Lithium phosphorodifluoridate

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

other: Bioconcentration test of chemical substances in fish and shellfish (Yakushokuhatsu No. 1121002, Heisei 15.11.13 Seikyoku No.2, Kanpokihatsu No. 031121002)

Version / remarks:

November 21, 2003; latest revision, April 1, 2005

Principles of method if other than guideline:

To evaluate the bioaccumulation potential of the test substance for notification under Chemical Substances Control Law of Japan

GLP compliance:

yes

Radiolabelling:

no

Details on sampling:

- Sampling intervals/frequency for test organisms: On days 2, 7, 14, 21 and 28 of the exposure period
- Sampling intervals/frequency for test medium samples: On days 0, 2, 7, 14, 21, 28 of the exposure period
- Sample storage conditions before analysis: Not stored

- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods):
Test media samples
High concentration level - a portion of the test water was diluted ten-fold with purified water (dilution ratio 1:10), Next, 0.5mL of the diluted solution was collected using an auto sampler vial, mixed with 0.5mL of acetonitrile (dilution 2:2). The samples were analysed with LC/MS
Low concentration level - collected 0.5 mL of the test water in an auto-sampler vial, mixed with 0.5mL of acetonitrile (dilution ratio 1:1). The samples were analysed with LC/MS

Test organism samples
Four fish taken from each concentration level, analysed two at a time: Two fish weighed, cut into pieces, added Acetonitrile 30mL, purified water 10mL, homogenised for 5 minutes at approx 800 rpm, filtered with suction through filter paper (S-60) on Buchner. Acetonitrile-water layer obtained, residue was resuspensed in acetonitrile and re-filtered and acetonitrile-water layer was combined with 1st filtration. Diluted to 100 mL with purified water. 1mL aliquot was removed and diluted to 10mL with 50% acetonitrile/purified water, passed through Sep-Pak C8 cartridge. 1 mL of the filterate was taken and diluted to 10 mLwith 50% acetonitrile/purified water. Samples then analysed using LC/MS analysis.

Vehicle:

no

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

Each feed solution was supplied to mixing glass tube with metering pump for feed solution, and diluted to each nominal concentration by mixing with dilution water delivered with metering pump for dilution water, then poured into each test chamber.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Niikura Fish Farm, 1217, Shimoya, Isehara-city, Kanagawa, Japan
- Age at study initiation (mean and range, SD): Less than a year after hatching
- Length at study initiation (length definition, mean, range and SD): 8 +/- 4 cm
- Weight at study initiation (mean and range, SD): approx. 2.5 g
- Lipid content at test initiation (mean and range, SD): 4.9% (n=3 4.5-5.6%)
- Description of housing/holding area: Aquarium with flowing water
- Feeding during test
- Food type: Babygold(R)
- Amount: amount 2% of fish weight
- Frequency: every day

ACCLIMATION
- Acclimation period: =/>2 weeks
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: amount 2% of fish weight
- Feeding frequency: once a day
- Health during acclimation (any mortality observed): Mortality during 1 week before testing: <5%

Route of exposure:

aqueous

Justification for method:

aqueous exposure method used for following reason: required by national regulations

Test type:

flow-through

Water / sediment media type:

natural water

Total exposure / uptake duration:

28 d

Hardness:

not specified

Test temperature:

24 +/- 2°C

pH:

6.0 - 8.5

Dissolved oxygen:

>60% of the saturation

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria , 50 L
- Aeration: Continuous
- Type of flow-through: continuous flow-through dilution system
- Renewal rate of test solution (frequency/flow rate): 800L/day
- No. of organisms per vessel: 28
- No. of vessels per concentration (replicates): 1
- No. of vessels per control / vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Yokohama municipal tap water treated with activated charcoal, subsequently added aqueous solution of sodium thiosulphate for dechlorination was used.
- Metals: less than detection limits
- Chlorine: <0.02 mg/L
- Alkalinity: 38 mg/L
- Conductance: 117 µS/cm
- Holding medium different from test medium: No
- Intervals of water quality measurement: checked every 6 months

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: approx. 16 hours light/day
- Light intensity:Hf fluorescent lamp (wavelength 400-700 nm)

RANGE-FINDING / PRELIMINARY STUDY
- based on 96h LC50 values from acute fish toxicity test

Nominal and measured concentrations:

Nominal: 0.2 mg/L and 0.02 mg/L

Reference substance (positive control):

no

Details on estimation of bioconcentration:

The bioconcentration factor (BCF) is calculated by the following equation

BCF = Cf at each point / Cw for the point

The steady state bioconcentration factor (BCFss) is calcuated by the following:

BCFss = mean Cf at the steady-state / mean Cw at the steady-state

Where Cf - concentration of the test substance iin test fish (ug/g)
Cw - concentration of the test substance in test wtaer (mg/L)

Steady state: steady state is decided to be reached, when three successive BCF at intervals of 48 hours or longer are within 20% of each other. Cf and Cw are calculated from the last three successive data. When the obtained BCF are all less than 100, it is considered that steady state is reached.

