Sodium 6,14-diethyl-1,1,1,2,2,18,18,19,19,19-decafluoro-8,12-dioxo-10-({[1-(2,2,3,3,3-pentafluoropropoxy)butan-2-yl]oxy}carbonyl)-4,7,13,16-tetraoxanonadecane-9-sulfonate

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 MAR 2012 - 08 JAN 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study performed according to OECD TG 305 (draft 2011)

Qualifier:

according to guideline

Guideline:

OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)

Version / remarks:

Draft 2011

Deviations:

no

Principles of method if other than guideline:

Minimised test acc. to OECD 305 draft (2011) with one exposure concentration, only.

GLP compliance:

yes (incl. QA statement)

Remarks:

Gewerbeaufsichtsamt Hildesheim, 20 October 2010

Radiolabelling:

no

Details on sampling:

- Sampling intervals/frequency for test organisms:
Test Group: 4 fish on uptake phase days 14 and 28
Control Group: 2 fish on uptake phase day 28
- Sampling intervals/frequency for test medium samples:
Test Group: on days -1, -2 (prior to start of experiment), 0, 7, 14 and 28
Control Group: on days 0 and 28
- Sample storage conditions before analysis: The fish were weighed and stored at ca. -18 °C until analysis, if necessary.
The test item concentration and the control were stabilised by factor 2 with acetonitrile directly after sampling. Further dilutions (factor 2 or factor 4) were made with a 50:50-mixture of HPLC water and acetonitrile.

Vehicle:

no

Details on preparation of test solutions, spiked fish food or sediment:

Tap water of local origin was used for holding and testing. The water was filtered on activated charcoal and aerated.
Water parameters:
pH-value: 6.0 - 8.5
Hardness: 10 - 250 mg CaCO3 / L

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Strain: Salmonidae
- Source: Forellenzucht Harkenbleck, Steinbrink 41, 30966 Hemmingen, Germany.
- Age at study initiation: 26 weeks
- Length at study initiation: 8 ± 4 cm
- Weight at study initiation: 2 g
- Weight at termination: 5 g (SD 0.98)
- Method of breeding: -
- Health status: healthy
- Feeding during test: The fish were fed daily in two feedings.
- Food type: Forellenzuchtfutter 0.8 -1.2 mm Teichcenter Rieger
- Amount: The given feed amount (1.5 % of body weight) was documented. The amount of food was adjusted accordingly to the weights of sacrificed fish to account for growth during the experiment.
- Frequency: daily

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions (same as test or not): same
- Type and amount of food: Forellenzuchtfutter 0.8 -1.2 mm Teichcenter Rieger
- Feeding frequency: daily
- Health during acclimation (any mortality observed): Only rainbow trout from a pool with at least 12 days of acclimation and mortality < 5 % within the last 7 days before the study started were used in the test.

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

28 d

Hardness:

39 - 55 mg CaCO3 / L

Test temperature:

14.8 - 15.2°C (control group); 14.9 - 15.2°C (test group)

pH:

7.11 - 7.80 (control group); 7.22 - 7.80 (test group)

Dissolved oxygen:

93 - 99 % (control group); 97 - 100 % (test group)

TOC:

1.95 mg/L (mean control; range 1.26 - 2.38), 1.84 mg/L (mean test vessel; range 1.28 - 2.35)

Details on test conditions:

TEST SYSTEM
- Test vessel: aquaria
- Type: covered by glass tops
- Material, size, headspace, fill volume: glass, 175 L
- Aeration: yes
- Renewal rate of test solution (frequency/flow rate): 25 L/h = 5 exchanges / day
- No. of organisms per vessel: 40 per group
- No. of vessels per concentration (replicates): one
- No. of vessels per control / vehicle control (replicates): one
- Biomass loading rate: 0.1 - 1 g fish (wet weight) per L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water
- Holding medium different from test medium: no
- Intervals of water quality measurement: three times per week in all test vessels.
- Intervals of test medium replacement: 5 exchanges/ day

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light / 8 hours dark
- Light intensity: 0.1 - 10 µmol /m^2/s

Nominal and measured concentrations:

0.2 mg/L (nominal) corresponding to 0.17 mg/L (mean measured concentration)

Reference substance (positive control):

no

Details on estimation of bioconcentration:

BASIS INFORMATION
- Measured concentrations in fish and media

BASIS FOR CALCULATION OF BCF
- Estimation software: GraphPad Prism, GRAPHPAD SOFTWARE, INC. + Excel, MICROSOFT CORPORATION
- Results: 19 and 20

Lipid content:

3.73 %

Time point:

start of exposure

Remarks on result:

other: mean

Lipid content:

5.14 %

Time point:

end of exposure

Remarks on result:

other: mean

Conc. / dose:

0.2 µg/L

Temp.:

15 °C

pH:

7.5

Type:

BCF

Value:

19 dimensionless

Basis:

whole body w.w.

Calculation basis:

other: calculated from uptake day 14

Conc. / dose:

0.2 µg/L

Temp.:

15 °C

pH:

7.5

Type:

BCF

Value:

20 dimensionless

Basis:

whole body w.w.

Calculation basis:

other: calculated from uptake day 28

Conc. / dose:

0.2 µg/L

Temp.:

15 °C

pH:

7.5

Type:

BCF

Value:

21 dimensionless

Basis:

normalised lipid fraction

Calculation basis:

other: calculated from uptake day 14

Conc. / dose:

0.2 µg/L

Temp.:

15 °C

pH:

7.5

Type:

BCF

Value:

19 dimensionless

Basis:

normalised lipid fraction

Calculation basis:

other: calculated from uptake day 28

Remarks on result:

not measured/tested

Remarks:

No depuration phase was carried out, since no significant uptake was observed. Thus, no depuration rates were calculated.

