4,4'-thiodiethylene hydrogen -2-octadecenylsuccinate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15th May 2013 - 30th September 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

Fish, Acute Toxicity Test stipulated in the "Testing Methods for New Chemical Substances" of
Japan (March 31, 2011; No.0331-7, Pharmaceutical and Food Safety Bureau, Minishy of Health,
Labour and Welfare; March 29, 2011, No.5, Manufacturing Industries Bureau, Ministry of
Economy, Trade and Industry; No. 110331009; April 2, 2012 partial revision, No. 0402-1,
Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; March 28, 2012,
No. 2, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No.
120402001, Environmental Policy Bureau, Ministry of the Environment, Japan)

GLP compliance:

yes

Specific details on test material used for the study:

Batch Ref: E00298-234-001

Physicochemical properties
Vapor pressure: 5.12E-39 mmHg (25°C)
Water solubility: Insoluble (visual)
Melting point: 32.5°C
Boiling point: 862°C (measurement pressure 760 mmHg)
Appearance: Brown gelled solid
Stability: Stable at ordinary temperature

Storage condition - The test sample was stored in a dark storage place at room temperature.

Identification and stability of test item under the storage condition - The infrared (IR) spectrum of the test item measured at this laboratory was confirmed to be identical to that provided by the sponsor. The stability of the test item was confumed by comparing the IR spectrum of the test item after the completion of the experiment in a storage condition with that before the start ofthe experiment.

Analytical monitoring:

yes

Details on sampling:

LC-MS/MS analysis (at the start and end of exposure)

Vehicle:

yes

Details on test solutions:

Required amount of the test sample was melted in a water bath of 40°C setup, was weighed and
added into the test vessel. The test sample added into the test vessel was melted again in a water
bath of 40°C setup and daubed onto the bottom of the test vessel with a magnetic stirrer. And then
dilution water was added to produce the 100 mg/L (nominal) of the test solution and stirred with a
magnetic stirrer for• 48 hours to prepare the test solution.

Test organisms (species):

Oryzias latipes

Details on test organisms:

Reason for selection of species: Species recommended in the test guideline
Supplier: CERI Kurume (in-laboratory production)
Size: Total length 2.3±1.2 cm
Allocation: Medaka was allocated at random to each test group.
Acclimation
Hatching date: March 25, 2013 (age at the start of exposure; five-month-old)
Water: Dechlorinated tap water
Acclimation type: Flow-through regime
Dissolved oxygen concentration: Not less than 80% of air saturation value
Temperature: 24±1°C
Photoperiod: 16-hour light/8-hour dark with morn light
Feed: Feed for fiy of carp crumble 2C (Nippon Formula Feed Mfg.)
Feeding amount and frequency: Amount corresponding to 2 % of body weight was fed everyday.
Use of medicament for external disinfection: None
Duration of acclimation: 75 days (June 26, 2013 - September 9, 2013)
Mortality of test groups: 0 % (during 7 days before the start of exposure)
Duration of no feeding: 24 hours before the start of exposure

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

30 mg/L

Test temperature:

25.5 degree C

pH:

7.8

Conductivity:

13 mS/m

Nominal and measured concentrations:

Nominal = 100 mg/L
Measured = 0.17mg/L (geometric mean)

Details on test conditions:

Type of test: Static regime (no renewal of test solution)
Exposure duration: 96 hours
Test concentration: A limit test around solubility in dilution water• (measured value in
preliminary study: 0.0830 mg/L)
The test concentration was decided from the result of preliminary study.
The results of the preliminary studies are shown in Additional data.
Control: Dilution water without the test item, which was treated in the same stirring manner as test solution
Replicate: 1 replicate/test level
Number of organisms: 7 fish/test level
Volume of test solution: Approximately 2.7 L/test level
Temperature of test solution: 24±1°C
Aeration: Conducted gently
pH adjustment: No adjustment
Irradiation condition: Room light, 16-hour light/8-hour dark
Feeding: No feeding

Reference substance (positive control):

no

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)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.17 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Details on results:

No mortality was observed in tested or control groups. No abnormal responses were observed

Reported statistics and error estimates:

The LC50 value was estimated as "> the test concentration" since the mortality over 50% was not observed at exposure level. The calculations of results were estimated based on a geometric mean of the measured concentrations of test item in test solution during exposure as the test concentration.

