Essential oil Schinus Terebinthifolius (Anacardiaceae) obtained from red berries by supercritical carbon dioxide extraction (indicative)

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2007

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Study conducted according to OECD Guideline 301 C but with deviations: no detail on material and methods

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Deviations:

yes

Remarks:

no detail on material and methods

Principles of method if other than guideline:

Not applicable

GLP compliance:

not specified

Specific details on test material used for the study:

No additional information

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- Concentration of sludge: 30 mg/L

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Details on study design:

No data

Preliminary study:

No data

Test performance:

No data

Key result

Parameter:

% degradation (O2 consumption)

Value:

>= 90 - <= 95

Sampling time:

28 d

Details on results:

- Indirect analysis (BOD): 91, 90 and 95%
- Direct analysis (GC): 100, 100 and 100%

Results with reference substance:

No data

None

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

Under the test conditions, alpha-pinene is readily biodegradable.

Executive summary:

In a ready biodegradation study performed according to OECD Guideline 301 C, alpha-pinene was tested at concentrations of 100 mg/L and the inoculum was activated sludge (30 mg/L). The degradation of the test material was assessed by the determination of the oxygen consumption.

At 100 mg/L test concentration, 90-95% degradation (biochemical oxygen demand) was reached in 28 days.

 

Under the test conditions, alpha-pinene is readily biodegradable.

 

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2015

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

This screening study was performed according to OECD Guideline 301 D but was not performed according to GLP as this study was originally conducted by R&D department to increase knowledge on the fate of the substance, not related to registration purposes. This study is well documented and is considered reliable with restrictions.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

This study was not performed according to GLP as this is a study conducted for global knowledge.

Specific details on test material used for the study:

No additional information

Oxygen conditions:

aerobic

Inoculum or test system:

other: two inoculum used: activated sludge and river water

Details on inoculum:

Two inoculums were used:
- Activated sludge was obtained from the wastewater treatment plant Nieuwgraaf in Duiven, The Netherlands. This plant treats predominantly domestic wastewater. The activated sludge was preconditioned to reduce the endogenous respiration rates. To this end, 400 mg Dry Weight (DW)/L of activated sludge was aerated for one week. The sludge was diluted to 2 mg DW/L and distributed over the appropriate biological oxygen demand (BOD) bottles (van Ginkel and Stroo, 1992).
- River water was sampled from the Rhine near Heveadorp, The Netherlands. The river water was aerated for 7 days 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 spiked with mineral salts of the nutrient medium was used undiluted.

Duration of test (contact time):

56 d

Initial conc.:

2 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium: The nutrient medium of the Closed Bottle test contained per liter of deionized water: 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.4 mg Na2HPO4•2H2O, 22.5 mg MgSO4•7H2O, 27.5 mg CaCl2, and 0.25 mg FeCl3•6H2O. Ammonium chloride was omitted from the medium to prevent nitrification.
- Solubilising agent (type and concentration if used): None
- Test temperature: 22-24 °C
- Suspended solids concentration: 2 mg DW/L
- Continuous darkness: Yes
- Deionized water containing no more than 0.01 mg/L copper was prepared in a water purification system.
- Other: None

TEST SYSTEM
- Culturing apparatus: 0.3 L Biological oxygen demand (BOD) bottles with glass stoppers
- Number of culture flasks/concentration: 3 bottles containing only inoculum (control) and 3 bottles for test substance with the inoculum
- Method used to create aerobic conditions: No data
- Method used to create anaerobic conditions: Not applicable
- Measuring equipment: The dissolved oxygen concentrations were determined electrochemically using an oxygen electrode (WTW Trioxmatic EO 200) and meter (WTW OXI 530) (Retsch, Ochten, The Netherlands). The pH was measured using a EUTECH instruments pH meter. The temperature was measured and recorded with a thermo couple connected to a data logger.
- Test performed in closed vessels: Yes
- Test performed in open system: No

CONTROL AND BLANK SYSTEM
- Inoculum blank: Yes

OTHERS:
- The dry weight of the inoculum was determined by filtrating 50 mL of the activated sludge over a pre-weighed 12 µm cellulose nitrate filter. This filter was dried for 1.5 hours at 104 °C and weighed after cooling. The dry weight was calculated by subtracting the weighed filters and by dividing this difference by the filtered volume.
The biological oxygen demand (BOD) mg/mg of the test compound was calculated by dividing the oxygen consumption by the concentration of the test substance in the closed bottle. The biodegradation was calculated as the ratio of the biochemical oxygen demand (BOD) to the theoretical oxygen demand (ThOD).

