[No public or meaningful name is available]

Bioaccumulation: aquatic / sediment

Administrative data

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 12 May to 21 August 2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP compliant; auxiliary solvents used but otherwise adequate for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Details on test material:

Purity 98.5%

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable

Radiolabelling:

no

Sampling and analysis

Details on sampling:

(1) Observation of test fish
Condition of test fish was observed visually twice a day (once a day at holiday).

(2) Flow rate of test water
Flow rate of stock solution and dilution water were measured with graduated cylinder and recorded once a day.

(3) Temperature of test water
Temperature of test water was measured with alcohol thermometer and recorded once a week.

(4) Concentration of dissolved oxygen in test water
Concentration of dissolved oxygen in test water was measured with dissolved oxygen probe and recorded once a week.

(5) pH of test water
pH of test water was measured with pH meter thrice in experimental period.

(6) Cleaning of test tank
In experimental period, excreta of carp, dirt on test tank, etc. were removed about once a day.

Analysis of test water and fish
Analysis of the test item in test water and test fish was performed with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis.

Frequency of analysis

(1) Test water analysis
The test water of each level was analyzed once before first analysis of test fish and at the same time as analysis of test fish thereafter. The number of each sample was one.

(2) Test fish analysis
The test fish of Level 1 and 2 was analyzed five times in duration of exposure. Four fish were taken out at each sampling time and divided into two groups, then both were analyzed individually*
The control fish was analyzed before the experimental starting and after the experimental completion. Four fish were taken out at each sampling time and divided into two groups, and then each was analyzed individually. In addition, two fish were taken out and three groups (two fish per group) were used for measurement of lipid contents.

* Because one fish was too small to take out the stored sample for the measurement of lipid content, two fish a group were employed.

Test solutions

Vehicle:

yes

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

Preparation of stock solution:
The test sample and Hydrogenated Castor Oil (HCO-40) (10 times amount of it) were dissolved in N,N-dimethylformamide to prepare 500 mg/L stock solution.

Preparation of test solutions:

Level 1
500 mg/L solution of the test item was prepared in the same way as described above. 40.0 mg/L stock solution was then prepared from this solution by dilution with N,N-dimethylformamide.

Level 2
500 mg/L solution of the test item was prepared in the same way as described above. 4.00 mg/L stock solution was then prepared from this solution by dilution with N,N-dimethylformamide.

Control
HCO-40 was dissolved in N,N-dimethylfornamide to prepare 400 mg/L stock solution.

Test concentrations:

Test concentrations of the test item were decided as follows. The control was set as a blank test.

Level 1: 1 µg/L
Level 2: 0.1 µg/L

Test organisms

Test organisms (species):

Cyprinus carpio

Details on test organisms:

Species: Carp (Cyprinns carpio)
Reason for selection : The previous data conducted with this species can be compared and the size of this species is adequate for handling.

Supplier: Kurume Laboratory Chemicals Evaluation and Research Institute, Japan
Date of hatch: December 21, 2007
Starting date of acclimatization: April 1, 2008

Conditions for acclimatization
Period: After rearing to test fish size in a receiving aquarium, the fish were transferred to an acclimatizing aquarium and acclimatized there after the external disinfection. The fish showing any abnormality during this period were removed and the remainder was reared for 22 days in a flow through system at the temperature of 25 ± 2°C. The fish were checked for health conditions and transferred to test tanks. Thereafter the fish were reared at the same temperature in the flow through system for 28 days, following the external disinfection.

External disinfection
The external disinfection in the acclimatizing aquarium was carried out in an aqueous solution containing 50 mg/L OTC for fisheries and 7 g/L sodium chloride for 24 hours. The external disinfection in test tanks was carried out in an aqueous solution containing 50 mg/L OTC for fisheries and 7 g/L sodium chloride for 24 hours.

Length: 6.7 - 11.2 cm

Lot No.: TFC-080401

Age: Yearling fish

Feeding
Feed: Feed for fry of carp
Composition:
Proteins content ≥43.0 %
Lipid content ≥3.0 %
Manufacturer: Nippon Formula Feed Mfg. Co., Ltd.
Feeding amount and interval: Amount corresponding to about 2 % of total body weight was fed twice a day in halves (once a day in all at holiday). The fish were starved for 24 hours before sampling.

