Boron, (benzenemethanamine)trifluoro-, (T-4)-, 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 each loading rate WAF test group from the bulk test preparation at 0 hours and from the pooled replicates (replicates R1 – R4) at 48 hours for quantitative analysis. Samples were stored frozen prior to analysis.
Duplicate samples were taken 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:

no

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 loading rates to be used in the definitive test were determined by a preliminary range-finding test.
In the range-finding test Daphnia magna were exposed to a series of nominal loading rates of 1.0, 10 and 100 mg/L.
Nominal amounts of test item (5.0, 50 and 500 mg) were each separately added to the surface of 5 liters of test water to give the 1.0, 10 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the 1.0, 10 and 100 mg/L loading rate WAFs.
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 150 mL test and control vessel contained 100 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 loading rate WAF 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.

Definitive Test
Based on the results of the range-finding test the following loading rates were assigned to the definitive test: 6.25, 12.5, 25, 50 and 100 mg/L.
Nominal amounts of test item (31.3, 62.5, 125, 250 and 500 mg) were each separately added to the surface of 5 liters of test water to give the 6.25, 12.5, 25, 50 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. The aqueous phase or WAF was removed by mid-depth siphoning (the first 75-100 mL discarded) to give the 6.25, 12.5, 25, 50 and 100 mg/L loading rate WAFs. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present.
The concentration and stability of the test item in the test preparations were verified by chemical analysis at 0 and 48 hours (see Annex 5).

Exposure Conditions
As in the range-finding test 150 mL glass beakers containing approximately 100 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. The test preparations were not renewed during the exposure period.

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:

EL50

Effect conc.:

57 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

NOELR

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOELR

Effect conc.:

50 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 1.0 mg/L loading rate WAF, however, immobilization was observed at 10 and 100 mg/L loading rate WAF.
Based on this information loading rates of 6.25, 12.5, 25, 50 and 100 mg/L were selected for the definitive test.
Chemical analysis of the test preparations at 0 hours showed measured test concentrations to range from 0.56 to 92 mg/L. There was no significant change in the measured concentrations at 48 hours indicatingthat the test item was stable under test conditions.
Definitive Test
Chemical Analysis of Test Loading Rates
Chemical analysis of the freshly prepared 12.5, 25, 50 and 100 mg/L loading rate WAF test preparations at 0 hours showed measured test concentrations to range from 8.7 to 86 mg/L. Chemical analysis of the aged test preparations at 48 hours showed measured test concentrations to range from 9.3 to 86 mg/L.
The dissolved test item may have been one or several components of the test item. Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.
Immobilization Data

Analysis of the immobilization data by Probit analysis using Linear Maximum-Likelihood regression at 24 and 48 hours based on the nominal loading rates gave the following results:
Time (h) EL50 (mg/L Loading Rate WAF) 95% Confidence limits (mg/L Loading Rate WAF)
24 >100 Not possible to determine
48 57 Not possible to determine
The No Observed Effect Loading rates after 24 and 48 hours exposure were 100 and 25 mg/L loading rate WAF, respectively. Correspondingly, the Lowest Effect Loading rate at 48 hours was considered to be 50 mg/L loading rate WAF.

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:

Exposure of the freshwater invertebrate Daphnia magna to the Boron, (benzenemethanamine)trifluoro-,(t-4)-, reaction products with bu glycidyl ether has been investigated and gave the following effect levels for 48 h exposure - EL50=57 mg/l Loading Rate WAF, NOEL = 25 mg/l Loading Rate WAF, LOEL=50 mg/l Loading Rate WAF.

Executive summary:

Exposure of the freshwater invertebrate Daphnia magna to the Boron, (benzenemethanamine)trifluoro-,(t-4)-, reaction products with bu glycidyl ether has been investigated and gave the following effect levels for 48 h exposure - EL50=57 mg/l Loading Rate WAF, NOEL = 25 mg/l Loading Rate WAF, LOEL=50 mg/l Loading Rate WAF.

Description of key information

Exposure of the freshwater invertebrate Daphnia magna to the Boron, (benzenemethanamine)trifluoro-,(t-4)-, reaction products with bu glycidyl ether has been investigated and gave the following effect levels for 48 h exposure - EL50=57 mg/l Loading Rate WAF, NOEL = 25 mg/l Loading Rate WAF, LOEL=50 mg/l Loading Rate WAF.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Remarks:

48 h exposure - EL50=57 mg/l Loading Rate WAF

Effect concentration:

57 mg/L