Reference substance:

not specified

Preliminary study:

None

Test performance:

The validity of the test is demonstrated by oxygen concentrations >0.5 mg/L in all bottles during the test period. The pH of the media was 7.3±0.2 (activated sludge) and 8.0±0.2 (river water).

Key result

Parameter:

% degradation (O2 consumption)

Value:

68

Sampling time:

28 d

Remarks on result:

other: activated sludge

Key result

Parameter:

% degradation (O2 consumption)

Value:

67

Sampling time:

28 d

Remarks on result:

other: river water

Details on results:

AKZO093 was readily biodegradable as demonstrated by the >60% result at day 28.

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

The test item should be regarded as readily biodegradable according to this test.

Executive summary:

This study was performed according to the OECD Guideline 301 D, to determine the biodegradability of the test substance by the Closed Bottle Test.

The test material was exposed to activated sewage sludge and river water micro-organisms at a concentration of 2 mg/L with culture medium in closed bottles in the dark at 22-24 °C for 7, 14, 21, 28, 42 and 56 days. The degradation of the test material was assessed by the measurement of oxygen consumption.

 

The validity of the test is demonstrated by oxygen concentrations > 0.5 mg/L in all bottles during the test period.

 

Test substance	Inoculum	Biodegradation at day (%)
		7	14	21	28	42	56
	Sludge	18	49	58	68	80	79
	River	29	48	61	67	70	71

 

The test item undergoes 68 and 67 % degradation in activated sludge and river, respectively, after 28 days, in the test conditions. Thus, the test item should be regarded as readily biodegradable according to this test.

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Study period:

2017-03-30 to 2017-03-30

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

results derived from a (Q)SAR model, with limited documentation / justification

Justification for type of information:

1. SOFTWARE
OASIS CATALOGIC v.5.11.19

2. MODEL (incl. version number)
CATALOGIC Kinetic 301F v.13.16 (June, 2016)

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
C/C1\CC/C=C(C)CCC2C(/C=1)C2(C)C

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
See attached QMRF

5. APPLICABILITY DOMAIN
See attached QPRF

6. ADEQUACY OF THE RESULT
See attached QPRF

Qualifier:

according to guideline

Guideline:

other: Reach Guidance on QSAR - R.6

Deviations:

no

Principles of method if other than guideline:

QSAR, CATALOGIC Kinetic 301F v.13.16 within LMC OASIS CATALOGIC v5.11.19.
More details are given in QMRF/QPRF attached to the dossier.

GLP compliance:

no

Specific details on test material used for the study:

No additional information

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Aerobic microorganisms: activated sludge taken from a treatment plant or laboratory-scale unit receiving predominantly domestic sewage. Inoculum may be pre-conditioned to the experimental conditions, but not pre-adapted to the test substance. Pre-conditioning consists of aerating activated sludge in mineral medium or secondary effluent for 5-7 days at the test temperature.

Duration of test (contact time):

28 d

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Not applicable

Preliminary study:

Not applicable

Test performance:

Not applicable

Key result

Parameter:

% degradation (O2 consumption)

Value:

56

Sampling time:

28 d

Remarks on result:

not readily biodegradable based on QSAR/QSPR prediction

Details on results:

The outcome of the model for bi-cyclogermacrene is not considered fully suitable to support the assessment of its biodegradability potential. The chemical fulfils the general properties requirements (log Kow, molecular weight and water solubility) but is out of the interpolation structural space and out of the metabolic domain.
The model indicates that bi-cyclogermacrene achieves 56% biodegradation (BOD) after 28 days, not exceeding the 60 % pass level set by the OECD Guidelines for testing of Chemicals (301) for ready biodegradability of a substance. It can be concluded that bi-cyclogermacrene is not readily biodegradable.
For additional informations, see the attached QPRF.

