Bis[2-[2-(1-methylethyl)-3-oxazolidinyl]ethyl] hexan-1,2-diylbiscarbamate

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:

2012-03-12 to 2012-03-16

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:

(1992)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: "Alga growth inhibition test, Daphnia sp. acute immobilisation test, and fish acute toxicity test” Japanese notification, Yakushokuhatsu 0331 No.7, Heisei 23.03.29 Seikyoku No.5, Kanpokihatsu No.110331009, March 31, 2011

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Based on the results of the range-finding test, the concentration for the definitive test was set 100 mg/L.
- Sampling method: Preparation method of test solution: The test substance was added to the dilution water and stirred with a magnetic stirrer for 48 hours, and then filtrated through a filter.
- Sample storage conditions before analysis: During the study, the test substance was stored in a desiccator in the laboratory (storage condition: room temperature, dark, nitrogen-sealed).
The infrared absorption spectrum of the test substance was measured after the end of exposure. The spectrum obtained was consistent with that measured before the start of exposure, indicating that the test substance was stable under the storage conditions.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Preparation of stock solution of the test substance - Inversion; Loading of the test substance - stirring with a magnetic stirrer for 48 hours (room temperature, approx. 24 °C)
- Eluate: Preparation of stock solution of the test substance: Acetone; Loading of the test substance: Acetone is dried up under nitrogen flow and Dechlorinated water is added to final volume.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): 1000 mg test substance was solved in 20 mL Acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): Stock solution: 50000 mg/L; final test solution: 100 mg/L
- Evidence of undissolved material: no

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka
- Source: Tsunashima Fishing Co. (18-1 Tsunashima-nishi 5-chome, Kouhoku-ku, Yokohama, Kanagawa, Japan)
- Age at study initiation: A year or less after hatching
- Length at study initiation: 2.39 cm (1.99 – 2.70 cm)
- Weight at study initiation: 0.135 g (0.089 – 0.182 g)
- Feeding during test: none
- Food type: TetraMin®, TetraWerke
- Feeding amount: 2 % of the body weight/day (approx.) (until 24 hours before exposure)

ACCLIMATION
- Acclimation period: March 5, 2012 – March 12, 2012
- Acclimation conditions: same as test
Water: Dechlorinated water
Procedure: Flow-through (Biomass loading: 1 g fish/L/day or less)
Temperature: 24 ± 1 °C
Dissolved oxygen concentration (D.O.): 80 % or more of the saturation
pH: 6.0 – 8.5
Light: Fluorescent light, 16-hour light (1000 lux or less) / 8-hour dark
- Type and amount of food: see above
- Health during acclimation (any mortality observed): Mortality during 7 days before the test less than 5 %

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

The total water hardness was 53 mg CaCO3/L.

Test temperature:

min. 23.8 °C - max. 24.1 °C

pH:

pH min. 7.2 - max. 8.6

Dissolved oxygen:

min. 5.8 mg/L - max. 7.6 mg/L

Nominal and measured concentrations:

Nominal concentration: 100 mg/L
Mean value of 4 measured samples: 101 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 5-L glass vessel (cover: clear polyvinyl chloride (PVC) plate)
- Type: closed, semi-static
- Material, size, headspace, fill volume: PVC, 5.0 L/vessel
- Aeration: min. 5.8 mg/L max. 7.6 mg/L
- Renewal rate of test solution: every 24 h
- No. of organisms per vessel: 1 vessel/test group; 10 fish/test group
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- No. of vessels per vehicle control: 1
- Biomass loading rate: 1.0 g fish/L or less

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated water processed by treating Yokohama city tap water with an activated charcoal filter and sodium thiosulfate was used as the dilution water
- Total organic carbon: Analytical method: Total organic carbon (TOC) analyzer
- Intervals of water quality measurement: Measurements of the temperature, dissolved oxygen, pH and observation of the appearance of the test solutions were performed once a day. Renewal of the test solutions was performed by transferring the fish to freshly prepared test solution.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: Fluorescent light, 16-hour light / 8-hour dark
- Light intensity: 1000 lux or less

EFFECT PARAMETERS MEASURED
Test fishes were observed at 3, 24, 48, 72 and 96 hours after the start of exposure.
LC50: concentration at which 50 % of the fish die during the exposure period

TEST CONCENTRATIONS
- Range finding study: range-finding test 1: Stirring for 1 minute: 5 fish/test group, test water: 2.5 L; Solvent: Tetrahydrofuran, 100 mg/L (constant volume) range-finding test 2: Stirring for 48 hours: substance was added to the dilution water and stirred with a magnetic stirrer for 48 hours and then filtrated through a filter.
- Test concentrations: 100 mg/L
- Results used to determine the conditions for the definitive study: Based on the results of the range-finding test, the concentration for the definitive test was set as 100 mg/L in consideration of effect of the test substance on other test species (algae and daphnids).

