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 microorganisms

Administrative data

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20-aug-2007 to 29-aug-2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and 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:

EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration 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:

no

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The test substance is non-soluble in water in concentrations needed for the test. The water solubility is 102.2 mg/L, therefore direct dosing of test substance was used. The substance was weighed into reaction container where 200 mL of water was introduced.

Test organisms

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

The activated sludge was collected two days before the day of testing. After the sample collection the sludge was washed with potable water for 0.5 hour and subsequently decanted for 0.5 hour. This procedure was repeated three times. Further the sludge was modified by addition of 50 mL of cultivation medium per 2 L of diluted sludge suspension at permanent aeration till the day of test. The dry weight was determined from 10 mL of sludge suspension after 0.5 hour sedimentation. Before the test the sludge was suspended in water up to concentration about 4000 mg of sludge dry weight per litre. The pH adjustment to 6.0 was carried out. In that way modified sludge suspension was aeraed untill the use.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Test conditions

Test temperature:

20 ± 2 °C

pH:

Start: 7.11 - 7.12
End: 7.62 - 7.88

Dissolved oxygen:

Control: 3.58 mg O2/L per 10 minuten
Highest dose level (1000 mg/L): 3.83 mg O2/L per 10 minuten

Nominal and measured concentrations:

Nominal concentrations: 100, 180, 320, 560 and 1000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: BOD bottles with a volume of 280 mL
- Aeration: filtered pressured air, airflow 0.8 L/min
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2
- Biomass loading rate: sludge dry weight: 1.76 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionised water


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- The rate of oxygen consumption for each concentration and control (respiration rate, R) was calculated from the linear parts of the graphs recording oxygen concentrations decline between 6.5 and 2.5 mg/L.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Preliminary study
- Test concentrations: 46, 100, 220, 460 and 1000 mg/L
- Results used to determine the conditions for the definitive study: 100, 180, 320, 560 and 1000 mg/L

Reference substance (positive control):

yes

Remarks:

3.5-dichlorphenol

Results and discussion

Effect concentrations

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Results with reference substance (positive control):

Results with reference substance valid? yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

All tested concentrations provided no significant inhibition of respiration rate, and the respiration inhibition of activated sludge caused by the test substance SP100 C6, the EC50 exceeded 1000 mg/L.