Oxirane, 2,2'-[methylenebis[(2,6-dimethyl-4,1-phenylene)oxymethylene]]bis-

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:

13 July 1995 to 06 October 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: "Method for Testing the Degree of Accumulation of Chemical Substances in Fish Body" stipulated in the "Testing Methods for New Chemical Substances"

Version / remarks:

July 13, 1974, Kanpogyo No.5, Planning and Coordination Bureau, Environment Agency, Yakuhatu No.615, Pharmaceutical Affairs Bureau, Ministry of Health and Welfare, and 49 Kikyoku No.392, Basic Industries Bureau, Ministry of International Trade and Industry, Japan

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 305 C (Bioaccumulation: Test for the Degree of Bioconcentration in Fish)

Version / remarks:

1981

GLP compliance:

yes

Radiolabelling:

not specified

Details on sampling:

- Test water analysis was performed twice a week.
- Test fish analysis was performed every two weeks (2nd, 4th, 6th and 8th week)
- Control fish analysis was performed before the initiation and the termination of exposure

Vehicle:

yes

Remarks:

HCO-20 and HCO-40

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

PREPARATION AND APPLICATION OF TEST SOLUTION
- HCO-20 and HCO-40 were used as dispersants
- Method: The test material and HC0-20 (20 times amount of the test material) and HC0-40 (20 times amount of the test material) were dissolved in acetone. After acetone was evaporated from it, ion-exchanged water was added to obtain 1000 mg/L of stock solution.
- Controls: HC0-20 and HC0-40 were dissolved with ion-exchanged water to obtain 800 mg/L of stock solution of each dispersant.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Length at study initiation: average 9.9 cm
- Weight at study initiation: average 25.2 g
- Lipid content at test initiation: average 3.5 %
- Health status: At reception, fish showing visible abnormal signs were removed. Then fish were reared according to flow through system for 2 days after an external disinfection.
- Feeding during test: yes
- Food type: Pelleted feed for carp.
- Amount: An amount corresponding to about 2% of the total body weight of the test fish was fed twice a day by halves. On the day of fish sampling feed was not supplied.

ACCLIMATION
- Acclimation period: After fish keeping, an external disinfection to exterminate parasite was carried out. The fish were transferred to the acclimatisation tank and acclimatised following the second external disinfection. During the period, abnormal fish were removed. Fish were reared according to flow through system at temperature of 25 ± 2°C for 56 days. Then the fish were transferred to test tanks and reared again at the same temperature according to flow through system for 42 days, after further external disinfections.

Route of exposure:

aqueous

Justification for method:

aqueous exposure method used for following reason: The test material was confirmed to be stable.

Test type:

flow-through

Water / sediment media type:

natural water

Remarks:

Underground water pumped up from the grounds of the testing laboratory

Total exposure / uptake duration:

8 wk

Test temperature:

25 ± 2°C

pH:

Initiation: 7.8 - 7.9
Termination: 7.8 - 8.0

Dissolved oxygen:

Initiation: 7.6 mg/L
Termination: 6.6 - 6.9 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass tank (100 litres in volume)
- Renewal rate of test solution: 1155 litres/day (2 mL/min for the stock solution and 800 mL/min for the dilution water)
- No. of organisms per vessel: 11 for the test material doses and 5 for the control
- 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: Underground water was pumped up from the ground of the testing laboratory. The quality of dilution water was confirmed to meet the ministerial ordinance of the Ministry of Public Welfare (December 21, 1992), the water quality criteria for fisheries (Shadanhozin Nihon Suisansigen Hogokyokai, March 1983), and OECD Guideline for Testing of Chemicals 210, Fish, Early-life Stage Toxicity Test

RANGE-FINDING / PRELIMINARY STUDY
- Considering the result of the preliminary test for 48-hr LC50 and analytical detection limit of the test material, the test concentrations of the test material were decided as: 0.1 and 0.01 mg/L.

