Triazine derivative

Bioaccumulation: aquatic / sediment

Administrative data

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study initiation date: April 21, 2008; Study completion date: June 27, 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP and in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not effect the quality of the relevant results.

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

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

Appearance: Yellow powder
Stability: Stable
Storage: Dark storage place at room temperature.

Radiolabelling:

no

Sampling and analysis

Details on sampling:

- Sampling intervals/frequency for test organisms:
The test fish of Level 1 and 2 were analysed five times in the duration of exposure. Four fish were taken out at each sampling time and divided into two groups, both both fish were analysed individually.
The control fish were analysed before the experiment started and after the experiment completion.

- Sampling intervals/frequency for test medium samples:
The test water of each level was analysed once before the first analysis of test fish and at the same time as analysis of the test fish thereafter.

Test solutions

Vehicle:

no

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

ACUTE TOXICITY TEST:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)

Preparation of stock solution:
The test sample was dissolved in ion-exchanged water to prepare 9.46 g/L stock solution.
Test concentration set at 200 mg/L and control.

BIOCONCENTRATION TEST:
Preparation of stock solution:
Level 1 - 757 mg/L stock solution was prepared by dissolving test sample in ion-exchanged water.
Level 2 - 75.7 mg/L stock solution was prepared by dissolving test sample in ion-exchanged water.
Test concentrations were set as:
Level 1 - 1.89 mg/L
Level 2 - 0.189 mg/L.
The control was set as a blank test.

Test organisms

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM FOR AXCUTE TOXICITY TEST
- Common name: Orange-red killifish (Oryzias Latipes)
- Source: Kurume Laboratory Chemicals Evaluation and Research Institute, Japan
- Length at study initiation (length definition, mean, range and SD): average 3.1 cm
- Weight at study initiation (mean and range, SD): average 0.24 g

ACCLIMATION
The fish were checked visually and those with any abnormalities were removed.
The remainder were transferred to an acclimatizing aquarium and acclimatized there after the external disinfection. The fish showing any abnormalities during this period were removed and the remainder was reared for 13 days in a flow-through system at the temperature of 25 ± 2°C.

TEST ORGANISM FOR BIOCONCENTRATION TEST
- Common name: Carp
- Strain: Cyprinus carpio
- Source: Kurume Laboratory Chemicals Evaluation and Research Institute, Japan
- Age at study initiation (mean and range, SD): Yearling fish
- Length at study initiation (length definition, mean, range and SD): 5.3 - 8.2 cm


ACCLIMATION
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. Fish showing any abnormality during this period were removed and the remainder was reared for 20 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 26 days, following the external disinfection.

Feeding:
Feed - feed for fry of carp
Composition - Proteins content >43.0%
- Lipid content >3.0%
Feeding amount and interval - amount corresponding to about 2% of total body weight was fed twice a day in halves.
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:

28 d

Test conditions

Hardness:

Total hardness of dilution water: 15 mg/l

Test temperature:

Acute toxicity test:
At initial exposure: 24.4°C
Before renewal of test water: 25.0°C

Bioconcentration test:
Level 1: 24.0 - 24.3°C
Level 2: 24.2 - 24.6°C
Control: 24.2 - 24.4°C

pH:

Acute toxicity test:
pH of test water at initial exposure: 8.1
pH of test water before renewal of test water: 8.1 - 8.3

Bioconcentration test:
Level 1: 8.0, 8.2
Level 2: 8.0, 8.1
Control: 8.0, 8.1

Dissolved oxygen:

Acute toxicity test:
At initial exposure: 8.1 mg/L
Before renewal of test water: 7.7 mg/L

Bioconcentration test:
Level 1: 8.0 - 8.2 mg/L
Level 2: 8.0 - 8.2 mg/L
Control: 8.0 - 8.2 mg/L

TOC:

Analytical results if dilution water gives the measured Total Organic Carbon content of 0.3 mg/L

Details on test conditions:

ACUTE TOXICITY TEST SYSTEM
- Test tank: Round glass vessel
- Volume of test water: 4 L / level
- Number of fish: 10 / level
- Duration of exposure: 96 hours
- Exposure method: Semi-static system (Renewal of test water at every 24 hours).
- Aeration: The test tank was supplied with air during the exposure.

BIOCONCENTRATION TEST SYSTEM
- Supply of test water: Flow-through system used.
- Test tank: 70 L glass tank.
- Flow rate of test water (Level 1 and 2): 2.0 mL/min for stock solution and 800 mL/min for dilution water, 1155 L/day of test water were supplied.
- Flow rate of test water (control): 800 mL/min dilution water, 11522 L/day of test water were supplied.
- Stock solution bottle: 25 L glass bottle (Frequency of renewal, 1 time/week).
- 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 - 28; Control - 12
- Duration of exposure: 28 days. A stead-state has been reached after 28 days.