Results with reference substance:

Not applicable

Prediction percentage of biodegradation after 28d:

%BOD [28d] = 0.56 ± 0.00914

Concomitant predictions: Not ready degradable

Primary half-life < 1 day

Ultimate half-life = 23.79 days

Start day of Calc. 10 days window = 3

%BOD calc at 10 days window = 0.4534

Classification of Calc. 10 days window = Not Ready

Validity criteria fulfilled:

not applicable

Interpretation of results:

not readily biodegradable

Conclusions:

Bi-cyclogermacrene is predicted to be not readily biodegradable, however the outcome of this prediction needs to be taken with caution because the substance falls outside the metabolic domain of the model even though a large part of bi-cyclogermacrene fragments are recognized by the model (80%).

Executive summary:

The ready biodegradability of bi-cyclogermacrene has been investigated using the QSAR model CATALOGIC Kinetic 301F v.13.16 within LMC OASIS CATALOGIC v5.11.19. This model allows to predict the ready biodegradability of a substance in an OECD 301F study.

The outcome of the model for bi-cyclogermacrene is not considered fully suitable to support the assessment of its biodegradability potential.

The model indicates that bi-cyclogermacrene achieves 56% biodegradation (BOD) after 28 days, not exceeding the 60 % pass level set by the OECD Guidelines for testing of Chemicals (301) for ready biodegradability of a substance. It can be concluded that bi-cyclogermacrene is not readily biodegradable.

Bi-cyclogermacrene is predicted to be not readily biodegradable, however the outcome of this prediction needs to be taken with caution because the substance falls outside the metabolic domain of the model even though a large part of bi-cyclogermacrene fragments are recognized by the model (80%).

Reference 4

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Study period:

2017-04-12 to 2017-04-12

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

results derived from a (Q)SAR model, with limited documentation / justification

Justification for type of information:

1. SOFTWARE
OASIS CATALOGIC v.5.11.19

2. MODEL (incl. version number)
CATALOGIC Kinetic 301F v.13.16 (June, 2016)

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
CC(=C)C1CC(C(C)(C)O)CCC1(C)C=C

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
See attached QMRF

5. APPLICABILITY DOMAIN
See attached QPRF

6. ADEQUACY OF THE RESULT
See attached QPRF

Qualifier:

according to guideline

Guideline:

other: Reach Guidance on QSAR - R.6

Deviations:

no

Principles of method if other than guideline:

QSAR, CATALOGIC Kinetic 301F v.13.16 within LMC OASIS CATALOGIC v5.11.19.
More details are given in QMRF/QPRF attached to the dossier.

GLP compliance:

no

Specific details on test material used for the study:

No additional information

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Aerobic microorganisms: activated sludge taken from a treatment plant or laboratory-scale unit receiving predominantly domestic sewage. Inoculum may be pre-conditioned to the experimental conditions, but not pre-adapted to the test substance. Pre-conditioning consists of aerating activated sludge in mineral medium or secondary effluent for 5-7 days at the test temperature.

Duration of test (contact time):

28 d

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Not applicable

Preliminary study:

Not applicable

Test performance:

Not applicable

Key result

Parameter:

% degradation (O2 consumption)

Value:

34

Sampling time:

28 d

Remarks on result:

not readily biodegradable based on QSAR/QSPR prediction

Details on results:

The outcome of the model for elemol is not considered fully suitable to support the assessment of its biodegradability potential. The chemical fulfils the general properties requirements (log Kow, molecular weight and water solubility) but is out of the interpolation structural space and out of the metabolic domain.
The model indicates that elemol achieves 34% biodegradation (BOD) after 28 days, not exceeding the 60 % pass level set by the OECD Guidelines for testing of Chemicals (301) for ready biodegradability of a substance. It can be concluded that elemol is not readily biodegradable. This may be due to the fact the model failed to simulate metabolism.
For additional informations, see the attached QPRF.

