Condensation products of Dihydrofuran-2,5-dione, mono-alkenyl derivs. with polyethylene amine and acetic anhydride

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:

July 10, 2009 -January 12, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with GLP standards

Qualifier:

equivalent or similar to guideline

Guideline:

other: Yakushokuhatsu No.1121002, Heisei 15.11.13 Seikyoku No.2, Kanpokihatsu No.031121002, November 21, 2003; the latest revision, November 20, 2006

GLP compliance:

yes

Details on sampling:

The bioconcentration test system was set up in duplicate for different concentrations and two groups of carp were kept in water containing the test substance. In addition, the system was also set up for control and a group of carp was kept in water without the test substance.
From the result of the preliminary acute toxicity test with Medaka, the 50% lethal concentration (LC50) for 96 hours of the test substance was >20 mg/L. Therefore the bioconcentration test was accordingly conducted at exposure concentration levels of 0.2 and 0.02 mg/L, less than a hundredth and a thousandth of the 96-hour LCso

Test organisms (species):

Cyprinus carpio

Details on test organisms:

Acute Toxicity Test:

Species: Medaka (Oryzias latipes)
Source: Tunashima Fishing Corp. 18-1 Tunashima-nishi 5-chome, Kouhoku-ku, Yokohama-city, Kanagawa, Japan
Pre-acclimation: At the start of acclimation, visual observation was made and abnormal fish was removed. The remaining normal fish were
reared in an aquarium with flowing water.
Body length: 2 ± 1 cm
Body weight: Approximately 0.2 g
Acclimation: Aquarium number: C-12, Water temperature: 24 ± 1°C, Period: June 23, 2009 to July 13, 2009, Feeding: The food (Tetramin® from Tetra Werke) was given in an amount of 2% of the fish weight everyday. (Feeding was discontinued during the 24 hours before testing.)
Mortality during the 1 week before testing: <5%
Medication during acclimation: None

Bioconcentration Test Method

Species: Carp (Cyprinus carpio)
Source: Niikura Fish Farm, 1217, Shimoya, Isehara-city, Kanagawa, Japan
Pre-acclimation: At the start of acclimation, visual observation was made and abnormal fish were removed. The remaining normal fish were then reared in an aquarium with flowing water.
Total length: 8 + 4 cm
Body weight: Approximately 6 g
Age: About a year after hatching
Acclimation : Aquarium number: A-6, Water temperature: 24 ± 2°C, Period: June 16, 2009 to July 24, 2009, Food: Babygold® from Kyorin , Feeding: The food was given in an amount of 1.5% of the fish weight everyday, Mortality during the 1 week before testing: <5%
Medication during acclimation: None

Route of exposure:

aqueous

Total exposure / uptake duration:

62 d

Hardness:

no data

Test temperature:

24±2°C

pH:

6.0 — 8.5

Dissolved oxygen:

6.9-8.3 mg/L

TOC:

no data

Salinity:

no data

Details on test conditions:

- Supply of test water: Continuous flow-through dilution system
- Exposure level (nominal concentrations of the test substance)
High concentration level: 0.2 mg/L (Vehicle concentration, Tetrahydrofuran: 50 ppm [v/v])
Low concentration level: 0.02 mg/L (Vehicle concentration, Tetrahydrofuran: 50 ppm [v/v])
Control: 0 mg/L (Vehicle concentration, Tetrahydrofuran: 50 ppm [v/v])
- Supply of dilution water: 400 L/day (Turnover rate: 8 times/day)
- Supply of feed solution: 20 mL/day
-Number of fish (at the start of the exposure period):
High concentration level: 36 fish in 50-L test water (>1 L/day/g)
Low concentration level: 36 fish in 50-L test water (>1 L/day/g)
Control: 12 fish in 50-L test water (>1 L/day/g)
- Water temperature: 24±2°C
- Dissolved oxygen: >60% of the saturation (>5 mg/L at 24°C)
- Aeration: Continuous
- pH: 6.0 — 8.5
- Lighting: Approximately 16 hr/day
(Hf fluorescent lamp with wavelengths of 400-700 nm)
- Exposure period: 62 days

Nominal and measured concentrations:

Test substance in test water (nominal concentrations)
High concentration level: 0.2 mg/L
Low concentration level: 0.02 mg/L
Control: 0 mg/L

Vehicles in test water
High concentration level: Tetrahydrofuran 50 ppm (v/v)
Low concentration level: Tetrahydrofuran 50 ppm (v/v)
Control: Tetrahydrofuran 50 ppm (v/v)

Details on estimation of bioconcentration:

Calculation of Bioconcentration Factors
The bioconcentration factor (BCF) was calculated by the following equation:
BCF. = Cf. Cw,n
The steady-state bioconcentration factor (BCFss) is calculated by the following
equation:
BCFss = Cf,s Cw,s
Where
BCF,,: The bioconcentration factor after n days, BCFss: The steady-state bioconcentration factor, Cf: Concentration of the test substance in test fish after n days (p.g/g); Mean concentration of the test substance in test water over n days (mg/L), Cts: Mean concentration in test fish at steady-state (p.g/g), Cw,s: Mean concentration in test water at steady-state (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. Cts and Cw,s are calculated from last three successive data.
When the obtained BCF are all less than 100, it is also considered that steady-state is reached. In this case, three successive BCF are not
necessary within 20% of each other. The BCFss is not calculated by above equation and BCFss is just declared "< (maximum BCF value) "or
< (maximum BCF value).

