2-propenoic acid, 2-methyl-, 4-benzoylphenyl ester

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09.04.2014 to 28.05.2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-propenoic acid, 2-methyl-, 4-benzoylphenyl ester

EC Number:

611-390-2

Cas Number:

56467-43-7

Molecular formula:

C17H14O3

IUPAC Name:

2-propenoic acid, 2-methyl-, 4-benzoylphenyl ester

impurity 1

Reference substance name:

(4-benzoylphenyl) acetate

Cas Number:

13031-44-2

Molecular formula:

C15H12O3

IUPAC Name:

(4-benzoylphenyl) acetate

Test material form:

solid

Specific details on test material used for the study:

97.14 % 4-(Methacryloyloxy)benzophenone
1.046 % Benzophenone acetat
1.066 % 4-Hydroxybenzophenon
Expiry Date: May 26, 2014
Storage: At room temperature (20 ± 5 °C), under dark and dry conditions

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
- Sampling method:
Duplicate samples from the freshly prepared test media of all test concentrations and the control were taken at the start of the test and at day 3.
For the determination of the stability of the test item under the test conditions, respectively the maintenance of the test item concentrations during the test period, samples were taken in duplicate out of all test media and the control at day1 and at the end of the test (96 hours).
However, from the test concentrations of nominal 45.5 and 100 mg test item/L duplicate samples were taken after 48 hours additionally, because all fish were dead.
All test medium samples were taken from the approximate centre of the aquaria.
- Sample storage conditions before analysis:
All samples were stored in a freezer (≤ - 10 °C), protected from light until analysis was performed.

Test solutions

Vehicle:

no

Details on test solutions:

Test Concentrations:: Filtrates of nominal 100, 45.5, 20.7 and 9.39 mg test item/L. Additionally, a control was tested in parallel.
Control: In the control, test water was used without addition of the test item.
Dosage of Test Item:
The test item was not well soluble in test water. To consider the
impurities of the test item all concentrations were prepared
individually by suspending the following amounts of test item in test
water:
For day 0: 103.9 mg in 11065 mL
228.6 mg in 11043 mL
501.7 mg in 11026 mL
1101.5 mg in 11015 mL

For day 1: 104.1 mg in 11068 mL
228.1 mg in 11019 mL
501.4 mg in 11020 mL
1101.1 mg in 11011 mL
For day 2: 104.1 mg in 11068 mL
29.0 mg in 11063 mL

For day 3: 104.3 mg in 11108 mL
228.9 mg in 11058 mL
The suspensions were stirred for 24 hours in the dark to dissolve as much test item as possible. Then, the undissolved test item was separated by filtration (0.65 + 0.45 μm Cellulose Acetate Filter). The test media were prepared just before introduction of the test fish (=start of the test and test medium renewal on day 1, 2 and 3).

Appearance of the Test Itemin Test Medium:
The appearance of the test item in the test medium was observed at least once every day in all freshly prepared and aged test media of all test concentrations.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

Species: Rainbow trout (Oncorhynchus mykiss)
Age and Size: Juveniles;
The mean body length of the fish* in the test was 5.34 cm ± 0.28 cm (Mean ± SD), the mean body wet weight 1.39 ± 0.21 g (Mean ± SD). * 10 fish from the test fish batch were measured before the start of the test
Sex: Male and female
Origin: The test fish were obtained from Forellenzuchtbetrieb Tautenhahn, 98646 Trostadt, Germany.
Holding Conditions: All fish were obtained and held in the laboratory for at least 12 days before the start of the test. They were held in water of the quality to be used in the test for at least seven days immediately before testing under the following conditions:
Light: 16 hours photoperiod daily
Temperature: 13 - 17 °C
Oxygen concentration: at least 80 % of the air saturation value
Feeding: three times per week or daily until 24 hours before the test was started

During the last 7 days prior to the start of the test no fish died in the test fish batch. Therefore the mortalities in the fish batch were below 5 % and the fish batch was accepted.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

2.5 mmol/L (= 250.0 mg/L) as CaCO3

Test temperature:

13 to 14 °C

pH:

7.6 to 7.9

Dissolved oxygen:

88 to 100 %

Nominal and measured concentrations:

9.39 mg/l (nominal), 0.29 mg/l (measured)
20.7 mg/l (nominal), 0.755 mg/l (measured)
45.5 mg/l (nominal), 1.47 mg/l (measured)
100 mg/l (nominal), 2.21 mg/l (measured)

Details on test conditions:

Type and Size: 12 litre glass aquaria with 11 litre test medium
Identification: Each test unit was uniquely identified with serial number and treatment.
Test Conditions
Test Environment: Controlled environmental room Measurement of pH-Values, Dissolved Oxygen and Water
Temperature: The water temperature, pH-values and the dissolved oxygen concentrations were determined daily in the freshly prepared and aged test media of each treatment group.
Water Temperature: 13 to 14 °C
pH-Values of Test Water: 7.6 to 7.9
Dissolved Oxygen
Concentrations: 88 to 100 % of the air saturation value (see Table 6)
Light Regime: 16 h light : 8 h dark; 30 min dawn/dusk period was provided
Light Intensity: 540 to 980 lux
Aeration of the Test Water: The test media were slightly aerated during the test.
Recording: Test conditions were recorded with suitable instruments and documented in the raw data.

Pre-Experiments: Pre-experiments were performed to determine the solubility of the test item in test water and to select suitable methods for the preparation of a stock solution and the dosage of the test item into the test media.
Range-Finding Test: For the determination of the test concentrations a range-finding test was performed. However, according to the OECD Series on testing and assessment No. 23: "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", 2000 concentrations in excess of the solubility were not tested.
No GLP compliance in pre-experiments and the range-finding test

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.399 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

14 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

In the control and at the concentration of nominal 9.39 mg test item/L, all fish survived until the end of the experiment and no signs of intoxication occurred. At nominal 20.7, 45.5 and 100 mg test item/L all fish were dead at test end after 96 hours of exposure.

Reported statistics and error estimates:

The LC50, LC20 and LC10 at the observation times were calculated by Probit analysis. The NOEC, the LOEC, the LC0 and the LC100 were determined directly from the raw data.
The software used to perform the statistical analysis was ToxRat Professional, Version 3.2.1, ToxRat Solutions GmbH.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Observed mortality of unfed Rainbow Trout (Oncorhynchus mykiss) exposed to Benzophenone methacrylate for 96 h

Initial loding rate	Geometric mean measuerd concentration	Mortality
mg test item /L	mg test item /L	0 h	2 h	24 h	48 h	72 h	96 h
		# mort	# mort	# mort	# mort	# mort	# mort
control	control	0	0	0	0	0	0
9.39	0.209	0	0	0	0	0	0
20.7	0.755	0	0	0	2	6	7
45.5	1.47	0	0	0	7	7	7
100	2.21	0	0	5	7	7	7
LC50 [mg/l] based on initial loading rate		n.d.	n.d.	92.2	22.3	18.0	14.0
LC50 [mg/l] based on geometric mean measured concentrations		n.d.	n.d.	2.12	0.801	0.592	0.399
95 % CI		n.d.	n.d.	n.d.	n.d.	n.d.	n.d.

 

# mort: Number of dead fish

n.d.: not determinable

Values in parenthesis refer to geometric mean measured concentrations

CI.: Confidence interval Values refer to initial loading rates.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the test results the 96-hour LC50 of Benzophenone methacrylate for Rainbow Trout (Oncorhynchus mykiss) was determined to be 0.399 mg test item/L based on geometric mean measured concentrations.

Executive summary:

 Acute Toxicity of Benzophenone methacrylate to Rainbow Trout (Oncorhynchus mykiss) was tested in a 96-hour Semi Static Test acc. OECD 203 (GLP compliant study based on the Commission Regulation (EC) No 440/2008, C.1, 2008).

Filtrates of nominal 100, 45.5, 20.7 and 9.39 mg test item/L, corresponding to the geometric mean measured concentrations of 2.21, 1.47, 0.755 and 0.209 mg test item/L were tested. Additionally, a control was tested in parallel. Based on the test results the 96-hour LC50 of Benzophenone methacrylate for Rainbow Trout (Oncorhynchus mykiss) was determined to be 14.0 mg test item/L based on initial loading rates, corresponding to 0.399 mg test item/L based on geometric mean measured concentrations. The LC0 was determined to be 9.39 mg test item/L based on initial loading rates, corresponding to 0.209 mg test item/L based on geometric mean measured concentrations.