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Environmental fate & pathways

Bioaccumulation: aquatic / sediment

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Reference
Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 SEP 2010 - 8 NOV 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)
Version / remarks:
before 2 Oct 2012
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: “Bioaccumulation test of chemical substance in fish " stipulated in “the Order Prescribing the Items of the Test Relating to the New Chemical Substance”
Deviations:
no
GLP compliance:
yes
Radiolabelling:
not specified
Details on sampling:
- Sampling intervals/frequency for test organisms: day 7, 14, 21, 26, 28
- Sampling intervals/frequency for test medium samples: day 7, 14, 21, 26, 28

Test water:
・Sampling: 200ml of Concentration I and II
・Placed them still for a while and let the floating stuffs settle.
・Measurement: 100ml of Concentration I and II (by graduated cylinder)
・Moved into the 200mL-separating funnel
← Washed inside of the cylinder with a small amount of Ultra-purewater
← Dichloromethane
・Shaker (dial7, 5 mis)
・Placed (until the 2 layers become transparent)
- Upper layer (water) - Bottom layer (Dichloromethane)
- Bottom layer (Dichloromethane) - Upper layer (water)


Filtrate
・Evaporator (water bath 35℃)
・Dried by air spray
・Ultra-pure water / THF/Acetonitrile
・Quantification
Concentration I: 25ml
Concentration II: 5ml

Quantified solution
HPLC

Analyzing test water of control group (at the beginning of the test) was in accordance with the method for Concentration II.

Fish body
Test fish (2 fish paired)
・Measurement: full length, body weight
・Chopping
・Mover into the homogenizer cup
・Homogenize (9000 r.p.m, 6 mins)
・Washing
・Put the washing fluid into the homogenizer cup
・Moved the content into the centrifuge tube
・Washing
・Moved the washing fluid into the centrifuge tube
・Adjusted the weight of centrifuge tube with a small amount of Acetonitrile
・Centrifugation (9000 r.p.m, about 10℃, 15 mins)

Supernatant Residue
・Filtration (Glass filter GC50 / Advantech Toyo
・Filtrated the washing fluid and put into the previous measuring flask
・Waited until the temperature of the solution becomes room temperature
・Quantification 100ml (Acetonitrile)

Quantified solution
・2.5ml taken by whole pipette
・Moved into the100mL-Egg-plant flask
・Evaporator (water bath 40℃)
・Dried by air spray


Sep-Pak Florisil
・Conditioning
・Elution
・Taken in the 50mL-Egg-plant flask
・Evaporator (water bath 40℃)
・Dried by air spray
・Quantification 1ml

Quantified solution
HPLC

The flow rate into the Sep-Pak Florisil was set about 3ml/min.
Vehicle:
yes
Details on preparation of test solutions, spiked fish food or sediment:
0.15 mg/L(1st concentration) and 0.015 mg/L(2nd concentration) of Tween 80 was used as solubilising agent
Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name:carp
Route of exposure:
aqueous
Test type:
flow-through
Water / sediment media type:
natural water: freshwater
Total exposure / uptake duration:
28 d
Hardness:
not specified
Test temperature:
not specified
pH:
not specified
Dissolved oxygen:
not specified
TOC:
not specified
Salinity:
not specified
Conductivity:
not specified
Details on test conditions:
TEST SYSTEM
- No. of organisms per concentration (replicates): 50
- No. of organisms per control / vehicle control (replicates): 15


PRELIMINARY STUDY (Acute toxicity test)
- Test concentrations: 31 mg/L (limit test)
- Results used to determine the conditions for the definitive study: LC50 (96hr) > 31 mg/L(Limit test), Danio rerio, solubilizing agent: Tween 80, 100 mg/L
Nominal and measured concentrations:
0.05 mg/L (1st concentration), 0.005 mg/L (2nd concentration)
0.0457 - 0.0520 mg/L, 0.00468 - 0.00502 mg/L
Reference substance (positive control):
not specified
Details on estimation of bioconcentration:
BASIS INFORMATION
- Measured/calculated logPow: >4.7
Lipid content:
4.7 %
Time point:
start of exposure
Lipid content:
4.6 %
Time point:
end of exposure
Conc. / dose:
0.15 mg/L
Type:
BCF
Value:
>= 3 - <= 20 dimensionless
Basis:
whole body w.w.
Time of plateau:
28 d
Calculation basis:
steady state
Conc. / dose:
0.005 mg/L
Type:
BCF
Value:
>= 25 - <= 31 dimensionless
Basis:
whole body w.w.
Time of plateau:
28 d
Calculation basis:
steady state
Details on results:
Recovery rate (average)
Recovery from water (%): Concentration I: 94.5%
Concentration II: 94.2%
Recovery from fish (%): 86.2%

Concentration test results

 

 Uptake period

 Day 7

 Day 14

 Day 21

 Day 26  Day 28

 Concentration I

 Test substance

concentration in

water (mg/L)

