3-hydroxy-p-anisaldehyde

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See 'Attached justification'.

Reason / purpose for cross-reference:

read-across source

GLP compliance:

no

Key result

Parameter:

% degradation (CH4 evolution)

Value:

72

Sampling time:

28 d

Remarks on result:

other: Read-across from analogue substance.

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

Based on the available information for the read-across approach, the target substance is deemed readily biodegradable.

Executive summary:

The study of the ready biodegradability of the analogue substance vanillin was performed following a serum-bottle variation of the Hungate technique for growing anaerobic bacteria adapted from Miller and Wolin (1974), similar to OECD 311 (non-GLP). Ten serum bottles containing 300 mg/L test item each were inoculated with 10%(v/v) seed from a laboratory anaerobic digester fed primary settled sewage sludge and incubated under anaerobic conditions in the dark at 35ºC. A negative control was run in parallel, and benzoic acid could serve as a positive control. The amount of biodegradation was measured by the determination of methane evolution at frequent intervals over a 28 day period. After 28 days, the biodegradation of the analogue substance reached 72% and therefore, it is readily biodegradable. Based on the read-across approach, the target substance is deemed readily biodegradable.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1979.

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 311 (Anaerobic Biodegradability of Organic Compounds in Digested Sludge: Measurement of Gas Production)

Principles of method if other than guideline:

- Principle of test: A serum-bottle variation of the Hungate technique for growing anaerobic bacteria was adapted from Miller and Wolin (see attached background material).
- Short description of test conditions: serum bottles closed with a butyl rubber stopper with a crimped metal seal were flushed with oxygen-free gas for 20 min before addition of the inoculum. Cultures were incubated in the dark at 35ºC, and maintained by regular replacement of one-fifth of the culture volume with fresh, pre-reduced media.
- Parameters analysed / observed: production of gas.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): vanillin
- Molecular formula: C8H8O3
- Molecular weight: 152.1473
- Smiles notation (if other than submission substance): COc1cc(C=O)ccc1O
- InChl (if other than submission substance): InChI=1/C8H8O3/c1-11-8-4-6(5-9)2-3-7(8)10/h2-5,10H,1H3

Oxygen conditions:

anaerobic

Inoculum or test system:

sewage, domestic, non-adapted

Details on inoculum:

The medium was inoculated with 10%(v/v) seed from a laboratory anaerobic digester fed primary settled sewage sludge on a 15-day detention time and incubated in the dark at 35ºC.

Duration of test (contact time):

28 d

Initial conc.:

300 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

CH4 evolution

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: Prereduced, defined medium contained the following (per liter): resazurin, 0.001 g; (NH4)2PO4, 0.04 g; NH4Cl, 0.2 g; MgCl2.6H20, 1.8 g; KCI, 1.3 g; MnCl2.4H20, 0.02 g; CoCl2.6H20, 0.03 g; H3BO3, 0.0057 g; CaCl2.2H20, 0.0027 g; Na2MoO4. 2H20, 0.0025 g; ZnCl2, 0.0021 g; FeCl2.4H20, 0.368 g; NaHCO3, 2.64 g; Na2S.9H20, 0.5 g; and 1% (vol/vol) vitamin solution. Oxygen was removed from the medium by boiling, followed by the addition of the sodium sulfide reducing agent. A C/N/P molar ratio of 100:15:1 was provided.
- Test temperature: 35ºC
- pH: The medium was buffered at pH 7.0 by a bicarbonate-CO2 system with a gas atmosphere of 30% CO2 and 70% N2.
- pH adjusted: yes
- Aeration of dilution water: no
- Continuous darkness: yes
- Other: Acclimated stock cultures were maintained in serum CHO bottles by regular replacement of one-fifth of the culture volume with fresh, prereduced media.

TEST SYSTEM
- Culturing apparatus: serum bottles closed with a butyl rubber stopper with a crimped metal seal.
- Number of culture flasks/concentration: 10
- Method used to create anaerobic conditions: Oxygen was removed from the medium by boiling, followed by the addition of the sodium sulfide reducing agent. Serum bottles were flushed (500 ml/min) with oxygen-free gas for 20 min before the inoculated medium was added.
- Measuring equipment: Gas composition was determined on a Fisher-Hamilton model 29 gas partitioner. Gas production and substrate concentration were corrected for background levels by subtracting values measured in a control culture which contained no aromatic substrate.
- Test performed in open system: no.

