Toluene-4-sulphonohydrazide

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:

2021

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:

also in accordance with Guidelines for Testing of Chemicals, Section 3.3 "Fish, Acute Toxicity Test", issued by the NIER (National Institute of Environmental Research), Korea.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Product Name: CELLCOM-H
Chemical Name: p-Toluenesulfonylhydrazide
CAS No.1576-35-8
EC No.216-407-3

Purity 99.5 wt.%
Batch/Lot No.202102001

Storage conditions: Room temperature, Protect from moisture.
Expiry date: February 28, 2022

- Solubility: This test item was soluble in dilution water and it is possible to prepare 100 mg/L of test solution by direct addition of test item (see details in the analytical report).

- Stability: Stability test was performed at 50 mg/L of test solution under test condition (without test organisms). The concentration decreased to 12.2% of initial concentration after 96hours. Therefore, semi-static system (24 hours renewal) was used in this study(see details in the analytical report).

OTHER SPECIFICS
- Other relevant information needed for characterising the tested material, e.g. if radiolabelled, adjustment of pH, osmolality and precipitate in the culture medium to which the test chemical is added:

Analytical monitoring:

yes

Remarks:

ANALYSIS OF THE TEST SOLUTION

Details on sampling:

An analysis was performed in groups of the control and 3.1, 6.3, 12.5, 25, 50 mg/L test solution at 0 hour (at the start), at 24hours (before/after renewal of test solutions), at 48 hours(before/after renewal of test solutions), at 72 hours (before/after renewal of test solutions) and at 96 hours (at the end) of the test.

Vehicle:

yes

Details on test solutions:

Dilution water source: Tap water was passed through a membrane filter (5, 1, 0.2 μm) to remove particulate matter and then a high-grade activated carbon filter to remove any organic contaminants.
The components of reconstituted water as dilution water for fish acclimation and toxicity test was as follows:
294.0 mg/L CaCl2·2H2O
123.3 mg/L MgSO4·7H2O
63.0 mg/L NaHCO3
5.5 mg/L KCl
Deionized water

A range-finding test was conducted at the nominal concentrations of 1, 10 and 100 mg/L. Based on the results of the range-finding test (0, 25 and 100% of mortality were observed in 1, 10 and 100 mg/L), the definitive test was performed using control and a nominal concentration of 3.1, 6.3, 12.5, 25, 50 mg/L by semi-static system.

503mg of test item (purity: 99.5%) was dissolved in 5000 mL of dilution water to prepare 100 mg/L of test solution. 50, 25, 12.5, 6.3 and 3.1 mg/L of test solutions were prepared by serial dilution. Only the dilution water was used for control.

Approximately 4000 mL of each test solution was transferred to the 7 L volume of glass tank to use for toxicity test. All test solution was clear and no remarkable phenomenon was observed.

Definitive test was carried out under semi-static systemwith 24 hours renewal of test solution.

Test organisms (species):

Oryzias latipes

Details on test organisms:

SOURCE: Fish culture room, Ecotoxicology research group, Gyeongnam Branch Institute, Korea Institute of Toxicology (KIT)

CULTERING METHOD: Fish were cultured under 16 h light: 8 h dark with 30-minutedawn and dusk transition periods at 23-27°C (water temperature) in the fish culture room of the Ecotoxicology research group, Gyeongnam Branch Institute, Korea Institute of Toxicology (KIT).Fish were fed daily with brine shrimp nauplii (MyungSun Co., Ltd., Korea) in the morning and Tetramin flake (Tetra®, Germany) in the afternoon.

ACCLIMATION:
The stock of fish (58 fish) was acclimated to test conditions 7 days before the limit test and feeding was not provided within 24hours of test initiation. The water temperature was 24.1-24.3°C and DO was 8.14-8.32mg/L. No mortality was observed and all fish were healthy during acclimation period.

Fish biomass loading was no greater than 0.8g fresh weight per liter of test solution for this test.
Size: 2.0±1.0 cm
Control fish were measured at the end of the exposure period. The mean total length and body weight were 1.5±0.1cm (mean±SD) and 0.027±0.005g (mean±SD), respectively.

Each glass tank with 4000mLof test solution and 7 fish per each concentration were used for definitive test. Mortality and toxicity behavior were observed for 96 hour.

