4,4'-oxydi(benzenesulphonohydrazide)

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016.08.25-2016.11.11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Guideline study (OECD 210) according to GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)

Version / remarks:

July 26, 2013

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

CAS No. 80-51-3
EC No. 201-286-1
KIT Code No.K-5450
Batch/Lot No. 201512001
Appearance: White fine powder
Purity: 99.5-100 %
Expiry date: December, 2016
Storage conditions: Room temperature/ Protect from moisture and light

Analytical monitoring:

yes

Details on sampling:

The concentrations of 4,4’-Oxybis(benzenesulfonyl hydrazide) (OBSH) in the test solution were verified using a HPLC method (see Analytical Report).
An analysis of the test solution was performed in the controls and each exposure concentration on before starting test, at the start, and on day 3, 10, 17, 24, 31, 38 and 45 (at the end) during the study.
A portion (5 mL) of the test solution was taken from each replicate (control and all test concentrations) on 1 day before starting the test, on the day of egg addition (Day 0), Day 3, 10, 17, 24, 31, 38 and 45. All samples were treated according to the recovery test before analysis.

Vehicle:

yes

Details on test solutions:

Dimethylforamide (DMF) was applied as a solvent

Test organisms (species):

Oryzias latipes

Details on test organisms:

Ricefish (Oryzias latipes)
Source: Fish culture room, Environmental Toxicology and Chemistry Research Center, Gyeongnam Department of Environmental Toxicology and Chemistry, Korea Institute of Toxicology (KIT)
Fertilized eggs, approximately 1-hour-old (1-2 cells stage) at the start of the study.

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

45 d

Remarks on exposure duration:

Experimental Starting Date August 25, 2016 Experimental Completion Date: November 11, 2016

Post exposure observation period:

not included

Hardness:

Day 0: Control mean value 33 mg/L (31-35)
Day 45: Control mean value 37.5 mg/L (37-38)

Test temperature:

ranging from 24.0-25.1 C

pH:

ranging from 7.25-7.88

Dissolved oxygen:

ranging from 7.51-8.80 mg/L (91-105% of Air Saturation Value),

Salinity:

NA

Conductivity:

NA

Nominal and measured concentrations:

Nominal: control, solvent control, 0.1; 0.31; 0.98; 3.13; 10 mg/L
meassured concentration: ND; ND; 0.09; 0.30; 1,01; 3.28; 10;22 mg/L

Details on test conditions:

The definitive test employed five test concentrations, together with a dilution water control and solvent control (100 µL DMF/L)
The concentrations were control, solvent control and nominal concentrations of 0.1, 0.31, 0.98, 3.13, 10 mg/L (factor: 3.2 from highest concentration) based on preliminary acute toxicity test as range-finding test.
The test vessel was Teflon aquaria, with approximate dimensions of 16.0 × 7.0 × 9.0 cm (length, width, height) fitted with a surface overflow (7.0 cm above the base), to hold an approximate volume of 800 mL of exposure medium. At the start of study, each test vessel contained one egg container (4 cm diameter), fitted with mesh ends to permit flow of the test solution in the vessel.
Twenty fertilized eggs were placed into each egg container, giving a total of 80 eggs per treatment group. Developing embryos were exposed under continuous flow conditions to the test or control media for a period of 45 days (approximately 15 days pre-hatch plus 30 days post-hatch). The egg containers were removed from the test vessels, when all of the eggs had hatched.

Reference substance (positive control):

not specified

Duration:

45 d

Dose descriptor:

EC50

Remarks:

hatching rate success

Effect conc.:

4.36 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

4.36 mg/L (1.44-25.9)

Duration:

45 d

Dose descriptor:

LOEC

Remarks:

Hatching rate succes

Effect conc.:

3.28 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Duration:

45 d

Dose descriptor:

NOEC

Remarks:

Hatching rate success

Effect conc.:

1.01 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Duration:

45 d

Dose descriptor:

LC50

Remarks:

post hatch survival

Effect conc.:

0.65 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

0.65 mg/L (0.45-0.89)