SAMPLING
- Sampling frequency: daily
- Sampling method: the volume of gas produced by 250-ml serum-bottle cultures was mesured daily by displacement of the plunger lubricated with water in a 20-ml glass syringe fitted with a 20-gauge needle. The bottle was tilted slightly while the syringe was held horizontally to minimize the effect of the plunger weight.

CONTROL AND BLANK SYSTEM
- Inoculum blank: a control culture which contained no aromatic substrate was used.

Reference substance:

benzoic acid, sodium salt

Key result

Parameter:

% degradation (CH4 evolution)

Value:

72

Sampling time:

28 d

Details on results:

Mass balances on the conversion of organic CO2 to CH4 were based upon the stoichometry of the Buswell equation.
The test item showed a degradation of 0% at 12 ± 1 days (acclimation period / lag), and reached a degradation of 72 ± 1.4% at 16 ± 1.1 days after lag phase.

Results with reference substance:

Benzoic acid reached a 91 ± 7.8% after 26 days (n = 5).

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

The target substance is degradable under strict anaerobic conditions, with a conversion of 72 ± 1.4% in 28d.

Executive summary:

The study of the ready biodegradability of the test substance was performed following a serum-bottle variation of the Hungate technique for growing anaerobic bacteria adapted from Miller and Wolin (1974), similar to OECD 311 (non-GLP). Ten serum bottles containing 300 mg/L test item each were inoculated with 10%(v/v) seed from a laboratory anaerobic digester fed primary settled sewage sludge and incubated under anaerobic conditions in the dark at 35ºC. A negative control was run in parallel, and benzoic acid could serve as a positive control. The amount of biodegradation was measured by the determination of methane evolution at frequent intervals over a 28 day period. After 28 days, the biodegradation of the test substance reached 72%. Therefore, it can be deemed readily biodegradable.

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See 'Attached justification'.

Reason / purpose for cross-reference:

read-across source

Key result

Parameter:

% degradation (test mat. analysis)

Value:

62.52

Sampling time:

6 d

Remarks on result:

other: Read-across from analogue substance.

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

The biodegradation of the vanillin contained in a fraction of the waste produced by the olive-oil extraction industry by A. terreus was 62.5% after 6 days. Based on the available information for the read-across approach, the target substance is expected to be readily biodegradable.

Executive summary:

The biodegradation of the analogue substance vanillin in the waste produced by the olive-oil extraction industry by B. pumilus and A. terreus was studied (non-GLP study). Microorganisms were cultured with 20 to 100% alpechin solutions in 5L reactors at 28ºC under aerobic conditions, and the COD, BOD and phenolics content were measured at regular intervals. The test item reached a 62.5% degradation after 6 days. Based on the available information for the read-across approach, the target substance is expected to be readily biodegradable.

Reference 4

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: The biodegradation of p-vanillin in 20-100% alpechin solutions by B. pumilus and A. terreus was calculated by COD and BOD. Degradation was calculated for each of the compounds contained in alpechin.
- Short description of test conditions: Cultures of B. pumilus and A. terreus were incubated in a 5 L batch reactor at 28ºC for 6 days.
- Parameters analysed / observed: The chemical oxygen demand (COD) was measured by the methods of Rodier (1985) and Capitán Valley (1987). The biochemical oxygen demand (BOD) was measured by the methods of Sauter & Stoub (1990), AOAC Official Methods of Analysis (15th ed.). The phenol content was measured by HPLC (UV-VIS detector).

GLP compliance:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Fraction of alpechin, the waste produced by the olive-oil extraction industry, detected by HPLC and confirmed by TLC, containing 0.52% p-vanillin. Samples were diluted to concentrations of 20, 40, 60, 80 and 100%, using Minimum Maintenance Medium (MMM) (Janshekar et al., 1981).
- Source and lot/batch No.of test material: Jimena Osuna SA, collected from the outlet of the last centrifuge.
- Physical and chemical properties of the alpechin used: pH 5.26, COD 212.25g/L, BOD 179.45g/L, Total phenolic compounds 0.7% (0.52% of which is p-vanillin), Total solids 55.59g/L, Volatile solids 44.46 g/L, Non-volatile solids 11.13 g/L, Settled solids 7.75 g/L, Suspended solids 2.44 g/L, Dissolved solids 45.40 g/L.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: stored in sterile flasks at -20ºC util use.

