Vanillin

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to microorganisms, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles.

Qualifier:

equivalent or similar to guideline

Guideline:

other: Microtox test

Deviations:

not specified

GLP compliance:

not specified

Analytical monitoring:

not required

Vehicle:

yes

Details on test solutions:

To avoid any errors of preparation of test solution, at least two different initial solutions were used.

- Method: According to Beckman Instruments Inc. method (1982).
- Chemical name of vehicle: solvent : methanol
- Concentration of vehicle in test medium: initial solution: 5%
- Evidence of undissolved material: no

Test organisms (species):

Photobacterium phosphoreum

Details on inoculum:

- Culture supplier: Lyophilised Microtox regeant
- Method of cultivation, preparation of inoculum for exposure, initial biomass concentration: as specified in the Beckman Instruments Inc. method (1982).

Test type:

static

Water media type:

brackish water

Total exposure duration:

15 min

Remarks on exposure duration:

reading at 5 min also

Test temperature:

15°C

Salinity:

2% NaCl

Nominal and measured concentrations:

no data

Details on test conditions:

TEST SYSTEM
- Test vessel: Microtox open glass tubes
- No. of vessels per concentration (replicates): at least 3
- No. of vessels per control (replicates): at least 3
- No. of vessels per vehicle control (replicates): at least 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: de-ionised water

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 or 5
- Spacing factor for range finding study: 10

Reference substance (positive control):

not specified

Duration:

5 min

Dose descriptor:

EC50

Effect conc.:

0.38 mmol/L

EC50 at 5 min = 0.38 mmol/l = 57.8 mg/l
Test performed with methanol as solvent (5 %)

Conclusions:

Vanillin is harmful for Photobacteriumphosphoreum in the conditions of the test.

Reference 2

Endpoint:

toxicity to microorganisms, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study seems to be well conducted.

Principles of method if other than guideline:

no data

GLP compliance:

not specified

Analytical monitoring:

no

Details on test solutions:

no data

Test organisms (species):

anaerobic sludge

Details on inoculum:

no data

Test type:

not specified

Water media type:

not specified

Total exposure duration:

49 h

Details on test conditions:

Specific methanogenic activity measurements were performed in 0.3 dm3 glass serum flasks. Granular sludge (1 g VSS/dm3, from a full scale
reactor treating distillery
Wastewater, not acclimated to the toxicants) was transferred to 0.1 l of the basal medium, and acetate was added from a neutralized at pH7 stock
solution to obtain a final concentration of 2 g COD/l. Subsequently, the flasks were sealed and shaked at 30 +/- 2 deg C .
After 1 day of incubation, the acetate concentration was measured and replenished to obtain 2 g COD/l. The required amount of inhibitory
compound was added.
After 2 days of exposure, the acetate concentration was replenished to 2 g COD/l, and the bottles were reincubated for 1 h prior to the
determination of the methane production rate. The methane composition in the head space content of each serum flask was determined periodically during the subsequent 4 to 5 h.

Reference substance (positive control):

not specified

Duration:

49 h

Dose descriptor:

EC50

Effect conc.:

1 800 mg/L

Basis for effect:

other: methane production inhibition

Duration:

49 h

Dose descriptor:

other: EC80

Effect conc.:

1 880 mg/L

Basis for effect:

other: methane production inhibition

Details on results:

no data

Reference 3

Endpoint:

toxicity to microorganisms, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles.

Principles of method if other than guideline:

no data

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

not specified

Details on test solutions:

- Controls: Medium + yeast cells

Analytical monitoring: not required because stock solutions were freshly prepared for each new test (at least 3 tests)

Test organisms (species):

Saccharomyces cerevisiae

Details on inoculum:

- Inoculum source: fresh baker's yeast was obtained daily from a local distributor. Viability checked with growth-time relationship.
- Method of cultivation: Wickerham medium (malt extract, yeast extract, peptone, glucose and salts)
- Initial biomass concentration: 150 mg/10 ml

Test type:

static

Water media type:

freshwater

Total exposure duration:

210 h

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass Erlenmeyer sealed with cotton plug and shaked
- No. of organisms per vessel: 10x10E+8/ml
- No. of vessels per concentration (replicates): at least 3
- No. of vessels per control (replicates): at least 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: distilled water

OTHER TEST CONDITIONS
- Adjustment of pH: pH adjusted to 5.5

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Gowth rate measured with Coulter counter

TEST CONCENTRATIONS
- Spacing factor for test concentrations: No data
- Range finding study: No data
Reference substance (positive control): No data

Reference substance (positive control):

not specified

Duration:

210 min

Dose descriptor:

EC50

Effect conc.:

179 mg/L

Basis for effect:

growth inhibition

Conclusions:

Vanillin is not harmful for Saccharomyces cerevisiae in the conditions of the test.

Reference 4

Endpoint:

toxicity to microorganisms, other

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Reliable biodegradation study whose result has been determined to be relevant for STEP micro-organisms hazard assessment.

Reason / purpose for cross-reference:

reference to same study

Guideline:

other: no data

GLP compliance:

not specified

Duration:

14 d

Dose descriptor:

IC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks on result:

other:

Remarks:

Result from a OECD test 301C, no inhibitory effect on the activity of activated sludge microorganisms at the tested concentration of 100 mg/L since the biodegradation reached 93% within 14 days

No more data on results except that Vanillin has no inhibitory effects on the activity of activated sludge microorganisms.

