3-[(phenylcarbamoyl)amino]phenyl 4-methylbenzenesulfonate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 November 2017 - 1 December 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

17 July 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Storage conditions: The test material was stored in a dark storage place at room temperature.
- Identification and stability under storage conditions: The IR spectrum of the test material measured at the test facility was confirmed to be the identical to that provided by the sponsor. Additionally, the IR spectrum of the test material after the completion of the experiment was the same as that before the start of the experiment, therefore indicating that the test material was stable under the storage conditions.

Analytical monitoring:

yes

Remarks:

HPLC analysis (at the start and end of exposure)

Details on sampling:

- The test solution for analysis (ca 200 mL) was taken out from the middle layer of the test vessel.
- Concentrations: The determination limit of the test material was the lowest concentration of the standard solution (0.0500 mg/L) within the range of the calibration confirmed. Therefore, the determination limit of the test material in the test solution was 0.100 mg/L in consideration of pretreatment.
- Sampling method: HPLC

Vehicle:

yes

Remarks:

Dilution water

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- The test material (0.280 g) and dilution water (2.8 L) were mixed to produce the nominal concentration of 100 mg/L and they were stirred with a magnetic stirrer for 48 hours to prepare the test solution.

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka
- Age at study initiation: two months old
- Length: total length: 2.3 ± 1.2 cm

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions: Flow-through regime
- Feeding frequency during acclimation: 3 % of body weight was fed every day. Feed withdrawal 24 hours before the start of exposure.
- Health during acclimation (any mortality observed): 0 % mortality

FEEDING DURING TEST
- No feeding during test.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

32 mg/L

Test temperature:

23.6 - 24.2 °C

pH:

7.8 - 8.0

Dissolved oxygen:

At least 80 % of air saturation value. 8.5 - 8.7 mg/L

Conductivity:

13 mS/m

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured: All of the measured concentrations of the test material in the test solution were below the determination limit (< 0.100 mg/L).

Details on test conditions:

TEST SYSTEM
- Test vessel: 3L glass tank (diameter: 16 cm, depth 16 cm)
- Fill volume: 2.6 L
- Type: closed (transparent plastic lid)
- Aeration: Conducted gently
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated tap water
- Total organic carbon: < 0.5 mg/L
- Suspended solid: < 1 mg/L
- Chloride ion: 9.3 mg/L
- Alkalinity: 36 mg/L
- Intervals of water quality measurement: Measured regularly.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: Room light, 16-hour light/8-hour dark

EFFECT PARAMETERS MEASURED
- Mortality and visible abnormality: 3, 24, 48, 72 and 96 hours after the start of exposure. A fish was considered as dead if the observable motion (motion of mouth and opercula etc.) was not observed and touching of the caudal peduncle with a glass rod produced no reaction. In addition, surviving organisms at the end of exposure were anaesthetised by cooling with ice water and were euthanised by freezing in a freezer.
- Total length and body weight of test organism: The test organisms anaesthetised by cooling with ice water in the control group were used for measuring total length and body weight after the end of exposure.
- Appearance of test solution: Observed at the start and end of exposure.
- Dissolved oxygen concentration, pH and temperature: Measured at the start of exposure and every 24 hours.
- Concentration of test material in test solution: Measured at the start and end of exposure.

TEST CONCENTRATIONS
- Range finding study: The solubility of the test material in dilution water was tested using 100 mg/L as the nominal concentration. The test material and dilution water were mixed and stirred. After settlement for 1 hour, the middle layer was taken out and filtered with a membrane filter (GV, 0.22 μm pore size, Merck Millipore) by suction. The filter was treated with the test solution of about 100 mL beforehand. The solubility of the test material in dilution water was below the determination limit (< 0.100 mg/L).
- Test concentrations: Therefore, a limit test around the solubility in dilution water was conducted with a nominal test material concentration of 100 mg/L.

CALCULATION METHOD OF LC50
- The LC50 was estimated as "> solubility in dilution water", since the mortality over 50 % was not observed in the exposure level and the concentrations of the test material in the test solutions were below the determination limit. In addition "LL50 (Median Lethal Loading) > 100 mg/L (nominal loading concentration)" was also described as a reference.

VALIDITY OF TEST
- The test was considered valid if the following conditions were met:
1) the mortality in the control should not exceed one fish.
2) Dissolved oxygen concentration must be at least 60 % of the air saturation value at the water temperature throughout exposure duration.