OTHER SPECIFICS:
Details on properties of test surrogate or analogue material (migrated information):
- Name of test material (as cited in study report): vanillin
- Molecular formula: C8H8O3
- Molecular weight: 152.1473
- Smiles notation (if other than submission substance): COc1cc(C=O)ccc1O
- InChl (if other than submission substance): InChI=1/C8H8O3/c1-11-8-4-6(5-9)2-3-7(8)10/h2-5,10H,1H3

Oxygen conditions:

aerobic

Remarks:

Aspergillus terreus

Inoculum or test system:

activated sludge, adapted

Details on inoculum:

- Source: A. terreus obtained from the Microbiology Department of the Faculty of Pharmacy of the University of Granada.
- Method of cultivation: a culture of A.terreus, grown on a malt-extract-agar slope, produced spores which were then suspended in a saline solution (1-9 × 10 s viable spores per ml). For each dose, two test tubes containing 15 ml were inoculated with 1"5 ml of spore suspension and incubated at 28°C for 72 h with constant shaking. Each tube was diluted with MMM to a volume of 150ml and incubated in two 500-ml flasks in the same manner as above. These two 150-ml samples were combined and added to alpechin to a volume of 3000 ml in the 5-1itre batch reactor and incubated.
- Incubation: 6 days at 28°C.

Duration of test (contact time):

6 d

Initial conc.:

0.52 other: % of total phenolic compounds.

Based on:

test mat.

Parameter followed for biodegradation estimation:

other: COD, BOD.

Parameter followed for biodegradation estimation:

test mat. analysis

Details on study design:

TEST CONDITIONS
- Composition of medium: minimal maintenance medium, containing 1.6g bipotassium phosphate, 0.5g monopotassium phosphate, 1.25g ammonium sulphate, 1g ammonium nitrate, 0.5g magnesium sulphate pentahydrate, 0.25g sodium chloride, 0.025g ferric chloride hexahydrate, 0.01g calcium chloride in 1000 ml distilled water.
- Additional substrate: no.
- Test temperature: 28ºC
- pH: 5.26
- pH adjusted: no
- Aeration of dilution water: steady flow of air (720mL/min)
- Continuous darkness: not specified.

TEST SYSTEM
- Culturing apparatus: Assays in 5L batch reactor, stirring rate 200 rpm.
- Number of culture flasks/concentration: 2

SAMPLING
- Sampling frequency: samples were taken at regular intervals.

Reference substance:

not specified

Key result

Parameter:

% degradation (test mat. analysis)

Value:

62.52

Sampling time:

6 d

Details on results:

The biodegradation of the alpechin fraction corresponding to p-vanillin reached a 62.5% biodegradation after 6 days. The time of maximum degradation was day 4.

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

The biodegradation of the vanillin contained in a fraction of the waste produced by the olive-oil extraction industry by A. terreus was 62.5% after 6 days. Based on this result, the substance is expected to be readily biodegradable.

Executive summary:

The biodegradation of vanillin in the waste produced by the olive-oil extraction industry by B. pumilus and A. terreus was studied (non-GLP study). Microorganisms were cultured with 20 to 100% alpechin solutions in 5L reactors at 28ºC under aerobic conditions, and the COD, BOD and phenolics content were measured at regular intervals. The test item reached a 62.5% degradation after 6 days. Based on this result, the substance is expected to be readily biodegradable.

Reference 5

Endpoint:

biodegradation in water: screening tests

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See 'Attached justification'.

Reason / purpose for cross-reference:

read-across source

Key result

Remarks on result:

other: degradable by benthic microorganisms as a sole carbon source.

Remarks:

Read-across from analogue.

Interpretation of results:

inherently biodegradable

Conclusions:

Vanillin can be utilized as a sole carbon source by micro-organisms of both dry mud and sandy sediments of an eutrophic lake. Based on the available information for the read-across approach, the target substance is expected to be readily biodegradable.

Executive summary:

A study on the capability of utilizing different aromatic compounds by benthic microorganisms isolated from two types of bottom sediment of an eutrophic lake was performed with vanillin. These microorganisms were cultured with 0.01% test item as a sole carbon source for 6 days at 26ºC, and colonial development was assessed by comparison of the cultures with control plates containing no carbon source. Under test conditions, the test item could be used as the sole carbon source. Therefore, the test item is expected to be readily biodegradable. Based on the available information for the read-across approach, the target substance is expected to be readily biodegradable.

Reference 6

Endpoint:

biodegradation in water: screening tests

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1970.

