Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
3rd to 7th February 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Propietary study conducted according to guidelines and GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
e-Caprolactone, colourless liquid, batch no. WA 2809/0791. Purity: >99%. Solubility: >1000 mg/l.
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No information available

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
At test initiation and at 48 hours and 96 hours after initiation, and when all fish in an aquarium had died, five samples of each test solution (5 x 0.5ml) were taken and combined to one final sample. This was done in duplicate. Control solutions were not sampled after 48 hours. To each sample 625µl acetonitrile was pipetted to bring the sample to mobile phase composition. The samples were analysed directly with HPLC.

Test solutions

Vehicle:
yes
Details on test solutions:
6.0g e-Caprolactone was dissolved in 6l ISO-water. The solution was stirred during addition and for 10 minutes after. Samples of 1500, 750, 375, 187 and 93ml were made up to 3l with ISO-water. The remaining portion of 3l stock solution was used for the highest concentration. Final concentrations were: 1000, 500, 250, 125, 62, 31 and 0mg/l. The ISO-water was prepared and aerated at least 24 hours prior to use.

Test organisms

Test organisms (species):
Poecilia reticulata
Details on test organisms:
Guppies (Poecilia reticulata), obtained from a commercial supplier (RASBORA, The Netherlands). The guppies arrived 3 weeks prior to the start of the main study. They were held in a plastic container of 40x60x40cm containing approximately 80l reconstituted ISO-water, which was continuously filtered and aerated. The aquaria were kept in a room with a temperature of 21±1oC. Fish were fed troutfeed and waterfleas daily.
The guppies used in the test were 2.1-3.2cm long, and the mean weight was 0.28g per fish.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
Total exposure duration was 96 hours

Test conditions

Hardness:
~250mg/l
Test temperature:
21.3-22.5oC
pH:
7.5-7.8
Dissolved oxygen:
7.3-8.4 mg/l
Salinity:
Not relevant - freshwater test
Nominal and measured concentrations:
Nominal concentrations; 0 (control), 31, 62, 125, 250, 500, 1000 mg/l (see below for a table listing the measured concentrations).
Details on test conditions:
One test aquarium was used per concentration, each aquarium contained 10 fish. The test aquaria were placed in a climate chamber where the temperature was maintained at 22±1oC. The fish were not fed during the test. The test solutions were aerated during the test. The light regime was 16h light and 8h dark.
Reference substance (positive control):
yes
Remarks:
potassium bichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
900 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: CL could not be determined because no data between 0 or 10 and 100% mortality were available
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
480 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: CL could not be determined because no data between 0 or 10 and 100% mortality were available
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
280 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: CL could not be determined because no data between 0 or 10 and 100% mortality were available
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
280 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: CL could not be determined because no data between 0 or 10 and 100% mortality were available
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
250 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
None of the fish in the control solution died in 96 hours. Mortality was only seen at concentrations of 250mg/l (10% mortality at 96 hours), 500mg/l (100 % mortality at 72 hours) and 1000mg/l (100% mortality at 24 hours). Fish that died at the higher concentrations showed uncontrolled movement and hypoactivity before death.
4h after exposure to 1000mg/l e-Caprolactone, 2 fish showed uncontrolled movement, 7 were less active and 1 was dead.
24h after exposure to 500mg/l e-Caprolactone, 2 fish out of 10 showed uncontrolled movement.
48h after exposure to 500mg/l e-Caprolactone, 3 fish showed uncontrollable movement, and 7 were dead.
No effects were observed at the lower concentrations.
Results with reference substance (positive control):
An acute toxicity study is conducted yearly at the laboratory with P. reticulata, using potassium bichromate as the reference substance. The most recently conducted test was conducted during November 1990. The 96h EC50 was 135 mg/l with a 95% confidence interval of 115-150 mg/l.
Reported statistics and error estimates:
The probit analysis model was used to determine the LC50 value from the percentage mortality and nominal test concentration. Because no data between 0 or 10 and 100% mortality were available, no confidence limits could be defined.

Any other information on results incl. tables

The NOEC was determined to be 250mg/l. The minimum concentration causing 100% mortality was 500mg/l.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of e-Caprolactone to guppies (Poecilia reticulata) was tested according to OECD guideline 203 at nominal test concentrations of 1000, 500, 250, 125, 62, 31 and 0 mg/l in ISO-water. The 96 hour LC50 was 280 mg/l. The NOEC for mortality and other effects was 250 mg/l. The minimum concentration causing 100% mortality was 500mg/l. The 96 hour LD50 was 280mg/l.
Executive summary:

The acute toxicity of e-Caprolactone to guppies (Poecilia reticulata) was tested according to OECD guideline 203 at nominal test concentrations of 1000, 500, 250, 125, 62, 31 and 0 mg/l in ISO-water. The 96 hour LC50 was 280 mg/l. The NOEC for mortality and other effects was 250 mg/l.