Boronic acid, (4-ethylphenyl)-

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:

2007-10-29 to 2008-02-25

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:

adopted July 17, 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 4.27, 9.39, 20.66,45.45 and 100.0 mg/L

- Sampling method: Samples were collected for analysis at the start (0-h) and end (96-h) of the experiment.

- Sample storage conditions before analysis: All samples were frozen after the sampling and stored intermediate in an icebox.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
Before exposure of the fish, the test medium (reconstituted water and test material) was freshly prepared: The calibrated flask with test material and vehicle, reconstituted water, was treated in an ultrasonic device for 1 hour. Subsequently, the preparation was stirred with a magnetic stirrer for further 23 hours. Thereafter, the formulation was given through a nutsch filter (pore size >10 - <16 µm). The filtrate was used for the study.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Source: West Aquarium
- Size: 2.0 + 1.0 cm
- Weight at study initiation: 6.8 g (control group) and 6.6 to 7.0 g (test material groups)

ACCLIMATION
- Acclimation period: > 14 days
- Acclimation conditions: same as test
- Type and amount of food during acclimation: Tetra Min® diet, daphnia, artemia or frozen mosquito larva were supplied daily.
- Health during acclimation (any mortality observed): the mortality of the test fish batch was lower than 5%.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

15° dH (about 267 mg/L CaCO3)

Test temperature:

24 to 25 °C

pH:

7.76 – 8.4

Dissolved oxygen:

> 60 % oxygen saturation throughout the study

Nominal and measured concentrations:

Nominal conc.: 4.27, 9.39, 20.66, 45.45 and 100.00 mg/L;
Mean measured: 93.7 - 98.2 % at the start and 301.9 - 107.4 % at the end of the 96 hours exposure period.

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria
- Type: open
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water
- Ca/mg ratio: 4:1
- Culture medium different from test medium: no


OTHER TEST CONDITIONS
- Adjustment of pH: no


EFFECT PARAMETERS MEASURED: fish were examined at the termination of test for sub-lethal and lethal effects


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

45.45 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.:

15.66 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % CI: 11.20 - 21.90 mg/L

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

4.27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

For the test material, the following LC50 values with the 95 % confidence intervals based on the nominal concentrations for zebrafish were determined:

24-h LC50: 30.64 mg/L (22.43 - 41.86 mg/L)
48-h LC50: 30.64 mg/L (22.43 - 41.86 mg/L)
72-h LC50: 30.64 mg/L (22.43 - 41.86 mg/L)
96-h LC50: 15.66 mg/L (11.20 - 21.90 mg/L)

For the test material, the following LC0 and LC100 values based on the nominal concentrations for zebrafish were determined:

24-h LC0: 20.66 mg/L
48-h LC0: 20.66 mg/L
72-h LC0: 20.66 mg/L
96-h LC0: 4.27 mg/L

24-h LC100: 45.45 mg/L
48-h LC100: 45.45 mg/L
72-h LC100: 45.45 mg/L
96-h LC100: 45.45 mg/L

Sublethal observations / clinical signs:

The summaries of the observations at different time points are given in the following tables.

Control group

Symptom	Observed fish / time point
	3h	24 h	48 h	72 h	96 h
No abnormalities detected	7	7	7	7	7
Deadfish	0	0	0	0	0

 

Concentration: 4.27 mg/L

Symptom	Observed fish / time point
	3h	24h	48 h	72 h	96 h
No abnormalities detected	7	7	7	7	7
Dead fish	0	0	0	0	0

 

Concentration: 9.39 mg/L

Symptom	Observed fish / time point
	3h	24h	48 h	72 h	96 h
Fish lying on side/back	0	0	0	1	0
Fish near the bottom of the aquarium	0	0	0	3	6
No abnormalities detected	7	7	7	3	0
Dead fish	0	0	0	0	1

 

Concentration: 20.66 mg/L

Symptom	Observed fish / time point
	3h	24h	48 h	72 h	96 h
Fish lying on side/back	0	0	0	4	1
Abdominal Position	0	0	6	2	0
Fish near the bottom of the aquarium	4	5	6	1	1
No abnormalities detected	3	2	1	0	0
Dead fish	0	0	0	0	5

 

Concentration: 45.45 mg/L

Symptom	Observed fish / time point
	3h	24h	48 h	72 h	96 h
Fish lying on side/back	4	0	0	0	0
No abnormalities detected	0	0	0	0	0
Dead fish	3	7	7	7	7

 

Concentration: 100.00 mg/L

Symptom	Observed fish / time point
	3h	24 h	48 h	72 h	96 h
No abnormalities detected	0	0	0	0	0
Dead fish	7	7	7	7	7

 

Mortality

Nominal concentration [mg/L]	Mortality
	24 h	48 h	72 h	96 h
Control group	0/7	0/7	0/7	0/7
4.27	0/7	0/7	0/7	0/7
9.39	0/7	0/7	0/7	1/7
20,66	0/7	0/7	0/7	5/7
45.45	7/7	7/7	7/7	7/7
100.00	7/7	7/7	7/7	7/7

Deaths occurred from 9.39 up to 100.00 mg/L and started 3 hours after exposure.

