3-hydroxy-p-anisaldehyde

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

yes

Remarks:

exposure of 6 or 11 days.

Principles of method if other than guideline:

Determination of growth stimulation or inhibition by measurement of biomass optical density and chlorophyll content after exposure to the test substance.

GLP compliance:

no

Remarks:

study not intended for regulatory purposes.

Specific details on test material used for the study:

OTHER SPECIFICS:
Details on properties of test surrogate or analogue material:
- Name of test material (as cited in study report): vanillin
- Molecular formula (if other than submission substance): C8H8O3
- Molecular weight (if other than submission substance): 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

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 25, 50, 100, 150, 200, 300 mg/L or 0, 60, 300 mg/L.
- Sampling method: Culture samples from specific cultivation days were taken and centrifuged.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 0, 25, 50, 100, 150, 200, 300 mg/L were present in 160 ml growth media.

Test organisms (species):

Chlorella sp.

Details on test organisms:

TEST ORGANISM
- Common name: Chlorella sp.
- Source (laboratory, culture collection): Culture Collection of Baltic Algae (CCBA)
- Method of cultivation: Cultivated in a modified Bold’s Basal Medium (BBM), in 500 ml round flasks with an initial culture volume of 160 ml, in an incubator (Certomat® BST) equipped with a light source (fluorescents lamps, 5 X 18 W) and a shaker (110 rpm). The culture was incubated in day/night cycles (16h/8h) during the period of 6 days.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

11 d

Nominal and measured concentrations:

0, 25, 100, 150, 200 and 300 mg/L (nominal) in one test and 0, 60, 300 mg/L (nominal) in another test.

Details on test conditions:

TEST SYSTEM
- Test vessel: 500 ml round flasks
- Material, size, headspace, fill volume: size 500 ml, fill volum 160 ml

GROWTH MEDIUM
- Standard medium used: Bold's Basal Medium
- Detailed composition if non-standard medium was used: K2HPO4 (0.075 g/L), KH2PO4 (0.175 g/L), MgSO4.7H2O (0.075 g/L), NaNO3 (0.25 g/L), CaCl2.2H2O (0.025 g/L), NaCl (0.025 g/L), Na2EDTA.2H2O (0.025 g/L), FeCl3 (6 mg/L), H3BO3 (1 mg/L), ZnSO4.7H2O (1.4 mg/L), MnSO4.H2O (0.16 mg/L), CuSO4.5H2O (0.16 mg/L), CoCl2.6H2O (0.065 mg/L), Na2MoO4.2H2O (0.135 mg/L).

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h light / 8h dark cycles
- Light intensity and quality: 5 x 18 W fluorescent lamps

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: spectrophotometer
- Clorophyll measurement: Samples of culture aliquots were taken and added to laboratory tubes and centrifugated to remove media. After removing media, pure methanol was added to tubes. When the extraction was complete, aliquots were centrifuged to separate cells from solvent because floating cells cause interferences during chlorophyll measurement. Chlorophyll a and b in obtained extracts were measured spectrophotometrically.
- Biomass density measurement: Optical density of Chlorella biomass in media with different vanillin concentration was determined by taking aliquots of the culture and measuring absorbance (T80+ UV/VIS Spectrometer PG Instruments Ltd) at wavelength 530 nm (OD530). Measurement days were: 0 (after inoculation), 2, 3, 4, 5, 6. There was no measurement after the first day of cultivation as it was time necessary for microalgae to adapt to new growth conditions. Optical density of the culture in the presence of vanillin was compared with the control culture to which no vanillin was added.

TEST CONCENTRATIONS
- Test concentrations: 0, 25, 100, 150, 200 and 300 mg/L or 0, 60, 300 mg/L.

Reference substance (positive control):

no

Key result

Duration:

11 d

Dose descriptor:

EC50

Effect conc.:

> 200 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

Results obtained from present research and literature data suggest that vanillin can be metabolized by microalgae.
- Exponential growth in the control (for algal test): For concentrations of 25, 100, 150, and 200 mg/L, there was an initial step of inhibition, and a second step of growth recovery, taking longer with higher doses. 50 mg/L yielded no inhibition, but growth stimulation after 3 days. 300 mg/L resulted in the growth inhibition and no growth recovery.
- Observation of abnormalities (for algal test): no

Validity criteria fulfilled:

not specified

Conclusions:

The EC50 of the test item on chlorella sp. was found to be greater than 200 mg/L.