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 101 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

QUALITY AND APPEARANCE OF TEST SOLUTIONS
- Temperatures: from 23.8 to 24.1 °C
- Dossolved oxygen: from 5.8 to 7.6 mg/L
- pH: from 7.2 to 8.6
- Appearance: In both the control and the test solutions appeared color-less, and any suspended substances, floating substances and precipitated substances were not observed.

CONCENTRATIONS IN TEST SOLUTIONS
TOC mearsured: control - (0 hour) and before renewal at 24 hours were 37.5 and 32.3 mg/L (dilution water quality (0.4 mg/L)) maybe acetone was not completely dried ; period 24h - 96h measured conc. 0.822 – 2.69 mg/L
TOC (percent of nominal): in Conc.1 (100 mg/L nominal) ranged from 87.2 to 116 %

OBSERVATIONS OF TEST SPECIES
In both the control and Conc. 1, no dead fish and no abnormal symptoms in living fish were observed. Although the high concentrations of TOC were observed in the control during 0 to 24 hours, it was judged that the fish tested was not affected by these high TOC concentrations.

Validity criteria fulfilled:

yes

Conclusions:

The LC50 of the test item was determined to be >101 mg/L.

Executive summary:

The study assessing the acute toxicity of the test item Incozol 4 to fish was conducted with Medaka (Oryzias latipes) according to OECD guideline no. 203. Based on a range-finding test a limit dose of 100 mg/L was chosen for the main test. The control and test group were observed at 0, 24, 48, 72, 96 hours. In one 5 L vessel were 10 fishes/test group. To maintain the concentration of the test substance, test solutions were renewed every 24-hour (semi-static).

In accordance with the guideline the temperatures ranged from 23.8 to 24.1 °C, the dissolved oxygen ranged from 5.8 to 7.6 mg/L, and pH ranged from 7.2 to 8.6. In both the control and test group, the test solutions appeared colourless, and any suspended substances, floating substances and precipitated substances were not observed in the test solutions during exposure. The mean measured concentration (mean of sum of the test substance and the transformation products) of the test group was 101 mg/L, being almost equivalent to the nominal concentration of 100 mg/L. In both the control and the test group, no dead fish and no abnormal symptoms in living fish were observed. Therefore, the LC50 of the test item Incozol 4 is >101 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1988-09-05 to 1988-09-09

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: Letale Wirkung beim Zebrabaerbling - Brachydanio rerio -LC 0, LC 50, LC 100; 48 - 96 Stunden Umweltbundesamt Berlin Stand: Mai 1984 SOP-Nr.4.002

Deviations:

not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not specified

GLP compliance:

not specified

Analytical monitoring:

no

Details on sampling:

No analytical monitoring

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test item has been diluted in purified water and mixed at 8000 rpm in an Ultra-Turrax.
- Eluate: Purified Leverkusener water
- Controls: Purified Leverkusener water
- Evidence of undissolved material: The test substance sedimented at the ground and wall of the test vessel respectively was swimming at the surface. Undissolved precipitates were decanted at the Ultra-Turrax-valve.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Brachydanio rerio
- Strain: Hamilton - Buchanan
- Source: Westaquarium / Bad Lauterberg
- Age at study initiation: 4 months
- Length: After death the end of the study 2.5 - 3.5 cm
- Weight: After death the end of the study 2.382 - 3.190 g

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

13.4° dH

Test temperature:

20.6 °C

pH:

6.5

Dissolved oxygen:

Measured oxygen concentrations 0 - 96 h
Control: 8.4 - 9.2 mg/L
Main Test: 7.6 - 9.4 mg/L

Nominal and measured concentrations:

Nominal concentrations: 50, 70.7, 100, 141, 200, 283, 400, 566 mg/L
No measured values.

Details on test conditions:

TEST SYSTEM
- Test vessel: Test aquarium 300x135x200 mm
- Type: open
- Material fill volume: 5 L test medium
- Aeration: Yes
- No. of organisms per vessel: 10
- No. of vessels per concentration: 1
- No. of vessels per control: 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Purified water
- Ca/mg ratio: 63.4 mg/L :11.7 mg/L
- Culture medium different from test medium: No

EFFECT PARAMETERS MEASURED
- Every 24 hrs : Oxygen (mg/L), pH , temperature (°C), mortality

TEST CONCENTRATIONS
- Range finding study: Pre test LC0: 70.7 mg/L LC100: 200 mg/L
- Test concentrations: 50, 70.7, 100, 141, 200, 283, 400, 566 mg/L (nominal concentration)

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

70.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

199.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

566 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Mortality after 0, 24, 48, 72, 96 h (10 fishes/dosis)

Control: 0
50 mg/L: 0
70.7 mg/L: 0
100 mg/L: 96h 1 fish
141 mg/L: 24h 4 fishes
200 mg/L: 24h 3 fishes
283 mg/L: 24h 8 fishes
400 mg/L: 24h 9 fishes
566 mg/L: 24h 10 fishes