If BCF values are smaller than 100 in all points and three successive mean BCF at intervals of 48 hours or longer are not within 20% each other, the BCFss is not calculated by above equation and BCFss is just declared /= maximum BCF value"

The lowest detectable amount of the test substance in fish was <0.83 µg/g corresponding to 4.2 and 42 times of the high and low concentration levels, respectively. It was indicated that BCF would obtained when the test substance in test water was concentrated by factors of approx. 4 and 42 for high and low concentration levels, respectively. However, these factors might vary depending on the variation of test substance concentration in water, fish weight used for analysis and / or recovery.

Lipid content:

4.9 %

Time point:

start of exposure

Lipid content:

5.8 %

Time point:

end of exposure

Conc. / dose:

0.2 mg/L

Temp.:

24 °C

Type:

BCF

Value:

< 4 dimensionless

Basis:

whole body w.w.

Calculation basis:

steady state

Conc. / dose:

0.02 mg/L

Temp.:

24 °C

Type:

BCF

Value:

< 39 dimensionless

Basis:

whole body w.w.

Calculation basis:

steady state

Details on results:

- Mortality of test organisms: <10% in control and treated fish
- Behavioural abnormalities: None
- Observations on feeding behavior: none

Table 1: Concentration of the test substance in the test fish and bioconcentration factor (BCF)

 High concentration level

		W1	W2	A	B	C	D	E	F	G
Exposure period		Fish Weight (g)		Peak Area		Conc. in final solution	Conc. in fish body	Mean conc. in water	BCF	Mean BCF
Day	No.	No.1	No.2	sample	Std	mg/L	μg/g	mg/L
2	1	2.71	3.88	ND	202732	<0.0005	<0.77	0.197	<4	NA
2	2	3.56	3.59	ND	202732	<0.0005	<0.71	0.197	<4
7	1	3.16	4.3	ND	188596	<0.0005	<0.68	0.197	<4	NA
7	2	3.88	3.65	ND	188596	<0.0005	<0.67	0.197	<4
14	1	4.74	3.14	ND	227889	<0.0005	<0.64	0.197	<4	NA
14	2	2.86	4.21	ND	227889	<0.0005	<0.72	0.197	<4
21	1	3.19	3.76	ND	126863	<0.0005	<0.73	0.197	<4	NA
21	2	4.66	3.57	ND	126863	<0.0005	<0.62	0.197	<4
28	1	3.91	4.01	ND	235523	<0.0005	<0.64	0.197	<4	NA
28	2	4.06	4.31	ND	235523	<0.0005	<0.61	0.197	<4

 

Table 2: Concentration of the test substance in the test fish and bioconcentration factor (BCF)

 Low concentration level

		W1	W2	A	B	C	D	E	F	G
Exposure period		Fish Weight (g)		Peak Area		Conc. in final solution	Conc. in fish body	Mean conc. in water	BCF	Mean BCF
Day	No.	No.1	No.2	sample	Std	mg/L	μg/g	mg/L
2	1	3.21	3.66	ND	202732	<0.0005	<0.74	0.0203	<37	NA
2	2	3.34	3.07	ND	202732	<0.0005	<0.79	0.0203	<39
7	1	3.33	4.26	ND	188596	<0.0005	<0.67	0.0200	<34	NA
7	2	2.88	3.78	ND	188596	<0.0005	<0.76	0.0200	<39
14	1	4.03	3.44	ND	227889	<0.0005	<0.68	0.0200	<34	NA
14	2	3.87	3.61	ND	227889	<0.0005	<0.68	0.0200	<34
21	1	2.88	4.60	ND	126863	<0.0005	<0.68	0.0199	<35	NA
21	2	4.21	4.57	ND	126863	<0.0005	<0.5	0.0199	<29
28	1	3.62	3.14	ND	235523	<0.0005	<0.75	0.0199	<38	NA
28	2	3.66	3.23	ND	235523	<0.0005	<0.74	0.0199	<37

 

 

Concentration of std. solution (Cstd)	0.01	mg/L
Final volume (FV)	10	mL
Sampling factor (SF1)	100
Sampling factor (SF2)	10
Recovery (R)	98.7%	%

 

Equations:

C=A÷B×Cstd

D=A÷B×Cstd×(FV/1000)×SF1×SF2÷(W1+W2)÷(R/100)×1000

F=A÷B×Cstd×(FV/1000)×SF1×SF2÷(W1+W2)÷(R/100)×1000÷E

G=(F1+F2)÷2

 

ND : Not detected, <5% of std.

NA : Not available

Validity criteria fulfilled:

yes

Remarks:

Test conditions were in line with test method.

Conclusions:

BCF at steady state where <4 at the high concentration and <39 at the low concentration.

It is concluded that the bioaccumulation potential of this test substance in fish is low under the test conditions.

Description of key information

It is concluded that the bioaccumulation potential of this test substance in fish is low under the test conditions. The BCF values were calculated to be <4 and <39 in the high and low concentrations tested, respectively. Therefore no BCF value is used for the chemical safety assessment.