Nominal and measured concentrations:

Nominal: 0.1 and 0.01 mg/L
Measured:
Level 1 (0.1 mg/L nominal)
- 2 weeks: 0.0747 mg/L
- 4 weeks: 0.0754 mg/L
- 6 weeks: 0.0763 mg/L
- 8 weeks: 0.00765 mg/L

Level 2 (0.01 mg/L nominal)
- 2 weeks: 0.00747 mg/L
- 4 weeks: 0.00739 mg/L
- 6 weeks: 0.00738 mg/L
- 8 weeks: 0.00737 mg/L

Reference substance (positive control):

not specified

Details on estimation of bioconcentration:

Calculation of bioconcentration factor (BCF)
- Test material: In the results of determinations of the test material in test fish, all the measurements were lower than the minimum limit of determination.
- Converted product 1: BCF was calculated by:
Level 1 and Level 2: K = (P x A (t) / A (std) / B x D x C / G –E) /F x 100
A: Peak area (µV·sec) [A(std): Standard solution, A(t): Sample]
B: Ratio of portion used for analysis 100/100
C: Final volume (100 mL)
D: Dilution factor
E: Average concentration of blank in analysis of control 0 µg/g
F: Recovery rate 84.3 %
G: Weight of test fish (g)
H: Average concentration of test material in test water (mg/L)
J: BCF J = K/H
P: Concentration of test material in standard solution 1.50 mg/L

Control: E = P x A (t) / A (std) / B x C
A: Peak area (µV·sec) [A(std): Standard solution, A(t): Sample]
B: Ratio of portion used for analysis 100/100
C: Final volume (100 mL)
G: Weight of test fish (g)
I: Concentration of blank in test fish (µg/g) I = E / G
P: Concentration of test material in standard solution 1.50 mg/L

- Converted product 2: In the results of determinations of the converted product 2 in test fish, all the measurements were lower than the minimum limit of determination.

- From the minimum limit of determination of the test material and the converted products in the test fish obtained, it is possible that the calculation of BCF is provided when the BCF is higher than the following figures:
Level 1: 2.3
Level 2: 24

Key result

Conc. / dose:

0.1 mg/L

Temp.:

25 °C

pH:

7.9

Type:

BCF

Value:

<= 2.3 dimensionless

Basis:

not specified

Remarks on result:

other: Test material (main product)

Key result

Conc. / dose:

0.01 mg/L

Temp.:

25 °C

pH:

7.9

Type:

BCF

Value:

<= 24 dimensionless

Basis:

not specified

Remarks on result:

other: Test material (main product)

Key result

Conc. / dose:

0.1 mg/L

Temp.:

25 °C

pH:

7.9

Type:

BCF

Value:

>= 103 - <= 326 dimensionless

Basis:

not specified

Remarks on result:

other: Converted product 1

Key result

Conc. / dose:

0.01 mg/L

Temp.:

25 °C

pH:

7.9

Type:

BCF

Value:

>= 115 - <= 218 dimensionless

Basis:

not specified

Remarks on result:

other: Converted product 1

Key result

Conc. / dose:

0.1 mg/L

Temp.:

25 °C

pH:

7.9

Type:

BCF

Value:

<= 2.3 dimensionless

Basis:

not specified

Remarks on result:

other: Converted product 2

Key result

Conc. / dose:

0.01 mg/L

Temp.:

25 °C

pH:

7.9

Type:

BCF

Value:

<= 24 dimensionless

Basis:

not specified

Remarks on result:

other: Converted product 2

Details on results:

TEST MATERIAL CONCENTRATIONS
Measured concentrations of the test material in test water at time periods during the study:
Level 1 (0.1 mg/L nominal)
- 2 weeks: 0.0747 mg/L
- 4 weeks: 0.0754 mg/L
- 6 weeks: 0.0763 mg/L
- 8 weeks: 0.00765 mg/L

Level 2 (0.01 mg/L nominal)
- 2 weeks: 0.00747 mg/L
- 4 weeks: 0.00739 mg/L
- 6 weeks: 0.00738 mg/L
- 8 weeks: 0.00737 mg/L
Overall, the mean of each exposure level was maintained at more than 70% of nominated level.

BIOCONCENTRATION FACTORS
The BCF values for the test material and its converted products were as follows:
Test material (main product): Level 1 ≤ 2.3 and Level 2 ≤ 24
Converted Product 1: Level 1: 103-326 and Level 2: 115-218
Converted Product 2: Level 1 ≤ 2.3 and Level 2 ≤ 24

TEST ANIMAL OBSERVATIONS
- No abnormality in behaviour and appearance of the test fish were observed.

