3-Fluorphenylboric acid

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

Study period:

06 Jul 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

At the start of the exposure and at the renewal (24 and 48 h), samples of the freshly prepared test item concentrations were taken and analyzed. At renewal and at the end of the exposure (24 and 48 h), sampled of the 24 h old media were taken directly fron the test vessels and analyzed.

Vehicle:

no

Details on test solutions:

Stock solution:
A stock solution (100 mg/L of the test item were weighed out) was freshly prepared with dilution water prior to the start of the exposure (0 h) and prior to the renewal of the test solutions (24 h). The stock solution was mixed thoroughly by manual agitation.

Test concentrations:
5 test item concentrations in a geometirc series with a separation factor of 2, prepared by dilution of the stock solution of 100 mg/L with dilution water were tested as follows: 6.25, 12.5, 25.0, 50.0 and 100 mg/L.

Control:
Dilution water without test item incubated under the same semi-static conditions as the test group.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna STRAUS (Clone 5)
- Source: Institut für Wasser-, Boden und Lufthygiene (WaBoLu), Berlin, Germany
- Age: < 24 h

FEEDING DURING TEST
- Food type: 5 x per week ad libitum with a mix of unicellular green algae

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

0 h: 171 mg CaCO3/L
24 h: 173 mg CaCO3/L

Test temperature:

20.2 - 20.4 °C

pH:

0 h: 7.42
24 h: 7.44

Dissolved oxygen:

0 h: 8.88 mg/L
24 h: 8.96 mg/L

Conductivity:

0 h: 437 µS/cm
24 h: 428 µS/cm

Nominal and measured concentrations:

Nominal concentrations: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
Mean measured concentrations:6.68 - 13.1 - 25.9 - 52.1 - 103.5 mg/L (details see Table 2 below)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 (ID) x 7 (H) cm)
- Type: loosely covered with watch glasses
- Material, size, fill volume: Glass beakers, 50 mL capacity, 20 mL
- Aeration: no
- Renewal rate of test solution: The test solutions were renewed after 24 h. For this purpose, a second set of test vessels was filled with the freshly prepared test solutions and the daphnids were transferred by pipette.
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: Culture Medium Elendt M4 according to OECD 202, Annex 3 (2004)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 1500 Iux

EFFECT PARAMETERS MEASURED
Immobilization was determined in all groups after 24 and 48 hours. An animal was considered immobile, if it was not able to swim in the water phase within 15 seconds after gentle agitation of the test vessel.

RANGE-FINDING STUDY
- Test concentrations: 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: yes, EC50 between 10 and 100 mg/L

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

35.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

50.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

26.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The study was performed according to OECD Guideline 202 (2004). The validity criteria were fulfilled:
• In the control group, no daphnids were immobilized or showed any signs of disease or stress, e.g. discoloration or unusual behavior such as trapping on the surface of the water, during the 48-hour test period (required: not more than 10% of the daphnids immobilized in the control).
• The dissolved O2 concentration in the 24-hours old media at the end of the exposure intervals was > 7.97 mg/L (required: a 3 mg/L at the test end of the exposure intervals) in the test vessels of all tested concentration levels and the control.

Results with reference substance (positive control):

The percentage of immobility for the reference item potassium dichromate (SIGMA-ALDRICH, batch number MKBV0900V, purity 99.0%, CAS RN 7778-50-9) was determined after 24 hours from 2017-07-11 to 2017-07-12. The EC50 value was determined to be 1.31 mg/L, which is in the valid range (0.6 - 2.4 mg/L)

Reported statistics and error estimates:

The EC10- and the EC50-values after 24 hours were calculated by sigmoidal dose-response regression with the software GraphPad Prism5. The respective 95% confidence limits were calculated from the standard error and the t-distribution. All calculations were carried out from the best-fit values with the software GraphPad Prism5. The EC10- and the EC50-values after 48 hours were calculated by linear dose-response regression with the software GraphPad PrismS. Since only 0 and 100% immobilization occurred, only the highest test item concentration resulting in 0% immobilization and the lowest concentration level resulting in 100% immobilization were used for calculation of the EC10 and the EC50 and as their 95% confidence limits.

