Registration Dossier

Administrative data

Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March 15, 2012 to Sep 21, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
other: METI Guideline: Concentration Test on Chemical Substances in Fish
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)
Deviations:
no
Principles of method if other than guideline:
SPAN 20 used as solubilizer
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Radiolabelling:
no

Sampling and analysis

Details on sampling:
- Sampling intervals/frequency for test organisms during uptake phase: days 0, 7, 14, 21, 23, 28
- Sampling intervals/frequency for test medium samples during uptake phase: days 0, 7, 14, 21, 23, 28
- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods):
Water: Extraction with CH2Cl2 with further HPLC analysis.
Fish: two fish per sampling homogenised, and extracted with acetonitril/n-hexane. The analysis was performed using HPLC.

- Sampling intervals/frequency for test organisms and test medium during depuration phase: sampling days 2, 6, 9, 14, 21

Test solutions

Vehicle:
yes
Details on preparation of test solutions, spiked fish food or sediment:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Tween 80
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)): 0.16 and 0.016 mg/l
Dose 1: Test item: 0.04 mg/L , Vehicle: 0.16 mg/L
Dose 2: Test item: 0.004 mg/L , Vehicle: 0.016 mg/L
Vehicle Control: test item: - , Vehicle: 0.16 mg/L

Test organisms

Test organisms (species):
Cyprinus carpio
Details on test organisms:
Common carp Cyprinus carpio (Lot No. 120316)
Supplier: Commercial Breeding Farm (Japan)
Dates purchased Mar 16, 2012
Feeding and acclimatization:
After receiving the fish at the Institute of Ecotoxicology Co., Ltd., those with a total length of approx. 6.9 ± 0.3 cm were selected.
Mean bodyweight 3.00 ± 0.29 g
Mean total length 6.9 ± 0.3 cm
Mortality of fish during the experiment
High exposure level 0%
Low exposure level 0%
Control 0%

Lipid contents (by chloroform/methanol extraction method)
Before experiment 4.8% (Mean value, n=2)
After experiment 4.3 % (Mean value, n=2)

Study design

Route of exposure:
aqueous
Test type:
flow-through
Water / sediment media type:
natural water: freshwater
Total exposure / uptake duration:
28 d
Total depuration duration:
21 d

Test conditions

Test temperature:
26.6 ° +/- 0.2 °C
pH:
7.7 +/- 0.1
Dissolved oxygen:
DO oftest water: 7.24 ± 0.2 mg/L
Salinity:
According to guideline
Details on test conditions:
After acclimatisation period, the fish (Cyprinus carpio) were transferred into the test media under flow through conditions. Exposure concentrations were analytically verified at days 0, 7, 14, 21, 23 and 28. The analytical values at nominal concentrations of 0.04 and 0.004 mg/L were maintained at about 100% using solubiliser, respectively. At days 0, 7, 14, 21, 23 and 28 two fish each were removed from each test group. Body weight was determined. The fish were homogenised and extracted using organic solvents. The extract was analysed with HPLC. Due to the high bioaccumulation potential, a depuration phase of 21 days was required.
Nominal and measured concentrations:
nominal concentration 0.04 mg/L:
analytically measured concentrations:

day 0: 0.0365 mg/L
day 7: 0.0374 mg/L
day 14: 0.0367 mg/L
day 21: 0.0424 mg/L
day 23: 0.0422 mg/L
day 28: 0.0389 mg/L


nominal concentration 0.004 mg/L
analytically measured concentrations:

day 0: 0.00388 mg/L
day 7: 0.00339 mg/L
day 14: 0.00383 mg/L
day 21: 0.00412 mg/L
day 23: 0.00362 mg/L
day 28: 0.00411 mg/L
Details on estimation of bioconcentration:
according to guideline

Results and discussion

Bioaccumulation factoropen allclose all
Conc. / dose:
0.04 mg/L
Temp.:
26 °C
pH:
7.7
Type:
BCF
Value:
3 350 dimensionless
Basis:
normalised lipid fraction
Time of plateau:
14 d
Calculation basis:
steady state
Conc. / dose:
0.004 mg/L
Temp.:
26.6 °C
pH:
7.7
Type:
BCF
Value:
3 241 dimensionless
Basis:
not specified
Time of plateau:
14 d
Calculation basis:
steady state
Depuration
Elimination:
yes
Parameter:
DT50
Depuration time (DT):
4.3 d
Rate constants
Rate constant:
overall depuration rate constant (d-1)
Value:
0.16
Details on kinetic parameters:
Steady state was reached within 14 days
Depuration phase was necessary.
Details on results:
- Mortality of test organisms: no effects
- Behavioural abnormalities:no effects
- Observations on body length and weight:no effects
- Other biological observations: no effects
- Bound residues forming a plateau: no
- Mortality and/or behavioural abnormalities of control: no
- Loss of test substance during test period: no
- Results with vehicle control: no effects

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The test material showed a significant bioconcentration potential with a mean BCFss of 3295.