2-Propanamine, compd. with boron trifluoride, reaction products with Bu glycidyl ether

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Samples were taken from the control and the 100 mg/L test group from the bulk test preparation at 0 hours and from the pooled replicates at 48 hours for quantitative analysis. All samples were stored frozen prior to analysis. Duplicate samples were taken at 0 and 48 hours and stored frozen for further analysis if necessary.

Vehicle:

no

Details on test solutions:

Reconstituted water (ISO medium) was used for the range-finding.. Elendt M7 media, as used for the culturing, was used for the definitive test.

Reconstituted Water – Elendt M7 Medium

Ingredient
Final Concentration (mg/L)
H3BO3
0.715
MnCl2.4H2O
0.090
LiCl
0.077
RbCl
0.018
SrCl2.6H2O
0.038
NaBr
0.004
Na2MoO4.2H2O
0.016
CuCl2.2H2O
0.004
ZnCl2
0.013
CoCl2.6H2O
0.010
KI
0.0033
Na2SeO3
0.0022
NH4VO3
0.00058
Na2EDTA.2H2O
0.625
FeSO4.7H2O
0.249
CaCl2.2H2O
293.8
NaHCO3
64.8
MgSO4.7H2O
123.3
Na2SiO3.9H2O
10
KCl
5.8
NaNO3
0.274
K2HPO4
0.184
KH2PO4
0.143
Thiamine hydrochloride
0.075
Cyanocobalamine (vitamin B12)
0.0010
D(+) biotin (vitamin H)
0.00075
The pH of the prepared media was 7.9 ± 0.3 and stored at approximately 21 ºC.

Elendt M7 media, as used for the culturing, was used for the definitive test

Reconstituted Water – ISO Medium
Ingredient
Final Concentration (mg/L)
CaCl2.2H2O
294
MgSO4.7H2O
123
NaHCO3
65
KCl
5.8

Test organisms (species):

Daphnia magna

Details on test organisms:

The test was carried out using first instar Daphnia magna derived from in-house laboratory cultures.
Adult daphnids were maintained in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room maintaining the water temperature at 18 to 22 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

See section with details on test solution

Test temperature:

18-22 C

pH:

See section with details on test solution

Dissolved oxygen:

See section with details on test solution

Salinity:

See section with details on test solution

Conductivity:

See section with details on test solution

Details on test conditions:

Range-finding Test
The test concentration to be used in the definitive test was determined by a preliminary range-finding test.
In the range-finding test Daphnia magna were exposed to a series of nominal test concentrations of 0.10, 1.0, 10 and 100 mg/L.
A nominal amount of test item (50 mg) was dissolved in test water with the aid of ultrasonication for approximately 5 minutes and the volume adjusted to 500 mL to give the 100 mg/L test concentration from which a series of dilutions was made to give further test concentrations of 0.10, 1.0 and 10 mg/L.
Each prepared concentration was inverted several times to ensure adequate mixing and homogeneity.
In the range-finding test 10 daphnids were placed in each test and control vessel and maintained in a temperature controlled room maintaining the water temperature at 18 to 22 °C with a maximum deviation of ±1 °C with a photoperiod of 16 hours light and 8 hours darkness for a period of 48 hours with 20 minute dawn and dusk transition periods. Each 250 mL test and control vessel contained 200 mL of test media and was covered to reduce evaporation. After 24 and 48 hours the number of immobilized daphnids were recorded.
The control group was maintained under identical conditions but not exposed to the test item.
A sample of each test concentration was taken for chemical analysis at 0 and 48 hours in order to determine the stability of the test item under test conditions. All samples were stored frozen prior to analysis. Only concentrations within the range to be used for the definitive test were analyzed.
Definitive Test
Based on the results of the range-finding test a "Limit test" was conducted at a concentration of 100 mg/L to confirm that at the maximum concentration given in the OECD/EC Test Guidelines, no immobilization or adverse reactions to exposure were observed.
A nominal amount of test item (200 mg) was dissolved in test water with the aid of ultrasonication for approximately 5 minutes and the volume adjusted to 2 liters to give the 100 mg/L test concentration.
The prepared concentration was inverted several times to ensure adequate mixing and homogeneity.
The concentration and stability of the test item in the test preparation was verified by chemical analysis at 0 and 48 hours