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

unsuitable test system

Qualifier:

no guideline available

Principles of method if other than guideline:

After 6 days incubation with the test item at 26ºC, colonial development was assessed by comparison of the cultures with control plates containing no carbon source. Plates containing ferrous gluconate (100mg/L) were used as check on the viability of the inoculum.

GLP compliance:

no

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material:
- Name of test material (as cited in study report): vanillin
- Molecular formula: C8H8O3
- Molecular weight: 152.1473
- Smiles notation (if other than submission substance): COc1cc(C=O)ccc1O
- InChl (if other than submission substance): InChI=1/C8H8O3/c1-11-8-4-6(5-9)2-3-7(8)10/h2-5,10H,1H3

Oxygen conditions:

aerobic

Inoculum or test system:

other: benthic microorganisms form an eutrophic lake (unable to grow on a medium without carbon source)

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Samples of bottom deposite were taken in May, July and August 1969 and March 1970 at two sampling sites located in the southern part of the eutrophic lake Jeziorak. Lake Jeziorak is the 6th largest lake of Poland (27km long, maximal width 2.7km, average depth 5.0m). Samples were collected from a depth of 2.5m (sampling site No.1) and 0.6-0.8m (sampling site No.2, sandy, located in the litoral zone) by means of the Eckman's grab provided with an additional opening arrangement at the top. The upper part of the sediment was transferred aseptically to sterile bottles and maintained in ice containers until analysed. The time between sampling and plating did not exceed 6-8 hours.
- Method of cultivation: Dilutions of the mud samples were made with sterile lake water and plating was made with the iron-peptone agar (Ferrer, Stapert, Sokolski, 1963). Following an incubation period of 14 days at 26ºC, all colonies from a plate or a representative section thereof were picked and stab cultures made with semisolid agar of the same compositoin (4.0g agar /L).
- Storage conditions: The cultures maintained in this medium were kept at 4ºC until studied.
- Storage length: 6-8 h.
- Preparation of inoculum for exposure: Before testing the aromatic compounds utilization, each strain was grown for 5 days at 26ºC in 4.0mL of the liquid iron-peptone medium. The cells were then harvested by centrifugation, washed twice with phosphate buffer (pH 7.0) and resuspended in 1mL of sterile phosphate buffer. This suspension was used for inoculation of media in studies on the ability of utilization of selected aromatics as the sole carbon source by the isolated strains of microorganisms. The Stevenson's (1967) method was applied for this purpose. The plating medium consisted of the Bacto Yeast Nitrogen Base (Difco) without aminoacids. Microorganisms able to grow in this medium (without carbon source) were discarded.

Duration of test (contact time):

6 d

Initial conc.:

0.01 other: %

Based on:

test mat.

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

other: colonial development

Details on study design:

TEST CONDITIONS
- Composition of medium: iron-peptone medium.
- Additional substrate: no.
- Test temperature: 26ºC

TEST SYSTEM
- Culturing apparatus: plates

SAMPLING
- Sampling frequency: after 5 and 6 days incubation.
- Sampling method: sample culture growth was compared with control plates.

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes, samples containing no carbon source.
- Other: plates containing 100 mg/L ferrous gluconate were used as check on the viability of the inoculum.

Reference substance:

other: ferrous gluconate

Reference substance:

benzoic acid, sodium salt

Key result

Remarks on result:

other: degradable by benthic microorganisms as a sole carbon source.

Interpretation of results:

other:

Conclusions:

Vanillin can be utilized by micro-organisms of both dry mud and sandy sediments of an eutrophic lake. Therefore, the test item is expected to be readily biodegradable.

Executive summary:

A study on the capability of utilizing different aromatic compounds by benthic microorganisms isolated from two types of bottom sediment of an eutrophic lake was performed with vanillin. These microorganisms were cultured with 0.01% test item as a sole carbon source for 6 days at 26ºC, and colonial development was assessed by comparison of the cultures with control plates containing no carbon source. Under test conditions, the test item could be used as the sole carbon source. Therefore, the test item is expected to be readily biodegradable.

Reference 7

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From August 11th to November 12th, 1993.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Version: July 17, 1992.

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Lot No. GA01
- Purity: 99.1%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: stored in a cool dark place.
- Stability under test conditions: Stable (checked by IR before start of the test).

