Registration Dossier

Administrative data

Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11.12.1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: German national standard method with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: German standard guideline: DIN 38412 part 27 (Draft)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): p-Fluoro-w-chloroacetophenone
- Analytical purity: > 99%
- Physical state: solid
- Lot/batch No.: 23473/86/2

Sampling and analysis

Analytical monitoring:
no

Test organisms

Test organisms (species):
Pseudomonas putida
Details on inoculum:
- Laboratory culture: The test strain of Pseudomonas putida DSM 50026 used is obtained at regular intervals from DSM (German Collection of Microorganism) and is kept on agar slants for further culturing in the Ecotoxicology Laboratory of BASF AG Ludwigshafen .
- Method of cultivation:
- pre-culture
-vessel: Erlenmeyer flask
-volume: 500 mL
-inoculating density: streak with inoculating loop
-incubation time: 16 ± 2 h with shaking
-adjustment of the bacteria suspension: Centrifuge at 6000 rpm for 10 min, decant supernatant, resuspend bacteria in buffer solution and centrifuge again as stated above. Repeat washing and centrifuging. Resuspend bacteria with as little buffer solution as possible.

- Pretreatment: The concentration of the glucose stock solution is 49.54 g/100 mL water ; dissolution time 2 hours. 2 mL of this solution are added to each test and control mix.

- Initial biomass concentration: 240 - 400 TE/F (Turbidity unit/Formazin)

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
30 min

Test conditions

Test temperature:
21 ± 1 °C
Details on test conditions:
TEST SYSTEM
-vessel: Erlenmeyer flask
-volume: 500 mL
-inoculating density: streak with inoculating loop
-incubation time: 16 ± 2 h with shaking
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

OTHER TEST CONDITIONS
- Light intensity: daylight

EFFECT PARAMETERS MEASURED:
measurement of the oxygen content in the medium: 2 - 5 minutes (continuously)
cell counting: determination of the optical density of the pre-culture as validity criterion
pH-measurement: at the start of the test in all concentrations


Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
30 min
Dose descriptor:
EC10
Effect conc.:
400 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Duration:
30 min
Dose descriptor:
EC50
Effect conc.:
970 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate
Duration:
30 min
Dose descriptor:
other: EC90
Effect conc.:
10 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Remarks:
respiration rate

Applicant's summary and conclusion

Validity criteria fulfilled:
yes