Pentyl 2-cyano-(3-(6-methoxybenzothiazol-2-ylimino)-2,3-dihydro-1H-isoindol-1-ylidene)acetate

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:

From Jul. 23, 2004 to Oct. 25, 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

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

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.031111002, November 21, 2003)

Deviations:

no

GLP compliance:

yes

Radiolabelling:

no

Details on sampling:

- Sampling intervals/frequency for test organisms: Day 5, 14, 28, 42 and 60 of the exposure period
- Sampling intervals/frequency for test medium samples: Day 0, 5, 14, 21, 28, 35, 42, 49 and 60 of the exposure period
- Sample storage conditions before analysis: Yes

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: Municipal tap water was treated with activated charcoal. The dechlorinated water was aerated and maintained at 24 °C. This dilution water was checked half-yearly and confirmed to suit aquatic living resources.
- Controls: Yes, dilution water
- Chemical name of vehicle: Dimethylformamide (DMF)
- Concentration of vehicle in test medium: Control - 21 ppm DMF; 0.01 mg/L test substance - 25 ppm DMF; 0.1 mg/L test substance - 21 ppm DMF
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Niikura Fish Farm, 1211, Shimoya, Isehara-city, Kanagawa, Japan
- Age at study initiation (mean and range, SD): About 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 5 g

ACCLIMATION
- Acclimation period: ≥ 2 wk
- Acclimation conditions (same as test or not): Same
- Type and amount of food: Baby gold, 2 % of the fish weight
- Health during acclimation (any mortality observed): Mortality= < 5 %

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

60 d

Hardness:

59 mg/L as CaCO3

Test temperature:

24±2 °C

pH:

6.0-8.5

Dissolved oxygen:

≥60% of the saturation (≥ 5 mg/L at 24 °C)

TOC:

No data

Salinity:

No data

Details on test conditions:

TEST SYSTEM
- Aeration: Continuous
- Type of flow-through (e.g. peristaltic or proportional diluter): Continuous flow-through diluter system
- No. of organisms per vessel: 44 fish in 50 L test water in treatment group, 28 fish in 50 mL test water in Control group
- No. of vessels per concentration (replicates): Three
- No. of vessels per control / vehicle control (replicates): Three


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Yokohama municipal tap water treated with activated charcoal
- Chlorine: <0.02 mg/L
- Alkalinity: 44 mg/L
- Ca/mg ratio: 4.22/1
- Conductance: 156 µ S/cm



OTHER TEST CONDITIONS
- Photoperiod: 16 h/day
- Light intensity: Hf fluorescent lamp with wavelengths of 400-700 nm

Nominal and measured concentrations:

Nominal: 0 (HCO-40 4 mg/L, Dimethylformamide=21 ppm (v/v)), 0.01 (HCO-40 0.4 mg/L, Dimethylformamide=25 ppm (v/v)), 0.1 mg/L (HCO-40 4 mg/L, Dimethylformamide=21 ppm (v/v))

Reference substance (positive control):

not specified

Details on estimation of bioconcentration:

BASIS FOR CALCULATION OF BCF
BCF=Cf/Cw
steady-state bioconcentration factor (BCFss)= BCFss= Cfs/Cws

Lipid content:

ca. 5.3 %

Time point:

start of exposure

Remarks on result:

other: (n=3; 4.9-5.7 %)

Lipid content:

ca. 4 %

Time point:

end of exposure

Remarks on result:

other: (n=3, 3.7-4.3 %)

Type:

BCF

Value:

ca. 38 dimensionless

Basis:

whole body w.w.

Time of plateau:

60 d

Calculation basis:

steady state

Remarks on result:

other: Conc.in environment / dose:0.1 mg/L nominal

Type:

BCF

Value:

ca. 236 dimensionless

Basis:

whole body w.w.

Time of plateau:

60 d

Calculation basis:

steady state

Remarks on result:

other: Conc.in environment / dose:0.01 mg/L nominal

Type:

BCF

Value:

>= 19 - <= 42 dimensionless

Basis:

whole body w.w.

Time of plateau:

60 d

Remarks on result:

other: Variation of individual time-specific BCF values

Remarks:

Conc.in environment / dose:0.1 mg/L nominal

Type:

BCF

Value:

>= 94 - <= 269 dimensionless

Basis:

whole body w.w.