Results with reference substance:

Not applicable

Prediction percentage of biodegradation after 28d:

%BOD [28d] = 0.34 ± 0.00738

Concomitant predictions: Not ready degradable

Primary half-life < 1 day

Ultimate half-life = 1m 15d

Start day of Calc. 10 days window = 6

%BOD calc at 10 days window = 0.2489

Classification of Calc. 10 days window = Not Ready

Validity criteria fulfilled:

not applicable

Interpretation of results:

not readily biodegradable

Conclusions:

Elemol is predicted to be not readily biodegradable by the present model based on the large portion of the structure which is correctly predicted (i.e. >80% of the atom centered fragments). However, this conclusion may be due to the fact that the simulator of metabolism failed to simulate the mineralization of elemol.

Executive summary:

The ready biodegradability of elemol has been investigated using the QSAR model CATALOGIC Kinetic 301F v.13.16 within LMC OASIS CATALOGIC v5.11.19. This model allows to predict the ready biodegradability of a substance in an OECD 301F study.

The outcome of the model for elemol is not considered fully suitable to support the assessment of its biodegradability potential.

The model indicates that elemol achieves 34% biodegradation (BOD) after 28 days, not exceeding the 60 % pass level set by the OECD Guidelines for testing of Chemicals (301) for ready biodegradability of a substance.

Elemol is predicted to be not readily biodegradable by the present model based on the large portion of the structure which is correctly predicted (i.e. >80% of the atom centered fragments). However, this conclusion may be due to the fact that the simulator of metabolism failed to simulate the mineralization of elemol.

Reference 5

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

This study was not performed according to international guideline and is not GLP as this is a publication conducted for global knowledge.

Specific details on test material used for the study:

No additional information

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Fresh activated sludge from a biological wastewater treatment plant treating predominantly domestic sewage (City of Geneva, Peney-Dessous, Switzerland) was used. The sludge was collected in the morning, washed three times in the mineral medium (by centrifuging at 1000 g for 10 min, discarding the supernatant and resuspending in mineral medium) and kept aerobic until being used on the same day. The dry weight of suspended solids was determined by taking two 50-ml samples of the homogenized sludge, evaporating water on a steam bath, drying in an oven at 1058C to 1108C for 2 h and weighing the residue.

Duration of test (contact time):

60 d

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

MINERAL MEDIUM:
The water used during the present study was ultrapure water, containing less than 5 ppb total organic carbon, produced by a Millipore Direct-Q 3 ultraviolet purification system, or equivalent. A mineral medium as described in the OECD 301 Guideline was prepared. The pH was measured and if necessary adjusted to 7.4 +/- 0.2 with phosphoric acid or potassium hydroxide.

BIODEGRADATION ASSAY:
Ready biodegradability tests were conducted at the laboratories of Givaudan in Vernier, Switzerland. The method used was the one described under No. 301 F in the OECD Guidelines for Testing of Chemicals. The respirometers used during the present study were Sapromat D 12, made by J. M. Voith. Glass capillaries were introduced between the test flasks (500 ml nominal volume) and the oxygen generators to limit losses of the volatile test substance. A reference compound, sodium benzoate, was run in parallel to confirm the viability of the bacterial inoculum. As suggested in the OECD 301 F method, the toxicity of the test substance for the inoculum was checked by testing a mixture of test substance (100 mg/L) and reference substance (100 mg/L) (i.e., both at the same test concentration as in the individual solutions). If the respirations in the toxicity control are lower than those of the flasks containing mineral medium, reference substance (100 mg/L), and inoculum, the test substance can be assumed to be inhibitory to the inoculum used. All determinations were carried out in duplicate.
Sesquiterpene test substance samples (25 mg, corresponding to 100 mg/L in a 250 ml flask) were weighed in small aluminum boats and added directly to the test flasks of the Sapromat, whereas reference substance samples (sodium benzoate) were added as 1.0 ml of a 25.0 mg/ml solution in mineral medium. All flasks were filled with 250 ml of mineral medium. Samples of test or reference substance, or both in the case of the toxicity control, were added. Then, a volume of suspended sludge corresponding to 30 mg/L dry weight (as determined) was added. Because of the low solubility of the test substances and in order not to remove any floating undissolved test substance from the test medium by dipping a glass electrode in it, the pH of each flask was not measured but assumed to be the same as the mineral medium. Neutral test substances (even sodium benzoate, which is slightly alkaline) have previously been shown not to affect the pH of the medium by more than
0.1 pH unit. Approximately 2 g soda lime was placed in an attachment of the stopper. The flasks were closed and placed in the water bath of the Sapromat. After temperature and pressure equilibration, the oxygen-meters of the instrument were set to zero (time zero of the experiment). Each day, the oxygen consumption of each flask was recorded and correct temperature and stirring were checked. At the end of the test period, the pH of each flask was measured and found to be in the range of pH 7.4 to 7.7 for all test substance flasks.

