Caryophyllene, Reaction product with Acetic anhydride and acetic acid

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

16 September 1999 - 19 October 1999

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

VETYNAL
Lot No. : 9000349063
Purity : GLC profile matches standard

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Fresh activated sludge from a biological waste water treatment plant treating predominantly domestic sewage (City of Geneva, Aïre) was used.

The sludge is collected in the morning, washed three times in the mineral medium (by centrifuging at 1000 g for 10 minutes, discarding the supernatant and resuspending in mineral medium) and kept aerobic until being used on the same day.

Duration of test (contact time):

32 d

Initial conc.:

30 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Water

The water used during this study is deionised water containing less than 10 mg/l dissolved organic carbon.


Preparation of the flasks

Test substance samples (30 mg, corresponding to 30 mg/l in a 1000-ml flask) are weighed in small aluminium boats and added directly to the test flasks of the BI-1000, whereas the reference substance sample (sodium benzoate) is added as 1.0 ml of a 100 mg/ml solution in mineral medium.

All flasks are filled with 1000 ml of mineral medium. Samples of test or reference substances are added. Then, a volume of suspended sludge corresponding to 100 mg dry weight (as determined from 3.7, generally 15 to 40 ml) is added. Except when the test substance has an acid or alkaline character, the pH of each flask is not measured but assumed to be the same as the mineral medium, in order not to remove any floating undissolved test substance from the test medium by dipping a glass electrode in it. Neutral test substances, even sodium benzoate, were shown not to affect the pH of the medium by more than 0.1 pH unit. About 5 g of soda lime is placed in an attachment of the stopper, the flasks are closed and placed in the water bath of the BI-1000. After temperature and pressure equilibration, the measuring cells are started (time zero of the experiment).

Performance of the test


The oxygen consumption of each flask is automatically recorded by the BI-1000 during the whole test duration. Correct temperature and stirring are regurlarly checked visually.

At the end of the test period, the pH of each flask is measured.

Reference substance:

benzoic acid, sodium salt

Remarks:

Purity : Minimum 99.0%

Parameter:

% degradation (O2 consumption)

Value:

37

Sampling time:

28 d

Key result

Parameter:

% degradation (O2 consumption)

Value:

42

Sampling time:

32 d

Results with reference substance:

Degradation of sodium benzoate is 77 % after 7 days and 88 % after 14 days, widely in excess of the validity criterium: the activity of the inoculum is thus verified and the test is considered valid.

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

VETYNAL undergoes 37 % biodegradation after 28 days (and 42 % after 32 days) in the test conditions.

Thus, VETYNAL should be regarded as not inherently but partially biodegradable according to this test.

Executive summary:

VETYNAL undergoes 37 % biodegradation after 28 days (and 42 % after 32 days) in the test conditions.

 

Thus, VETYNAL should be regarded as not inherently but partially biodegradable according to this test,

with 37% of the starting Parent material completely mineralised in to CO2 and H2O by Day 28, increasing to 42% by Day 32.

Reference 2

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

February 04 - April 11, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name (as stated in the report): VETYNAL Extra
Batch: VE00375495
Expiration date: June 07, 2017
Purity: Conform

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Fresh activated sludge from a biological waste water treatment plant treating predominantly domestic sewage (Bois-de-Bay, Satigny, Switzerland) was used.

The sludge is collected in the morning, washed three times in the mineral medium (by centrifuging at 1000 g for 10 minutes, discarding the supernatant and resuspending in mineral medium) and kept aerobic until being used on the same day.

Duration of test (contact time):

67 d

Initial conc.:

30 mg/L

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Water

The water used during this study is ultrapure water, containing less than 5 ppb total organic carbon, produced by using a Millipore Direct-Q 3 UV purification system.


Preparation of the flasks

Test substance samples (13.47 mg, corresponding to 30.0 mg/l in 449 ml of test volume) are weighed in small aluminium boats and added directly to the test flasks of the Oxitop that were previously charged with 432 ml of mineral medium. Then, 17.1 ml of suspended sludge, diluted to a concentration of 2.62 g/l dry matter, corresponding to 44.8 mg dry weight is added. Flasks containing sludge only (100 mg/l) are charged with 432 ml of mineral medium. Then, 17.1 ml of suspended sludge, diluted to a concentration of 2.62 g/l dry matter, are added.

Reference substance samples (~43.70 mg, corresponding to 100 mg/l in 437 ml of test volume) are weighed in small aluminium boats and added directly to the test flasks of the Oxitop that were previously charged with 432 ml of mineral medium. Then, 5.0 ml of suspended sludge, diluted to a concentration of 2.62 g/l dry matter, corresponding to 13.1 mg dry weight is added. Flasks containing sludge only (30 mg/l) are charged with 432 ml of mineral medium. Then, 5.0 ml of suspended sludge, diluted to a concentration of 2.62 g/l dry matter, are added.

Except when the test substance has an acid or alkaline character, the pH of each flask is not measured but assumed to be the same as the mineral medium, in order not to remove any floating undissolved test substance from the test medium by dipping a glass electrode in it. Neutral test substances, even sodium benzoate, were shown not to affect the pH of the medium by more than 0.1 pH unit. Two sodium hydroxide pellets are placed in the quivers on top of the bottle, and the flasks are closed tightly with the measuring heads. The flasks are allowed to equilibrate to the test temperature. The measurement is started by programming the measuring unit of the Oxitop test flasks, and the test flasks are placed in the temperature controlled cupboard of the Oxitop system. After temperature equilibration, the controller of the instrument starts the data acquisition (time zero of the experiment).

Performance of the test

Everyday the oxygen consumption of each flask is recorded and correct temperature and stirring are checked.