Executive summary:

A study on the effect of vanillin on algae growth was perfomed, by a method similar to OECD 201 (non-GLP). Cultures of Chlorella sp. were exposed to various concentrations of test item ranging from 0 (control) to 300 mg/L, and biomass density and chlorophyll content were measured at regular intervals. 50 and 60 mg/L yielded no inhibition, but growth stimulation after 3 days, which would suggest that vanillin can be metabolized by microalgae. At concentrations up to 200 mg/L, there was an initial step of inhibition, and a second step of growth recovery, taking longer with higher doses. 300 mg/L resulted in the growth inhibition and no growth recovery. Under test conditions, the EC50 of the test item on chlorella sp. was found to be greater than 200 mg/L.

Reference 2

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1954.

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:

Algal growth of six algal cultures was visually determined after exposure to 2 mg/L of test item at periods of 3, 7, 14, and 21 days.

GLP compliance:

no

Specific details on test material used for the study:

OTHER SPECIFICS:
Details on properties of test surrogate or analogue material:
- Name of test material (as cited in study report): vanillin
- Molecular formula (if other than submission substance): C8H8O3
- Molecular weight (if other than submission substance): 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

Analytical monitoring:

no

Details on sampling:

- Concentrations: 2 ppm

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Sterile, double strength medium was inoculated with a small amount (one-fifteenth to one-thirtieth by volume) of an actively growing culture of the alga.A 7.5 ml. portion of this inoculated medium was then combined with a like quantity of distilled water containing 4 ppm by weight, in the case of solids, or 0.0004 percent by volume in the case of liquids, of the chemical to be tested. This resulted in a final chemical concentration of 2 p.p.m. (or 0.0002%) in normal strength medium and with an inoculum of one-thirtieth to one-sixtieth the volume of the original algal culture.
- Controls: normal strength culture medium without test item.

Test organisms (species):

other: Cylindrospermum licheniforme, Mycrocystis aeruginosa, Schenedesmus obliquus, Chlorella variegata, Gomphonema parvulum , Nitzschia palea.

Details on test organisms:

TEST ORGANISM
- Common name: blue-green algae, green algae, diatoms.
- Strain: Cylindrospermum licheniforme B. and F. and Microcystis aeruginosa Ktz. representing the blue-green algae (Myxophyceae), Scenedesmus obliquus (Turp.) Ktz. and Chlorella variegata Beijerinck representing the green algae (Chlorophyceae), and Gomphonema parvulum (Ktz.) V. H. and Nitzschia palea (Ktz.) W. Smith representing the diatoms (Bacillarieae).
- Method of cultivation: All cultures were unialgal and selected for their ability to produce rapid and uniform growth under laboratory conditions.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

0 d.

Test temperature:

22ºC.

Nominal and measured concentrations:

2ppm (0.0002%), nominal.

Details on test conditions:

TEST SYSTEM
- Test vessel: 25 ml Erlenmeyer flasks
- Initial cells density: 125000 cells per ml on average.

GROWTH MEDIUM
- Standard medium used: no
- Detailed composition if non-standard medium was used: The culture medium approximated Gerloff's modification of Chu No. 10 with the amount of nitrate doubled (Palmer and Maloney, 1953).

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: distilled water.
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Photoperiod: illuminated culture room

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: visual determination. Growth was compared with that in the control flasks containing 15 ml of normal strength inoculated culture medium but no test chemical. The amount of visible algal growth was recorded at specified intervals for a total of 21 days (at periods of 3, 7, 14, and 21 days for each of the six cultures of algae).

Reference substance (positive control):

yes

Remarks:

Colloidal silver.

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Validity criteria fulfilled:

yes

Conclusions:

The test item was found to be non toxic to algae, with a NOEC ≥ 2 mg/L.