Reference substance:

benzoic acid, sodium salt

Preliminary study:

None

Test performance:

See details below

Key result

Parameter:

% degradation (O2 consumption)

Value:

19

Sampling time:

28 d

Remarks on result:

other: Not readily biodegradable

Key result

Parameter:

% degradation (O2 consumption)

Value:

23

Sampling time:

60 d

Remarks on result:

other: degradation curve had not reached a plateau at the end of the test + evidence of primary degradation

Details on results:

Germacrene D achieved only 23% biodegradation in the prolonged test (60 days). The germacrene D sample contained 12.7% of b-caryophyllene, which achieved 70% biodegradation in a previous 301F test. The level of biodegradation seen for the germacrene D sample is higher than one would expect for the b-caryophyllene content indicating that some biodegradation of germacrene D must have occurred. The degradation curves for germacrene D had still not reached a plateau at day 63 to 65 showing that biodegradation was still progressing albeit very slowly.

Analysis of the test flasks at the end of the germacrene D experiment showed that the sesquiterpene test substance had disappeared from the test medium. No germacrene D was recovered from the test flasks at the end of the test. Because glass capillaries were used to link the test flasks to the oxygen generator, simple evaporation from the test system is unlikely and the disappearance of the test substance can be regarded as evidence for primary biodegradation. The b-caryophyllene present in the germacrene D sample is also not present in the cyclohexane extract of the germacrene D test flask. However, two new small peaks occur at retention times 12.5 and 15.0 min that may be metabolites from primary biodegradation or products of abiotic reactions.

Because primary biodegradation is predicted for Germacrene D, it is pertinent to check the PBT-characteristics of expected metabolites. The most plausible long-lived or stable metabolites generated by Catalogic and their estimated log KOW are all <4.5, which means that they do not fulfil the REACH B/vB screening criterion and, in turn, are not potential PBT or vPvB degradation products. Because germacrene D is predicted to undergo complete primary degradation they have been assigned as borderline non-persistency case.

Results with reference substance:

The curves obtained with the reference substance alone and with sesquiterpene and reference substance show no toxic effect of the sesquiterpene to the microorganisms at the test concentration (100 mg/L).

See attached document in "Attached full study report"

Validity criteria fulfilled:

not applicable

Interpretation of results:

other: Not readily biodegradable but complete primary degradation

Conclusions:

Although germacrene D only achieved 23% ultimate biodegradation after 60 days, specific analysis at the end of the test did indicate complete primary degradation. Specific analysis showed that no parent sesquiterpene remained at the end of the test period, indicating primary degradation and possibly also loss due to abiotic reactions.

Executive summary:

Sesquiterpenes are ubiquitous in essential oils but an assessment of their environmental behavior is still required for their use as components of natural fragrance ingredients and oral care flavors. Persistency plays a key role in hazard and risk assessment, but the current knowledge on the biodegradation of sesquiterpenes in the aquatic environment is limited. This could have important consequences for the persistent, bioaccumulative and toxic (PBT) assessment of essential oils because most of the sesquiterpene components have a log KOW of >4.5 and are identified as potentially bioaccumulating according to REACH screening criteria. In the present study, a persistency screening assessment was conducted on 11 cyclic sesquiterpenes selected from 10 different families of sesquiterpenes characterized by their carbon skeleton. Aerobic biodegradation was measured in a standard or prolonged Organisation for Economic Co-operation and Development (OECD) 301F Manometric Respirometry test for ready biodegradability. Although germacrene D only achieved 23% ultimate biodegradation after 60 days, specific analysis at the end of the test did indicate complete primary degradation. In addition, the degradation curve had not reached a plateau at the end of the test. Given that the shape of the biodegradation curves indicates poor bioavailability and ready biodegradability tests are very stringent, it is expected that all the sesquiterpenes tested in the present study would be degraded under environmental conditions.