Time of plateau:

60 d

Remarks on result:

other: Variation of individual time-specific BCF values

Remarks:

Conc.in environment / dose:0.01 mg/L nominal

Details on kinetic parameters:

Not investigated

Metabolites:

Not investigated

Results with reference substance (positive control):

Not applicable

Details on results:

- Mortality of test organisms: No
- Behavioural abnormalities: No
- The individual time-specific bioconcentration factors (BCF) of the test substance varied between 19 and 42 at the high concentration level, and from 94 to 277 at the low concentration level.
- BCF at steady state (BCFss) was 38 at the high concentration level and 236 at the low concentration level (in the study report itself an erroneous BCFss of 244 is documented, due to a calculation error for the mean aqueous test substance concentration at the steady state)

Reported statistics:

No data

The nominal concentration in bioconcentration test was confirmed to be less than a hundredth and a thousandth of the 96-h LC50.

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the bioconcentration factors (BCF) of the test substance were 19-42 at high concentration level, and 94 -277 at low concentration level. BCF at steady state (BCFss) were 38 at high concentration level and 236 at low concentration level.

Executive summary:

The study was conducted evaluate the potential of the test substance to accumulate in the tissues of the carp in accordance with "Test Method Relating to New Chemical Substances Bioconcentration test of chemical substances in fish and shellfish, (Yakushokuhatsu No.1121002, Heisei 15.11.13 Seikyoku, No.2, Kanpokihatsu No.031111002, November 21, 2003)" and in compliance with GLP.

Fish were exposed to nominal concentrations 0, 0.01 and 0.1 mg/L of test substance in test water using continuous flow-through diluter system (800 L/day) for 60 d.

Prior to the bioconcentration test, an acute toxicity test with Medaka was conducted in accordance with the OECD guideline 203 to confirm that the nominal concentrations of the bioconcentration test were less than a hundredth and a thousandth of the 96 h LC50. Concentrations of the test substance in the test water were not measured, and any effects were evaluated on the basis of nominal concentrations. No mortality occurred at the limit test concentration hence the 96-h LC50 of the test substance was >10 mg/L.

HPLC was used for the analysis of test substance in test media and test fish.

The lipid content was determined to be 5.3 and 4.0 % at the start and end of the experiment, respectively.

Steady state was reached at day 28 in both the low and the high test concentration. Under the test conditions, the steady state (BCFss) was 38 at the high concentration level and 236 at the low concentration level.

Description of key information

The steady-state bioconcentration factor (BCFss) was 38 at the high concentration level and 236 at the low concentration level. In an conservative approach, the higher values is adopted for the chemical safety assessment.

Key value for chemical safety assessment

BCF (aquatic species):

236 dimensionless

Additional information

The study was conducted evaluate the potential of the test substance to accumulate in the tissues of the carp in accordance with the Test Method Relating to New Chemical Substances Bioconcentration test of chemical substances in fish and shellfish, (Yakushokuhatsu No.1121002, Heisei 15.11.13 Seikyoku, No.2, Kanpokihatsu No.031111002, November 21, 2003) and in compliance with GLP.

Fish were exposed to nominal concentrations of 0.01 and 0.1 mg/L of test substance in test water, and a blank control, using continuous flow-through diluter system (800 L/d) for 60 days.

Prior to the bioconcentration test, an acute toxicity test with Medaka was conducted in accordance with the OECD guideline 203 to confirm that the nominal concentrations of the bioconcentration test were less than a hundredth and thousandth of the 96 h LC50. Concentrations of the test substance in test water were not measured in this acute toxicity test hence any effects were evaluated on the basis of nominal concentrations. There was not mortality at the limit test concentration of 10 mg/L, therefore the 96-h LC50 of the test substance was determined to be > 10 mg/L.

In the bioaccumulation test, HPLC was used for the analysis of test substance in test media and test fish. The lipid content was determined to be 5.3 and 4.0 % at the start and end of the experiment, respectively.

Steady state was reached after 28 days at both the low and the high test concentration. The BCF at steady state (BCFss) was 38 at the high concentration level and 236 at the low concentration level (Oshioka, 2004).

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