N,N’-di[3-(p-toluene sulfonyl)oxy]phenyl urea

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 305 (Bioaccumulation in Fish: Aqueous and Dietary Exposure) -I: Aqueous Exposure Bioconcentration Fish Test

Qualifier:

according to guideline

Guideline:

other: I: Aqueous Exposure Bioconcentration Fish Test in "Method for Testing the degree of Accumulation of Chemical Substances in Fish Body as stipulated in the "Testing Methods for New Chemical Substance" of Japanese Substances Control Law

Version / remarks:

March 31, 2011 Yakushokuhatsu 0331 No. 7; Heisei 23.03.29 Seikyoku No. 5; Kampoki hatsu No. 110331009; Latest revision March 29, 2018, Yakuseihatsu 0329 No. 13, 20180326 Seikyoku No. 1; Kampokihatsu No. 1803293)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Radiolabelling:

no

Details on sampling:

- Sampling intervals/frequency for test organisms: every 2 days and final analysis was conducted at 28 days
- Sampling intervals/frequency for test medium samples: once before uptake phase and then every 2 days (same as test organisms)
- Sample storage conditions before analysis: not specified
- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods):

TEST WATER: Aliquots of the test water from each tank (500 mL for Level1, 500 mL control) and pre-treated for LC-MS/MS analysis as follows: Addition of Formic acid (500 µL); Column Chromatography*
Eluate made up to 2mL with acetonitrile

TEST ORGANISMS:
Step 1: Body weight and length measured, chopped to pieces, acetonitrile 30 mL added, homogenised for 1 minute, washed with approx. 5 mL acetonitrile, centrifuged (7000xg, 5 minutes).
Step 2: Supernatant passed through absorbent cotton = filtrate
Step 3: Fill up to 50mL (acetonitrile), 5mL transferred to 250mL ion-exchanged water, addition of Formic acid (500 µL); Column Chromatography*
Eluate made up to 2mL with acetonitrile

* Condition of column chromatograph
Sep-Pak Plus tC2
Conditionings:
Acetonitrile. approx. 10mL
Ultrapure water. Approx. 10 mL
Loading: Whole volume of the solution was loaded
Elution: Elute Acetonitrile 1.8 mL
Eluate used for analysis

Vehicle:

yes

Remarks:

Tetrahydrofuran

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

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test sample (100 mg) and HCO-40 (5.0g) were dissolved in tetrahydrofuran to prepare a 1000mg/L test item solution (100mL). This solution (2 mL) was taken out and diluted with tetrahydrofuran to prepare the stock solution of 4 mg/L as the test item and 200 mg/L as HCO-40 (500 mL).
- Controls: HCO-40 (600 mg/) was dissolved in tetrahydrofuran to prepare 200 mg/L stock solution of HCO-40 (3L).
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): HCO-40 and tetrahydrofuran
- Evidence of undissolved material (e.g. precipitate, surface film, etc): not reported

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Common carp
- Source: In-house laboratory
- Age at study initiation (mean and range, SD): Yearling fish
- Length at study initiation (length definition, mean, range and SD): 6.8 - 7.2 cm
- Weight at study initiation (mean and range, SD): minimum fish weight was more than two-thirds of the maximum weight
- Weight at termination (mean and range, SD): not specified
- Lipid content at test initiation (mean and range, SD): 5%
- Health status: Mortality during the acclimatisation was less than 5%
- Feeding during test
- Food type: Feed for coloured carp (protein content: >30%; lipid content >4%)
- Amount: 2% of total body weight was fed
- Frequency: The daily dose of feed was split into 2 feeding (one per day during weekend and holidays). The test fish were starved for 24 hours before sampling.

ACCLIMATION
- Acclimation period: 89 days
- Acclimation conditions (same as test or not): 25 +/-2°C 
- Type and amount of food: same as above
- Feeding frequency: same as above
- Health during acclimation (any mortality observed):Mortality during the acclimatisation was less than 5%

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water

Total exposure / uptake duration:

28 d

Hardness:

Test Item = 18.8 mgCaCO3/L
Control = 19.2 mgCaCO3/L

Test temperature:

Test Item = 25.2 - 25.4°C 
Control = 24.9 - 25.2°C 

pH:

Test Item; control = 7.7 - 7.9

Dissolved oxygen:

