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

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:

2012-03-14 to 2012-03-16

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:

(2004)

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 maximum 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:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: National Institute for Environmental Studies, Japan
- Age at study initiation: Juveniles less than 24-hour-old (female)
- Method of breeding: In house breeding
- Feeding during test: None
- Amount: 20 daphnids/test group (5 daphnids/vessel)

ACCLIMATION
- Acclimation period: Not applicable
- Acclimation conditions: same as test
- Type and amount of food: Chlorella vulgaris (unicellular green algae); 6 mg C (organic carbon) / 2 L/day
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): No

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

242 mg CaCO3/L

Test temperature:

20 ± 1 °C

pH:

pH 6.0 – 9.0

Dissolved oxygen:

>3 mg/L 60 % or more of saturation
min 8.4 - max 8.8 mg/L

Nominal and measured concentrations:

nominal concentrations: 10, 18, 32, 56, 100 mg/L
measured concentrations: 8.57, 15.5, 27.8, 48.4 and 87.1 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 100-mL glass beaker (Cover: Teflon® sheet)
- Type: Closed, semi-static
- Material, size, headspace, fill volume: Glass see above
- Renewal rate of test solution: 24 h
- No. of organisms per vessel: 5 daphnids/vessel ; 4 vessels/test group; 20 daphnids/test group
- No. of vessels per concentration: 4
- No. of vessels per control: 1
- No. of vessels per vehicle control: 1
- Biomass loading rate: Biomass loading: 1 daphnid or less/80 mL (25 daphnids or less/2 L)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: Fluorescent light 16-hour light / 8-hour dark
- Light intensity: 800 lux or less

EFFECT PARAMETERS MEASURED
Observation of the test animals: Immobility to swim within 15 second; abnormal behavior or appearance
Frequency: 24-hour (before renewal) and 48-hour exposure
Test solution in one vessel of each test group: measured items: Temperature, dissolved oxygen, pH, and appearance of the test solution
Frequency: Two sets

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study: Conc. 100 mg/L (10 daphnids/test group (5 daphnids/vessel, 2 replicates/test group))
Results of the range-finding test 1 (stirring for 1 minute) solvent control Tetrahydrofuran
Results of the range-finding test 2 (stirring for 48 hours) solvent dilution water filtrated through a filter (Qualitative filter paper No. 2, Advantec Toyo Kaisha, Ltd. (equivalent to JIS P3801 type 2, retention particle size: 5 μm)

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 87.1 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.:

> 87.1 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

15.5 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

15.5 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Measurements of Test Conditions and Observations of Test Solutions
Temperature: 20 ± 1 °C,
Dissolved oxygen: 3 mg/L or more
pH: 6.0 – 9.0
Appearance: test solutions of all the test groups- no suspended substances, no floating substances and no precipitations were observed, and the color was colourless.

Concentrations in Test Solutions
In Conc. 1 – 5, the mean measured concentrations (mean of sum of the test substance and the transformation products) were 8.57, 15.5, 27.8, 48.4 and 87.1 mg/L, respectively, which were higher than 80 % of the nominal concentrations.

Observation of Test Animals
In the control, the percentage of immobilisation was 0 % at the end of exposure, and no daphnids were observed floating on the surface of the water.
In Conc. 1 - 5, the percentage of immobilisation was 0, 0, 5, 5, and 40 %, respectively.
Immobilised daphnids were observed in the test solution prepared by stirring for 48 hours, while not observed in the test solution prepared by stirring for 1 minute. It was estimated that immobility observed in the definitive test was caused by water-soluble transformation products.

Validity criteria fulfilled:

yes

Conclusions:

The acute immobilisation of the test item Incozol 4 to the freshwater invertebrate Daphnia magna has been investigated. The 48-hour EC0 value was determined to be 15.5 mg/L. The EC50 value has been estimated to be >87.1 mg/L.

Executive summary:

The acute immobilisation test of the test item Incozol 4 to Daphnia magna was conducted, according to OECD guideline no. 202 and the Japanese notification,Yakushokuhatsu 0331 No.7, Heisei 23.03.29 Seikyoku No.5, Kanpokihatsu No.110331009, March 31, 2011. During a period of 48 hours the immobilisation of the daphnids in different substance concentrations (nominal 10, 18, 32, 56, 100 mg/L) were examined. In a preliminary range-finding test, a relationship between stirring time after adding the test substance to the dilution water and effect on daphnids was examined. As a result, immobilised daphnids were observed in the test solution prepared by stirring for 48 hours. It is assumed that immobility observed in the definitive test was caused by water-soluble transformation products. The highest concentration with 0 % immobility (EC0) was examined to be 15.5 mg/L (mean measured concentration). The lowest concentration with 100 % immobility (EC100) was not observed. Therefore the Median Effect Concentration (EC50) was estimated to be >87.1 mg/L.

Description of key information

The acute immobilisation of the test item Incozol 4 to the freshwater invertebrate Daphnia magna has been investigated. The 48-hour EC0 value was determined to be 15.5 mg/L. The EC50 value has been estimated to be >87.1 mg/L.

Additional information

The acute immobilisation test of the test item Incozol 4 to Daphnia magna was conducted, according to OECD guideline no. 202 and the Japanese notification,Yakushokuhatsu 0331 No.7, Heisei 23.03.29 Seikyoku No.5, Kanpokihatsu No.110331009, March 31, 2011. During a period of 48 hours the immobilisation of the daphnids in different substance concentrations (nominal 10, 18, 32, 56, 100 mg/L) were examined. In a preliminary range-finding test, a relationship between stirring time after adding the test substance to the dilution water and effect on daphnids was examined. As a result, immobilised daphnids were observed in the test solution prepared by stirring for 48 hours. It is assumed that immobility observed in the definitive test was caused by water-soluble transformation products. The highest concentration with 0 % immobility (EC0) was examined to be 15.5 mg/L (mean measured concentration). The lowest concentration with 100 % immobility (EC100) was not observed. Therefore the Median Effect Concentration (EC50) was estimated to be >87.1 mg/L.