2,3-dihydroxypropyl methacrylate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental start date 18 September 2017. Experimental completion date 04 November 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

please see any other information on materials and methods section

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

please see any other information on materials and methods section

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Range-finding Test
A sample of each test concentration was taken for chemical analysis at 0 and 24 hours in order to determine the stability of the test item under test conditions. All samples were stored frozen prior to analysis. Only concentrations within the range to be used for the definitive test were analyzed.

Definitive Test
Water samples were taken from the control and 100 mg/L test solution at 0, 24, 48 and 72 hours from fresh media and at 24, 48, 72 and 96 hours from old media for quantitative analysis. The 0, 24, 48 and 72-Hour samples were stored frozen prior to analysis, whereas the 96-Hour samples were analyzed on the day of sampling.
Duplicate samples at 0 hours (fresh media), 24, 48 and 72 hours (fresh and old media) and 96 hours (old media) were taken and stored frozen for further analysis if necessary.

Vehicle:

no

Details on test solutions:

Range-finding Test
A nominal amount of test item (2500 mg) was dispersed in 1000 mL of test water to give a 2500 mg/L stock solution which was inverted several times to ensure adequate mixing and homogeneity. A series of dilutions was made to give the test concentrations of 1.0, 10 and 100 mg/L. The prepared test concentrations were stirred for approximately 1 minute with a flat bladed stirrer to ensure homogeneity.

Definitive Test
A nominal amount of test item (2500 mg) was dispersed in 1000 mL of test water to give the 2500 mg/L stock solution and was inverted several times to ensure adequate mixing and homogeneity. A dilution was made to give the test concentration of 100 mg/L. The test concentration was stirred with a flat bladed mixer for approximately 1 minute to ensure homogeneity.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

The test was carried out using juvenile common carp (Cyprinus carpio). Fish were obtained from White Water Fisheries, Ripley, Derbyshire, UK and maintained in-house since 26 June 2017. Fish were maintained in a glass fiber tank with a "single pass" water renewal system. Fish were acclimatized to test conditions from 23 October 2017 to 30 October 2017. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

total hardness of approximately 140 mg/L as CaCO3

Test temperature:

Temperature was maintained at approximately 20 °C to 22 °C
measured using a Hanna Instruments HI 93510 digital thermometer

pH:

7.6 - 8.4
there were no treatment related differences for or pH.
measured using a Hach Flexi handheld meter

Dissolved oxygen:

10.5 - 8.1 mg O2/L
there were no treatment related differences for oxygen concentration.
measured using a Hach Flexi handheld meter

Nominal and measured concentrations:

Range finding test
Nominal 1.0, 10 and 100 mg/L
Chemical analysis of the 100 mg/L test solution at 0 hours showed a measured test concentration of 98 mg/L. The measured concentration in the corresponding old media at 24 hours was 99 mg/L indicating that the test item was stable under test conditions.

Definitive Test
Nominal 100 mg/L
Analysis of the fresh test preparations at 0 and 72 hours showed measured test concentrations of 121 and 97 mg/L. Measured concentrations in the corresponding expired test solutions at 24 and 96 hours were 120 and 98 mg/L indicating that the test item was stable under the test conditions.

Details on test conditions:

Range-finding Test
The test concentration to be used in the definitive test was determined by a preliminary range-finding test.
In the range-finding test fish were exposed to a series of nominal test concentrations of 1.0, 10 and 100 mg/L. The test item was dissolved directly in test water.
In the range-finding test three fish were placed in each test and control vessel, the test vessels were then covered to reduce evaporation and maintained in a temperature controlled room maintaining the water temperature at 20 to 24 °C with a maximum deviation of ±1 °C with a photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours. Each 25 to 30 liter test and control vessel contained 20 liters of test media and was covered to reduce evaporation. After 1, 3, 6, 24, 48, 72 and 96 hours any mortalities or sub-lethal effects of exposure were determined by visual inspection of the test fish.
The control group was maintained under identical conditions but not exposed to the test item.