Basis for effect:

mortality

Duration:

45 d

Dose descriptor:

LOEC

Remarks:

Post hatch survival

Effect conc.:

0.3 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

45 d

Dose descriptor:

NOEC

Remarks:

Post hatch survival

Effect conc.:

0.09 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

45 d

Dose descriptor:

LC50

Remarks:

Overall survival

Effect conc.:

0.42 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

0.42 mg/L (0.28-0.57)

Basis for effect:

mortality

Duration:

45 d

Dose descriptor:

LOEC

Remarks:

Overall survival

Effect conc.:

0.3 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

45 d

Dose descriptor:

NOEC

Remarks:

Overall survival

Effect conc.:

0.09 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

The arithmetic mean was used to calculate overall concentration during test period because all measured concentration within 80-120% of nominal concentration. The arithmetic mean measured concentrations were 0.09, 0.30, 1.01, 3.28 and 10.22 mg/L for 0.1, 0.31, 0.98, 3.13, 10 mg/L of nominal concentration during test period.

Results with reference substance (positive control):

No data

Reported statistics and error estimates:

The data of hatching rate (hatching success) and normal survival (post hatching survival), normality (overall survival), length and weight at the end of the study were subjected to statistical analysis.
The toxicity value (LCX or ECX or ICX), LOEC and NOEC for each response was analyzed using CETIS Version 1.8.7. 15 (Tidepool Scientific Software, USA) by comparisons of each treatment group to the solvent control group.

In the present study, the solvent control was used for all subsequent comparisons because there were no significant differences between dilution water control and solvent control (by Equal Variance t Two-sample Test and Wilcoxon Rank Sum Two-Sample Test). A parametric test was used to determine if data were normally distributed and whether variances were homogeneous across treatments. If these assumptions were violated, non-parametric tests were used to determine the LOEC and NOEC.
The toxicity effects (ECX or LCX or ICX with 95% confidence limits) of each response of treatment groups compared to the solvent control group were analyzed by Linear regression and linear Interpolation method. The LOEC and NOEC were analyzed by Dunnett Multiple Comparison Test

Validity criteria fulfilled:

yes

Conclusions:

4,4’-Oxybis(benzenesulfonyl hydrazide) (OBSH) had effects on the hatching success depending on expose concentrations.

(95% confidence limit) LC/EC/IC50 LOEC NOEC
Hatching success rate 4.36 (1.44-25.9) 3.28 1.01
Post-hatching survival 0.65 (0.45-0.89) 0.30 0.09
Overall survival 0.42 (0.28-0.57) 0.30 0.09

Executive summary:

A 45 days laboratory study was conducted with the test substance 4,4’-Oxybis(benzenesulfonyl hydrazide) (OBSH), applying OECD guideline no 210 and following GLP. The results showed that the test substance had effects on the hatching successd epending on expose concentrations.

      

Hatching success (rate) ranged from 41.3 to 86.3% in the exposure groups compared to 90.0 and 92.5% for the control and solvent control groups. Up to 11 days,  more than 80% of embryos survived at any concentration employed in the test (0.1 to 10 mg/L), however from 12 days, number of dead embryos (absence of heart-beat or body movement) increased depending on concentrations. Finally, the survival rate of embryos was 90, 92.5, 86.3, 72.5, 63.8, 43.8 and 41.8% for control, solvent control, 0.09, 0.30, 1.01, 3.28 and 10.22 mg/L (mean measured concentrations), respectively.

 

 

	LC/EC/IC50 (95% confidence limit)	LOEC	NOEC
Hatching success rate	4.36 (1.44-25.9)	3.28	1.01
Post-hatching survival	0.65 (0.45-0.89)	0.30	0.09
Overall survival	0.42 (0.28-0.57)	0.30	0.09

Description of key information

A reliable long-term toxicity study (45 d) to fish has been carried out according to OECD 210, resulting in a NOEC (mortality) of 0.09 mg/L (arithmetic mean).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

NOEC

Remarks:

mortality

Effect concentration:

0.09 mg/L