t-Butyl (2S,2aS,5aR)-1-azabicyclo[3.3.0] oct-2-carboxylate monooxalate salt

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006-10-30 to 2007-04-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline and GLP, well documented, only a slight deviation from guideline. As the objectives of the definitive test were met, this has no impact on the integrity or validity of the study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

temperature deviation: In the definitive test, the temperature of the contents of the additional vessel was recorded to range from 20.2 to 24.2oC. However, the room temperature was recorded as 19.6 to 21.9 deg C over the same period. As the objectives of

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

yes

Remarks:

temperature deviation: In the definitive test, the temperature of the contents of the additional vessel was recorded to range from 20.2 to 24.2oC. However, the room temperature was recorded as 19.6 to 21.9 deg C over the same period. As the objectives of

GLP compliance:

yes (incl. QA statement)

Remarks:

Department of Health of the Government of the United Kingdom

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): VRT-126028
- Physical state: White
- Analytical purity: 99.3 area %
- Lot/batch No.: 25414
- Expiration date of the lot/batch: May 2008
- Storage condition of test material: Room temperature

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All concentrations
- Sampling method: 2 10 mL samples were taken from the freshly prepared control and test media. After 48 h, the contents of the test vessels from each group were pooled, and further samples taken for analysis.
- Sample storage conditions before analysis: On each occasion, one sample from each set was analyzed immediately; the remaining samples were frozen in case further analysis was required.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 200 mg of the test substance was dispersed in 2L Elendt M4 medium in a volumetric flask, treated with ultrasound for 5 min and then filtered through 0.2 um cellulose nitrate. The filtrate was either used directly at the highest test concentration (100 mg/L), or diluted with additional M4 for the lower concentrations.
- Eluate: Not applicable
- Differential loading: Not applicable
- Controls: Elendt M4 media
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution): Not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Not applicable

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Source: In-house culture, originally from the Institute National de Recherche Chimique Appliqué, France
- Age at study initiation (mean and range, SD): <24 h
- Weight at study initiation (mean and range, SD): Not applicable
- Length at study initiation (length definition, mean, range and SD): Not applicable
- Valve height at study initiation, for shell deposition study (mean and range, SD): Not applicable
- Peripheral shell growth removed prior to test initiation: Not applicable
- Method of breeding: Parthanogenesis
- Feeding during test
- Food type: Not fed during test
- Amount: Not applicable
- Frequency: Not applicable


ACCLIMATION
- Acclimation period: No
- Acclimation conditions (same as test or not): Not applicable
- Type and amount of food: Not applicable
- Feeding frequency: Not applicable
- Health during acclimation (any mortality observed): Not applicable


QUARANTINE (wild caught)
- Duration: Not applicable
- Health/mortality: Not applicable

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

None

Test conditions

Hardness:

268 mg/L

Test temperature:

The temperature ranged from 20.1 -20.3 at time 0 and 21.1-21.4 at time 48 h

pH:

The pH ranged from 7.36 – 7.71 SU at time 0 and 7.43 – 7.84 SU at time 48 h

Dissolved oxygen:

The DO ranged from 98 - 100% of air saturation value at time 0 and 99 - 102% at time 48h

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal- Control, 6.25, 12.5, 25, 50, and 100 mg/L
Measured- Control (< 1 mg/L, none detected) 5.22, 10.5, 22.9, 43.9, 89.6 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed-loosely covered
- Material, size, headspace, fill volume: glass dishes, 100 mL fill volume
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): None
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): Not applicable
- Biomass loading rate: 20 mL media per organism


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 test media
- Total organic carbon: Not applicable
- Particulate matter: Not applicable
- Metals: Not applicable
- Pesticides: Not applicable
- Chlorine: Not applicable
- Alkalinity: 33 mg/L
- Ca/Mg ratio: Not applicable
- Conductivity: Not applicable
- Culture medium different from test medium: No
- Intervals of water quality measurement: Daily


OTHER TEST CONDITIONS
- Adjustment of pH: yes
- Photoperiod: 16 h light 8 h dark
- Light intensity: no data


EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Immobility, 24 and 48 h


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: Not applicable
- Range finding study
- Test concentrations: 1, 10, and 100 mg/L
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 89.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 45% immobilization at 48 h

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

22.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No additional observations

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Statistical analysis was performed using SAFEStat LD50 application, SAS 8.2. The NOEC was derived by direct inspection of the data on the immobility of the animals.

Any other information on results incl. tables

Not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:

not applicable

Conclusions:

The EC50 of the test substance to D. magna is > 89.6 mg/L mean measured concentration. The NOEC is 22.9 mg/L.

Executive summary:

Not applicable