Neither feed and nor aeration was provided during the test period.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

294.0 mg/L CaCl2·2H2O

Test temperature:

24.0-24.8ºC

pH:

7.67-7.97

Dissolved oxygen:

7.43-8.33mg/L

Nominal and measured concentrations:

Nominal concentrations (mg/L): Control (0), 3.1, 6.3, 12.5, 25, 50
Mean measured concentrations (mg/L): Control (0), 2.29, 4.50, 9.67, 19.0, 37.9

Details on test conditions:

ENVIRONMENTAL CONDITION:
Water temperature was 24.0-24.8ºC (Table 1), dissolved oxygen was 7.43-8.33mg/L (88-101% of Air Saturation Value, Table 2) and pH was 7.67-7.97(Table 3) during the test. The light intensity ranged from 826to 832lux.

The photoperiod was 16 hours of light and 8 hours of dark, including 30-minute dawn and dusk transition periods. Light intensity was measured at the start and the end of the test. Neither feed and nor aeration was provided during the test period.

The fish were inspected at 2, 5 and 24 hours after exposure and then all vessels with living fish were inspected twice per day (morning/afternoon) until exposure termination. Mortalities were recorded and any abnormalities in appearance or behavior shown by surviving fish were summarized daily. The criteria of death were absence of respiratory movement and lack of responses to physical stimulation. Dead fish were removed when observed.

EXPOSURE DESIGN:
Based on the results of the range-finding test(0, 25 and 100% of mortality were observed in 1, 10 and 100 mg/L), a definitive test was performed using control and a nominal concentration of 3.1, 6.3, 12.5, 25, 50 mg/L by semi-static system. Each glass tank with 4000mLof test solution and 7 fish per each concentration were used for definitive test. Mortality and toxicity behavior were observed for 96 hours.
Neither feed and nor aeration was provided during the test period

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: See details on test solutions
- Total organic carbon: 0.5~1.8
- Particulate matter: Not detected
- Metals: No information
- Pesticides: Not detected
- Chlorine: Not detected
- Alkalinity: No information
- Ca/mg ratio: No information
- Intervals of water quality measurement: Water quality parameters of pH, dissolved oxygen and temperature were measured in each test vessel at 0, 24, 48, 72, and 96 hours of the test.

Reference substance (positive control):

yes

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.21 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Results with reference substance (positive control):

The 96hr-LC50 value (95% confidence limits) of Potassium dichromate (K₂Cr₂O₇) to Oryzias latipes were determined 185mg/L (153-224mg/L).

Reported statistics and error estimates:

The 24, 48, 72, and 96-hour LC50values and associated 95% confidence limits was calculated using CETIS software (Version 1.8.7.15 (Tidepool Scientific Software, USA). Binomial/Graphical method and Spearman-Karber method were applied to obtain the lethal concentrations.

Validity criteria fulfilled:

yes

Conclusions:

Short-term toxicity to fish, conducted according to OECD Guideline no. 203 and GLP, with test substance toluene-4-sulphonohydrazide and test organism ricefish (Oryzias latipes), showed a 96hr-LC50 (95% confidence limits) value expressed in terms of the time-weighted mean concentration of 3.21mg/L (2.29-4.50mg/L).

Executive summary:

The study was conducted to assess the acute toxicity effect of toluene-4-sulphonohydrazide to ricefish, Oryzias latipes under semi-static test conditions for a period of 96 hours.

The study was conducted in accordance with the OECD Guideline No. 203 “Fish., Acute Toxicity Test” (June18, 2019)

Range-finding test was conducted at the nominal concentrations of 1, 10 and 100 mg/L by static system. 0, 25 and 100% of mortality were observed in 1, 10 and 100 mg/L of test solution, respectably.

Definitive test was performed with a control and a nominal concentration of 3.1, 6.3, 12.5, 25 and 50 mg/L for 96 hours and 7 fish were exposed for each concentration without replicate.

During 96 hour of study period, nomortality and no toxic symptoms were observed in control groupsand 3.1 mg/L of test solution (mean measured concentration: 2.29 mg/L). However, mortality showed a dose response relationship and all fish were dead in 6.3, 12.5, 25 and 50 mg/L of test solutions (mean measured concentration: 4.50, 9.67, 19.0, 37.9mg/L).

The toxicity are expressed as the median lethal concentration (LC50)

(Unit: mg/L, time-weighted mean concentration)

Exposure time	48-hour	96-hour
LC50 values	8.98	3.21
95% confidence limit	6.86-11.8	2.29-4.50

Description of key information

One short-term (96 hrs) toxicity study with fish have been carried out (OECD 203), showing a LC50 3.2 mg/L. 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

3.2 mg/L

Additional information

A short-term (96 hrs) toxicity study with fish have been carried out (OECD 203), showing a LC50 of 3.2 mg/L