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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 July 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Confirmatory analysis is inadequate for test solutions
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
A sample of 1 ml was taken from the old and new test solutions of the highest four concentrations on each day of the study. These samples were qualitatively analyzed by HPLC. As Capa-203 consists of a mixture of components the chromatogram measured at 218 nm shows a number of peaks.

As no samples of the pure components (i.e. analytical standards) of Capa-203 were available no quantitative conclusions could be drawn.
The chromatogram of freshly prepared and 24 hours old solutions were only compared qualitatively.

From each freshly prepared clear saturated solution (stock solution) a sample of 50 ml was taken for determination of the total organic carbon content (TOC). These determinations were done by Centrilab, Soest, The Netherlands. To avoid precipitation the solutions were diluted 1:1 with ISO-water before shipment.
Vehicle:
no
Details on test solutions:
The test was conducted using semi-static conditions (media renewal every 24 hours).

In order to simulate an accidental spill a saturated solution of Capa-203 was made by adding 6.5 g Capa-203 to 6.5 L of ISO-water under stirring by an ultra-turrax to provide a nominal stock solution at 1 g/L. Stirring was continued for 10 minutes which resulted in a turbid emulsion.

A clear solution was obtained by filtration through a nylon membrane filter (Gelman, Nylaflo, 0.45 pm) under vacuum. Dilutions were made by making up 1500, 750, 375,187 and 95 mL of the saturated solution to 3 L with ISO-water.

A control with ISO-water was included in the test.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
The zebra fish used on the study originated from a commercial supplier, Ruinemans, Montfoort, The Netherlands. The batch of fish were received on the 20 May 1992 and held in a plastic container 40 x 60 x40 cm containing ca. 80L of reconstituted ISO-water which was continuously filtered and aerated. The aquaria were kept in a room at 21 ± 1˚C.

The fish were fed trout feed and Daphnia (waterfleas).

Stock fish held at 21 ± 1˚C test conducted at ca. 23˚C.

The fish used in the final test were 2.3 - 2.9 cm long and had a mean weight of 0.25 g per fish.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
Not applicable
Hardness:
Hardness of ISO reconstituted water ca. 250 mg/L.
Test temperature:
The temperature of the test solutions varied between 22.0 and 23.8 °C.
pH:
The pH of the test solutions ranged from 7.3 to 8.0 during the test, except in one aquarium with dead fishes where the pH was 6.6.
Dissolved oxygen:
The dissolved oxygen concentration was between 5.3 and 9.5 mg/L.
In the aquarium with dead fishes (140 mg/L) the O2 concentration dropped to 1.8 mg/L.
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal concentrations: 0, 19, 38 , 75, 150, 300 and 600 mg/L.
Measured concentrations: 0, 18, 35, 70, 140, 289 and 560 mg/L (Based on TOC analysis of initial saturated solution and calculation).
As % nominal: NA, 95, 92, 93, 93, 96 and 93%.
Details on test conditions:
The initial saturated stock was prepared to simulate and accidental spill.

Test media reconstituted ISO water:
Deionised water
CaCl2.2H2O - 294 mg/L
MgSO4. 7H2O - 123 mg/L
NaHCO3 - 64 mg/L
KCl - 5.8 mg/L
pH 7.8

Test solutions were aerated during the test and the light regime was 16 h light and 8 h dark.
Reference substance (positive control):
yes
Remarks:
Potassium bichromate
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
72 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
other: Total organic carbon
Basis for effect:
mortality
Remarks on result:
other: 35 and 140 mg/L
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
140 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
other: Total organic carbon
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
35 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
other: Total organic carbon
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
<= 18 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
other: Total organic carbon
Basis for effect:
behaviour
Details on results:
All fishes exposed to 280 and 560 mg Capa-203/1 were dead within 3 hours, and were removed from the aquaria. All fishes exposed to 140 mg/L died within 24 hours, but stayed in the aquarium during this whole period which influenced the pH and O2 concentration of the test solution.
40% mortality occurred after 96 hours when zebra fishes were exposed to 70 mg Capa-203/L. At lower concentrations no mortality was observed.
As the mortality in the controls is below 10%, the study meets the criterion.

