2-hydroxy-3-phenoxypropyl methacrylate

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07-09 August 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

13 April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-hydroxy-3-phenoxypropyl methacrylate

EC Number:

240-994-5

EC Name:

2-hydroxy-3-phenoxypropyl methacrylate

Cas Number:

16926-87-7

Molecular formula:

C13H16O4

IUPAC Name:

2-hydroxy-3-phenoxypropyl 2-methylprop-2-enoate

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 171512
- Expiration date of the lot/batch: 23 December 2019

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15-25ºC, below 70RH%), protected from humidity (tigh closed container)

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Final dilution of a dissolved solid, stock liquid or gel: Stock solution 100.00 mg/L

Sampling and analysis

Analytical monitoring:

yes

Remarks:

Analytical measurement was performed at the applied test concentration levels and from the control at the beginning and at the end of the experiment. The samples were analysed by HPLC system with UV detection method.

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: Untreated control - the dilution water (ISO-medium) was used without addition of the test item. Reference control - Potassium dichromate
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ISO-medium (according to OECD 202)
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): Stock solution - 100.00 mg/L
Final soltions (nominal) - 6.25, 12.5, 25.0, 50.0, 100.00 mg/L

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain/clone: Daphnia Magna
- Age at study initiation (mean and range, SD): <24 hours at the begining of the test
- Source: Istvan Szent University

ACCLIMATION
- Acclimation period: No acclimatization because the water used was similar to the culture water

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

The reconstituted water (ISO medium) had a total hardness of 245 mg/L (as CaCO3).

Test temperature:

The water temperature was measured at the start of the experiment and 24-hour intervals thereafter in each test vessel at all concentration levels. The test temperature was in the range of 20.2 – 20.7°C measured in the test vessels.
The additionally measured temperature in the climate chamber was between 19.7 and 21.0ºC.

pH:

The pH of the test solution was not adjusted and not varied by more than 1.5 units in any one test. The pH was measured at the start and at the end of the experiment in each test vessel at all concentration levels and was in the range of 7.50 – 7.72.

Dissolved oxygen:

The dissolved oxygen concentration was measured in each test vessel at all concentration levels at the start and at the end of the test and was in the range of 8.1 – 8.6 mg/L.

Nominal and measured concentrations:

Nominal: 6.25, 12.5, 25.0, 50.0, 100.00 mg/L
Measured: 6.67, 13.20, 25.75, 51.25, 102.50 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers
- Type (delete if not applicable): open

RANGE-FINDING STUDY
- Test concentrations: 0.1, 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: Because significant immobility was observed at the highest concentration level (100.0 mg/L) during the preliminary range-finding test, five test concentrations in a geometric series (using factor 2.0) and one control were used in the main test under static conditions (based on analytical results indicated that a static test was feasible).

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Twenty animals, divided into four groups (glass beaker) of five animals each were used at the test concentrations and for the control as well. The 24 and 48 hours EC50 values of the test item could not be calculated due to the slight effects observed. Consequently, the 24 and 48 hours EC50 and the 48h EC100 values of the test item were determined directly from the raw data.For the determination of the LOEC and NOEC, the immobilization at the test concentrations was tested on significant differences to the control values by Dunnett’s Test using TOXSTAT software. All validity criteria were met during this study.

Reported statistics and error estimates:

The 24 and 48 hours EC50 values of the test item could not be calculated due to the slight effects observed. Consequently, the 24 and 48 hours EC50 and the 48h EC100 values of the test item were determined directly from the raw data.For the determination of the LOEC and NOEC, the immobilization at the test concentrations was tested on significant differences to the control values by Dunnett’s Test using TOXSTAT software.

Any other information on results incl. tables

Validity

 

There was no immobilization in the control group and the dissolved oxygen concentration at the end of the test in control and test vessels was more than 3 mg/L.

All validity criteria were within acceptable limits and therefore the study is considered as valid.

Concentration of the test item

The following nominal concentrations were tested: 6.25, 12.5., 25.0, 50.0 and 100.0 mg/L.

