Fatty acids, linseed-oil, reaction products with 2-amino-2-(hydroxymethyl)-1,3-propanediol and formaldehyde

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:

2017-20-11 - #

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:

17 July 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

30 May 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: As the test item is an UVCB substance, a water-accommodated fraction (WAF) was pre-pared for the test. This was done by mixing the nominal load of 5.17 g/L resp. 5.5 mL/L test item (based on a density of 0.94 g/mL stated in the MSDS) with the corresponding amount of tap water and stirring for 24 hours, assuming to achieve the maximum water solubility of the substance. The lower phase was used unfiltered as test solution.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Species: Danio rerio HAMILTON-BUCHANAN
- Source: in-house breeding (originally from Umweltbundesamt)
- Age: sexually immature young fish, length 2 +/- 1 cm
- Vessels: polyethylene aquaria
- Medium: chlorine-free tap water
- Photo period: 12/12 hours, using neon tubes
- Temperature 23 +/- 2 °C

ACCLIMATION
- Acclimation period: Before being used for the test, the fish were kept for at least 12 days under test conditions. 24 hours before the start of the test, the test fish were no longer fed.

FEEDING DURING TEST
- Feeding: three times a day with warm-water fishfood, totaling about 1-2 % of body weight per day.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

0.71 mmol/L (from Drinking Water Analysis: March 2017, Wasserwerk Kirrweiler)

Test temperature:

21.8 - 22.8 °C

pH:

The pH was 7.9 - 8 in the control and 7.8 - 7.9 in the test item treatment.

Dissolved oxygen:

The dissolved oxygen concentration was 7.4 - 8.7 mg/L in the control and 6.6 - 8.2 mg/L in the test item treatment

Conductivity:

174 µS/cm (from Drinking Water Analysis: March 2017, Wasserwerk Kirrweiler)

Nominal and measured concentrations:

Control: -
Test item: measured: t(0h) = 10.84 mg/L, t(48h) 0.93 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria, containing 7 L
- Aeration: accomplished with glass tubes
- Medium renewal: none
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 1fish/L
- The vitality of the fish was assessed after 24 h, 48 h, 72 h and 96 h exposure


TEST MEDIUM / WATER PARAMETERS
- Test medium: Good quality drinking water
- Measurement interval of water parameter: The pH, conductivity and O2-concentrations were measured daily. The temperature was measured continuously.

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: 12 h light: 12 h dark
- Light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Limit test
- Range finding study: yes
- In a non-GLP pre-test, no toxicity was observed. Therefore, the test design of a Limit test was chosen.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 3.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 3.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

> 3.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Results with reference substance (positive control):

- Mortality of control: As no fish died until the end of the exposure, the mortality in the control as well as in the test item treatment was 0 %.

Reported statistics and error estimates:

As no effect on the test organisms was observable, no statistical data evaluation was performed.

Sublethal observations / clinical signs:

Results of analytical determination

At the beginning and at the end of the test, the content of the test item in the test solution was determined using DOC-determination. The correlation between nominal and measured concentration was weak because of the limited solubility of the test item. At the end of the test the measured DOC concentration in the test solution was much lower than at the beginning. This might be caused by the presence of the test organism and adsorption or ingestion of dissolved test item. Therefore, the geometric mean of the measured concentrations was used for the determination of the results. Geometric mean is calculated by multiplication of n participating concentrations and taking the n^throot.

Criteria of validity

- The mortality in the blank control may not exceed 1 fish at the end of the test. No mortality occurred in the blank control.

- The dissolved oxygen concentration must be at least 60% throughout the test. The concentration of dissolved oxygen was at least 6.6 mg/L or 76 % throughout the test

- The pH-value in the test solutions should not vary by more than 1 unit during the test. The highest variation was 0.1 units.

Validity criteria fulfilled:

yes

Conclusions:

The toxicity of the test item against Danio rerio was tested in a study conducted in accordance of the OECD guideline 203 and GLP. The study was performed using one concentration containing 5.17 g/L nominal concentration (3.18 mg/L measured concentration). No toxic effects were observed.

Executive summary:

In a 96-h acute toxicity study, according to the OECD guideline 203, Danio rerio (zebrafish) were exposed to VOELOFA Monomer at one concentration containing 5.17 g/L nominal concentration (3.18 mg/L measured concentration) for 96 h. 

The test solution was prepared by moderately stirring for 24 hours and it is assumed that the test conditions represent the maximum solubility of the test item in the present test medium.

At the beginning and at the end of the test, the content of the test item in the test solution was determined using DOC-determination. The measured concentrations were very low and correlation between nominal and measured concentration was weak because of the the limited solubility of the test item. At the end of the test the measured DOC concentration in the test solution was much lower than at the beginning. This might be caused by the presence of the test organism and adsorption or ingestion of dissolved test item. The presence of small colloids of undissolved test item in the test solution can be excluded as considering the density of 0.94 g/mL, the presence of undissolved test item would result in a thin layer of undissolved test item on the surface of the test solution which was not the case here.

The geometric mean of the measured concentrations was used for the determination of the results.

No observations were made which might cause doubts concerning the validity of the study outcome. All validity criteria were met.

The result of the test is considered valid.

As no effect on the test organisms was observable, no statistical data evaluation was performed.

Based on the results of this study, VOELOFA Monomer does not have to be classified as hazardous to fish in accordance with Section 4.1.2.6., Annex I of Regulation 1272/2008/EC.

This toxicity study is classified as acceptable and satisfies the guideline requirement for fish acute toxicity studies with D. rerio.

Results:

96h-NOEC ≥ 3.18 mg/L

96h-LOEC > 3.18 mg/L

96h-EC50 > 3.18 mg/L

Endpoint(s) Effected: mortality

Description of key information

The toxicity of the test item against Danio rerio was tested in a study conducted in accordance of the OECD guideline 203 and GLP. The study was performed using one concentration containing 5170 mg/L nominal concentration (3.18 mg/L measured concentration). No toxic effects were observed.

Additional information

In a 96-h acute toxicity study, according to the OECD guideline 203, Danio rerio (zebrafish) were exposed to VOELOFA Monomer at one concentration containing 5170 mg/L nominal concentration (3.18 mg/L measured concentration) for 96 h. The test solution was prepared by moderately stirring for 24 hours and it is assumed that the test conditions represent the maximum solubility of the test item in the present test medium. No observations were made which might cause doubts concerning the validity of the study outcome. All validity criteria were met. The result of the test is considered valid.