Sub-lethal observations were identified at all test concentrations during the test, the sub lethal effects were uncontrolled and hypoactivity.
Results with reference substance (positive control):
Once a year an acute toxicity test with Brachydanio rerio and the reference substance, potassium bichromate, is conducted. The most recent test (as of the date when the current study was performed) was conducted in April 1992. The 96 h EC50 found in this reference test was 142 mg/L (study number C.REF.51.006b).
Reported statistics and error estimates:
The percentage mortality was plotted against the test concentration using a probit analysis model. From the results it was concluded that the 96-h LC50 was 72 mg/L, with confidence limits of 35 and 140 mg/L.

Fish mortality and sub-lethal observation during the study are presented below.

Calculated test concentrations (mg/L)

Number of surviving zebra fish per test vessel

 

% mortality

0 h

3 h

24 h

48 h

72 h

96 h

0 (control)

10

10

10

10

10

10

0

18

10

10

10

104)

104)

105)

0

35

10

10

10

103)

103)

106)

0

70

10

101)

7

63)

63)

63)

40

140

10

12)

0

0

0

0

100

280

10

0

0

0

0

0

100

560

10

0

0

0

0

0

100

1) one fish showed uncontrolled movement

2) one fish showed hypo activity

3) all fishes showed uncontrolled movement and hypo activity

4) 5 fishes showed uncontrolled movement and hypo activity

5) 2 fishes showed uncontrolled movement and hypo activity

6) 4 fishes showed uncontrolled movement and 10 fishes showed hypo activity

Validity criteria fulfilled:
yes
Conclusions:
Zebra fish were exposed to Capa-203 at calculated concentrations of 18, 35, 70, 140, 280 and 560 mg/L in a semi-static 96 -hour acute toxicity test according to OECD 203 guidelines. The test design included an untreated control comprising of ISO dilution water only.
The 96-hour LC50 determined was 72 mg Capa-203/L (based calculated concentrations).
Executive summary:

Zebra fish were exposed to Capa-203 at calculated concentrations of 18, 35, 70, 140, 280 and 560 mg/L in a semi-static 96-hour acute toxicity test according to OECD 203 guidelines. The test design included an untreated control comprising of ISO dilution water only.

The 96-hour LC50 determined was 72 mg Capa-203/L (based calculated concentrations).

This test was conducted in accordance with the 1984 version of the OECD 203 guideline. The current guideline of 1994 states that there must be evidence that the concentration of the substance being tested has been satisfactorily maintained and preferably it should be at least 80% of the nominal concentration throughout the test. This reported study was conducted prior to the issue of the 1994 OECD 203 document and therefore samples of test media have not been determines using an appropriate analytical method.

Description of key information

The key study for acute toxicity of 2-Oxepanone, polymer with 1,4-butanediol to fish found a 96-hour LC50 of 72 mg/L.

Key value for chemical safety assessment

LC50 for freshwater fish:
72 mg/L

Additional information

Zebra fish were exposed to Capa-203 at calculated concentrations of 18, 35, 70, 140, 280 and 560 mg/L in a semi-static 96-hour acute toxicity test according to the OECD 203 guideline. The test design included an untreated control comprising of ISO dilution water only.

The 96-hour LC50 determined was 72 mg Capa-203/L (based on calculated concentrations).

This test was conducted in accordance with the 1984 version of the OECD 203 guideline. The current guideline of 1994 states that there must be evidence that the concentration of the substance being tested has been satisfactorily maintained, and preferably it should be at least 80% of the nominal concentration throughout the test. This reported study was conducted prior to the issue of the 1994 OECD 203 document and therefore samples of test media have not been determines using an appropriate analytical method.