Oxygen conditions:

aerobic

Inoculum or test system:

mixture of sewage, soil and natural water

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Sludge samples were collected from the following 10 locations: Fucho River Treatment Plant (Sapporo City, Hokkaido), Nakahama treatment plant (Osaka prefecture Osaka city), Kitakami River (Ishinomaki-shi, Miyagi), Yoshino River (Tokushima City, Tokushima Prefecture), Hiroshima Bay (Hiroshima-shi, Hiroshima), Shiba processing plant (Kashima-gun, Ibaraki prefecture), Ochiai processing plant (Shinjuku-ku, Tokyo), Shinano river (Niigata prefecture Nishikanbara gun), Lake Biwa (Otsu City, Shiga Prefecture), Dokai Bay (Kitakyushu City, Fukuoka Prefecture)
- Preparation of inoculum for exposure: Each 500 ml of the filtrate of the sludge collected in the aforementioned locations was mixed with 5 L of the filtrate of the former activated sludge which had been subjected to the test, in order to get 10g, and pH was adjusted to 7.0 ± 1.0.

Duration of test (contact time):

14 d

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

DOC removal

Parameter followed for biodegradation estimation:

TOC removal

Parameter followed for biodegradation estimation:

test mat. analysis

Details on study design:

TEST CONDITIONS
- Composition of medium: Glucose, peptone, monopotassium phosphate were dissolved in dechlorinated water so as to be 5 (w/v) %, respectively, and pH adjusted to 7.0 ± 1.0 with sodium hydroxide.
- Test temperature: 25 ± 2 ºC.
- pH: 7.0 ± 1.0
- pH adjusted: yes
- Aeration of dilution water: yes
- Suspended solids concentration: The concentration of suspended solids in activated sludge was 5000 mg/L, determined according to JIS K 0102-1986.
- Continuous darkness: yes

TEST SYSTEM
- Number of culture flasks/concentration: 3 flasks per test item sample, plus a flask for the blank control and a flask for the reference substance control.
- Measuring equipment: Closed Oxygen Consumption Measurement Device (Okura Denki Kuiguchi Mekyu), Data processing device manufactured by Asahi Kenki Industry Co., Ltd.)
- Test performed in closed vessels due to significant volatility of test substance: no.
- Details of trap for CO2 and volatile organics if used: soda lime, No.1 (first class reagent manufactured by Wako Pure Chemical Industries, Ltd.)

SAMPLING
- Sampling frequency: analysis were performed at the end of the test.

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes

Reference substance:

aniline

Remarks:

Lot SC-2726 (Showa Chemical Industry Co., Ltd.)

Key result

Parameter:

% degradation (DOC removal)

Value:

>= 96 - <= 98

Sampling time:

14 d

Key result

Parameter:

% degradation (TOC removal)

Value:

>= 97 - <= 100

Sampling time:

14 d

Key result

Parameter:

% degradation (test mat. analysis)

Value:

100

Sampling time:

14 d

Results with reference substance:

The degree of decomposition of aniline from BOD after 7 days and 14 days was 65% and 81%, respectively. Therefore, it was confirmed that the test conditions were valid.

The status of the test solutions at the start and the end of the test was the following:

	Test solution	Situation	pH
Start	(Water + test substance) system	No insoluble matter was observed.	1) 5.3
	(Sludge + test substance) system	No insoluble matter was observed.	2) 6.8 3) 6.8 4) 6.8
End	(Water + test substance) system	No insoluble matter was observed.	5) 5.8
	(Sludge + test substance) system	Growth of sludge was observed.	6) 7.1 7) 7.2 8) 7.2

The analysis results after 14 days are as follows.

		(Water + test item) system	(Sludge + test item) system			Theoretical amount (mg)
		1	2	3	4
BOD#		0.4	51.6	52.5	51.6	53.7
D O C residual	mg	18.7	0.6	0.0	0.0	18.9
	%	99	3	0	0
Remaining test substance (HPLC)	mg	29.1	0.0	0.0	0.0	30
	%	97	0	0	0

# (Sludge + test substance) system was expressed by subtracting the value of sludge blank system.

## The residual rate (%) was calculated based on the following formula, and one digit after the decimal point was rounded and expressed as an integer

Residual rate (%) =   (Residual amount (mg))/(Theoretical quantity (mg))*100

The results obtained in the assay were:

	Biodegradation (%)
	2	3	4
BOD	96	98	96
TOC	97	100	100
HPLC	100	100	100

Validity criteria fulfilled:

yes

Remarks:

The result with reference substance was within the expected values.

