Oligomerisation products of 2,2'-[(1-methylethylidene)bis(4,1-phenyleneoxymethylene)]bisoxirane with acrylic acid and lauric acid

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:

July 30, 2012 to October 15, 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 1: Static Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Samples for possible analysis were taken from all test concentrations with surviving fish and the control according to the schedule below. In addition, samples were taken after 48 hours of exposure from the test concentrations in which all the fish had died.
Frequency: at t=0 h, t=24 h and t=96 h
Volume: 2 ml
Storage : samples were stored in a freezer until analysis.

Additionally, reserve samples of 2 ml were taken from all test solutions for possible analysis. If not already used, these samples are stored in a freezer for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis

Vehicle:

no

Details on test solutions:

The batch of RD04/201 tested was a clear colourless viscous liquid and was not completely soluble in test medium at the loading rates prepared. Weighing of test substance and preparation of test concentrations was performed under dimmed light.
All test solutions were prepared separately and started with loading rates ranging from 10 to 100 mg/l in test medium with a temperature of 60°C. Two days of magnetic stirring at room temperature was applied to reach maximum solubility of the test substance in the test medium. The resulting aqueous mixtures were left to stabilize for two hours where after the clear and colourless WAFs were siphoned off and used for testing.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp (Cyprinus carpio, Teleostei, Cyprinidae)
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands
- Length at study initiation (length definition, mean, range and SD): Final test: 2.9 ± 0.3 cm
- Weight at study initiation (mean and range, SD): Final test: 0.81 ± 0.19 g
- Feeding during test: no feeding
ACCLIMATION
- Acclimation period: At least 12 days after delivery
- Acclimation conditions (same as test or not): same as the test
- Feeding: Daily with pelleted fish food
- Health during acclimation (any mortality observed): In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

180 mg/l expressed as CaCO3

Test temperature:

Between 21.0 and 22.3 °C

pH:

Between 7.5 and 8.3

Dissolved oxygen:

Between 5.9 and 9.0 mg/l O2

Nominal and measured concentrations:

Nominal concentrations: WAFs prepared at loading rates of 10, 18, 32, 56 and 100 mg/l
Measured concentrations:. Actual test concentrations were based on the major component of the test substance. Analysis of the samples taken at the start of the final test showed measured concentrations that were all higher than the highest concentration measured during the range-finding test, which can be related to the differences in preparation of the WAFs (see section 6.4). These concentrations remained relatively stable during the first 24 hours of exposure (79-86% of initial) but the test concentrations with surviving fish decreased during the remainder of the test (67-73% of initial after 48 hours of exposure and
44-47% of initial at the end of the test period).

The actual concentration in the WAF prepared at 18 mg/l was slightly lower than the actual concentration in the WAF prepared at 10 mg/l. This result was confirmed by fish mortality at these two concentrations because at the end of the test period 6 fish exposed to the 10 mg/l WAF had died while only 1 fish exposed to the 18 mg/l WAF had died. For the determination of the effect parameters, average exposure concentrations were calculated and used in ascending order, i.e. 2.9, 3.3, 5.3, 6.2 and 14 mg/l.
.

Details on test conditions:

TEST SYSTEM
- Test vessel: 8 litres, all-glass, containing 7 litres of test solution
- Aeration: Aeration was introduced after 2 days of exposure and maintained until the end of the test.
- No. of organisms per vessel:7 per concentration
- No. of vessels per concentration (replicates):1
- No. of vessels per control (replicates):1
- Biomass loading rate: 0.81 g fish/litre, i.e. 7 fish per 7 litres of test medium
- Feeding:No feeding from 24 hours prior to the test and during the total test period

TEST MEDIUM / WATER PARAMETERS
Adjusted ISO medium, formulated using RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following composition:
CaCl2.2H2O: 211.5 mg/l
MgSO4.7H2O: 88.8 mg/l
NaHCO3: 46.7 mg/l
KCl: 4.2 mg/l
- Intervals of water quality measurement: Dissolved oxygen content pH and temperature, Daily in all vessels with surviving fish, beginning at the start of the test (day 0).
OTHER TEST CONDITIONS
- Adjustment of pH: no
- Illumination:16 hours photoperiod daily under dimmed light
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :LC50 t=24, 48, 72, 96 h
TEST CONCENTRATIONS
- Final study: control and WAFs prepared at loading rates of 10, 18, 32, 56 and 100 mg/l

Reference substance (positive control):

yes

Remarks:

pentachlorophenol (PCP)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

5.9 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

4.2 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% confidence interval 3.3-5.3 mg/l