 

Test concentrations were analytically determined at the start and at the end of the experiment. The corresponding measured geometric mean test item concentrations were: 6.67, 13.20, 25.75, 51.25 and 102.50 mg/L.

As the analytically measured concentrations deviated not more than 20 per cent from the nominal, the biological results are related to the nominal test item concentrations.

 

Analytically measured concentrations are shown in Table 3.

 

Table 3: Calculation of exposure concentrations

Nominal Concentration (mg/L)	Measured concentrations (mg/L)		Geometric mean
	0 h/start	48h/end
Control	n.d.	n.d.	-
6.25	6.58	6.76	6.67
12.5	13.1	13.3	13.20
25.0	26.1	25.4	25.75
50.0	51.6	50.9	51.25
100.0	102.7	102.3	102.50

n.d. - not detected

IMMOBILISATION

The number of immobilised animals and the percentage of immobility were determined at the 24^th and 48^th hour (Table 4).

In addition to immobility, no abnormal behaviour or appearance of test animals was detected.

 

Table 4: Number and percentage of immobilised animals

Nominal Concentration (mg/L)	Number of treated animals	Immobilised animals
		24 hours		48 hours
		number	percent	number	prcent
Control 6.25 12.5 25.0 50.0 100.0	20 20 20 20 20 20	0 0 0 0 0 0	0 0 0 0 0 0	0 0 0 0 0 3*	0 0 0 0 0 15

*: statistically significantly different compared to the control values (Dunnett’s Test; a= 0.05)

Data of immobility in each test vessel are detailed in Appendix 2.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Acute toxicity of PHPM was assessed with Acute immobilisation test on Daphnia magna, over an exposure period of 48 hours in a static system.

All validity criteria were met during this study.

Under the conditions of this Daphnia magna acute immobilisation study the observed endpoints for the effect of PHPM were the followings:

The 24h EC50 value: > 100.0 mg/L (nominal)
The 48h EC50 value: > 100.0 mg/L (nominal)
The 48h EC100 value: > 100.0 mg/L (nominal)
The 48h No-Observed Effect Concentration (NOEC): 50.0 mg/L (nominal)
The 48h Lowest Observed Effect Concentration (LOEC): 100.0 mg/L (nominal)

Executive summary:

Acute toxicity of PHPM on Daphnia magna was assessed in an acute immobilisation test, over an exposure period of 48 hours in a static test system.

Because significant immobility was observed at the highest examined concentration level during the preliminary range-finding test, five test concentrations in a geometric series (factor 2.0) and one control group were tested in the definitive test under static conditions.

 

The test concentrations were analytically determined at the start and at the end of the experiment.

The nominal concentrations of test item used in the main experiment were: 6.25, 12.5, 25.0, 50.0 and 100.0 mg/L.

The corresponding measured geometric mean test item concentrations were: 6.67, 13.20, 25.75, 51.25 and 102.50 mg/L.

As the measured concentrations deviated not more than 20 per cent from the nominal in all cases, biological results are based on thenominal test item concentrations.

Twenty animals, divided into four groups (glass beaker) of five animals each were used at the test concentrations and for the control as well.

The 24 and 48 hours EC₅₀ values of the test item could not be calculated due to the slight effects observed. Consequently, the 24 and 48 hours EC₅₀ and the 48h EC₁₀₀ values of the test item were determined directly from the raw data.

For the determination of the LOEC and NOEC, the immobilization at the test concentrations was tested on significant differences to the control values by Dunnett’s Test using TOXSTAT software.

All validity criteria were met during this study.

 

Under the conditions of this Daphnia magna acute immobilisation study the observed endpoints for the effect of PHPM were the followings:

 

The 24h EC₅₀value:                > 100.0 mg/L (nominal)

The 48h EC₅₀value:                > 100.0 mg/L (nominal)

The 48h EC₁₀₀value:               > 100.0 mg/L (nominal)

The 48h No-Observed Effect Concentration (NOEC):          50.0 mg/L (nominal)

The 48h Lowest Observed Effect Concentration (LOEC):   100.0 mg/L (nominal)