TEST MEDIUM / WATER PARAMETERS
- Origin: Groundwater from the premises of testing laboratory.
The dilution water was analysed and measured and it was confirmed that the water met the requirements of the required standards.

Nominal and measured concentrations:

Acute toxicity test:
Nominal: 200 mg/l and control

Bioconcentration test:
Test concentrations were set as follows. The control was set as a blank test
Nominal concentrations:
Level 1: 1.89 mg/L
Level 2: 0.189 mg/L

Measured concentrations:
Level 1: 1.77 - 1.88
Level 2: 0.172 - 0.184

Reference substance (positive control):

no

Details on estimation of bioconcentration:

Calculable BCF:
On the basis of the minimum determination limit of the test item (calculated as 0.10 mg/L for Level 1 and 0.010 mg/L for Level 2), BCF can be obtained when BCF exceeds the following. The average concentration of the test item in test water obtained from all the analysed samples was used to calculate the following calculable BCF.
Level 1 - 1.1
Level 2 - 11

Results and discussion

Lipid content

Lipid content 1

Lipid content:

4.94 %

Time point:

start of exposure

Lipid content 2

Lipid content:

6.07 %

Time point:

end of exposure

Bioaccumulation factor

Bioaccumulation factor 1

Type:

BCF

Value:

<= 1.1 dimensionless

Basis:

whole body w.w.

Time of plateau:

28 d

Calculation basis:

steady state

Remarks on result:

other: Conc.in environment / dose:1.89 mg/L (Level 1)

Bioaccumulation factor 2

Type:

BCF

Value:

<= 11 dimensionless

Basis:

whole body w.w.

Time of plateau:

28 d

Calculation basis:

steady state

Remarks on result:

other: Conc.in environment / dose:0.189 mg/L (Level 2)

Results with reference substance (positive control):

Not applicable.

Details on results:

- Mortality of test organisms: No mortality was observed.
- Behavioural abnormalities: No abnormality in behavior or appearance was noted.

Any other information on results incl. tables

Results:

Concentration of the test item in test water:

The measured concentrations of the test item in test water are shown in the table below.

Each concentration of the test item was maintained at more than 91% of each nominated concentration. The variation of the concentrations of the test item was within ± 20% of the average measured concentrations.

Measured concentrations of test item in test water (unit: mg/L)

Level	After 1 day	After 8 days	After 13 days	After 16 days	After 20 days	After 28 days	Average
1	1.79	1.77	1.86	1.88	1.80	1.81	1.82
2	0.181	0.172	0.180	0.179	0.179	0.184	0.179

Bioconcentration Factors:

BCFs are shown in the table below.

BCFs of the test item were not more than 1.1 at Level 1 and not more than 11 at Level 2.

Level	After 8 days	After 13 days	After 16 days	After 20 days	After 28 days
1	≤1.1 ≤1.1	≤1.1 ≤1.1	≤1.1 ≤1.1	≤1.1 ≤1.1	≤1.1 ≤1.1
2	≤11 ≤11	≤11 ≤11	≤11 ≤11	≤11 ≤11	≤11 ≤11

BCFs at a steady-state (BCFss)

Because the test item in all test fish at the last three analyses were not more than the minimum determination limit of the test item, BCFss was not calculated.

However, because all BCFs were less than 100, it was evaluated that a steady-state was reached after 28 days.

Result of Acute Toxicity Test:

LD₅₀: > 200 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

BCFs of the test item were not more than 1.1 at Level 1 and not more than 11 at Level 2.

Executive summary:

Bioconcentration study of C-Y9 in carp

Test conditions:

Acute toxicity test:

Test fish: Orange-red killifish (Oryzia latipes)

Duration of exposure: 96 hours

Exposure method: Semi-static system (Renewal of test water at every 24 hours).

Bioconcentration test:

Test fish: Carp (Cyprinus carpio)

Nominal concentrations of test item: High exposure level (Level 1) - 1.89 mg/L

Low exposure level (Level 2) - 0.189 mg/L

Duration of exposure: 28 days

Exposure method: Continous flow system

Analytical method: Liquid chromatography-mass spectometry

Results:

96 -hour LC₅₀ value: >200 mg/L

Bioconcentration factors:

Level 1: <= 1.1

Levle 2: <= 11