Key result

Remarks on result:

other: see tabulated results below

Details on results:

During the test period, there was no abnormality in shape of the body or in swimming and eating behavior in the fish at all the concentration levels throughout the test period. Therefore, it was confirmed that the test fish had been reared appropriately.

Component	High Concentration level	Low Concentration Level
A	BCFSS 56 (49-75*1)	BCFSS <90 (<59 -90*2)
B	BCFSS < 87 (<60 -87*2)	BCF < 667 (< 623 -<667 *3)
C	BCFSS< 39 (<7 -39*2)	BCFSS <64 (< 62 -<64 *2)
D	BCFSS< 39 (3 -39*2)	BCFSS < 33 (<31 -<33 *2)

*1: The variation of mean BCF at the last three concecutive mesurement (over 48 hrs) were confirmed to fall within ±20%, the concentration factor was considered to reach the steady-state during 62 days of exposure period. *2: The variation of mean BCF at the three consecutive measurements were not confirmed to fall within ±20%. However all of the BCF values during the exsposure period were less than 100, thus the BCF were considered to reach the steady-state. *3: The variation of mean BCF at the last three consecutive measurements (over 48 hrs) of component B were not confirmed to fall within ±20%, and did not juge to reach the steady-state. Therefore the BCF value was not calculated.

Validity criteria fulfilled:

yes

Conclusions:

Bioconcentration factors were determined for the test substance that is a multicomponent substance-High concentration level - A= BCFSS 56 (49-75*1), B= BCFSS < 87 (<60-87*2), C=BCFSS < 39 (<7-39*2) , D=BCFSS < 39 (3-39*2) and Low concentration level - A=BCFSS < 90 (<59-90*2), B=BCF < 667 (<623-<667*3), C=BCFSS < 64 (<62-<64*2), D=BCFSS < 33 (<31-<33*2)

Executive summary:

This study was conducted to evaluate the bioaccumulation potential of the test substance for notification under the Chemical Substances Control Law of Japan. The bioconcentration test system was set up in duplicate for different concentrations ( 0.2 mg/L, 0.02 mg/L and control ) and two groups of carp were kept in water containing the test substance. During the exposure period, the concentrations of the test substance in water and fish were measured periodically. The bioconcentration factors (BCF) calculated from the results were used to evaluate bioaccumulation potential of the test substance in tissues of fish. The test substance was multi component substance (A,B,C,D), so the measureable 4 components (m/z: 504.4, 520.4, 782.6, 810.6) were analyzed and the bioaccumulation potential was judged. The BCF of a test substance in test fish were as follow: High concentration level - A= BCFSS 56 (49-75*1), B= BCFSS < 87 (<60-87*2), C=BCFSS < 39 (<7-39*2) , D=BCFSS < 39 (3-39*2) and Low concentration level - A=BCFSS < 90 (<59-90*2), B=BCF < 667 (<623-<667*3), C=BCFSS < 64 (<62-<64*2), D=BCFSS < 33 (<31-<33*2) where BCFSS: The steady-state bioconcentration factor, *: Range of BCF and judgmental reason

*1: The variation of mean BCF at the last three concecutive mesurement (over 48 hrs) were confirmed to fall within ±20%, the concentration factor was considered to reach the steady-state during 62 days of exposure period.

*2: The variation of mean BCF at the three consecutive measurements were not confirmed to fall within ±20%. However all of the BCF values during the exsposure period were less than 100, thus the BCF were considered to reach the steady-state.

*3: The variation of mean BCF at the last three consecutive measurements (over 48 hrs) of component B were not confirmed to fall within ±20%, and did not juge to reach the steady-state. Therefore the BCF value was not calculated.

Description of key information

Bioconcentration  factors were determined for the test substance that is a multicomponent substance-High concentration level - A= BCFSS 56 (49-75*1), B= BCFSS < 87 (<60-87*2), C=BCFSS < 39 (<7-39*2) , D=BCFSS < 39 (3-39*2) and Low concentration level - A=BCFSS < 90 (<59-90*2), B=BCF < 667 (<623-<667*3), C=BCFSS < 64 (<62-<64*2), D=BCFSS < 33 (<31-<33*2)

Additional information

A GLP study was conducted to evaluate the bioaccumulation potential of the test substance for notification under the Chemical Substances Control Law of Japan. The bioconcentration test system was set up in duplicate for different concentrations ( 0.2 mg/L, 0.02 mg/L and control ) and two groups of carp were kept in water containing the test substance. During the exposure period, the concentrations of the test substance in water and fish were measured periodically. The bioconcentration factors (BCF) calculated from the results were used to evaluate bioaccumulation potential of the test substance in tissues of fish. The test substance was multi component substance (A,B,C,D), so the measureable 4 components (m/z: 504.4, 520.4, 782.6, 810.6) were analyzed and the bioaccumulation potential was judged. The BCF of a test substance in test fish were as follow: High concentration level - A= BCFSS 56 (49-75*1), B= BCFSS < 87 (<60-87*2), C=BCFSS < 39 (<7-39*2) , D=BCFSS < 39 (3-39*2) and Low concentration level - A=BCFSS < 90 (<59-90*2), B=BCF < 667 (<623-<667*3), C=BCFSS < 64 (<62-<64*2), D=BCFSS < 33 (<31-<33*2)