of Good Laboratory Practice. tert-Butyl peroxyneodecanoate did cause a slight reduction in the endogenous respiration only at day 7. The test substance is therefore considered to be non-inhibitory to the inoculum. tert-Butyl peroxyneodecanoate was biodegraded by 63% at day 28 in the OECD 301 Closed Bottle test. The substance should therefore be classified as readily biodegradable. The test is valid as shown by an endogenous respiration of 1.3 mg/L and by the complete degradation of the reference compound, sodium acetate. Sodium acetate was degraded by 85% of its theoretical oxygen demand after 14 days. Finally, the most important criterion was met by oxygen concentrations >0.5 mg/L in all bottles during the test period.

Description of key information

The ready biodegradability of the test item was assessed according to OECD guideline 301D (2018). The test item was biodegraded 63 % at day 28 in the Closed Bottle test. Therefore, it can be concluded that the test item is readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable

Type of water:

freshwater

Additional information

In order to assess the biotic degradation of the test item, a ready biodegradability test was performed which allows the biodegradability to be measured in an aerobic aqueous medium. The ready biodegradability was determined in the Closed Bottle test performed according to slightly modified OECD, EU and ISO Test Guidelines, and in compliance with the OECD principles of Good Laboratory Practice. tert-Butyl peroxyneodecanoate did cause a slight reduction in the endogenous respiration only at day 7. The test substance is therefore considered to be non-inhibitory to the inoculum. tert-Butyl peroxyneodecanoate was biodegraded by 63% at day 28 in the OECD 301 Closed Bottle test. The substance should therefore be classified as readily biodegradable. The test is valid as shown by an endogenous respiration of 1.3 mg/L and by the complete degradation of the reference compound, sodium acetate. Sodium acetate was degraded by 85% of its theoretical oxygen demand after 14 days. Finally, the most important criterion was met by oxygen concentrations >0.5 mg/L in all bottles during the test period.