Registration Dossier

Administrative data

Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2004-12-14 to 2005-01-11
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study performed under GLP and according to OECD Guideline 305. Even though Tween 80 was used as solubilizer, the test is regarded as valid since the lower test concentration is below the water solubility.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report Date:
2005

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)
Version / remarks:
adopted on 14 June, 1996
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Radiolabelling:
no

Sampling and analysis

Details on sampling:
- Sampling intervals/frequency for test organisms: on day 7, 14, 21, 23, and 28
- Sampling intervals/frequency for test medium samples: on day 0, 7, 14, 21, 23, and 28
- Sample storage conditions before analysis: not applicable
- Details on sampling and analysis of test organisms and test media samples:
Test organisms: sampling on the day of analysis; sample preparation: mincing, homogenization, extraction of test substance to receive a fixed volume solution; HPLC analysis of the generated solution
Test media: preparation of the test water with the aim of generating an organic layer and to receive a fixed volume of test sample; HPLC analysis of the generated solution; pH, DO (adjusted to be approximately 7 mg/L) and water temperature in each aquarium were measured and recorded on the same days when water analyses were performed.

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)
- Method: preparation of a stock solution with dispersant and solvent
- Controls: A 250 mg/L aqueous solution of the dispersant was diluted with the test water supply and then continuously supplied to a test aquarium.
- Chemical name of vehicle:
Dispersant: Polyoxyethylenesorbitan monooleate (Tween 80);
Solvent: Methanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)):
Stock solution: the maximum tween 80 amount was 5-times of the test substance;
Final test solution: the high concentration level with 0.5 mg/L test substance had a dispersant concentration of 2.5 mg/L and the low concentration level with 0.05 mg/L test substance a dispersant concentration of 0.25 mg/L

Test organisms

Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name: common carp
- Source: Kitamura Fish Farm (388, Gunchiku 12-bancho, Yatsushiro-shi, Kumamoto)
- Length at study initiation (lenght definition, mean, range and SD): Mean length: 6.2 ± 0.2 cm
- Weight at study initiation (mean and range, SD): Mean bodyweight: 2.6 ± 0.3 g
- Feeding during test: No feed was given on the day before sampling of the test fish.
- Food type: Mixed carp pellets (Suimmy) with crude protein 37.0% or greater and crude fat 2.5% or greater (manufacturer: Nippon Pet Food Co., Ltd.)
- Amount: The test fish were fed pellets in the amount of ca. 2% per bodyweight/day.

ACCLIMATION
- Acclimation period: 7 days before the study
- Acclimation conditions (same as test or not): yes; temperature: 25 ± 2.0°C
- Type and amount of food: Mixed carp pellets (Suimmy) with crude protein 37.0% or greater and crude fat 2.5% or greater (manufacturer: Nippon Pet Food Co., Ltd.); The test fish were fed pellets in the amount of ca. 2% per bodyweight/day.
- Health during acclimation (any mortality observed): less than 5 %

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

Hardness:
No data.
Test temperature:
24.7 ± 0.7°C
pH:
pH 7.1 ± 0.2
Dissolved oxygen:
7.4 ± 0.2 mg/L
TOC:
The total organic carbon was confirmed to be 10 mg/L or less:
High exposure level 1.518 mg/ L
Low exposure level 0.449 mg/ L
Control (dispersant) group 1.454 mg/L
Salinity:
No data.
Details on test conditions:
Test aquarium:
"Flow-through system"
Made of glass
Volume 100 L (water volume 50 L)
Flow-through system 300 mL/min (432 L/day)
Aquarium No. 1, No. 2 and No. 3 were used for concentration level 1 at a high exposure level, concentration level 2 at a low exposure level and control group, respectively.
Reference substance (positive control):
not specified
Details on estimation of bioconcentration:
Calculation of the bioconcentration factor (BCF) and the bioconcentration factor at steady state (BCFss):

BCF = Concentration of the test substance in fish (µg/g)/ Concentration of the test subsance in aquarium (mg/L)

BCFss = Mean concentration in fish in the steady-state (µg/g) / Mean concentration in aquarium in the steady-state (mg/L)

Results and discussion

Lipid contentopen allclose all
Lipid content:
4.7 %
Time point:
start of exposure
Lipid content:
5 %
Time point:
end of exposure
Bioaccumulation factoropen allclose all
Type:
BCF
Value:
>= 7 - <= 12 L/kg
Basis:
total lipid content
Time of plateau:
28 d
Calculation basis:
steady state
Remarks on result:
other: range of BCFss obtained for the main and minor constituents (total of 13) of the test item
Remarks:
Conc.in environment / dose:0.5 mg/L
Type:
BCF
Value:
>= 10 - <= 18 L/kg
Basis:
total lipid content
Time of plateau:
28 d
Calculation basis:
steady state
Remarks on result:
other: range of BCFss obtained for the main and minor constituents (total of 13) of the test item
Remarks:
Conc.in environment / dose:0.05 mg/L

Any other information on results incl. tables

The bioconcentration factors (BCFs) of the present study of all components were less than 50 folds both at the high (0.5 mg/L) and low exposure level (0.05 mg/L).

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The test item does not significantly accumulate in organisms.