Remarks on result:

not measured/tested

Remarks:

No significant uptake of the test item in fish was observed

Details on results:

- Mortality of test organisms: no
- Behavioural abnormalities: no
- Observations on body length and weight: no
- Other biological observations: no abnormalities
- Organ specific bioaccumulation: no
- Bound residues forming a plateau: no
- Mortality and/or behavioural abnormalities of control: no abnormalities
- Loss of test substance during test period: no
- Results with vehicle control: no abnormalities

Mean concentrations determined in fish were 3.2 ± 0. 7 μg/g on day 14 of the uptake phase and 3.4 ± 1.1 μg/g on day 28. These values resulted in lipid normalized concentrations of 3.6 ± 0.8 and 3.3 ± 1 .1 μg/g body weight.

No significant uptake was observed in fish. All measured concentrations in fish were below 50 μg/g fish (the value which would have triggered a depuration phase). The highest concentration was found in fish on day 28 of the uptake phase with 4.98 μg/g.

Validity criteria fulfilled:

yes

Conclusions:

The bioconcentration factors (at any time and lipid normalised, calculated on day 28) determined in this study with the test item were 20 and 19, respectively. The result is below the regulatory value of concern regarding bioaccumulation (< 500).

Executive summary:

Study Design

A minimised bioaccumulation study with rainbow trout (Oncorhynchus mykiss) was conducted to determine the bioconcentration potential of the test material. The GLP-study was conducted according to OECD Guideline 305 (Draft 2011).

A flow-through test with 2 groups (one control group and one exposure group) and an exposure (uptake phase) of 28 days was carried out. The depuration phase was waived, since the measured concentration in fish was < 50 µg / g fish

Analytical verification:
The concentration of the test material was determined via LC-MS/MS analysis. Four fish were sampled on uptake phase days 14 and 28 from the exposure  group. Two fish were sampled from the control group on uptake phase day 28. Samples of the aqueous phase were taken and analysed on uptake phase days -1, -2, 0, 7, 14 and 28 from the exposure group and on uptake phase day 0 and 28 from the control group. The lipid content of the fish was determined at the start and end of the study using 2 fish for the exposure group and 1 fish of the control group.

Evaluation:
Fish growth rates have been calculated. No significant uptake of the test item in fish was observed. Therefore, no uptake / depuration rates and no bioconcentration factors (steady state and kinetic, respectively) have been calculated. Bioconcentration factors at any sampling date during the uptake phase have been calculated based on fish body weight.

Results

Uptake phase:
The aqueous concentrations were stable in the range of 74 - 96 % (Day 0 - 28) of the nominal values throughout the uptake phase. There was no increase of the test item concentrations observed in the fish. All the measured concentrations in fish were below 50 µg / g fish.

Depuration phase:
No depuration rates were calculated since no significant uptake was observed. No depuration phase was carried out.

Bioconcentration factors:
The bioconcentration factors (at any time and lipid normalised, calculated on day 28) determined in this study with the test item were 20 and 19, respectively. The result is below the regulatory value (< 500) of concern regarding bioaccumulation.

Validity:
The validity criteria regarding temperature, oxygen saturation, test item concentrations in water and mortality were fulfilled.

Conclusion

The bioconcentration factors (at any time and lipid normalised, calculated on day 28) determined in this study with the test item were 20 and 19, respectively. The result is below the regulatory value (< 500) of concern regarding bioaccumulation.

Description of key information

Key, bioaccumulation in fish, O. mykiss, OECD 305, GLP, 28 d, BCF = 20

Key value for chemical safety assessment

BCF (aquatic species):

20 dimensionless

Additional information

Study Design

A minimised bioaccumulation study with rainbow trout (Oncorhynchus mykiss) was conducted to determine the bioconcentration potential of the test material. The GLP-study was conducted according to OECD Guideline 305 (Draft 2011).

A flow-through test with 2 groups (one control group and one exposure group) and an exposure (uptake phase) of 28 days was carried out. The depuration phase was waived, since the measured concentration in fish was < 50 µg / g fish

Analytical verification:
The concentration of the test material was determined via LC-MS/MS analysis. Four fish were sampled on uptake phase days 14 and 28 from the exposure  group. Two fish were sampled from the control group on uptake phase day 28. Samples of the aqueous phase were taken and analysed on uptake phase days -1, -2, 0, 7, 14 and 28 from the exposure group and on uptake phase day 0 and 28 from the control group. The lipid content of the fish was determined at the start and end of the study using 2 fish for the exposure group and 1 fish of the control group.

Evaluation:
Fish growth rates have been calculated. No significant uptake of the test item in fish was observed. Therefore, no uptake / depuration rates and no bioconcentration factors (steady state and kinetic, respectively) have been calculated. Bioconcentration factors at any sampling date during the uptake phase have been calculated based on fish body weight.

Results

Uptake phase:
The aqueous concentrations were stable in the range of 74 - 96 % (Day 0 - 28) of the nominal values throughout the uptake phase. There was no increase of the test item concentrations observed in the fish. All the measured concentrations in fish were below 50 µg / g fish.

Depuration phase:
No depuration rates were calculated since no significant uptake was observed. No depuration phase was carried out.

Bioconcentration factors:
The bioconcentration factors (at any time and lipid normalised, calculated on day 28) determined in this study with the test item were 20 and 19, respectively. The result is below the regulatory value (< 500) of concern regarding bioaccumulation.

Validity:
The validity criteria regarding temperature, oxygen saturation, test item concentrations in water and mortality were fulfilled.

Conclusion

The bioconcentration factors (at any time and lipid normalised, calculated on day 28) determined in this study with the test item were 20 and 19, respectively. The result is below the regulatory value (< 500) of concern regarding bioaccumulation.