Reference 6

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1980

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Study conducted according to OECD Guideline 301 C but with deviations: no detail on material and methods

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Deviations:

yes

Remarks:

no detail on material and methods

Principles of method if other than guideline:

Not applicable

GLP compliance:

not specified

Specific details on test material used for the study:

No additional information

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- Concentration of sludge: 30 ppm

Duration of test (contact time):

14 d

Initial conc.:

100 other: ppm

Based on:

test mat.

Details on study design:

No data

Preliminary study:

No data

Test performance:

No data

Key result

Parameter:

% degradation (O2 consumption)

Value:

>= 41 - <= 98

Sampling time:

14 d

Details on results:

- Indirect analysis (BOD): 41, 81 and 98%
- Direct analysis (GC): 100%
- Direct analysis (TOC): 50, 73 and 81%

Results with reference substance:

No data

None

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

Under the test conditions, Limonene is readily biodegradable.

Executive summary:

In a ready biodegradation study performed according to OECD Guideline 301 C, Limonene was tested at concentrations of 100 ppm and the inoculum was activated sludge (30 ppm). The degradation of the test material was assessed by the determination of the oxygen consumption.

At 100 ppm test concentration, 41-98% degradation by biochemical oxygen demand was reached in 14 days.

 

Under the test conditions, Limonene is readily biodegradable.

 

Reference 7

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1987

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Study conducted according to OECD Guideline 301 C but with deviations: no detail on material and methods

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Deviations:

yes

Remarks:

no detail on material and methods

Principles of method if other than guideline:

Not applicable

GLP compliance:

not specified

Specific details on test material used for the study:

No additional information

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- Concentration of sludge: 30 ppm

Duration of test (contact time):

2 wk

Initial conc.:

100 other: ppm

Based on:

test mat.

Details on study design:

No data

Preliminary study:

No data

Test performance:

No data

Key result

Parameter:

% degradation (O2 consumption)

Value:

86

Sampling time:

2 wk

Details on results:

- Indirect analysis (BOD): 86%
- Direct analysis (GC): 99%
- Direct analysis (TOC): 87%

Results with reference substance:

No data

None

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

Under the test conditions, myrcene beta is readily biodegradable.

Executive summary:

In a ready biodegradation study performed according to OECD Guideline 301 C, myrcene beta was tested at concentrations of 100 ppm and the inoculum was activated sludge (30 ppm). The degradation of the test material was assessed by the determination of the oxygen consumption.

At 100 ppm test concentration, 86% degradation by biochemical oxygen demand was reached in 2 weeks.

 

Under the test conditions, myrcene beta is readily biodegradable.

 

Reference 8

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1987

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Study conducted according to OECD Guideline 301 C but with deviations: no detail on material and methods

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Deviations:

yes

Remarks:

no detail on material and methods

Principles of method if other than guideline:

Not applicable

GLP compliance:

not specified

Specific details on test material used for the study:

No additional information

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- Concentration of sludge: 30 ppm

Duration of test (contact time):

2 wk

Initial conc.:

100 other: ppm

Based on:

test mat.

Details on study design:

No data

Preliminary study:

No data

Test performance:

No data

Key result

Parameter:

% degradation (O2 consumption)

Value:

88

Sampling time:

2 wk

Details on results:

- Indirect analysis (BOD): 88%
- Direct analysis (GC): 100%
- Direct analysis (TOC): 89%

Results with reference substance:

No data

None

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

Under the test conditions, p-cymene is readily biodegradable.

