tert-butyl {(1S,3S)-3-{(S)- hydroxy[4-methoxy-3-(3-methoxypropoxy)phenyl]methyl}-4-methyl-1-[ (2S,4S)-5-oxo-4-(propan-2-yl)tetrahydrofuran-2-yl ]pentyl}carbamate

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

Study period:

14-sep-2007 to 27-sep-2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

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 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

Test material

Test material information

Constituent 1

Reference substance name:

tert-butyl N-[(1S,3S,4S)-4-hydroxy-4-[4-methoxy-3-(3-methoxypropoxy)phenyl]-1-[(2S,4S)-5-oxo-4-(propan-2-yl)oxolan-2-yl]-3-(propan-2-yl)butyl]carbamate

EC Number:

700-317-0

Cas Number:

934841-33-5

Molecular formula:

C30H49NO8

IUPAC Name:

tert-butyl N-[(1S,3S,4S)-4-hydroxy-4-[4-methoxy-3-(3-methoxypropoxy)phenyl]-1-[(2S,4S)-5-oxo-4-(propan-2-yl)oxolan-2-yl]-3-(propan-2-yl)butyl]carbamate

Details on test material:

- Name of test material (as cited in study report): SPP100 C6
- Substance type: White, fine-crystallic powder
- Physical state: Solid
- Stability under test conditions: Not indicated
- Storage condition of test material: Stored at ambient temperature
- Analytical purity: 99.2%
Significant impurities: Sum of all impurities by HPLC: 0.8%
Solvent content: Isopropyl acetate (CAS No. 108-21-4): 1.9% (w/w)
Methylcyclohexane (CAS 108-87-2): 5.8% (w/w)
- Lot/batch No.: HH-853.35Kr
- Expiration date of the lot/batch: December 2007

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

-Concentrations: highest and lowest test concentrations and the highest concentration without algae
- Sampling method: at t=0 h and t=72 h
- Sample storage conditions before analysis: Samples were analysed on the same day

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The water solubility of the test substance is 102.2 mg/L. Since the water solubility of the test substance is about 100 mg/L, the saturated solution in test medium was used as stock solution. 100 mg of the test substance was weighed into 1000 mL of test medium for the preliminary test and 110 mg of the test substance was weighed into 1000 mL of test medium for the full test. These mixtures were stirred on a shaft stirrer throughout three days, ultrasonicated for 15 minutes and filtered through 0.45 micron filter. The concentrations of solutions were obtained by dilution of the saturated solution with test medium.

Test organisms

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

TEST ORGANISM
- Common name: Desmodesmus subspicatus
- Strain: Brinkmann 1953/SAG 86.81
- Source (laboratory, culture collection): The Botanic Institute of the Czech Academy of Science, Trebo, 5.10.2006
- Age of inoculum (at test initiation): Preculturing of cells 3-days before start of the test
- Method of cultivation: The strain culture was preinoculated from the stock solution and cultivated in flasks with the test medium

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test conditions

Test temperature:

23 ± 2°C

pH:

Preliminary test:
At t=0 h: 7.8
At t=72 h: 8.3 - 8.8

Full test:
At t=0 h: 7.7 - 7.8
At t=72 h: 7.8 - 9.2

Nominal and measured concentrations:

preliminary test
Nominal concentrations: 1, 5, 10, 50 and 100% of a 0.45 µm filtered solution prepared at a loading rate of 100 mg/L
Determined concentrations: 0.97, 4.94, 9.88, 49.4 and 98.8 mg/L

Full test
Nominal solution concentrations: 56, 62, 68, 75, 83, 91 and 100% of a 0.45 µm filtered solution prepared at a loading rate of 110 mg/L
Determined concentrations: 62.8, 66.3, 72.7, 80.2, 88.7, 97.3 and 106.9 mg/L

Details on test conditions:

- Test vessel: 100 ml, all glass
- Type: open
- Material, size, headspace, fill volume: 150 ml, normal headspace, 50 ml
- Aeration: no
- Initial cells density: 10000 cells/ml
- Control end cells density: 2041667 cells/ml (full test)
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3 replicate of each test concentration without algae.
- No. of vessels per vehicle control (replicates): 3

GROWTH MEDIUM
- Standard medium used: yes

OTHER TEST CONDITIONS
- Sterile test conditions: no
- Adjustment of pH: no
- Photoperiod: continuous
- Light intensity and quality: TLD-lamps 36W/33, white light, wattage 36 Watt.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: The cell densities was evaluated microscopically at 24, 48 and 72 hours. The cell density was measured by direct counting of living cells in Burker's counting chamber.
- Other: 72 h NOErC, 72 h NOEbC, 72 h ErC50, 72 h EbC50


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.1
- Range finding study
- Test concentrations: Nominal concentrations: 1, 5, 10, 50 and 100% of a 0.45 µm filtered solution prepared at a loading rate of 100 mg/L
- Full test
- Nominal solution concentrations: 56, 62, 68, 75, 83, 91 and 100% of a 0.45 µm filtered solution prepared at a loading rate of 110 mg/L

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 106.9 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 106.9 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

- Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test): no
- Any stimulation of growth found in any treatment: no

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50:
The EC50 for growth rate reduction (ERC50: 0-72h) was 0.35 mg/l with a 95 % confidence interval ranging from 0.3 to 0.39 mg/l. The historical ranges for growth rate reduction lie between 0.28 and 0.38 mg/l. Hence, the ERC50: 0-72h for the present batch corresponds with this range.

The EC50 for yield inhibition (EYC50: 0-72h) was 0.67 mg/l with a 95 % confidence interval ranging from 0.6 to 0.75 mg/l. The historical ranges for yield inhibition lie between 0.67 and 1.06mg/l. Hence, the EYC50: 0-72h for the present batch corresponds with this range.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The test substance, SPP100 C6 did not reduce growth rate at any of the concentrations tested and the 72h-EC50 exceeded 100 mg/L.