- Behavioural abnormalities: Swimming behavior
- Observations on body length and weight : Yes

Observations after 2, 24, 48, 72, 96 hours of 10 animals/dosis
Swimming bavior, body length (range), weight (average):

Control: Normal swimming behavior, 2.7 - 3.5 cm, 2.752 g
50 mg/L: Normal swimming behavior, 2.5 - 3.5 cm, 2.636 g
70.7 mg/L: Normal swimming behavior, 2,7 - 3.5 cm, 2.465 g
100 mg/L: Normal swimming behavior, 2.5 - 3.2 cm, 2.443 g
141 mg/L: Normal swimming behavior, 2.7 - 3.5 cm, 2.967 g
200 mg/L: Normal swimming behavior, 2.5 - 3.5 cm, 2.382 g
283 mg/L: After 2h 9 fishess inert, lethargic swimming behavior, 2.8 - 3.5 cm, 2.770 g
400 mg/L: After 2h 7 fishes inert, lethargic swimming behavior, 1 fish motionless by contact uncontrolled swimming movement 2.8 - 3.5 cm, 3.190 g
566 mg/L: After 2h 3 fishes normal, 1 fish inert, lethargic swimming behavior, 2 fishes motionless by contact uncontrolled swimming movement after 24h all fishes died, 2.8 - 3.5 cm, 3.036 g

Validity criteria fulfilled:

yes

Conclusions:

The LC50 value was determined to be 199.2 mg/L.

Executive summary:

The acute fish toxicity test with the test item Incozol 4 was conducted with Brachydanio rerio according to "Letale Wirkung beim Zebrabaerbling - Brachydanio rerio -LC 0, LC 50, LC 100; 48 - 96 Stunden Umweltbundesamt Berlin Stand: Mai 1984" that is equivalent or similar to OECD 203. The fish were exposed for 96 h to the test item. Based on a pretest (LC0: 70.7 mg/L and LC100: 200 mg/L) the concentrations for the main test were set as followed: 50, 70.7, 100, 141, 200, 283, 400, 566 mg/L. During the exposure time the swimming behavior and the mortality were observed. At the concentration of 283 mg/L in 9 fishes inert, lethargic swimming behavior has been observed and the first mortality was observed at the concentration of 141 mg/L after 24 hours. All animals died at the concentration of 566 mg/L. Therefore the LC values were determined as followed:

LC0: 70.7 mg/L; LC100: 566 mg/L; LC50: 199.2 mg/L. The test item was considered to be toxic to Brachydanio rerio at a LC50 value of 199.2 mg/L.

Description of key information

In a study assessing the acute toxicity of the test item to fish the LC50 was determined to be >101 mg/L.

Additional information

Key Study

The study assessing the acute toxicity of the test item Incozol 4 to fish was conducted with Medaka (Oryzias latipes) according to OECD guideline no. 203. Based on a range-finding test a limit dose of 100 mg/L was chosen for the main test. The control and test group were observed at 0, 24, 48, 72, 96 hours. In one 5 L vessel were 10 fishes/test group. To maintain the concentration of the test substance, test solutions were renewed every 24-hour (semi-static).

In accordance with the guideline the temperatures ranged from 23.8 to 24.1 °C, the dissolved oxygen ranged from 5.8 to 7.6 mg/L, and pH ranged from 7.2 to 8.6. In both the control and test group, the test solutions appeared colourless, and any suspended substances, floating substances and precipitated substances were not observed in the test solutions during exposure. The mean measured concentration (mean of sum of the test substance and the transformation products) of the test group was 101 mg/L, being almost equivalent to the nominal concentration of 100 mg/L. In both the control and the test group, no dead fish and no abnormal symptoms in living fish were observed. Therefore, the LC50 of the test item Incozol 4 is >101 mg/L.

Supporting Study

The acute fish toxicity test with the test item Incozol 4 was conducted with Brachydanio rerio according to "Letale Wirkung beim Zebrabaerbling - Brachydanio rerio -LC 0, LC 50, LC 100; 48 - 96 Stunden Umweltbundesamt Berlin Stand: Mai 1984" that is equivalent or similar to OECD 203. The fish were exposed for 96 h to the test item. Based on a pretest (LC0: 70.7 mg/L and LC100: 200 mg/L) the concentrations for the main test were set as followed: 50, 70.7, 100, 141, 200, 283, 400, 566 mg/L. During the exposure time the swimming behavior and the mortality were observed. At the concentration of 283 mg/L in 9 fishes inert, lethargic swimming behavior has been observed and the first mortality was observed at the concentration of 141 mg/L after 24 hours. All animals died at the concentration of 566 mg/L. Therefore the LC values were determined as followed:

LC0: 70.7 mg/L; LC100: 566 mg/L; LC50: 199.2 mg/L. The test item was considered to be toxic to Brachydanio rerio at a LC50 value of 199.2 mg/L.