Exposure Conditions
As in the range-finding test 250 mL glass vessels containing approximately 200 mL of test preparation were used. At the start of the test five daphnids were placed in each test and control vessel at random, in the test preparations. Four replicate test and control vessels were prepared. The test vessels were then covered to reduce evaporation and maintained in a temperature controlled room maintaining the water temperature at 18 to 22 °C with a maximum deviation of ±1 °C with a photoperiod of 16 hours light (between 200 and 1200 Lux) and 8 hours darkness with 20 minute dawn and dusk transition periods. The daphnids were not individually identified, received no food during exposure and the test vessels were not aerated.
The control group was maintained under identical conditions but not exposed to the test item.

Test Organism Observations
Any immobilization or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure. The criterion of effect used was that daphnia were considered to be immobilized if they were unable to swim within 15 seconds after gentle agitation.

Water Quality Criteria
Water temperature was recorded daily throughout the test. Dissolved oxygen concentrations and pH were recorded at the start and termination of the test. The pH and dissolved oxygen concentration were measured using a Hach Flexi handheld meter whilst the temperature was measured using a Hanna Instruments HI 93510 digital thermometer.
The light intensity during the light period was measured using an ATP Instrumentation Lux meter.
The appearance of the test media was recorded daily.

Reference substance (positive control):

yes

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Range finding test
No immobilization was observed at the test concentrations of 1.0, 10 and 100 mg/L. Three daphnids were observed to be immobile in the 0.10 mg/L test preparation after 24 and 48 hours. This was considered to be due to natural causes given that no adverse effects to exposure were observed at the higher concentrations employed.
Based on this information, a single test concentration of four replicates, of 100 mg/L was selected for the definitive test. This experimental design conforms to a "Limit test" to confirm that at the maximum test concentration given in the OECD/EC Test Guidelines, no immobilization or adverse reactions to exposure were observed.
Chemical analysis of the 100 mg/L test preparations at 0 and 48 hours showed measured test concentrations of 53% and 55% of nominal respectively were obtained. Inspection of the data could find no cause for the low levels obtained; however, these results did demonstrate that the test item was stable over the test duration.

Definitive Test
Verification of Test Concentrations
Analysis of the test preparations at 0 and 48 hours showed measured test concentrations to be near nominal (97% and 90%) and so the results are based on nominal test concentrations.
Immobilization Data
Exposure of Daphnia magna to the test item gave EC50 values of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.
It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.
Sub-Lethal Effects
No sub-lethal effects of exposure were observed throughout the test.
Validation Criteria
The test was considered to be valid given that none of the control daphnids showed immobilization or other signs of disease or stress and that the oxygen concentration at the end of the test was equal to or greater than 3 mg/L in the control and test vessels.
Water Quality Criteria
The results of the water quality measurements are satisfactory. Temperature was maintained at 19 °C to 22 °C throughout the test, while there were no treatment related differences for oxygen concentration or pH.
Throughout the test the light intensity was observed to be in the range 460 to 516 Lux.
Observations on Test Item Solubility
At the start and throughout the test all control and test solutions were observed to be clear colorless solutions.

Results with reference substance (positive control):

A positive control (Envigo Study Number XT01HS) used potassium dichromate as the reference item at concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.
Exposure conditions for the positive control were similar to those in the definitive test.
The results from the positive control with potassium dichromate were within the normal range for this reference item*.

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the 2-Propanamine, compd. with boron trifluoride, reaction products with Bu glycidyl ether to the freshwater invertebrate Daphnia magna has been investigated using OECD 202 method and gave a 48-Hour EC50 value of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.

Executive summary:

The acute toxicity of the 2-Propanamine, compd. with boron trifluoride, reaction products with Bu glycidyl ether  to the freshwater invertebrate Daphnia magna has been investigated using OECD 202 method and gave a 48-Hour EC50 value of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.

Description of key information

The acute toxicity of the 2-Propanamine, compd. with boron trifluoride, reaction products with Bu glycidyl ether  to the freshwater invertebrate Daphnia magna has been investigated using OECD 202 method and gave a 48-Hour EC50 value of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L