[1,1'-Biphenyl]-2-carbonitrile, 4'-[(2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-en-3-yl)methyl]-

Toxicity to microorganisms

Administrative data

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 13 September 1996 to 27 November 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study. GLP compliance.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Directive 87/302/CEE - JOCE L 133 (30.05.800) - partie (N° L 133/118): Biodégradation - Boues activées : essai d'inhibition de la respiration

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Deviations:

yes

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-ene-3-yl)methyl)(1,1'-biphenyl)-2-carbonitrile

EC Number:

604-075-6

Cas Number:

138401-24-8

Molecular formula:

C25H27N3O

IUPAC Name:

4'-((2-butyl-4-oxo-1,3-diazaspiro[4.4]non-1-ene-3-yl)methyl)(1,1'-biphenyl)-2-carbonitrile

Details on test material:

- Code of test material (as cited in study report): SR47563
- Chemical name:4’-[(2-butyl-4-oxo-1,3-diaza-spiro[4.4]non-1-en-3-yl)methyl]- biphenyl]-2-carbonitrile
- Physical state: white-beige powder
- Lot/batch No.: 6R00003
- Solubility: 1.3E-3 g/L at 23˚C
- Stability: stable under normal conditions of use
- Moll mass: 385.5
- Molecular formular: C25H27N3O
- Melting point: 97˚C

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

- Concentrations: 100 - 5000 - 1000 - 2000 mg/l
- Sampling method: Substance directly added in the medium (adequate quantity)

Test solutions

Vehicle:

yes

Test organisms

Test organisms (species):

activated sludge, domestic

Details on inoculum:

- Preparation of inoculum for exposure: Decanted, washed and resuspension in an isotonic solution (0.5g NaCl/0.12g MgSO4,7 H2O for 1l of water)
- Pretreatment: Aeration in the test conditions
- Initial biomass concentration: 2.45g/l

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

180 min

Test conditions

Test temperature:

20 +/- 1°C

pH:

7.3

Nominal and measured concentrations:

Concentrations d'essai : solution saturée et 100 - 5000 - 1000 - 2000 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: 1L flasks for 500mL of medium
- Aeration: 0.5 to 1L/min
- No. of vessels per concentration (replicates):
- No. of vessels per control (replicates): 2

OTHER TEST CONDITIONS
- Light intensity: Darkness

Reference substance (positive control):

yes

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

30 min

Dose descriptor:

EC50

Effect conc.:

> 2 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Effect concentrations 2

Duration:

180 min

Dose descriptor:

EC50

Effect conc.:

> 2 000 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 (between 5 and 30mg/l)
- Relevant effect levels: EC50.30min=10.0mg/l, EC50.180min=9.3mg/l

Reported statistics and error estimates:

EC50

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this study, the substance did not affect respiration of micro organisms present in activated sludge at levels largely exceeding its solubility limit in water of ca. 1 mg/L (> 2000 mg/L).