 

LC₅₀ value

The LC₅₀ value is the calculated mean lethal concentration, which is expected to kill 50 percent of the test fish.

	24 h	48 h	72 h	96 h
Nominal LC50	30.64 mg/L	30.64 mg/L	30.64 mg/L	15.66 mg/L

 

Analytical results

Nominal concentration [mg/L]	Start (t = 0 hour)		End (t = 96 hours)
	Mean analytical concentration [mg/L]	%of nominal concentration	Mean analytical concentration [mg/L]	% of nominal concentration
4.27	4.00	93.7	4.35	101.9
9.39	9.06	96.7	9.95	106.0
20.66	20.09	97.2	22.02	106.6
45.45	44.62	982	48.81	107.4
100.00	97.90	97.9	106.46	106.5

 

Validity criteria fulfilled:

yes

Conclusions:

The test item was tested in an open static test system. Under the test conditions, the 96-h LC50 value for zebrafish was determined to be 15.66 mg/L (95 % CI: 11.20 - 21.90 mg/L).

Executive summary:

The acute toxicity (LC₅₀) of the test item on zebrafish, Danio rerio was determined in a study according to OECD TG 203 and EU method C1. For this purpose, 7 zebrafish per test item group and per control group were exposed over 96 hours, under defined conditions. The fish were observed for signs of toxicity or death for 96 hours. Zebrafish of the test material groups were exposed to a saturated aqueous test item solution of nominal 4.27, 9.39, 20.66, 45.45 and 100.00 mg/L in an open static system. The analytical determination of the test item concentration was performed using HPLC with UV-detection. Samples were collected at the start (0-h) and at the end (96-h) of the experiment. The limit of quantification of the analytical method was 0.01 mg/L. The analytically determined concentrations were between 93.7 - 98.2 % at the Start and 101.9 - 107.4 % at the end of the 96 hour exposure period. During the experimental phase of the study, the test material concentrations could be maintained at > 80 % of the initial concentrations. Therefore, the LC₅₀ values were calculated using the nominal concentrations. No signs of toxicity were seen in the control group and in the zebrafish exposed to nominal concentration of 4.27 mg/L. Signs of toxicity were observed at nominal concentrations ranging from 9.39 to 100.00 mg/L. They were observed immediately after exposure and lasted up to the end of the exposure. Signs of toxicity consisted of: fish lying on the side/back, fish near the bottom of the aquarium and abdominal position. Under the test conditions, the 96-h LC₅₀ value for zebrafish was determined to be 15.66 mg/L (95 % CI: 11.20 - 21.90 mg/L).

Description of key information

The test item was tested in an open static test system. Under the test conditions, the 96-h LC₅₀ value for zebrafish was determined to be 15.66 mg/L (95 % CI: 11.20 - 21.90 mg/L) (reference 6.1.1-1).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

15.66 mg/L

Additional information

The acute toxicity (LC₅₀) of the test item on zebrafish, Danio rerio was determined in a study according to OECD TG 203 and EU method C1. For this purpose, 7 zebrafish per test item group and per control group were exposed over 96 hours, under defined conditions. The fish were observed for signs of toxicity or death for 96 hours. Zebrafish of the test material groups were exposed to a saturated aqueous test item solution of nominal 4.27, 9.39, 20.66, 45.45 and 100.00 mg/L in an open static system. The analytical determination of the test item concentration was performed using HPLC with UV-detection. Samples were collected at the start (0-h) and at the end (96-h) of the experiment. The limit of quantification of the analytical method was 0.01 mg/L. The analytically determined concentrations were between 93.7 - 98.2 % at the Start and 101.9 - 107.4 % at the end of the 96 hour exposure period. During the experimental phase of the study, the test material concentrations could be maintained at > 80 % of the initial concentrations. Therefore, the LC₅₀ values were calculated using the nominal concentrations. No signs of toxicity were seen in the control group and in the zebrafish exposed to nominal concentration of 4.27 mg/L. Signs of toxicity were observed at nominal concentrations ranging from 9.39 to 100.00 mg/L. They were observed immediately after exposure and lasted up to the end of the exposure. Signs of toxicity consisted of: fish lying on the side/back, fish near the bottom of the aquarium and abdominal position. Under the test conditions, the 96-h LC₅₀ value for zebrafish was determined to be 15.66 mg/L (95 % CI: 11.20 - 21.90 mg/L).