[3R-(3α,3aβ,6α,7β,8aα)]-octahydro-3,6,8,8-tetramethyl-1H-3a,7-methanoazulen-5-yl acetate

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:

16th of February 2016 until 17th of March 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

In accordance with GLP

Qualifier:

according to guideline

Guideline:

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

Principles of method if other than guideline:

This test has been performed according to slightly modified EU, OECD and ISO Test Guidelines (OECD, 1992; EU, 1992; ISO, 1994).

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

natural water

Details on inoculum:

River water was sampled from the Rhine near Heveadorp, The Netherlands (11-02-2016). The nearest plant treating domestic wastewater biologically was 3 km upstream. The river water was aerated for 7 days before use to reduce the endogenous respiration (van Ginkel and Stroo, 1992). River water without particles was used as inoculum. The particles were removed by sedimentation after 1 day while moderately aerating.

Duration of test (contact time):

28 d

Initial conc.:

2 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, stocks and administration:
The river water used in the Closed Bottle test was spiked per liter of water; 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.3 mg Na2HPO4·2H2O, 22.5 mg MgSO4·7H2O, 27.5 mg CaCl2, 0.25 mg FeCl3·6H2O. Ammonium chloride was omitted from 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 top with aluminium foil and the content was mixed vigorously. Subsequently 0.2 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. 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 2.0 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 of the test. The pH of the medium at day 28 was 7.9 (test) and 8.0 (controls). Temperatures were within the prescribed temperature range of 22 to 24°C.

Reference substance:

acetic acid, sodium salt

Remarks:

purity > 99%

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 88% of its theoretical oxygen demand after 14 days. Degradation of sodium acetate was not inhibited by the test substance. 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:

73

Sampling time:

28 d

Parameter:

% degradation (O2 consumption)

Value:

64

Sampling time:

21 d

Parameter:

% degradation (O2 consumption)

Value:

46

Sampling time:

14 d

Parameter:

% degradation (O2 consumption)

Value:

13

Sampling time:

7 d

Details on results:

The substance is biodegraded by 73% at day 28 in the Closed Bottle test. Over 60% biodegradation was achieved in a period of 14 days immediately following the attainment of 10% biodegradation. Cedryl acetate therefore fulfilled the 14-day time window (10-day time window for other OECD 301 tests) criterion for ready biodegradable compounds and hence Cedryl acetate should be classified as readily biodegradable.

Results with reference substance:

The biodegradation percentage of the reference compound, sodium acetate, at day 14 was 46.

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 Cedryl acetate monoconstituent 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.

Table I Dissolved oxygen concentrations (mg/L) in the closed bottles.

Time (days)	Oxygen consumption (mg/L)		Biodegradation (%)
	Test substance	Acetate	Test substance	Acetate
0	0.0	0.0	0	0
7	0.7	4.1	13	76
14	2.6	4.6	46	85
21	3.6		64
28	4.1		73

Table II Oxygen consumption (mg/L) and the percentages biodegradation of the test substance, Cedryl acetate monoconstituent (BOD/ThOD) and sodium acetate (BOD/ThOD) in the Closed Bottle test

Time (days)	Oxygen consumption (mg/L)
	Ocs	Ot	Oc	Oa
0	8.8	8.8	8.8	8.8
	8.8	8.8	8.8	8.8
Mean (M)	8.8	8.8	8.8	8.8
7	8.2	7.6	8.2	4.0
	8.2	7.4	8.2	4.1
Mean (M)	8.2	7.5	8.2	4.1
14	8.1	5.5	8.2	3.6
	8.2	5.6	8.2	3.5
Mean (M)	8.2	5.6	8.2	3.6
21	7.8	4.1	7.6
	7.8	4.3	7.8
Mean (M)	7.8	4.2	7.7
28	7.4	3.3	7.5
	7.5	3.5	7.5
Mean (M)	7.5	3.4	7.5

OcsRiver water with nutrients and silica gel

OtRiver water with nutrients, test material (2.0 mg/L) and silica gel

OcRiver water with nutrients

OaRiver water with nutrients and sodium acetate (6.7 mg/L)

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

The substance is biodegraded by 73% at day 28 in the Closed Bottle test and should therefore be classified as readily biodegradable.

Executive summary:

In order to assess the biodegradation of Cedryl acetate, 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 2 mg/L of the substance for 28 days. Cedryl acetate did not cause a reduction in the endogenous respiration. Furthermore, the validity criteria of the test were met.

Cedryl acetate was biodegraded by 73% at day 28 in the standard Closed Bottle screening test and should therefore be classified as readily biodegradable.

Description of key information

In order to assess the biodegradation of Cedryl acetate, 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 2 mg/L of the substance for 28 days. Cedryl acetate did not cause a reduction in the endogenous respiration. Furthermore, the validity criteria of the test were met.

Cedryl acetate was biodegraded by 73% at day 28 in the standard Closed Bottle screening test and should therefore be classified as readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable