Toluene-4-sulphonohydrazide

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

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 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

(April 13, 2004)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Batch/Lot No.: 202102001
Purity (wt.%) 99.5
Expiry date: February 28, 2022

Analytical monitoring:

yes

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
Daphnia magna

Gravid adults were isolated 24 hours prior to initiation of the test. Young daphnids produced overnight (< 24 hours old) were used for testing.

SOURCE: Ephippia was obtained from MicroBioTests Inc. Belgium (imported date: May 19, 2019). Daphnia has been cultured successively for a generation in invertebrate culture room,
Environmental Toxicology Research Center, Gyeongnam Department of Environmental
Toxicology and Chemistry, Korea Institute of Toxicology (KIT).

CULTURING: Brood stock of Daphnia magna were cultured under 16 h light: 8 h dark with 30 min transition period at 18-22 ºC in glass beakers containing three liters of M4 medium. Cultures were fed daily with a suspension of Pseudokirchneriella subcapitata and the amount (3 mL/L vessel/day) of the supplied algae was range of 6 x 106-107 cell/mL. 1 mL/L of YCT (Yeast, Chlorophyll and Tetramine, NeoEnbiz, Korea) was feed twice in a week. Culture conditions ensured that reproduction was by parthenogenesis.

Food type and feeding during test: M4 medium was used as culturing and dilution water for toxicity test, and was prepared according to the method of OECD Guideline (2004).

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

238-242 mg/L

Test temperature:

21.3-21.9ºC

pH:

7.82-7.98

Dissolved oxygen:

8.21-8.63 mg/L (94-97of Air Saturation Value)

Nominal and measured concentrations:

Nominal concentrations (mg/L): Control (0), 0.6, 1.3, 2.5, 5.0, 10
Time-weighted mean concentrations (mg/L): Control (0), 0.565, 1.12, 2.24, 4.33, 8.45

The test concentration was determined using HPLC. The time-weighted mean concentrations were calculated as a geometric mean of measurements of test concentration at 0, 24 and 48 hours.

Details on test conditions:

TEST SYSTEM
Definitive test was carried out under semi-static system with 24- hours renewal of test solution.
20 daphnids, preferably divided into four groups of five daphnids each, were used for each test
concentration and control groups (dilution water).
100 mL of test solution was provided for each test vessel.
Neither feed and nor aeration was provided during the test period.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- Total organic carbon: 0.7~0.8 mg/L
- Particulate matter: Not detected
- Pesticides: Not detected
- Chlorine: Not detected
- Intervals of water quality measurement: Water quality parameters of pH, dissolved oxygen and temperature were measured in each surviving test vessel at 0, 24 and 48 hours of the test.

OTHER TEST CONDITIONS
- Photoperiod: The photoperiod in this study 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.
- Light intensity: The light intensity ranged from 1139-1189 lux.

RANGE-FINDING STUDY
- Test concentrations: The range-finding test was conducted at the nominal concentrations of 0.1, 1, 10 and 100 mg/L by static system.
- 5 daphnids were exposed in each test vessel and immobilization was observed
during 48 hours.
- Results used to determine the conditions for the definitive study: No immobilization was observed in 0.1, 1 mg/L and 100% immobilization was observed in 10, 100 mg/L of test solution.

Reference substance (positive control):

yes

Remarks:

The 24hr-EC50 value (95% confidence limits) of Potassium dichromate (K₂Cr₂O₇) to Daphnia magna were determined 1.70 mg/L (1.57-1.84 mg/L).

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.12 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

2.71 mg/L

95% CI:

2.41 - 3.06

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks:

The animals were considered to be immobilized if they were unable to swim for approximately 15 seconds after gentle agitation.

Details on results:

- Behavioural abnormalities: No remarkable phenomenon was observed in all test solutions throughout test period
- Mortality of control: 0 animals
- Other adverse effects control: 0 observations
- Immobilisation of control: 0 animals
- Abnormal responses: 0 observations
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: 0 observations

VALIDITY OF THE TEST:
- Immobilization of daphnia in the control: 0% (criteria: < 10%)
- Dissolved oxygen concentration at the end of the test in control and test vessel: 8.21-8.63 mg/L (criteria: ≥ 3 mg/L)

Results with reference substance (positive control):

The 24hr-EC50 value (95% confidence limits) of Potassium dichromate (K₂Cr₂O₇) to Daphnia magna were determined 1.70 mg/L (1.57-1.84 mg/L).

Reported statistics and error estimates:

EC50 value with 95% confidence limits and NOEC (No observed effected concentration) were
analyzed by CETIS Version 1.8.7.15 (Tidepool Scientific Software, USA). Binomial/Graphical
method and Spearman-Karber method were applied to obtain the effective concentrations and
Steel Many-One Rank Sum test and Dunnett Multiple Comparison Test were used for NOEC
in this study. EC50 and NOEC are expressed based on mean measured concentration.

OBSERVATIONS:

Cumulative immobilization of Daphnia magna

Nominal concentration (mg/L)	Time weighted mean concentration (mg/L)	No. of animals tested	Cumuative no. of immobilized dahpnids 24-hours (Total)	Cumuative no. of immobilized dahpnids 24-hours (%)	Cumuative no. of immobilized dahpnids 48-hours  (Total)	Cumuative no. of immobilized dahpnids 48-hours (%)
Control	NA	20	0	0	0	0
0.6	0.565	20	0	0	0	0
1.3	1.12	20	0	0	0	0
2.5	2.24	20	0	0	4	20
5.0	4.33	20	0	0	20	100
10	8.45	20	20	100	20	100

 

Symptoms of intoxication of the test item to Daphnia magna:

Nominal concentration (mg/L)	Time-weighted mean concentration (mg/L)	Symptoms of intoxication 24-hours	Symptoms of intoxication 48-hours
Control	NA	Normal (20)	Normal (20)
0.6	0.565	Normal (20)	Normal (20)
1.3	1.12	Normal (20)	Normal (20)
2.5	2.24	Normal (20)	Normal (15) Lethargic (1)
5.0	4.33	Normal (19) Swimming Carrying (1)	-
10	8.45	-	-

RESULTS

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

Exposure time	24-hour	48-hour
EC50 values	6.05	2.71
95% confidence limits	4.33-8.45	2.41-3.06
NOEC	4.33	1.12

Validity criteria fulfilled:

yes

Conclusions:

To assess the acute toxicity of toluene-4-sulphonohydrazide a study according to OECD Guideline No. 202 was conducted. The 48hr-EC50 value (95% confidence limits) expressed in terms of the time-weighted mean concentration of toluene-4-sulphonohydrazide to Daphnia magna were determined to 2.71 mg/L (2.41-3.06 mg/L) and NOEC was 1.12 mg/L.
mg/L) and NOEC was 1.12 mg/L.

Executive summary:

To assess the acute toxicity of toluene-4-sulphonohydrazide a study according to OECD Guideline No. 202 was conducted.

Range-finding test was conducted at the nominal concentrations of 0.1, 1, 10 and 100 mg/L by static system. No immobilization was observed in 0.1, 1 mg/L and 100% immobilization was observed in 10 and100 mg/L of test solution. . Therefore, the definitive test was performed with a control and nominal concentrations of 0.6, 1.3, 2.5, 5.0, 10 mg/L using a semi-static system. The definitive test was performed for 48 hours and 20 daphnids were exposed for each concentration with four replicates.

The time-weighted mean measured concentrations were 0.565, 1.12, 2.24, 4.33 and 8.45 mg/L (94.2, 86.2, 89.2, 86.6, 84.5 % of nominal concentration) for nominal concentrations of 0.6, 1.3, 2.5, 5.0 and 10 mg/L during test period.

During 48 hours of study period, no immobilization and toxic symptoms were observed in control, 0.6 and 1.3 mg/L of test solutions (time-weighted mean concentration: 0.565, 1.12 mg/L, respectively) and 20% immobilization was observed in 2.5 mg/L (time-weighted mean concentration: 2.24 mg/L). 100% immobilization (including dead) was observed in 5.0 and 10 mg/L of test solutions (time-weighted mean concentration: 4.33, 8.45 mg/L).

The 48hr-EC50 value (95% confidence limits) expressed in terms of the time-weighted mean concentration of toluene-4-sulphonohydrazide to Daphnia magna were determined to 2.71 mg/L (2.41-3.06 mg/L) and NOEC was 1.12 mg/L.
mg/L) and NOEC was 1.12 mg/L.

Description of key information

To assess the acute toxicity of toluene-4-sulphonohydrazide a study according to OECD Guideline No. 202 was conducted. The 48hr-EC50 value (95% confidence limits) expressed in terms of the time-weighted mean concentration of toluene-4-sulphonohydrazide to Daphnia magna were determined to 2.71 mg/L (2.41-3.06 mg/L) and NOEC was 1.12 mg/L.
mg/L) and NOEC was 1.12 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

2.71 mg/L