Phosphoric acid, 2-ethylhexyl ester, sodium salt

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:

from 2019-05-13 to 2019-07-18

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:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

LC/MS/MS

Details on sampling:

For determination of the test item concentrations, five replicate samples were taken from the test solution and five replicate samples from the control at the start and at the end of the test. The sampling was performed from different part of the aquariums.

Vehicle:

no

Details on test solutions:

The test solution was prepared by mechanical dispersion without using any solubilising agent. An amount of 0.5008 g test item was dissolved in 5008 mL ISO medium, resulting a nominal concentration of 100 mg/L. The test solution was freshly prepared in the testing laboratory just before introduction of fish (start of experiment).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish (Danio rerio)
- Source: Akvárium Magazin Kft. (Pasaréti Gyula), 1222 Budapest, Dévény u. 36., Hungary
- Age at study initiation: juveniles
- Length at study initiation: 2.00 - 2.50 cm
- Weight at study initiation: 0.163 - 0.197 g

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions: same as test condition
- Type and amount of food during acclimation: commercial diet for fish
- Feeding frequency during acclimation: at least three times per week until 24 hours before the treatment
- Health during acclimation (any mortality observed): no mortality occured in 7 days before the start of the test

FEEDING DURING TEST
- Food type: no feeding during test

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

249 mg/L (as CaCO3)

Test temperature:

21.8 - 22.3 °C

pH:

7.48 - 7.84

Dissolved oxygen:

63.2 - 86.9 %

Nominal and measured concentrations:

Nominal concentration: 100 mg/L
Measured concentration: at the start - 91 mg/L, at the end - 86 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: aquariums
- Type: open
- Material, size, headspace, fill volume: glass, 5 L testing solution
- Aeration: none, in the treatment group at the 72th hour slight aeration was applied to contain the dissolved oxygen level above 60 %
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.33 - 0.39 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium, prepared according to OECD 203
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: 16 h light : 8 h darkness

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 100 mg/L
- Results used to determine the conditions for the definitive study: no mortality occured during the exposure period

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

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:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

There was no mortality observed in the 10 fish in the control group or in the 10 fish exposed to the test item concentration of 100 mg/L. No sub-lethal effect was observed in the control and in the test item treated group during the study.
The 96-h No Observed Effect Concentration (NOEC) was determined to be 100 mg/L. The 96-h LC50, LC100 and the LOEC values were determined to be > 100 mg/L, the 96-h LC0 value was determined to be 100 mg/L based on nominal test concentration.

Sublethal observations / clinical signs:

Analytical Results

A single test item concentration of 100 mg/L (limit test) and a concurrent control were tested in the main test. The concentration of the test item was analytically determined in the test item solution and in the control at the start and at the end of the experiment. The mean of the measured concentrations of the test items main component was 91 mg/L at the start and 86 mg/L at the end of the test in the test item treated group. The measured test item concentration remained within ± 20 % of the nominal during the test period of 96 hours. Therefore, the biological results are based on the nominal test item concentration. Interfering component was not detected in the control samples.

Validity criteria fulfilled:

yes

Conclusions:

In this 96-hour acute toxicity test on Zebrafish (Danio rerio) the test item had no toxic effect on fish up to the limit concentration of 100 mg/L. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.

Executive summary:

The acute toxicity of the test item to fish (Zebrafish) was determined according to the OECD Guideline 203, EU Council Regulation No. 440/2008 method C.1 and EPA OPPTS 850.1075. A static test was performed as a limit test with a test concentration of 100 mg/L. The test item solution was prepared by mechanical dispersion – 0.5008 g test item were dissolved in 5008 mL ISO medium. Ten test organisms were exposed to the test concentration and the control, respectively, for 96 hours. The measured concentration of the test item at the start of the test was 91 mg/L and at the end of the test was 86 mg/L, respectively. The measured test item concentration remained within ± 20 % of the nominal during the test period of 96 hours. Therefore, the biological results are based on the nominal test item concentration. In conclusion, the test item had no toxic effect on fish up to the limit concentration of 100 mg/L. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.

Description of key information

In a 96-hour acute toxicity test on Zebrafish (Danio rerio) the test item had no toxic effect on fish up to the limit concentration of 100 mg/L. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.

Additional information

The acute toxicity of the test item to fish (Zebrafish) was determined according to the OECD Guideline 203, EU Council Regulation No. 440/2008 method C.1 and EPA OPPTS 850.1075. A static test was performed as a limit test with a test concentration of 100 mg/L. The test item solution was prepared by mechanical dispersion – 0.5008 g test item were dissolved in 5008 mL ISO medium. Ten test organisms were exposed to the test concentration and the control, respectively, for 96 hours. The measured concentration of the test item at the start of the test was 91 mg/L and at the end of the test was 86 mg/L, respectively.The measured test item concentration remained within ± 20 % of the nominal during the test period of 96 hours. Therefore, the biological results are based on the nominal test item concentration.In conclusion,the test item had no toxic effect on fish up to the limit concentration of 100 mg/L. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.