Sublethal observations / clinical signs:

see attached background document.

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour LC50 of the test item for Medaka was >100 mg/L (nominal).

Executive summary:

The objective of this study is to estimate the 96-hour median lethal concentration (LC50) by conducting an acute toxicity study of the Substance to Fish using the Medaka.

 

The Test Substance was melted in a water bath to produce a solution with a concentration of 100 mg/L (limit dose) and stirred for 48 hours. The 7 Fish were then exposed to the test solution in a static regime (no renewal) to the test solution for 96 hours.A control group using dilution water without the test item, treated in the same stirring manner was also conducted. Analysis of concentration of test item in test solution was measured at the start and end of the exposure by LC-MS/MS.

 

Mortality and visible abnormality were observed at 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.) were not observed and touching of the caudal peduncle with glass rod produced no reaction. 

 

No mortality was observed in tested or control groups. No abnormal responses were observed.

 

Based on the study, LC50 of the test item for Medaka was >100 mg/L (nominal) or > 0.17 mg/L (Measured – geometric mean).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13th October 1992 - 6th January 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Specific details on test material used for the study:

Description: Amber waxy solid
Storage Conditions: Room temperature in vented hood
Vehicle: None

Analytical monitoring:

yes

Details on sampling:

Samples were removed from each replicate chamber on Day 0 (—30 minutes after the addition of test material), Day 2 and Day 4 for Dissolved Organic Carbon (DOC) analysis.

Vehicle:

yes

Details on test solutions:

Treatments were prepared by adding the appropriate amount of test material to each replicate test chamber through the cylinder which was then chopped by the mixing apparatus before circulating through the dispersion system. Two replicate chambers were prepared per treatment and were covered with glass tank covers to prevent evaporation and/or volatilization. The study was performed in a temperature controlled water bath. The test material was continually dispersed throughout the study

Test organisms (species):

Cyprinodon variegatus

Details on test organisms:

Test System
cyprinodon variegatus - Lot 211

Method of Species Verification
Keyed using How to Know the Freshwater Fishes, Third Edition, Eddy, S. and J. C. Underhill, 1978., Wm. C. Brown Company Publishers

Justification for Selection of Test System
The sheepshead minnow, Cyprinodon variegatu.c historically has been used in safety evaluation and is a common test species for marine and estuarine toxicity studies.

Supplier
Aquatic Biosystems, Inc., Fort Collins, CO

Husbandiy and Acclimation
The sheepshead minnows were quarantined and observed for parasites and disease. Organisms were acclimated to test conditions for at least 12 days and held in dilution water for at least 7 days before the test. Fish were not treated for disease or parasites
before use in this study. Mortality occurring during acclimation period was insignificant. Fish were fed Tetramin® flake fish food.

Test type:

static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

0

Test temperature:

20.9 - 22.8 degree C

pH:

4.7-8.8

Dissolved oxygen:

7.5-8.2 mg/L

Salinity:

24 ppt

Nominal and measured concentrations:

Nominal = 0, 0.625, 1.25, 2.5, 5.0, 10 g/L
Measured DOC = 1.7063.918 mg/L

Details on test conditions:

Test Chamber / Volume
19L glass aquaria / IOL

Exposure Conditions
Mean test temperature: 22.0°C (sd = 1.3), continuously monitored.
Diurnal light: 16 hours light: 8 hours dark with a gradual intensity conversion between periods. Daylight intensity ranged from 66.83 to 69.58 footcandles during full daylight periods of the study.