Table 1: Immobilization rates after 24 and 48 h

Nominal test item concentration [mg/L]	IMMOBILIZATION [%]
	24 hours					48 hours
	Replicates					Replicates
	1	2	3	4	MV	1	2	3	4	MV
100	100	100	100	100	100	100% mortality after 24 hours
50.0	60	40	40	40	45	100	100	100	100	100
25.0	0	0	0	0	0	0	0	0	0	0
12.5	0	0	0	0	0	0	0	0	0	0
6.25	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

Table 2: Measured Concentrations and Percent of Nominal Concentration of the Test Item

Sampling date	2017-07-05 Fresh media, 0 hours		2017-07-06 Old media, 24 hours		2017-07-06 Fresh media, 24 hours		2017-07-07 Old media, 48 hours
Start of analysis	2017-07-05		2017-07-06		2017-07-06		2017-07-07
Nominal test item concentration [mg/L]	Art. 281382 (3-Fluoro-phenylboronic acid)
	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%
100	103	103	104	104	Not determined, due to 100% immobilization after 24 hours
50.0	51.1	102	49.9	100	51.7	103	55.6	111
25.0	25.4	101	26.1	104	25.7	103	26.6	106
12.5	12.6	101	12.8	102	13.4	107	13.7	109
6.25	6.48	104	6.49	104	6.84	109	6.91	111
Control	< LOQ		< LOQ		< LOQ		< LOQ

Meas. conc. = measured concentration of the test item, mean value of 2 injections, dilution factors taken into account

% = percent of the nominal concentration of the test item

LOQ = limit of quantification of the analytical method (0.1 mg/L of the test item)

 

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item, the 48 h EC50 for Daphnia magna was 35.4 mg/L (95% confidence limits: 25.0 - 50.0 mg/L).

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the test item were determined at the test facility according to OECD 202 (2004). The study was conducted under semi-static conditions over a period of 48 hours with 5 concentrations of the test item in the range of 6.25 to 100 mg/L, prepared in a geometric series with a separation factor of 2. All tested concentration levels were visually clear throughout the exposure. Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control. The concentrations of the test item were analytically verified via HPLC-DAD in fresh media of all concentration levels and in the control at the start of the exposure and at renewal (0 and 24 hours) and in 24-hours old media at renewal and at the end of the test (24 and 48 hours). The measured concentrations of the test item in fresh media at the start of the exposure and at renewal (0 and 24 hours) were in the range of 101 to 109% of the nominal values, and 100 to 111% in old media at renewal and at the end of the test (24 and 48 hours). The measured test item concentrations were all within ± 20% of the nominal concentrations. This indicates that the test item concentrations were successfully maintained for the duration of the test. Therefore, all effect concentrations are based on the nominal concentrations of the test item. The validity criteria of the test guidelines were fulfilled. The 48 h EC50 for Daphnia magna was 35.4 mg/L (95% confidence limits: 25.0 - 50.0 mg/L), the EC10 was 26.8 mg/L.

Description of key information

Based on the nominal concentrations of the test item, the 48 h EC50 for Daphnia magna was 35.4 mg/L (95% confidence limits: 25.0 - 50.0 mg/L) (reference 6.1.3-1).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

35.4 mg/L

Additional information

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the test item were determined at the test facility according to OECD 202 (2004). The study was conducted under semi-static conditions over a period of 48 hours with 5 concentrations of the test item in the range of 6.25 to 100 mg/L, prepared in a geometric series with a separation factor of 2. All tested concentration levels were visually clear throughout the exposure. Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control. The concentrations of the test item were analytically verified via HPLC-DAD in fresh media of all concentration levels and in the control at the start of the exposure and at renewal (0 and 24 hours) and in 24-hours old media at renewal and at the end of the test (24 and 48 hours). The measured concentrations of the test item in fresh media at the start of the exposure and at renewal (0 and 24 hours) were in the range of 101 to 109% of the nominal values, and 100 to 111% in old media at renewal and at the end of the test (24 and 48 hours). The measured test item concentrations were all within ± 20% of the nominal concentrations. This indicates that the test item concentrations were successfully maintained for the duration of the test. Therefore, all effect concentrations are based on the nominal concentrations of the test item. The validity criteria of the test guidelines were fulfilled. The 48 h EC50 for Daphnia magna was 35.4 mg/L (95% confidence limits: 25.0 - 50.0 mg/L), the EC10 was 26.8 mg/L (reference 6.1.3-1).