Executive summary:

A preliminary screening study on the effects of vanillin on freshwater algae growth was performed (non-GLP study), by exposing six algal cultures to 2mg/L of the test item for 21 days. The following algae were used: Cylindrospermum licheniforme, Mycrocystis aeruginosa, Schenedesmus obliquus, Chlorella variegata, Gomphonema parvulum, Nitzschia palea. Under test conditions, no effects were observed in any of the algae after 21 days. Therefore, the test item was found to be non toxic to algae at up to 2mg/L, the highest dose tested. Therefore, the NOEC of the test item in algae is ≥ 2 mg/L.

Reference 3

Endpoint:

toxicity to aquatic algae and cyanobacteria

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

Duration:

11 d

Dose descriptor:

EC50

Effect conc.:

> 200 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks on result:

other: Read-across from analogue.

Validity criteria fulfilled:

not specified

Conclusions:

Based on the available information for the read-across approach, the EC50 of the target substance on chlorella sp. is expected to be greater than 200 mg/L.

Executive summary:

A study on the effect of the analogue substance vanillin on algae growth was perfomed, by a method similar to OECD 201 (non-GLP). Cultures of Chlorella sp. were exposed to various concentrations of test item ranging from 0 (control) to 300 mg/L, and biomass density and chlorophyll content were measured at regular intervals. 50 and 60 mg/L yielded no inhibition, but growth stimulation after 3 days, which would suggest that vanillin can be metabolized by microalgae. At concentrations up to 200 mg/L, there was an initial step of inhibition, and a second step of growth recovery, taking longer with higher doses. 300 mg/L resulted in the growth inhibition and no growth recovery. Under test conditions, the EC50 of the test item on chlorella sp. was found to be greater than 200 mg/L. Based on the available information for the read-across approach, the EC50 of the target substance on chlorella sp. is expected to be greater than 200 mg/L.

Reference 4

Endpoint:

toxicity to aquatic algae and cyanobacteria

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

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Validity criteria fulfilled:

yes

Conclusions:

Based on the read-across approach, the target substance is expected to have a NOEC ≥ 2 mg/L in algae.

Executive summary:

A preliminary screening study on the effects of the analogue substance vanillin on freshwater algae growth was performed (non-GLP study), by exposing six algal cultures to 2mg/L of the test item for 21 days. The following algae were used: Cylindrospermum licheniforme, Mycrocystis aeruginosa, Schenedesmus obliquus, Chlorella variegata, Gomphonema parvulum, Nitzschia palea. Under test conditions, no effects were observed in any of the algae after 21 days. Therefore, the test item was found to be non toxic to algae at up to 2mg/L. Therefore, the NOEC ≥ 2 mg/L. Based on the available information for the read-across approach, the target substance is expected to have a NOEC ≥ 2 mg/L.

Description of key information

Weight of evidence: Based on the available information for the read-across approach, EC50 for the target substance is expected to be > 200 mg/L, and the NOEC > 2 mg/L.

Key value for chemical safety assessment

EC50 for freshwater algae:

200 mg/L

EC10 or NOEC for freshwater algae:

2 mg/L

Additional information

Weight of evidence: Based on the read-across approach, the target substance is found to be non toxic to algae, with an EC50 > 200 mg/L.

A study on the effect of the analogue substance vanillin on algae growth was performed, by methods similar to OECD 201 (non-GLP). Under test conditions, the IC50 for the test substance was > 200 mg/L. At lower concentrations, the test substance showed growth stimulation, which would suggest that vanillin can be metabolized by microalgae. Based on the read-across approach, the target substance is expected to have an EC50 > 200 mg/L. A preliminary screening study on the effects of vanillin on freshwater algae growth was performed (non-GLP), by exposing six algal cultures to 2 mg/L of the test item for 21 days. The following algae were used: Cylindrospermum licheniforme, Mycrocystis aeruginosa, Schenedesmus obliquus, Chlorella variegata, Gomphonema parvulum, Nitzschia palea. Under test conditions, no effects were observed in any of the algae after 21 days. Therefore, the test item was found to be non toxic to algae at up to 2 mg/L, the highest concentration tested. Based on the read-across approach, the target substance is expected to have a NOEC > 2 mg/mL in algae.