DISCUSSION
- Analysis of test water and test fish: Two converted products were detected through HPLC analysis during preliminary experiments. These peaks were identified as the converted product 1 (two epoxides of main product converted to dial form) and the converted product 2 (one epoxide of main product converted to dial form) by high performance liquid chromatograph-mass spectrometer. On the analysis of test fish, the quantitative analysis for the converted products were performed with the main product because there were no standard substances for these converted products. On the analysis of test water, the quantitative analysis for the converted products were not performed.
- Bioconcentration of the test material: The main product and the converted product 2 (same as by-product 2) were not detected in the analysis of test fish. The BCFs of the converted product 1 ranged from 103 to 326 at level 1 and from 115 to 218 at level 2. It is considered that the by-product 2 has no tendency of bioaccumulation because the structure of this substance is the same as that of the converted product 2 which was not detected in the analysis of test fish. The by-product 1 (dimer of the test material) was not detected in the present test.

Validity criteria fulfilled:

not specified

Conclusions:

Under the conditions of this study, the main product and the converted product 2 were not detected in the analysis of test fish. While the BCFs of the converted product 1 ranged from 103 to 326 at level 1 and from 115 to 218 at level 2.

Executive summary:

The bioaccumulation potential of the test material in fish was investigated in accordance with Japanese guidelines and in a study similar to that of OECD 305C. The test was performed under GLP conditions.

Cyprinus carpio were exposed to the test material for 8 weeks at concentrations of 0.1 and 0.01 mg/L (nominal) in a continuous flow through system.

Two converted products were detected through HPLC analysis during preliminary experiments. These peaks were identified as the converted product 1 (two epoxides of main product converted to dial form) and the converted product 2 (one epoxide of main product converted to dial form) by high performance liquid chromatograph-mass spectrometer. On the analysis of test fish, the quantitative analysis for the converted products were performed with the main product because there were no standard substances for these converted products. On the analysis of test water, the quantitative analysis for the converted products were not performed.

Bioconcentration of the test material: The main product and the converted product 2 (same as by-product 2) were not detected in the analysis of test fish. The BCFs of the converted product 1 ranged from 103 to 326 at level 1 and from 115 to 218 at level 2. It is considered that the by-product 2 has no tendency of bioaccumulation because the structure of this substance is the same as that of the converted product 2 which was not detected in the analysis of test fish. The by-product 1 (dimer of the test material) was not detected in the present test.

Under the conditions of this study, the main product and the converted product 2 were not detected in the analysis of test fish. The BCFs of the converted product 1 ranged from 103 to 326 at level 1 and from 115 to 218 at level 2.

Description of key information

Under the conditions of the study, the main product and the converted product 2 were not detected in the analysis of test fish. The BCFs of the converted product 1 ranged from 103 to 326 at level 1 and from 115 to 218 at level 2.

Additional information

The bioaccumulation potential of the test material in fish was investigated in accordance with Japanese guidelines and in a study similar to that of OECD 305C. The test was performed under GLP conditions. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

Cyprinus carpio were exposed to the test material for 8 weeks at concentrations of 0.1 and 0.01 mg/L (nominal) in a continuous flow through system.

Two converted products were detected through HPLC analysis during preliminary experiments. These peaks were identified as the converted product 1 (two epoxides of main product converted to dial form) and the converted product 2 (one epoxide of main product converted to dial form) by high performance liquid chromatograph-mass spectrometer. On the analysis of test fish, the quantitative analysis for the converted products were performed with the main product because there were no standard substances for these converted products. On the analysis of test water, the quantitative analysis for the converted products were not performed.

Bio concentration of the test material: The main product and the converted product 2 (same as by-product 2) were not detected in the analysis of test fish. The BCFs of the converted product 1 ranged from 103 to 326 at level 1 and from 115 to 218 at level 2. It is considered that the by-product 2 has no tendency of bioaccumulation because the structure of this substance is the same as that of the converted product 2 which was not detected in the analysis of test fish. The by-product 1 (dimer of the test material) was not detected in the present test.

Under the conditions of the study, the main product and the converted product 2 were not detected in the analysis of test fish. The BCFs of the converted product 1 ranged from 103 to 326 at level 1 and from 115 to 218 at level 2.