1,2,3,6-tetrahydrophthalimide

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant study conducted to international test methods

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1,2,3,6-tetrahydrophthalimide

EC Number:

201-602-8

EC Name:

1,2,3,6-tetrahydrophthalimide

Cas Number:

85-40-5

Molecular formula:

C8H9NO2

IUPAC Name:

2,3,3a,4,7,7a-hexahydro-1H-isoindole-1,3-dione

Details on test material:

- Name of test material (as cited in study report): THPI D4
- Physical state: Solid
- Analytical purity: 97.91%
- Lot/batch No.: STI4208324
- Expiration date of the lot/batch: 2009-11-21
- Stability under test conditions: Stable
- Storage condition of test material: Room temperature

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100 mg/L
- Sampling method: Direct sampling at test start and end
- Sample storage conditions before analysis: 6 +/- 2 Dec C

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solution prepared in dilution water (test medium)
- Eluate: Dilution water (test medium)
- Differential loading: No
- Controls: Dilution water (test medium)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)) Not applicable:
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None

Test organisms

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata
- Strain: Hindak, SAG 61.81
- Source (laboratory, culture collection): Sammlung von Algenkulturen, Pflanzenphysiologisches Institut der Universitat, D-37073 Gottingen
- Age of inoculum (at test initiation): 3 days
- Method of cultivation: Stock cultures prepared monthly on Z-agar


ACCLIMATION
- Acclimation period: 3 days
- Culturing media and conditions (same as test or not): Yes
- Any deformed or abnormal cells observed: No

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Test conditions

Test temperature:

22 - 24 Deg C

pH:

8.06 - 8.52

Nominal and measured concentrations:

Nominal - 100 mg/L
Measured - at test start: 101 mg/L at test end: 98.5 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Erlenmeyer flasks
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: Glass, 250mL, 100 mL
- Aeration: No
- Initial cells density: 1E+4 cells/mL
- Control end cells density: 1.83E+6 cells/mL
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: OECD media as described in test guideline
- Culture medium different from test medium: No


OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: No
- Photoperiod: Continuous
- Light intensity and quality: 60-120 microE/m-2/s-1, daylight fluorescent tubes


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Fluorimeter


TEST CONCENTRATIONS
- Spacing factor for test concentrations: Not applicable
- Justification for using less concentrations than requested by guideline: Limit test
- Range finding study
- Test concentrations: 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 4

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 5

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 6

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

- Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test):
- Unusual cell shape: None
- Colour differences: None
- Flocculation: None
- Adherence to test vessels: None
- Aggregation of algal cells: None
- Any stimulation of growth found in any treatment: Minor (8% increase over controls, expressd as biomass)
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50: (growth rate) = 0.97 (0.90-1.04) mg/L (biomass) = 0.48 (0.44-0.53) mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Substance does not inhibit the growth of freshwater green algae

Executive summary:

Substance does not inhibit the growth of freshwater green algae