(2S)-[[(benzyloxy)carbonyl]amino](cyclohexyl)acetic acid

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:

2006-10-31 to 2007-06-12

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, except for a small protocol temperature variation.

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:

yes

Remarks:

There were no deviations from the guideline(s) followed; however, the following deviations were noted from the protocol: the temperature varied by 1.4 deg C, rather than the 1 deg allowed in the protocol.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

There were no deviations from the guideline(s) followed; however, the following deviations were noted from the protocol: the temperature varied by 1.4 deg C, rather than the 1 deg allowed in the protocol.

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Depatment 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-126016
- Molecular formula (if other than submission substance): Not applicable
- Molecular weight (if other than submission substance): Not applicable
- Smiles notation (if other than submission substance): Not applicable
- InChl (if other than submission substance): Not applicable
- Structural formula attached as image file (if other than submission substance): Not applicable
- Substance type: No data
- Physical state: White powder
- Analytical purity: 99.8 area %
- Impurities (identity and concentrations): Not applicable
- Composition of test material, percentage of components: Not applicable
- Isomers composition: Not applicable
- Purity test date: No data
- Lot No.: 1-4107-6-02-02
- Batch no: 25719
- Expiration date of the lot/batch: 2008-02-01
- Stability under test conditions: No data
- Storage condition of test material: Room temperature
- Other: No data

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: Four 10 mL, mid vessel samples were taken from the control and test concentration at 0 and 72 h (fresh media) and at 24 and 96 h (expired media). On each ocassion, two samples from each batch were analyzed immediately; the others refrigerated in case further analysis was required.
- Sample storage conditions before analysis: Refridgeration

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 2.5 g of the test substance was dissolved in diluent water in a 5L flask. On days 0, 2, and 3, the contents were shaken and treated with ultrasound for one h before being poured into a test vessel containing 15L diluent water and further diluted to 25L. On day 1, the flask was treated with ultrasound for 80 min, and the whole 25L volume stirred for 1.5 h.
- Eluate: Not applicable
- Differential loading: Not applicable
- Controls: Diluent water only
- 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): No

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): Approximately 5 months
- Length at study initiation (length definition, mean, range and SD): Mean length of 10 random fish-6.43 cm
- Weight at study initiation (mean and range, SD): Mean weight of those ten fish- 2.54 g
- Method of breeding: Obtained from commercial breeder
- Feeding during test
- Food type: Not fapplicable
- Amount: Not applicable
- Frequency: Not applicable

ACCLIMATION
- Acclimation period: Approximately 3 months in test conditions prior to use
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: Commercial fish food (Nutra Fry 02) equivalent to between 1.25 and 1.5 % of the total weight of the fish in the holding tank.
- Feeding frequency: daily
- Health during acclimation (any mortality observed): No medication was given in the 14 days prior to the test, and mortalities were < 1 %.

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

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

Not applicable

Test conditions

Hardness:

139-140 mg/L as CaCO3

Test temperature:

14.6 +/- 1.4 deg C

pH:

7.23-7.98

Dissolved oxygen:

95-102% air saturation value

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal- Control and 100 mg/L
Mean Measured- Control and 106 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): Open
- Material, size, headspace, fill volume: Glass aquaria, 25L media, 16 cm deep
- Aeration: Supplimental aeration via narrow -bore glass tubes
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): Daily
- No. of organisms per vessel: 10
- 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/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water, was dechlorinated and softened, and 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
- 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-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: single concentration limit test
- 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.:

> 106 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

106 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: None
- Observations on body length and weight: None
- Other biological observations: None
- Mortality of control: None
- Other adverse effects control: None
- Abnormal responses: None
- 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: None

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

The NOEC was derived by direct inspection of the data, for lethal and treatment related effects.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Not applicable

Applicant's summary and conclusion

Conclusions:

The 96 h LC50 for the test substance to rainbow trout could not be calculated, but must be > 106 mg/L.

Executive summary:

Not applicable