 0.0457 0.0495   0.0520  0.0506  0.0476
   concentration rate

 8

10

 9

8

 8

12

 20

7

 6

3

 Concentration II

 Test substance

concentration in

water (mg/L)

 0.00477

 0.00502

 0.00494

 0.00494

 0.00468

 

 Concentration

rate

 <11

<11

 <11

25

 31

<12

 27

25

 <12

<12

Validity criteria fulfilled:
not specified
Conclusions:
During 28 days exposure period BCF values showed less than 50 fold through 28 days’ exposure period. Therefore 28 days after the start of exposure the concentration rate was regarded to reach steady state and the test was finished.
Bioconcentration factors (BCF) of the test substance in carp were 3 to 20 fold for the 1st concentration and 25 to 31 fold for the 2nd concentration. Hence, the substance is not regarded as bioaccumulative.
Executive summary:

The study was conducted in compliance with GLP regulations and in accordance with the OECD Test Guideline No.305 "Bioconcentration : Flow-through Fish test” (before 2nd October 2012).

10 fishes of Danio rerio (Zebra fish) were used for acute toxicity test and 100 mg/L of Tween 80 was used as solubilising agent. LC50(96hr) was defined as >31 mg/L (limit test).

115 fishes (50 fishes for each test concentration, 15 fishes for control) of Cyprinus carpio were used for the bioaccumulation test. Based on 96 hours LC50 value the tests were performed in each 0.05 mg/L (1st concentration), 0.005mg/L (2nd concentration) of the test substance concentration and 0.15 mg/L (1st concentration) 0.015 mg/L (2nd concentration) of Tween 80 was used as solubiliging agent. Analysis of fish was done on 7th, 14th, 21st, 26th and 28th days after the start of exposure.

Result

Concentration test result

 

Uptake period

7 day

14 day

21 day

26 day

28 day

1st concentration

Test substance concentration in water (mg/L)

0.0457

0.0495

0.0520

0.0506

0.0476

Concentration

rate

8

9

8

20

6

10

8

12

7

3

2nd concentration

Test substance concentration in water (mg/L)

0.00477

0.00502

0.00494

0.00494

0.00468

Concentration

rate

<11

<11

31

27

<12

<11

25

<12

25

<12

Average recovery rate from water was determined as 94.5 % (1st concentration) and 94.2 % (2nd concentration) and average recovery rate from fish was determined as 86.2 %.

Theoretical detection limit of BCF value was determined as 1 (1st concentration) and 12 (2nd concentration).

During 28 days exposure period BCF values showed less than 50 fold through 28 days’ exposure period. Therefore 28 days after the start of exposure the concentration rate was regarded to reach steady state and the test was finished.

Bioconcentration factors (BCF) of the test substance in carp were 3 to 20 fold for the 1st concentration and 25 to 31 fold for the 2nd concentration. Hence, the substance is not regarded as bioaccumulative.

Description of key information

Key, Bioconcentration, Cyprinus carpio, 28 d, OECD 305, GLP: BCF = 31, not bioaccumulative

Key value for chemical safety assessment

BCF (aquatic species):
31 dimensionless

Additional information

OECD 305

The study was conducted in compliance with GLP regulations and in accordance with the OECD Test Guideline No.305 "Bioconcentration : Flow-through Fish test” (before 2nd October 2012).

10 fishes of Danio rerio (Zebra fish) were used for acute toxicity test and 100 mg/L of Tween 80 was used as solubilising agent. LC50(96hr) was defined as >31 mg/L (limit test).

115 fishes (50 fishes for each test concentration, 15 fishes for control) of Cyprinus carpio were used for the bioaccumulation test. Based on 96 hours LC 50 value the tests were performed in each 0.05 mg/L (1st concentration), 0.005mg/L (2nd concentration) of the test substance concentration and 0.15 mg/L (1st concentration) 0.015 mg/L (2nd concentration) of Tween 80 was used as solubiliging agent. Analysis of fish was done on 7th, 14th, 21st, 26th and 28th days after the start of exposure.

Result

Concentration test result

 

Uptake period

7 day

14 day

21 day

26day

28 day

1st concentration

Test substance concentration in water (mg/L)

0.0457

0.0495

0.0520

0.0506

0.0476

Concentration

rate

8

9

8

20

6

10

8

12

7

3

2nd concentration

Test substance concentration in water (mg/L)

0.00477

0.00502

0.00494

0.00494

0.00468

Concentration

rate

<11

<11

31

27

<12

<11

25

<12

25

<12

Average recovery rate from water was determined as 94.5% (1st concentration) and 94.2% (2nd concentration) and average recovery rate from fish was determined as 86.2%.

Theoretical detection limit of BCF value was determined as 1 (1st concentration) and 12 (2nd concentration).

During 28 days exposure period BCF values showed less than 50 fold through 28 days’ exposure period. Therefore 28 days after the start of exposure the concentration rate was regarded to reach steady state and the test was finished.

Bioconcentration factors (BCF) of the test substance in carp were 3 to 20 fold for the1st concentration and 25 to 31 fold for the 2nd concentration.