Definitive Test
Based on the results of the range-finding test a "Limit test" was conducted at a concentration of 100 mg/L to confirm that at the maximum concentration given in the OECD/EC Test Guidelines, no mortalities or sub-lethal effects of exposure were observed.
As in the range-finding test, 25 to 30 liter glass exposure vessels containing 20 liters of test media were used for each control and test concentration. At the start of the test seven fish were placed in each test vessel at random, in the test preparations. The test vessels were then covered to reduce evaporation and maintained in a temperature controlled room maintaining the water temperature at 20 to 24 °C with a maximum deviation of ±1 °C with a photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours. The test vessels were aerated via narrow bore glass tubes. The fish were not individually identified and received no food during exposure.
The control group was maintained under identical conditions but not exposed to the test item.
A semi-static test regime was employed in the test involving a daily renewal of the test preparations to prevent the build up of nitrogenous waste products.

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:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Range-finding Test
The results showed no mortalities and no sub-lethal effects of exposure during the range-finding test.
Chemical analysis of the 100 mg/L test solution at 0 hours (see Annex 3) showed a measured test concentration of 98 mg/L. The measured concentration in the corresponding old media at 24 hours was 99 mg/L indicating that the test item was stable under test conditions.
Based on this information, a single test concentration of 100 mg/L was selected for the definitive test. This experimental design conforms to a "Limit test" to confirm that at the maximum test concentration given in the OECD/EC Test Guidelines, no mortalities or sub-lethal effects of exposure were observed.

Definitive Test
Verification of Test Concentrations
Analysis of the fresh test preparations at 0 and 72 hours showed measured test concentrations of 121 and 97 mg/L. Measured concentrations in the corresponding expired test solutions at 24 and 96 hours were 120 and 98 mg/L indicating that the test item was stable under the test conditions. The 0-Hour measured concentration was outside of the range 80 to 120%, however, as the measured concentration was only marginally outside of this range and as the test item was shown to be stable over the exposure period, it was considered justifiable to base the results on the nominal concentration.

Mortality Data
There were no mortalities in seven fish exposed to a test concentration of 100 mg/L for a period of 96 hours.
The NOEC was 100 mg/L.
It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.

Sub-Lethal Effects
There were no sub-lethal effects of exposure observed in the test.

Water Quality Criteria
Temperature was maintained at approximately 20 °C to 22 °C throughout the test, while there were no treatment related differences for oxygen concentration or pH.

Observations on Test Item Solubility
The test item preparations were observed to be clear colorless solutions throughout the test.

Sublethal observations / clinical signs:

Mortality Data

Inspection of the mortality data gave the following results:

Time (Hour)	LC50 (mg/L)
1	>100
3	>100
24	>100
48	>100
72	>100
96	>100

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item to the common carp (Cyprinus carpio) has been investigated and based on the nominal test concentrations gave a 96-Hour LC50 of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.
It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to common carp (Cyprinus carpio). The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (1992) No 203, "Fish, Acute Toxicity Test" referenced as Method C.1 of Commission Regulation (EC) No. 440/2008.

Methods

Following a preliminary range-finding test, seven fish were exposed to an aqueous solution of the test item, at a single concentration of 100 mg/L for a period of 96 hours at a temperature of approximately 20 ºC to 22 °C under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure in each test and control vessel were determined 1 and 3 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

Results

Analysis of the fresh test preparations at 0 and 72 hours showed measured test concentrations of 121 and 97 mg/L. Measured concentrations in the corresponding expired test solutions at 24 and 96 hours were 120 and 98 mg/L indicating that the test item was stable under the test conditions. The 0-Hour measured concentration was outside of the range 80 to 120%, however, as the measured concentration was only marginally outside of this range and as the test item was shown to be stable over the exposure period, it was considered justifiable to base the results of the nominal concentration.

Exposure of common carp (Cyprinus carpio) to the test item gave LC50 values based on the nominal test concentrations of greater than 100 mg/L. The No Observed Effect Concentration (NOEC) was 100 mg/L.

It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.

Description of key information

A study was performed to assess the acute toxicity of the test item to common carp (Cyprinus carpio). The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (1992) No 203, "Fish, Acute Toxicity Test" referenced as Method C.1 of Commission Regulation (EC) No. 440/2008.

Exposure of common carp (Cyprinus carpio) to the test item gave LC50 values based on the nominal test concentrations of greater than 100 mg/L. The No Observed Effect Concentration (NOEC) was 100 mg/L.

It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L