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

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006-11-02 to 2007-03-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study performed according to OECD Guideline 203 (Fish, Acute Toxicity Test) and EU Method C.1 (Acute Toxicity for Fish) without deviations.

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 203 (Fish, Acute Toxicity Test)

Deviations:

no

Remarks:

The testing facility indicated that the protocol was followed without deviation.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Remarks:

The testing facility indicated that the protocol was followed without deviation.

GLP compliance:

yes (incl. QA statement)

Remarks:

The 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 powder
- 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: Two 10 mL mid-vessel samples were taken from the control and test media at 0 and 72 h (fresh media) and at 24 and 96 hours (expired media) for analysis.
- Sample storage conditions before analysis: On each occasion, one of the samples was analyzed immediately and the other was stored in a freezer in case futher analysis was required.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 4 g of the test substance was dispersed in 5 L diluent water in a volumetric flask, shaken vigorously, and treated by ultrasound for 30 minutes. The solution was then filtered through 0.2 um cellulose nitrate, and 107, 235, 518, 1140, or 2500 mL aliquots added to the test vessels, and brought to volume with diluent water.
- Eluate: Not applicable
- Differential loading: Not applicable
- Controls: Diluent water
- 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.): Not applicable

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Strain: no data
- Source: commercial fish farm in the UK
- Age at study initiation (mean and range, SD): Approx. 4 months
- Length at study initiation (length definition, mean, range and SD): mean of 10 random fish- 6.53 cm
- Weight at study initiation (mean and range, SD): mean wet weight of those 10 fish- 2.86 g
- Method of breeding: young obtained from local supplier
- Feeding during test
- Food type: not fed during test
- Amount: Not applicable
- Frequency: Not applicable


ACCLIMATION
- Acclimation period: More than 4 months
- Acclimation conditions (same as test or not): yes
- Type and amount of food: Commercial trout chow (Nutra Fry 02), in an amount equal to 1% of the total wet weight of the fish in the holding tank.
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): Mortality <1% in the 14 days preceeding test initiation, with no medication given.


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

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Test conditions

Hardness:

146-150 mg/L

Test temperature:

13.4-14.4 deg C

pH:

7.73-7.98 SU

Dissolved oxygen:

85-95% of air saturation value

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal-Control, 4.27, 9.39, 20.7, 45.5, and 100 mg/L.
Measured (Geometric mean) Control, 4.09, 8.69, 18.6, 40.0, and 99.7 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass 20 L test solution, 14 cm deep
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): semi-static
- Renewal rate of test solution (frequency/flow rate): solution renewed daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): Not applicable
- Biomass loading rate: 1 g bodyweight /L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water, dechlorinated and softened, then carbon filtered. A portion was then passed through reverse osmosis, and the two portions mixed to produce the appropriate hardness.
- Total organic carbon: 0.87 mg/L
- Particulate matter: <3 mg/L
- Metals: trace
- Pesticides: <0.02 ug/L
- Chlorine: 0.13 mg/L
- Alkalinity: No data
- Ca/mg ratio: Ca-52 mg/L; Mg-2.3 mg/L
- Conductivity: 280 uS/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: Daily


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light, 8 h dark, with periods of subdued lighting at the beginning and end of each light phase.
- Light intensity: No data


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality and sublethal effects at 2, 4, 24, 48, 72, and 96 h.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2-3
- 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:

96 h

Dose descriptor:

LC50

Effect conc.:

63.2 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL 40-99.7 mg/L

Effect concentrations 2

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

8.69 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: At 18.6 mg/L and higher, hyperventilation, darkened pigmentation, lethargy, and loss of co-ordination were seen.
- Observations on body length and weight: no data
- Other biological observations: no data
- Mortality of control: none
- Other adverse effects control: no data
- Abnormal responses: One agressive fish in the 8.69 mg/L exposure at 24 h
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The precipitation seen in the range finding study was solved by filtering the solutions prior to test initiation.
- Effect concentrations exceeding solubility of substance in test medium: no

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

Sublethal observations / clinical signs:

Not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:

not applicable

Conclusions:

The LC50 of the test substance to rainbow trout is 63.2 mg/L, with 95% confidence limits of 40.0-99.7 mg/L, based on geometric means of the concentrations tested. The NOEC was 8.69 mg/L.

Executive summary:

Not applicable