Exposure Duration
96 hours

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 g/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No mortality occurred during the 96-hour test period at any concentration tested (10, 5, 2.5, 1.25 and 0.625 g/L)

Sublethal observations / clinical signs:

See attached background document

Validity criteria fulfilled:

yes

Conclusions:

No mortality occurred during the 96-hour test period; therefore the test material is not considered acutely toxic to sheepshead minnows for the specified period at the levels tested. LD50 is considered to be > 10 g/L

Executive summary:

A sheepshead minnow (Cyprinodon variegatus) test was performed to evaluate the acute toxicity of the Test Substance. The test was performed in a dispersion system. The dispersion system consists of 19.0L glass tanks containing 12cm long stainless steel cylinders (7.5cm diameter) in the center that house a stainless steel rod and propeller. The motorized stirrers were set at a speed in which an approximately one inch vortex was formed in each tank. The nominal treatment concentrations for the test were 10.0g/L, 5.0g/L, 2.5g/L, 1.25g/L, 0.625g/L and a laboratory dilution water control. Each treatment was prepared by adding the appropriate amount of test material to each replicate test tank. After allowing the test material to circulate through the dispersion system, samples were removed from each replicate for Dissolved Organic Carbon (DOC) analysis. The sheepshead minnows were exposed for a 96-hour period. No mortality occurred during the 96-hour test period; therefore the test material is not considered acutely toxic to sheepshead minnows for the specified period at the levels tested. LD50 is considered to be > 10 g/L

Description of key information

Two acute fish toxicity studies are available for the Substance. One study was conducted with a freshwater species, Medaka and the other with saltwater water species, Sheepshead Minnow. The acute fish toxicity study conclusions are summarised as follows:

Acute Fish 2013 (Medaka) - LL50 > 100 mg/L

Acute Fish 1993 (Sheepshead Minnow)- LL50 > 10000 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Marine water fish

Marine water fish

Effect concentration:

10 000 mg/L

Additional information

Acute Fish 2013 (Medaka):

The objective of this study is to estimate the 96-hour median lethal concentration (LC50) by conducting an acute toxicity study of the Substance to Fish using the Medaka.

 

The Test Substance was melted in a water bath to produce a solution with a concentration of 100 mg/L (limit dose) and stirred for 48 hours. The 7 Fish were then exposed to the test solution in a static regime (no renewal) to the test solution for 96 hours.A control group using dilution water without the test item, treated in the same stirring manner was also conducted. Analysis of concentration of test item in test solution was measured at the start and end of the exposure by LC-MS/MS.

 

Mortality and visible abnormality were observed at 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.) were not observed and touching of the caudal peduncle with glass rod produced no reaction. 

 

No mortality was observed in tested or control groups. No abnormal responses were observed.

 

Based on the study, LC50 of the test item for Medaka was >100 mg/L (nominal) or > 0.17 mg/L (Measured – geometric mean).

Acute Fish 1993 (Sheepshead Minnow):

A sheepshead minnow (Cyprinodon variegatus) test was performed to evaluate the acute toxicity of the Test Substance. The test was performed in a dispersion system. The dispersion system consists of 19.0L glass tanks containing 12cm long stainless steel cylinders (7.5cm diameter) in the center that house a stainless steel rod and propeller. The motorized stirrers were set at a speed in which an approximately one inch vortex was formed in each tank. The nominal treatment concentrations for the test were 10.0g/L, 5.0g/L, 2.5g/L, 1.25g/L, 0.625g/L and a laboratory dilution water control. Each treatment was prepared by adding the appropriate amount of test material to each replicate test tank. After allowing the test material to circulate through the dispersion system, samples were removed from each replicate for Dissolved Organic Carbon (DOC) analysis. The sheepshead minnows were exposed for a 96-hour period. No mortality occurred during the 96-hour test period; therefore the test material is not considered acutely toxic to sheepshead minnows for the specified period at the levels tested. LD50 is considered to be > 10 g/L