Reaction mass of 2-dodecylhexadecyl methacrylate and 2-Tetradecyloctadecyl 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:

August - October 2020.

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:

2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

The test material Reaction Mass of 2-Propenoic acid, 2-methyl-, 2- dodecylhexadecyl ester and 2-
Propenoic acid, 2- methyl-, 2-tetradecyloctadecyl ester was stored at room temperature in the dark. The batch number of the test item was F1-9K038C.

Analytical monitoring:

not required

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
In order to determine an appropriate test preparation method, an assessment was made of the material’s behaviour in seawater. A 1000 mg/l stock was prepared in filtered seawater, and the resulting mixture was stirred for one hour left to settle for one hour and its behaviour assessed. As the material was observed to be poorly soluble, then it was stirred again for approximately 19 hrs and then left to settle for one hour and its behaviour assessed. As the material was observed to be poorly soluble the material was prepared by Water Accommodated Fractions (WAFs). Test material stocks were made by weighing out an appropriate amount of test material, adding treated seawater and stirring for approximately 20h. The stocks were then left to settle for 1h and the mid column of the stock was siphoned off. 3L of the siphoned water fraction was added to the test vessel.

Test organisms (species):

Cyprinodon variegatus

Details on test organisms:

The test organism C. variegatus (common name Sheepshead minnow) were reared and maintained at the testing facility under flow-through conditions in appropriately sized tanks, containing 1μm filtered ultra-violet treated seawater and supplied with gentle aeration.
The fish were 76 days old, with a length ranging between 13.76-16.69 mm, and an average biomass of ca. 0.1 g. They were subject to a photoperiod of 16h of light daily, and fed food in flake form ad libitum at least twice daily.

Test type:

semi-static

Water media type:

saltwater

Remarks:

1μm filtered ultra-violet treated seawater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

Temperature was 25 °C ±2 °C, with difference between changeover tanks ≤2°C

pH:

6-8.5

Dissolved oxygen:

98-100(%)

Salinity:

36 ± 4‰

Details on test conditions:

TEST SYSTEM
Seven test fish were randomly selected for testing (in both testing and control conditions) and they were not fed within 24h of test commencement. Test solution was replaced after 48 hours. The maximum biomass loading rate was 0.8 g fish/L. Observation intervals were every 24 hours in a 96 hours time frame. At test termination control fish were measured to confirm the validity length range of between 10 to 20 mm.

LIMIT TEST
The limit test was conducted in a temperature controlled room to maintain a temperature of 25 ±2 ºC in the test medium. Five litre capacity glass beakers containing three litres of test media were used. The test material preparations were mixed in situ to ensure thorough dissolution before introduction of the test organisms.
All tanks received gentle aeration throughout the test duration. This limit test was conducted under semi-static conditions with a replacement of media at 48h. Water quality measurements were carried out prior to and following each renewal of media. The test duration was 96h with observation intervals every 24h, and final observations taken within 2h of the end of the test. All vessels were checked daily to monitor the condition of test animals. Animals which appeared immobile were gently prodded, and those which showed no response or opercular movement were recorded as dead. If all fish were alive after 96h of exposure it can be concluded that at the test concentration within the water phase the test material had no effect. The result was based on nominal concentrations of 1120 mg/L of the test material.

TEST MEDIUM / WATER PARAMETERS
Natural local seawater was pumped to holding tanks from Scapa Flow, Orkney (Latitude 58°50'8.02"N, Longitude 3° 8'10.80"W). The treatment process included sand filters to remove particulate matter, followed by a 1 μm filter. This water was heated to 20 ºC, UV treated, degassed and held at a quantity of 3000 to 5000 litres. The treated seawater was constantly pumped through the processing laboratory water system and trickle replenished as the water was used.
Water quality measurements were measured twice weekly and mortality observations were conducted daily. The raw water analysis showed Total Organic Carbon (TOC) of 27.0 mg/L, while metals and pesticides were all below detection levels.

TEST CONCENTRATION
The test concentration was derived from the test material EC/LC50 value between the most sensitive acute toxicity test species Pseudokirchneriella sp. and Daphnia magma. From allied studies the crustacean species Daphnia magma was noted to be more sensitive with an LC50 value of 1120mg/l.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 120 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Previous studies on the test material short-term effect on Daphnia and Algae.