Executive summary:

In a ready biodegradation study performed according to OECD Guideline 301 C, p-cymene was tested at concentrations of 100 ppm and the inoculum was activated sludge (30 ppm). The degradation of the test material was assessed by the determination of the oxygen consumption.

At 100 ppm test concentration, 88% degradation by biochemical oxygen demand was reached in 2 weeks.

 

Under the test conditions, p-cymene is readily biodegradable.

 

Description of key information

Based on a Weight of Evidence approach with data on constituents, the registered substance is considered readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable

Additional information

The major part of the mixture is composed by readily biodegradable constituents (see table below):

Constituents	CAS N°	% in the mixture*	Biodegradation result	Reference
Alpha-phellandrene	99-83-2	32.46	Considered readily biodegradable based on similar compounds like limonene or paracymene	Read-across with limonene or paracymene: Both substances have the same carbon skeleton. The only difference is the position of the double bonds. This is not expected to affect the sensitivity to biodegradation in a significant way.
Alpha-pinene	80-56-8	15.57	Readily biodegradable	MITI, 2007
Germacrene D	37839-63-7	10.13	Not readily biodegradable but complete primary degradation and therefore not persistent	Karen, 2010
Limonene	138-86-3	9.72	Readily biodegradable	MITI, 1980
Beta-phellandrene**	555-10-2	7.69	Readily biodegradable	ECHA disseminated dossier of the Reaction mass of 3-isopropyl-6-methylenecyclohexene and (4R)-1-methyl-4-(prop-1-en-2-yl)cyclohexene and (4S)-1-methyl-4-(prop-1-en-2-yl)cyclohexene: https://echa.europa.eu/es/registration-dossier/-/registered-dossier/13867/5/3/2   Composition of the test material: -       48.4% of dl-limonene; -       20.6% of beta phellandrene; -       9.8% of alpha terpinene; -       5.8% of gamma terpinene; -       4.5% of terpinolene.
Delta-3-carene	13466-78-9	6.89	Readily biodegradable	ECHA disseminated dossier of the substance: https://echa.europa.eu/registration-dossier/-/registered-dossier/2226/5/3/2
Elemol	639-99-6	1.82	Not readily biodegradable	QSAR, CATALOGIC Kinetic 301F v.13.16
Myrcene	123-35-3	1.62	Readily biodegradable	MITI, 1987
Paracymene	99-87-6	1.59	Readily biodegradable	MITI, 1987
Bicyclogermacrene	24703-35-3	1.59	Not readily biodegradable	QSAR, CATALOGIC Kinetic 301F v.13.16
Sabinene	3387-41-5	1.48	Readily biodegradable	Givaudan data*** OECD Guideline 301F,2010. Test material: 71.83% pure substance + 26.92% beta-pinene as the major secondary constituent. Biodegradation result: 76% biodegradation after 28 days within the 10-day window.
Beta-caryophyllene	87-44-5	1.24	Readily biodegradable	AkzoNobel, 2015
Beta pinene	127-91-3	1.0	Readily biodegradable	ECHA disseminated dossier of the substance: https://echa.europa.eu/registration-dossier/-/registered-dossier/15934/5/3/2

* these percentage values correspond to the typical legal entity composition provided by the registrant of the substance which consider all constituents > 1%

** Beta-phellandrene does not exist as a pure substance

*** Givaudan gave the approval to the registrant to refer to the result of their biodegradation study without including the robust study summary in the REACH registration dossier of the present mixture.

 

Based on a Weight of Evidence approach, the substance is considered readily biodegradable.

Indeed, 10 constituents representing 79.26% of the substance (corresponding to 85.41% of the known constituents of the mixture) are readily biodegradable.

In addition, one constituent representing 10.13% of the substance is not readily biodegradable but primarily degraded and therefore not persistent. Therefore, 11 constituents representing 89.39% of the substance (corresponding to 96.33% of the known constituents of the mixture) are not persistent in the environment.

Only 2 other constituents representing 3.41% of the substance (equivalent to 3.67% of the known constituents of the mixture) are considered as not readily biodegradable. Thus together with the 10.13% of Germacrene, 13.54% of the substance can be considered as not rapidly biodegradable and overall the substance may be considered readily biodegradable.