Registration Dossier

Administrative data

Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from August 16, 2009 to October 8, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

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
Specific details on test material used for the study:
Batch:Dl9202595
Purity: 99.8% (GC)
Radiolabelling:
no

Sampling and analysis

Details on sampling:
- Sampling intervals/frequency for test organisms: days 0, 7, 14, 21, 26, 28
- Sampling intervals/frequency for test medium samples: days 0, 7, 14, 21, 26, 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.

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):HCO-40
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)): 0.004 and 0.0004 mg/l
Dose 1: Test item: 0.002 mg/L , Vehicle: 0.004 mg/L
Dose 2: Test item: 0.0002 mg/L , Vehicle: 0.0004 mg/L
Vehicle Control: test item: - , Vehicle: 0.004 mg/L

Test organisms

Test organisms (species):
Cyprinus carpio
Details on test organisms:
Common carp Cyprinus carpio (Lot No. 90325)
Supplier Kitamura Carp Breeding Farm (12-388 Gunchiku, Yashiro-shi,
Kumamoto Prefecture, Japan)
Dates purchased March 25, 2009
Feeding and acclimatization:
After receiving the fish at the Institute of Ecotoxicology Co.,
Ltd., those with a total length of approx. 8.0 ± 4.0 cm were
selected (SOP 2.4). After the fish were treated with Elevage ®
( 4-[2-( 5-nitro-2-furanyl)ethenyl]benzoic acid, sodium salt) and
rock salt, the fish were raised. Fish wete selected again on
August 10, 2009 and the fish were transferred to a water-tank.
After another treatment with Elevage, the process of
acclimatization was started (SOP 2.3, SOP 2.5). Finally, fish
were acclimatized at water temperature 25 ± 2.0°C for 7 days
and it was confirmed that the mortality was 5% or less before
use in the test.
Mean bodyweight 3.19 ± 0.33 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 5.2% (Mean value, n=2)
After experiment 6.1 % (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

Test conditions

Test temperature:
25 ° +/- 0.3 °C
pH:
7.6 +/- 0.1
Dissolved oxygen:
DO oftest water: 7.5 ± 0.1 mg/L
TOC:
High exposure level: 0.6310 mg/L
Low exposure level: 0.6856 mg/L
Control ( dispersant added): 0.4615 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, 26 and 28. The analytical values show that the nominal concentrations of 0.002 and 0.0002 mg/L were maintained at about 100% using solubiliser, respectively. At days 0, 7, 14, 21, 26 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 low bioaccumulation potential, no depuration phase was required.
Nominal and measured concentrations:
nominal concentration 0.002 mg/L:
analytically measured concentrations:

day 0: 0.00174 mg/L
day 7: 0.00202 mg/L
day 14: 0.00196 mg/L
day 21: 0.00202 mg/L
day 26: 0.00198mg/L
day 28: 0.00205 mg/L

nominal concentration 0.0002 mg/L
analytically measured concentrations:

day 0: 0.000160 mg/L
day 7: 0.000206 mg/L
day 14: 0.000161 mg/L
day 21: 0.000183 mg/L
day 26: 0.000166 mg/L
day 28: 0.000169 mg/L
Details on estimation of bioconcentration:
according to guideline

Results and discussion

Bioaccumulation factoropen allclose all
Conc. / dose:
0.002 mg/L
Temp.:
25 °C
pH:
7.7
Type:
BCF
Value:
73 dimensionless
Basis:
normalised lipid fraction
Time of plateau:
7 d
Calculation basis:
steady state
Conc. / dose:
0 mg/L
Temp.:
25 °C
pH:
7.7
Type:
BCF
Value:
93 dimensionless
Basis:
not specified
Time of plateau:
7 d
Calculation basis:
steady state
Details on kinetic parameters:
Steady state was reached within 7 days
Due to the low uptake, no 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 no relevant bioaccumulation at concentrations at or above the water solubility. The steady state BCF values were at 73 and 93 (mean 82).