Interpretation of results:

readily biodegradable

Conclusions:

Under test conditions, the biodegradation of the test item is equal or higher than 96%. Therefore, the test item is readily biodegradable.

Executive summary:

A study on the ready biodegradability of the analogue substance vanillin was performed according to OECD 301C, modified MITI test (GLP study). The test item was introduced in triplicate at a concentration of 100 mg/L in serum bottles containing inoculum mixed from ten collection sites (mixture of sewage, soil, natural water), along with a reference control (aniline) and a blank control. Then, BOD, TOC and test item concentration (by HPLC-UV) were measured and the percent degradation was determined for each parameter. After 14 days, the test item had reached a biodegradation of 96 -98% (by BOD), 97 -100% (by TOC) and 100% (by HPLC). Therefore, the test item is readily biodegradable.

Reference 8

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See 'Attached justification'.

Reason / purpose for cross-reference:

read-across source

Key result

Parameter:

% degradation (test mat. analysis)

Value:

>= 96

Sampling time:

14 d

Remarks on result:

other: read-across from analogue.

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

Based on the available information for the read-across approach, the target substance is expected to be readily biodegradable.

Executive summary:

A study on the ready biodegradability of the analogue substance vanillin was performed according to OECD 301C, modified MITI test (GLP study). The test item was introduced in triplicate at a concentration of 100 mg/L in serum bottles containing inoculum mixed from ten collection sites (mixture of sewage, soil, natural water), along with a reference control (aniline) and a blank control. Then, BOD, TOC and test item concentration (by HPLC-UV) were measured and the percent degradation was determined for each parameter. After 14 days, the test item had reached a biodegradation of 96 -98% (by BOD), 97 -100% (by TOC) and 100% (by HPLC). Therefore, the test item is readily biodegradable. Based on the available information for the read-across approach, the target substance is expected to be readily biodegradable.

Description of key information

Weight of evidence. Based on the available information for the read-across approach, the target substance can be considered readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable

Additional information

Weight of evidence:

- A study on the ready biodegradability of the analogue substance vanillin was performed according to OECD 301C, modified MITI test (GLP study). The test item was introduced in triplicate at a concentration of 100 mg/L in serum bottles containing inoculum mixed from ten collection sites (mixture of sewage, soil, natural water), along with a reference control (aniline) and a blank control. Then, BOD, TOC and test item concentration (by HPLC-UV) were measured and the percent degradation was determined for each parameter. After 14 days, the test item had reached a biodegradation of 96 -98% (by BOD), 97 -100% (by TOC) and 100% (by HPLC). Therefore, the test item is readily biodegradable. Based on the available information for the read-across approach, the target substance is expected to be readily biodegradable.

- A study of the ready biodegradability of the analogue substance vanillin was performed following a serum-bottle variation of the Hungate technique for growing anaerobic bacteria adapted from Miller and Wolin (1974), similar to OECD 311 (non-GLP). Ten serum bottles containing 300 mg/L test item each were inoculated with 10%(v/v) seed from a laboratory anaerobic digester fed primary settled sewage sludge and incubated under anaerobic conditions in the dark at 35ºC. A negative control was run in parallel, and benzoic acid could serve as a positive control. The amount of biodegradation was measured by the determination of methane evolution at frequent intervals over a 28 day period. After 28 days, the biodegradation of the analogue substance reached 72% and therefore, it is readily biodegradable. Based on the read-across approach, the target substance is deemed readily biodegradable.

- In a study of the biodegradation of vanillin in the waste produced by the olive-oil extraction industry (no guideline, no GLP), the test substance was found to be readily biodegradable, reaching a 62.5% degradation in under 14 days (Martinez et al., 1993).

- In another study on the ability to biodegrade vanillin of bacterial and soil samples (method similar to OECD TG 302C, no GLP) , the extent of vanillin oxidation in soil suspension and in structural soil attained 50 -70% of the total oxygen consumption required for complete oxidation after 48h (Kunc et al., 1971).

- Finally, in a study on the capability of utilizing different aromatic compounds by benthic microorganisms isolated from two types of bottom sediment of an eutrophic lake (no guideline, no GLP), vanillin was found to be biodegradable when being used as a sole carbon source (Strzelczyk et al., 1972).

The analogue vanillin, which is an isomer of isovanillin and therefore shares the same functional groups with the substance, also has comparable values for the relevant molecular properties. Based on the available information for the read-across approach, the target substance isovanillin can be considered as readily biodegradable.