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

3.3 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% confidence interval 3.2 - 3.6 mg/l

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.1 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Behavioral abnormalities:
- Mortality of control: No mortality in control
- Abnormal responses: no abnormalities were observed
- Effect concentrations exceeding solubility of substance in test medium: yes

Results with reference substance (positive control):

Under the conditions of the present test with carp exposed to PCP, the 96h-LC50 was 0.25 mg/l based on nominal concentrations, with a 95% confidence interval between 0.20 and 0.41 mg/l. This effect was already reached within 24 hours of exposure

Reported statistics and error estimates:

The 24h, 72h and 96h LC50-values were determined using the maximum likelihood estimation method with the probits of the percentages of dead fish as function of the logarithms of the corresponding concentrations (Finney, D.J., 1971: Probit analysis, Cambridge University Press, Cambridge, U.K., 3rd edition).

The 48h-LC50 could not be determined using the maximum likelihood estimation method. This was because there was no concentration between the highest concentration (A) at which there was 0% mortality and the lowest concentration (B) at which 100% mortality occurred. Instead, the LC50 was calculated as (AB)½, with A and B being limits of the 95% confidence interval.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the present test RD04/201 induced lethal effects in carp at average exposure concentrations of 2.9 mg/L and higher.

The 96h-LC50 was 3.1 mg/L.

Executive summary:

A study was conducted to determined the acute toxicity to fish in accordance with OECD Guideline 203 and EU Method C.1 and ISO Method 7346-1, under GLP conditions. A static test was conducted. Owing to the poor water solubility of the test substance, all test solutions were prepared as water accomodated fractions (WAFs). Solutions were prepared with loading rates ranging from 10 to 100 mg/L in the test medium assisted by magnetic stirring at room temperatureand left to stabilize for two hours where after the clear and colourless WAFs were siphoned off and used for testing. Seven fish per concentration were exposed to WAFs prepared at loading rates of 0, 10, 18, 32, 56 and 100 mg/L. The total test period was 96 hours and samples for analytical confirmation of actual exposure concentrations were taken at the start, after 24 hours of exposure and at the end of the test. In addition, samples were taken after 48 hours of exposure from the test concentrations in which all fish had died. The test concentrations were based on the major components of the test substance. Analysis of the samples taken at the start of the test showed measured concentrations ranging from 4.2 to 16 mg/L. These concentrations remained relatively stable during the first 24 hours of exposure (79-86% of initial) but the test concentrations with surviving fish decreased during the remainder of the test (67-73% of initial after 48 hours of exposure and 44-47% of initial at the end of the test period). Six out of ten fish died at 10 mg/L and one out of ten at 18 mg/L. For the determination of the effect parameters average exposure concentrations were calculated. The study met the acceptability criteria prescribed by the protocol and was considered valid. Under the study conditions, the 96 h L(L)C50 of the test substance was determined to be 3.1 mg/L (Bouwman, 2013).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

3.1 mg/L

Additional information

A study was conducted to determine the acute toxicity to fish in accordance with OECD Guideline 203 and EU Method C.1 and ISO Method 7346-1, under GLP conditions. A static test was conducted. Owing to the poor water solubility of the test substance, all test solutions were prepared as water accommodated fractions (WAFs). Solutions were prepared with loading rates ranging from 10 to 100 mg/L in the test medium assisted by magnetic stirring at room temperature and left to stabilize for two hours where after the clear and colourless WAFs were siphoned off and used for testing. Seven fish per concentration were exposed to WAFs prepared at loading rates of 0, 10, 18, 32, 56 and 100 mg/L. The total test period was 96 hours and samples for analytical confirmation of actual exposure concentrations were taken at the start, after 24 hours of exposure and at the end of the test. In addition, samples were taken after 48 hours of exposure from the test concentrations in which all fish had died. The test concentrations were based on the major components of the test substance. Analysis of the samples taken at the start of the test showed measured concentrations ranging from 4.2 to 16 mg/L. These concentrations remained relatively stable during the first 24 hours of exposure (79-86% of initial) but the test concentrations with surviving fish decreased during the remainder of the test (67-73% of initial after 48 hours of exposure and 44-47% of initial at the end of the test period). Six out of ten fish died at 10 mg/L and one out of ten at 18 mg/L. For the determination of the effect parameters average exposure concentrations were calculated. The study met the acceptability criteria prescribed by the protocol and was considered valid. Under the study conditions, the 96 h L(L)C50 of the test substance was determined to be 3.1 mg/L (Bouwman, 2013).