Basis for effect:

mortality (fish)

Details on results:

After 96h exposure to Reaction Mass of 2-Propenoic acid, 2-methyl-, 2- dodecylhexadecyl ester and 2-Propenoic acid, 2- methyl-, 2-tetradecyloctadecyl ester, no mortalities were observed in the marine fish C. variegatus.

Results were checked against the following acceptance criteria:

Acceptance criterion 1: no mortality was assessed in the control group, which is within the specified limit of <10% (or one fish, if fewer than 10 control fish) at the end of the exposure.
Acceptance criterion 2: the dissolved oxygen concentration ranged between 98-100% during the exposure in all test vessels throughout the exposure, which is over the minimum specified limit of 60%.
Acceptance criterion 3: the average salinity in the the test systems was over the specified limit of 35‰ (36 ± 4‰). This was due an administrative oversight. However, this should not have had significant influence on the test results as control and test salinity measurements were within both ranges.

Sublethal observations / clinical signs:

Table 1. Number of test fish alive throughout the test

Nominal concentration (mg/L)	Time point (h)	Total alive (n)	Condition of surviving fish
1120	0	7	Active, mild
1120	24	7	Active, mild
1120	48	7	Active, mild
1120	72	7	Active, mild
1120	96	7	Active, mild

Table2. Number of control fish alive throughout the test

Nominal concentration (mg/L)	Time point (h)	Total alive (n)	Condition of surviving fish
control	0	7	Active, mild
control	24	7	Active, mild
control	48	7	Active, mild
control	72	7	Active, mild
control	96	7	Active, mild

Validity criteria fulfilled:

yes

Conclusions:

It can be concluded that Reaction Mass of 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2- methyl-, 2-tetradecyloctadecyl ester did not meet the criteria for classification as toxic to the aquatic environment according to Regulation (EC) N°1272/2008.

Executive summary:

The short-term toxicity effect on fish of the Reaction Mass of 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2- methyl-, 2-tetradecyloctadecyl ester was tested in accordance of the OECD Guideline 203 during a GLP compliant study. This test evaluates the acute toxicity of a chemical by measurement of its mortality effect on fish individuals. Visible abnormalities related to appearance and behaviour are also recorded. Where possible, the concentrations to kill 50% of the fish (LC50) are determined.

Since the substance was poorly soluble, Water Accomodated Fraction (WAF) were prepared.

In this study, the marine fish Cypridon variegatus are exposed to the test chemical at a concentration of 1120 mg/L for a period of 96 hours, under semi-static conditions. 

The results show no mortalities in the marine fish after 96 hours of exposure to the test chemical. Acceptance criteria were satisfied.

Hence, it can be concluded that Reaction Mass of 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2- methyl-, 2-tetradecyloctadecyl ester did not meet the criteria for classification as toxic to the aquatic environment according to Regulation (EC) N°1272/2008.

Description of key information

The short-term toxicity effect on fish of the Reaction Mass of 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2- methyl-, 2-tetradecyloctadecyl ester was tested in accordance of the OECD Guideline 203 during a GLP compliant study. This test evaluates the acute toxicity of a chemical by measurement of its mortality effect on fish individuals. Visible abnormalities related to appearance and behaviour are also recorded. Where possible, the concentrations to kill 50% of the fish (LC50) are determined.

Since the substance was poorly soluble, Water Accomodated Fraction (WAF) were prepared.

In this study, the marine fish Cypridon variegatus are exposed to the test chemical at a concentration of 1120 mg/L for a period of 96 hours, under semi-static conditions. 

The results show no mortalities in the marine fish after 96 hours of exposure to the test chemical. Acceptance criteria were satisfied.

Hence, it can be concluded that Reaction Mass of 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2- methyl-, 2-tetradecyloctadecyl ester did not meet the criteria for classification as toxic to the aquatic environment according to Regulation (EC) N°1272/2008.

Key value for chemical safety assessment

Marine water fish

Marine water fish

Dose descriptor:

LC50

Effect concentration:

> 1 120 mg/L