Study design

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

60 d

Total depuration duration:

0 d

Test conditions

Hardness:

NDA

Test temperature:

Temperature of test water
Level 1: 24.0-24.5°C
Level 2: 24.0-24.6°C
Control: 23.9-24.2°C

pH:

pH of test water
Level 1: 7.9-8.0
Level 2: 7.9-8.1
Control: 7.8-8.1

Dissolved oxygen:

Concentrations of dissolved oxygen in test water
Level 1: 7.5-8.1 mg/L
Level 2: 7.6-8.1 mg/L
Control: 7.7-8.1 mg/L

TOC:

NDA

Salinity:

NDA

Details on test conditions:

Conditions of test and circumstances:

Supply of test water: Flow-through system assembled at this laboratory was used.
Test tank: 70-L glass tank
Flow rate of test water: 0.04 mL/min for stock solution and 1600 mL/min for dilution water, 2304 L/day of test water, were supplied.
Stock solution bottle: 1 L glass brown bottle (Frequency of renewal 2 - 3 times/month)
Time of irradiation with light: Artificial light of white fluorescent lamp (14 hours/day)
Number of fish (at the beginning of exposure):
Level 1 and 2 29
Control 12
Duration of exposure: 60 days
Reason: A steady-state has been reached after 60 days

Dilution water for test:

Origin: Groundwater from the premises of Kurume Laboratory.

Water quality assessment:
The dilution water for test was taken out on July 1, 2008, and it was analyzed and measured. It was confirmed that the dilution water met the requirements of at least one of the following standards.
- Ministerial ordinance of the Ministry of Health, Labour and Welfare No.101 (Revised May 30, 2003)
- OECD Guidelines for Testing of Chemicals, Fish, Early-life Stage Toxicity Test (Guideline 210, July 17, 1992)
- Water quality criteria for fisheries (Japan Fisheries Resource Conservation Association, March 1983)
- Environmental Quality Standards for Water Pollutants No.14 (Revised February 22, 1999, Environment Agency)
- OECD Guidelines for Testing of Chemicals, Bioconcentration : Flow-through Fish Test (Guideline 305, June 14, 1996)

Nominal and measured concentrations:

Nominal concentrations:
Level 1: 1 µg/L
Level 2: 0.1 µg/L

Measured concentrations:
Level 1: 0.989 - 1.04 µg/L
Level 2: 0.101 - 0.106 µg/L

Reference substance (positive control):

no

Details on estimation of bioconcentration:

For details on how the BCF values were calculated, please see attachment "Bioconcentration carp study NX 8000J calculation of BCF values.pdf"

Results and discussion

Lipid content

Lipid content 1

Lipid content:

4.38 %

Time point:

start of exposure

Lipid content 2

Lipid content:

5.05 %

Time point:

end of exposure

Bioaccumulation factor

Bioaccumulation factor 1

Type:

BCF

Value:

77

Basis:

total lipid content

Time of plateau:

60 d

Calculation basis:

steady state

Remarks on result:

other: Conc.in environment / dose:1 µg/L

Bioaccumulation factor 2

Type:

BCF

Value:

150

Basis:

total lipid content

Time of plateau:

60 d

Calculation basis:

steady state

Remarks on result:

other: Conc.in environment / dose:0.1 µg/L

Details on kinetic parameters:

No data available

Metabolites:

No data available

Results with reference substance (positive control):

Not applicable

Details on results:

- Mortality of test organisms: None
- Behavioural abnormalities: None recorded
- Observations on body length and weight: No adverse effects on appearance noted
- Mortality and/or behavioural abnormalities of control: No data available
- Loss of test substance during test period: None. The test concentrations were very near nominal values. The variation of the concentrations of the test item was within a ± 20 % of the average of the measured concentrations.
- Results with vehicle control: No data available.

Reported statistics:

Values were rounded in accordance with JIS Z 8401:1999 rule B. The each value used for calculation was used without rounding on the way of the calculation.
The concentration values of the test item in test water and fish were rounded to 3 figures. BCFs values were rounded to 2 figures.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The bioconcentration factors at a steady state were calculated as being:
Level 1 (1 µg/L): 77
Level 2 (0.1 µg/L): 150