Validity criteria fulfilled:

not specified

Conclusions:

No inhibitory effect on the activity of activated sludge microorganisms.

Executive summary:

Vanillin was investigated for its ready biodegradability in an OECD 301C test over 14 days .
The percent biodegradation of the test item was calculated based on the biochemical oxygen demand (BOD) and reached 93% within the 14-day.
Consequently, Vanillin was found to be readily biodegradable under the test conditions within 14 days and had no inhibitory effect on the activity of activated sludge microorganisms at the tested concentration of 100 mg/L.

Reference 5

Endpoint:

toxicity to microorganisms, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles.

Principles of method if other than guideline:

Tetratox assay

GLP compliance:

not specified

Analytical monitoring:

not required

Vehicle:

yes

Details on test solutions:

- Method: dissolution of the substance in the test medium with vehicle
- Controls: Test medium + Inoculum
- Chemical name of vehicle: Solvent dimethyl sulfoxide (DMSO)
- Concentration of vehicle in test medium: Stock solution : < 0.75%

Test organisms (species):

Tetrahymena pyriformis

Details on inoculum:

- Laboratory culture: yes, strain GL-C
- Method of cultivation: growth medium mainly composed with proteose peptone (5g/l), D-glucose (5g/l), yeast extract (1g/l) and Tris-HCl (1.2g/l). The experiments were conducted with log-growth-phase cultures.
- Preparation of inoculum for exposure: no adaptation, the test medium was the growth medium
- Initial biomass concentration: around 2500 cells/ml

Test type:

static

Water media type:

freshwater

Total exposure duration:

40 h

Hardness:

no data

Test temperature:

27 +/- 1°C

pH:

7.40 (buffered with Tris-HCl)

Dissolved oxygen:

no data

Nominal and measured concentrations:

no data

Details on test conditions:

TEST SYSTEM
- Test vessel:250 ml glass Erlenmeyer flasks, sealed with
- No. of organisms per vessel: 2500 cell/ml
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2
- No of experiments: at least 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: sterile distilled water
- Ca/mg ratio: 50 mg/l
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: yes, pH buffered with Tris-HCl

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
The effect parameter measured was the impairment growth, with a spectrophotometric method (wavelength: 540 nm)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: No data
- Range finding study: Yes, at least 2 tests

Reference substance (positive control):

not specified

Duration:

40 h

Dose descriptor:

IC50

Effect conc.:

163 mg/L

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Details on results:

no data

Reported statistics and error estimates:

IC50 is calculated with the Finney probit analysis using percent control-normalized absorbance as the dependent variable and the toxicant concen-
tration (mg/L) as the independant variable.

Validity criteria fulfilled:

not specified

Conclusions:

Vanillin is not Harmful for Tetrahymena pyriformis in the conditions of the test.

Reference 6

Endpoint:

toxicity to microorganisms, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study meeting generally accepted scientific principles

Principles of method if other than guideline:

Microtox-test

GLP compliance:

not specified

Analytical monitoring:

no

Details on test solutions:

no data

Test organisms (species):

Photobacterium phosphoreum

Details on inoculum:

Photobacterium phosphoreum (T3 mutation) was supplied in the form of freeze-dried powder by the Insitute of Soil Science, the Chinese Academy of Sciences, Nanjing, P.R. China. Stock cultures were maintained on agar slants at 4°C. The culturebroth was yeast-tryptone-salts-glycerol, pH 7.0
+/- 0.5. The medium was heat sterilized.

Test type:

static

Water media type:

not specified

Total exposure duration:

15 min

Test temperature:

20°C

pH:

Near neutral

Details on test conditions:

no data

Reference substance (positive control):

not specified

Duration:

15 min

Dose descriptor:

EC50

Effect conc.:

100.5 mg/L

Details on results:

Log EC50= -3.180 mole/l

Description of key information

Vanillin was investigated for its ready biodegradability in an OECD 301C test over 14 days . The percent biodegradation of the test item was calculated based on the biochemical oxygen demand (BOD) and reached 93% within the 14-day. Consequently, Vanillin was found to be readily biodegradable under the test conditions within 14 days and had no inhibitory effect on the activity of activated sludge microorganisms at the tested concentration of 100 mg/L, which can be used as a NOEC value. 

Key value for chemical safety assessment

EC10 or NOEC for microorganisms:

100 mg/L

Additional information

Apart from the key study mentioned above, most of the available data are on single species.

The growth inhibition test on Tetrahymena pyriformis has been selected in supporting study and led to an IC50 (40H) = 163 mg/l (Schultz, 1997).

An EC50 (210 min) = 179 mg/l has been reported on Saccharomyces cerevisiae (Koch et al., 1993) and Microtox tests gave EC50 (5 min) = 57.8 mg/l and EC50 (15 min) = 100.5 mg/l (Cronin et al., 1981; Jin et al., 1999), however these data are not recommended for STP microorganisms risk characterisation.

The only data available on mixed inoculum concerned anaerobic sludge and revealed an EC50 (49H) = 1800 mg/l on methane production inhibition (Sierra-Alvarez & Lettinga, 1991).