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adult fish: sub(lethal) effects
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from August 16, 2009 to October 8, 2009
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
study performed according to OECD 305

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guidelineopen allclose all
according to guideline
other: OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)
according to guideline
other: METI Guideline: Concentration Test on Chemical Substances in Fish
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
Cas Number:
Molecular formula:
Specific details on test material used for the study:
Batch: Dl9202595
Purity: 99.8% (GC)

Sampling and analysis

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

Test solutions

Details on test solutions:
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

Test type:
Water media type:
Limit test:
Total exposure duration:
28 d

Test conditions

Test temperature:
25 ° +/- 0.3 °C
7.6 +/- 0.1
Dissolved oxygen:
DO of test water: 7.5 ± 0.1 mg/L
Nominal and measured concentrations:
nominal concentration 0.02 mg/L:
analytically measured concentrations:

day 7: 0.0185 mg/L;
day 14: 0.0213 mg/L
day 21: 0.0183 mg/L
day 24: 0.0188 mg/L
day 28: 0.0168 mg/L

nominal concentration 0.002 mg/L
analytically measured concentrations:

day 7: 0.00198 mg/L
day 14: 0.00198 mg/L
day 21: 0.00189 mg/L
day 24: 0.00198 mg/L
day 28: 0.00187 mg/L
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.

Results and discussion

Effect concentrationsopen allclose all
28 d
Dose descriptor:
Effect conc.:
0.002 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: concentration was above the water solubility limit stabilized with Tween 80
14 d
Dose descriptor:
Effect conc.:
0.002 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: concentration was above the water solubility limit stabilized with Tween 80

Applicant's summary and conclusion

Validity criteria fulfilled:
The test material showed no toxic potential at concentrations at or above the water solubility. The NOEC was 0.002 mg/L and thus higher than the limit of water solubility.The test material showed no relevant toxic potential at water saturated conditions.