At the end of the test period (normally 28 days), the pH of each flask is measured again.

Reference substance:

benzoic acid, sodium salt

Remarks:

purity: min. 99.0 %

Parameter:

% degradation (O2 consumption)

Value:

39

Sampling time:

28 d

Key result

Parameter:

% degradation (O2 consumption)

Value:

56

Sampling time:

67 d

Results with reference substance:

Degradation of sodium benzoate exceeded 40 % after 7 days and 65 % after 14 days: the activity of the inoculum was thus verified (validity criterion).

Validity criteria fulfilled:

yes

Interpretation of results:

other: inherently primarily biodegradable

Conclusions:

In the test conditions Vetynal Extra undergoes 39% biodegradation after 28 days (56% biodegradation after 67 days).

Vetynal Extra did not significantly inhibit the intrinsic respiration of the inoculum at the test concentration and was therefore considered to be non-toxic to the inoculum at the test concentration.

Vetynal Extra cannot be regarded as inherently and ultimately biodegradable according to this test. However, Vetynal Extra should be regarded as inherently primarily biodegradable according to this test.

Executive summary:

The Inherent Biodegradability of Vetynal Extra was determined by the Manometric Respirometry Test according to the OECD Guidelines for Testing of Chemicals, Method No. 302C.

In the test conditions Vetynal Extra undergoes 39% biodegradation after 28 days (56% biodegradation after 67 days).

 

Vetynal Extra did not significantly inhibit the intrinsic respiration of the inoculum at the test concentration and was therefore considered to be non-toxic to the inoculum at the test concentration.

Vetynal Extra cannot be regarded as inherently and ultimately biodegradable according to this test. However, Vetynal Extra should be regarded as inherently primarily biodegradable according to this test with 39% of the starting Parent material completely mineralised in to CO2 and H2O by Day 28, injcreasing to 56% by Day 67.

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 July 2015 - 19 October 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

EU Method C.4-D (Determination of the "Ready" Biodegradability - Manometric Respirometry Test)

Qualifier:

according to guideline

Guideline:

other: ECHA Guidance for the implementation of REACH, Guidance on information requirements and chemical safety assessment, Chapter R.7b: Endpoint specific guidance, November 2014 (version 2.0)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 835.3110 (Ready Biodegradability)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name (as stated in the report): VETYNAL Extra
Batch: VE00375495
Expiration date: June 07, 2017
Purity: Conform

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Fresh activated sludge from a biological waste water treatment plant treating predominantly domestic sewage (Bois-de-Bay, Satigny, Switzerland) was used.

The sludge is collected in the morning, washed three times in the mineral medium (by centrifuging at 1000 g for 10 minutes, discarding the supernatant and resuspending in mineral medium) and kept aerobic until being used on the same day.

Duration of test (contact time):

62 d

Initial conc.:

30 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Preparation of the flasks

Test substance samples (13.10 mg, corresponding to 30.0 mg/l in 437 ml of test medium) are weighed in small aluminium boats and added directly to the test flasks of the Oxitop. For reference substance samples 43.70 mg (corresponding to 100.0 mg/l in 437 ml of test medium) are weighed in small aluminium boats and added directly to the test flasks of the Oxitop.

Flasks are filled with 432 ml of mineral medium. Samples of test or reference substance are added. Then 5.00 ml of suspended sludge diluted to a concentration of 2.62 g/l dry matter is added. Except when the test substance has an acid or alkaline character, the pH of each flask is not measured but assumed to be the same as the mineral medium, in order not to remove any floating undissolved test substance from the test medium by dipping a glass electrode in it. Neutral test substances, even sodium benzoate, were shown not to affect the pH of the medium by more than 0.1 pH unit. Two sodium hydroxide pellets are placed in the quivers on top of the bottle, and the flasks are closed tightly with the measuring heads. The flasks are allowed to equilibrate to the test temperature. The measurement is started by programming the measuring unit of the Oxitop test flasks, and the test flasks are placed in the temperature controlled cupboard of the Oxitop system. After temperature equilibration, the controller of the instrument starts the data acquisition (time zero of the experiment).

Performance of the test

Everyday the oxygen consumption of each flask is recorded and correct temperature and stirring are checked.

At the end of the test period, the pH of each flask is measured again.

Reference substance:

benzoic acid, sodium salt

Remarks:

99% purity, concentration 100 mg/L

Key result

Parameter:

% degradation (O2 consumption)

Value:

12

Sampling time:

28 d

Parameter:

% degradation (O2 consumption)

Value:

41

Sampling time:

62 d

Results with reference substance:

Degradation of sodium benzoate exceeded 40% after 7 days and 65% after 14 days: the activity of the inoculum was thus verified (validity criterion).

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

Vetynal Extra undergoes 12% biodegradation after 28 days (41% after 62 days) in the test conditions.

Vetynal Extra did inhibit the intrinsic respiration of the inoculum at the test concentration at the beginning of the test. Toxicity towards the inoculum at test concentration can therefore not be excluded.

Vetynal Extra should be regarded as not readily biodegradable according to this test.

Executive summary:

The Ready Biodegradability of Vetynal Extra was determined by the Manometric Respirometry Test.

Vetynal Extra undergoes 12% biodegradation after 28 days (41% after 62 days) in the test conditions.

Vetynal Extra did inhibit the intrinsic respiration of the inoculum at the test concentration at the beginning of the test. Toxicity towards the inoculum at test concentration can therefore not be excluded.

Vetynal Extra should be regarded as not readily biodegradable according to this test. Significant mineralisation was, nonetheless, observed during the study suggesting a high rate of metabolism of the parent.

Key value for chemical safety assessment

Biodegradation in water:

inherently biodegradable