Bontron E-108

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 December 2001 to 21 December 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): S178207.
- Substance type: White powder.
- Physical state: Solid.
- Composition of test material, percentage of components: 91.4% w/w total ligand, 25.9% w/w free ligand and 69.1% w/w for the strength of S178207.
- Lot/batch No.: NBZ 0155/44.
- Storage condition of test material: At ambient temperature in the dark, in the container in which it was received.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Aqueous samples for analysis were taken from the centre of the test solutions. They were diluted with tetrahydrofuran and analysed. The excess test solutions were sampled at 0-h, and one replicate of the dilution water control, solvent control and each test concentration was sampled at 96-h.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution (0.32 g of the test substance in 10 ml of dimethylforamide (DMF). This stock solution was ultrasonicated for 15 minutes giving a white, cloudy homogeneous suspension. The test solutions were prepared from the stock solution by addition of appropriate volumes to 20 litre of dilution water and stirring throughly. The nominal concentrations of the test solutions were 0.32, 0.56, 1.0, 1.8 and 3.2 mg/l. A solvent control was similarly prepared by the addition of 2000 ul of DMF to 20 litres of dilution water, to give a solvent concentration of 100 ul/l. Equalising volumes of DMF were added, where applicable, to each of the test exposure solutions to give a final DMF concentration of 100 ul/l in each exposure vessel. The 0.32 mg/l test solution was clear and colourless, all higher test solutions were all observed to be white and cloudy.
- Controls: DMF and water.
- Chemical name of vehicle: DMF for the preparation of a stock solution, and water as media for the test.
- Concentration of vehicle in test medium (stock solution): 100 ul/l.
- Evidence of undissolved material: The resultant solutions were clear and colourless, all higher test solutions were all observed to be white and cloudy.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow Trout.
- Source: West Country Trout, Trafalgar Farm, Boscastle, Cornwall, UK on 26 September 2001.
- Feeding during test: The fish were not fed during the course of the test.


ACCLIMATION
- Acclimation period: Minimum of 7 days.
- Acclimation conditions (same as test or not): At the test temperature (15 ± 1°C).
- Type of food: Commercial fish food.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not applicable.

Test conditions

Hardness:

48.3 mg/l of CaCo3.

Test temperature:

15 ± 1°C.

pH:

7.44 - 7.80.

Dissolved oxygen:

6.04 mg/l.

Salinity:

< 1%.

Nominal and measured concentrations:

0.32, 0.56, 1.0, 1.8 and 3.2 mg/l (nominal), and 0.31, 0.54, 0.95, 1.7 and 3.1 mg/l (measured).

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: Single borosilicate glass vessels of 27.5 litre maximum capacity and 20 litre working capacity.
- Aeration: Yes,
- No. of organisms per vessel: 10.
- No. of vessels per concentration (replicates): 1.
- No. of vessels per control (replicates): 1.
- Biomass loading rate: 0.48 mg/l.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated tap water.
- Chlorine: As Cl2. Free residual of < 2 ug/l, combined residual of < 2 ug/l.
- Alkalinity: As CaCO3. 22 mg/l,
- Conductivity:225 uS/cm.
- Intervals of water quality measurement: Monitored every week day or once a week.


OTHER TEST CONDITIONS
- Adjustment of pH: No.
- Photoperiod: Photoperiod of 16 hours fluorescent light and 8 hours darkness with 20 minute dawn and dusk transition periods.


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations for mortalities and symptom of toxicity were made at 3, 24, 48, 72 and 96 hours.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

2.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 2.1 - 4.2 mg/l

Effect concentrations 2

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

1.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 1.4 - 2.3 mg/l

Effect concentrations 3

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

1.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0.92 - 1.8 mg/l

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0.86 - 1.7 mg/l

Effect concentrations 5

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

< 3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: general symptoms of toxicity

Details on results:

There was a single mortality observed at 0.32 mg/l. Several fish exhibited symptoms of toxicity such as quiescence, sounding, loss of balance, dark discolouration, irregular and laboured respiration and generally weak behaviour. However, there were no mortalities or symptoms observed in the fish at 0.56 mg/l.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

Not reported.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table Analytical Results ^a

Nominal conc (mg/l)	Measured conc (mg/l)		Mean measured conc over the test duration^b (mg/l)	Mean measured conc as % of nominal
	0 hour	48 hour
Dilution water control	< 0.041	< 0.041	< 0.041	-
Solvent control	< 0.041	< 0.041	< 0.041	-
0.32	0.33	0.29	0.31	97
0.56	0.58^c	0.50^d	0.54	96
1.0	0.99	0.90	0.95	95
1.8	1.7	1.6	1.7	94
3.2	3.2	3.0	3.1	97

a All measurements are quoted to two significant figures and percentages are quoted to the nearest integer.

b Calculated using the arithmetic mean of the 0 - 96 hour results

c Mean of triplicate analysis (0.58, 0.57, 0.58 mg/l)

d Mean of triplicate analysis (0.49, 0.51, 0.49 mg/l)

The limit of detection in the study was 0.041 mg/l.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test material to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LC50 value of 1.2 mg/l with 95% confidence limits of 0.86 – 1.7 mg/l. The No Observed Effect Concentration was< 0.32 mg/l.

Executive summary:

Introduction. A study was performed to assess the acute toxicity of the test material, S178207, to rainbow trout Oncorhynchus mykiss,17 to 21 December 2001.  The method followed that described in the OECD Guidelines for Testing of Chemicals (April 1984) No 203, " Fish, Acute Toxicity Test", also considered acceptable when compared to requirements of the relevant US Environmental Protection Agency guideline and EU guideline. 

Methods: Rainbow trout Oncorhynchus mykiss, batch ref 01/40A were obtained from West Country Trout, Trafalgar Farm, Boscastle, Cornwall,UK, and used for the test. The test conditions are dilution water control, solvent control and nominal concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/l of the test material at a temperature of 15 ± 1ºC. The duration of the study was 96 hours.

 

Results. The 24 hour LC 50 was 2.7 mg/l (95%CI 2.1 – 4.2), 48 hour LC50 was 1.7 mg/l (95%CI 1.4 – 2.3), 72 hour LC50 was 1.2 mg/l (95%CI 0.92 – 1.8) and 96 hour LC50 was 1.2 mg/l (95%CI 0.86 – 1.7) based on moving average angle of fish. The 96 hour NOEC based on mortality was < 0.32 mg/l. There was a single mortality observed at 0.32 mg/l. Several fish exhibited symptoms of toxicity such as quiescence, sounding, loss of balance, dark discolouration, irregular and laboured respiration and generally weak behaviour. However, there were no mortalities or symptoms observed in the fish at 0.56 mg/l.