Test item = 7.2 - 7.7 mg/L
Control = 7.4 - 7.8 mg/L

TOC:

Test item = 24.5 - 26.5 mg C/L
Control = 25.2 - 28.2 mgC/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 70L glass tank
- Aeration: yes
- Type of flow-through (e.g. peristaltic or proportional diluter): in-house design
- Renewal rate of test solution (frequency/flow rate): 0.04 mL/min for the stock solution and 800 mL/min for the dilution water, 1152 L/day of the test water were supplied to the test tank.
- No. of organisms per vessel: 26 fish (test item); 16 fish (control)
- No. of vessels per concentration (replicates): single
- No. of vessels per control / vehicle control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: groundwater sourced from laboratory premises
- Particulate matter: <1 mg/L
- Metals: all less than detection limits
- Pesticides: all less than detection limits
- Chlorine: <0.02 mg/L
- Alkalinity: 100 mg/L
- Ca/mg ratio: 15 mg/L
- Conductance: 280µS/cm

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 14h light / 10h dark
- Light intensity: - not stated - artificial light of white fluorescent lamp

RANGE-FINDING / PRELIMINARY STUDY
- Test concentrations: 1.0 mg/L acute toxicity fish test
- Results used to determine the conditions for the definitive study: no
- Other justification for choice of test concentrations: 1/10 of the water solubility of the test item in the test water.

Nominal and measured concentrations:

0.2µg/L (nominal)

Reference substance (positive control):

no

Details on estimation of bioconcentration:

BCF = Cr/Cw

BCF: Bioconcentration factor (L/kg)
Cr: Test item concentration in test fish (by subtracting FB) (ng/g)
Cw: Average test item concentration in test water during uptake phase (µg/L)
FB: Average concentration of fish blanks in the analysis of control (ng/g)

Lipid content:

5 %

Time point:

start of exposure

Lipid content:

4.25 %

Time point:

end of exposure

Conc. / dose:

0.2 µg/L

Type:

BCF

Value:

14 - 26 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks:

As all BCFs were less than 100 L/kg it was evaluated that steady state was reached after 28 days.

Rate constant:

growth rate constant (d-1)

Value:

0.012

Remarks on result:

other: Test Item

Rate constant:

growth rate constant (d-1)

Value:

0.014

Remarks on result:

other: Control

Details on results:

- Mortality of test organisms: none
- Behavioural abnormalities: none
- Observations on feeding behavior: not abnormal
- Observations on body length and weight: not abnormal
- Reproduction during test period: not applicable
- Organ specific bioaccumulation: no
- Mortality and/or behavioural abnormalities of control: None
- Loss of test substance during test period: Test substance remained within 20% of the nominal concentrations (analytically verified)

Reported statistics:

Not applicable

Table 1: Measured concentrations of the test item in the test water

Level	After 8 days	After 12 days	After 15 days	After 22 days	After 28 days	Average (Standard Deviation)
1	0.200	0.210	0.198	0.200	0.216	0.205 (0.0079)

Unit: µg/L

 

Table 2: Measured concentrations of the test item in test fish

Level	After 8 days	After 12 days	After 15 days	After 22 days	After 28 days
1	<2.9	0.210	0.198	0.200	0.216
	3.67	<2.9	<2.9	<2.9	<2.9

Unit: ng/g

 

Table 3: BCFs

Level	After 8 days	After 12 days	After 15 days	After 22 days	After 28 days
1	<14	21	18	26	17
	18	<14	<14	<14	<14

Unit: L/kg

Validity criteria fulfilled:

yes

Remarks:

Water temperature = 25 +/-2°C Dissolved oxygen was >60% of saturated concentration (8.1 mg/L) at 25°C The concentration of the test item in the test tank within 20% of the measured values No abnormalities/mortality observed with the test item or control

Conclusions:

The estimated BCF for the test item was <26 kg/L. The substance is not expected to bioaccumulate in the aquatic environment.

Description of key information

A study conducted in accordance with the national guidelines, Method for Testing the degree of Accumulation of Chemical Substances in Fish Body as stipulated in the "Testing Methods for New Chemical Substance" of Japanese Substances Control Law. The estimated BCF for the test item was <26 kg/L. The substance is not expected to bioaccumulate in the aquatic environment.

Key value for chemical safety assessment

BCF (aquatic species):

26 L/kg ww