Reference substance (positive control):

yes

Remarks:

Copper(II) sulphate pentahydrate

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: The LC50 was determined to be above the solubility of the test material in dilution water, which was below the determination limit of 0.100 mg/L.

Details on results:

OBSERATION AND MEASUREMENT OF TEST SOLUTION
- Appearance: The test solution in the exposure level at the start of exposure was suspended to white, and the floating substance and precipitates were also observed. The test solution in the exposure level at the end of exposure was clear and colourless, and the floating substance and precipitates were also observed.
- Condition of test solutions: The measured value of of dissolved oxygen concentration met the criterion for the validity of the test (at least 60 % of saturate concentration at the water temperature, saturated dissolved oxygen concentration (23 - 25 °C): 8.58 - 8.36 mg/L).
- Concentration of test material in test solution: All of the measured concentrations of the test material were below the determination limit (< 0.100 mg/L).

Results with reference substance (positive control):

The positive control study was conducted from 13 to 17 November 2017. The 96-hour LC50 was determined to be 0.33 mg/L (converted to copper(II) sulphate). This value was within the stipulated range (mean ± 2 S.D.: 0.14 - 0.90 mg/L, n=61) to background data at the Test Facility.

Reported statistics and error estimates:

LC50
- The 96 hour LC50 of the test material for Medaka was > the solubility of the test material in dilution water. (Reference value: The 96-hour LL50: > 100 mg/L (Nominal loading concentration)).

Sublethal observations / clinical signs:

MORTALITY
- Dead fish in the exposure level during exposure was not confirmed. Number of dead fish in the control at the end of the exposure was 0, which met the criterion for the validity of the test (i.e. not exceed one fish).

OBSERVED PERFORMANCE STATUS ETC.
- No abnormal response was observed in the control. No abnormal response was observed in the exposure level during exposure, based on the comparison with control organisms.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this study the 96-hour LC50 of the test material to Medaka was greather than the solubility of the test material in dilution water, which was below the determination limit of 0.100 mg/L. Therefore the 96-hour LC50 was determined to be > 0.100 mg/L. In terms of nominal test material concentration, the LC50 was > 100 mg/L.

Executive summary:

The acute toxicity of the test material on Medaka was investigated in accordance with the standardised guidelines OECD 203, under GLP conditions. 

This study was conducted as a limit test in order to confirm the effect of the test material on the test organisms at around the solubility of the test material in dilution water. As a result, no adverse effect on the test organisms was found in the definitive study. Since the concentration of the test material in the test solution during exposure was below the determination limit (< 0.100 mg/L), the dissolution concentration was not clear. However, no effects on the test organisms were observed, indicating that the test material exerted no acute effect on the test organisms at around the solubility in dilution water. The environmental conditions were within the suitable range. Therefore, it was concluded that this study complied with the applied test guidelines. 

Under the conditions of this study the 96-hour LC50 of the test material to Medaka was greater than the solubility of the test material in dilution water, which was below the determination limit of 0.100 mg/L. Therefore the 96-hour LC50 was determined to be > 0.100 mg/L. In terms of nominal test material concentration, the LC50 was > 100 mg/L.

Description of key information

Matsuura (2018)

The 96-hour LC50 of the test material to Medaka was greater than the solubility of the test material in dilution water, which was below the determination limit of 0.100 mg/L. Therefore the 96-hour LC50 was determined to be > 0.100 mg/L. In terms of nominal test material concentration, the LC50 was > 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 0.1 mg/L

Additional information

Matsuura (2018)

The acute toxicity of the test material on Medaka was investigated in accordance with the standardised guidelines OECD 203, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

This study was conducted as a limit test in order to confirm the effect of the test material on the test organisms at around the solubility of the test material in dilution water. As a result, no adverse effect on the test organisms was found in the definitive study. Since the concentration of the test material in the test solution during exposure was below the determination limit (< 0.100 mg/L), the dissolution concentration was not clear. However, no effects on the test organisms were observed, indicating that the test material exerted no acute effect on the test organisms at around the solubility in dilution water. The environmental conditions were within the suitable range. Therefore, it was concluded that this study complied with the applied test guidelines. 

Under the conditions of this study the 96-hour LC50 of the test material to Medaka was greather than the solubility of the test material in dilution water, which was below the determination limit of 0.100 mg/L. Therefore the 96-hour LC50 was determined to be > 0.100 mg/L. In terms of nominal test material concentration, the LC50 was > 100 mg/L.