Decahydrospiro[furan-2(3H),5'-[4,7]methano[5H]indene]

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:

17 October 2016 to 16 November 2016

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)

GLP compliance:

yes (incl. QA statement)

Oxygen conditions:

aerobic

Inoculum or test system:

natural water

Details on inoculum:

River water was sampled from the Rhine near Heveadorp, The Netherlands. The river water was aerated for 7 days before use to reduce the endogenous respiration. River water without particles was used as inoculum. The particles were removed by sedimentation after 1 day while moderately aerating. The river water was spiked with mineral salts. Ammonium chloride was not added to the river water to prevent nitrification.

Duration of test (contact time):

28 d

Initial conc.:

1 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Remarks:

as a percentage of ThOD

Details on study design:

Test bottles:
The test was performed in 0.30 L BOD (biological oxygen demand) bottles with glass stoppers.

Nutrients and stocks:
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. 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 cap and the content was mixed vigorously. Subsequently 0.10 g of silica gel with the test substance was added to the test bottles. The resulting concentration of test substance in the bottles was 1.0 mg/L. Next the bottles were filled with nutrient medium with inoculum and closed. Sodium acetate was added to the bottles using a stock solution of 1.0 g/L.

Test procedure:
Use was made of 10 bottles containing only river water, 10 bottles containing river water and silica gel, 10 bottles containing river water and silica gel with test substance, 6 bottles with river water and sodium acetate. The concentrations of the test substance, and sodium acetate in the bottles were 1.0 and 6.7 mg/L, respectively. Each of the prepared solutions was dispensed into the respective group of BOD bottles so that all bottles were completely filled without air bubbles. The zero time bottles were immediately analyzed for dissolved oxygen using an oxygen electrode. The remaining bottles were closed and incubated in the dark. Two duplicate bottles of all series were withdrawn for analyses of the dissolved oxygen concentration at day 7, 14, 21, and 28.

Test conditions:
The pH of the media was 8.0 at the start and the end of the test (control, control with silica gel and test). Temperatures were within the prescribed temperature range of 22 to 24°C.

Reference substance:

acetic acid, sodium salt

Remarks:

purity > 99%

Preliminary study:

A preliminary study was performed to assess the most suitable inoculum in the Closed Bottle test, using activated sludge and river water. Inhibition by the test substance of the endogenous respiration was detected during a period of at least 84 days. In this study the test substance was not biodegraded. For the main test it was recommended to use river water as inoculum and the lowest possible initial test substance concentration (1.0 mg/L) to reduce potential toxic effects.

Test performance:

The test is valid as shown by an endogenous respiration of 1.2 mg/L at day 28. Furthermore, the differences of the replicate values at day 28 were less than 20%. Sodium acetate was degraded by 83% of its theoretical oxygen demand after 14 days. Inhibition of the endogenous respiration of the inoculum by the test substance at day 7 was not detected. Therefore, no inhibition of the biodegradation due to the "high" initial test substance concentration is expected. Finally, the most important criterion was met by oxygen concentrations >0.5 mg/L in all bottles during the test period.

Key result

Parameter:

% degradation (O2 consumption)

Value:

3

Sampling time:

28 d

Details on results:

The substance is not biodegraded (3% at day 28) in the Closed Bottle test and should therefore not be classified as readily biodegradable.

Results with reference substance:

The biodegradation percentage of the reference compound, sodium acetate, at day 14 was 83.

Toxicity to inoculum:

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

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The substance is not biodegraded (3% at day 28) in the Closed Bottle test and should therefore not be classified as readily biodegradable.

Executive summary:

In order to assess the biodegradation of the test substance, a screening test was performed according to OECD TG 301D (Closed Bottle test) and under GLP conditions. In this study river water was exposed to 1.0 mg/L of the test substance for 28 days. Inhibition of the endogenous respiration of the inoculum by the test substance at day 7 was not detected. The test substance was biodegraded for 3% at day 28) in the Closed Bottle test and should therefore not be classified as readily biodegradable. The test is valid as shown by an endogenous respiration of 1.2 mg/L and by the complete degradation of the reference compound, sodium acetate. Sodium acetate was degraded by 83% of its theoretical oxygen demand after 14 days. The oxygen concentrations were >0.5 mg/L in all bottles during the test period.

Description of key information

In order to assess the biodegradation of the test substance, a screening test was performed according to OECD TG 301D (Closed Bottle test) and under GLP conditions. In this study river water was exposed to 1.0 mg/L of the test substance for 28 days. Inhibition of the endogenous respiration of the inoculum by the test substance at day 7 was not detected. The test substance was biodegraded for 3% at day 28) in the Closed Bottle test and should therefore not be classified as readily biodegradable. The test is valid as shown by an endogenous respiration of 1.2 mg/L and by the complete degradation of the reference compound, sodium acetate. Sodium acetate was degraded by 83% of its theoretical oxygen demand after 14 days. The oxygen concentrations were >0.5 mg/L in all bottles during the test period.

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed