Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Oct 2000 - Jan 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: According to OECD guideline under GLP
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Principles of method if other than guideline:
EC guideline Part 2 - Testing Methods, Part C.1
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Canrenone
- Analytical purity: 98.4%
- Lot/batch No.: 90515001

Sampling and analysis

Analytical monitoring:
yes

Test solutions

Vehicle:
no

Test organisms

Test organisms (species):
Pimephales promelas

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h

Results and discussion

Effect concentrations
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
7.7 - 15.2 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mortality

Any other information on results incl. tables

Table 1: Mortality of fish exposed to canrenone over 96 hours

 Dilution of canrenone    concentration [mg/L]    3 hours    6 hours    24 hours    48 hours    72 hours    96 hours  
0                     
 control (tap water)   0 0 0 0 0 0 0
 1:80   0,1 0 0 0 0 0 0
 1:40   0,3 0 0 0 0 0 0
 1:20   0,8 0 0 0 0 0 0
 1:10   1,5 0 0 0 0 0 0
 1:5   3,2 0 0 0 0 0 0
 1:2,5   7,7 0 0  1)    1)    1)    1)  
 Saturated solution (limit test)    15,2 (TOC analysis)    10 (after 20 min)    -   -   -   -   - 
 
 1) all fish (10 of 10) sacrified in moribund status  

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The LC50/96 hours of canrenone can be expected between 7.7 and 15.2 mg/L taking into account
that all fish died in a saturated solution (which was measured during the limit test to contain
15.2 mg/L). Although mortality is only expected at the saturated solution, the results of this
study showed that slight effects occurred at concentrations from 1.5 mg/L upwards and severe
effects at concentrations from 3.2 mg/L upwards.
Therefore, canrenone is toxic to fish, since effects were observed below 10 mg/L.
Executive summary:

The purpose of this study was to detennine the acute toxicity including the LCso/96 hours of canrenone (ZK 57587) to the fathead minnow Pimephales prome1as. Canrenone is an intermediate of the synthesis of Eplerenone, a carbolactone derivative. The study was conducted in accordance with the test guideline OECD 203, and the EC guideline Part 2 - Testing Methods, Part C.1. In a preceeding limit test, which is not reported in detail, all fish died in an saturated solution.

Ten fish (fathead minnow Pimephales promelas) were used for each test solution of canrenone and ten for the control group. The fish were exposed for a period of 96 hours under static conditions. Mortality and visible abnonnalities were recorded at approximately 3, 6, 24, 48, 72 and 96 hours. For the preparation ofthe test solution a suspension (1 liter) with a nominal Ioading of 800 mg/L canrenone was ultrasonified for 30 minutes. Thereafter the suspension was filied up to 8 liters and stirred for approximately 24 hours. This suspension was filtered through a glassfibre filter. After filtration, the resulting solution served as stock solution. It was diluted 1:2.5,1:5, 1:10, 1 :20, 1 :40, and 1 :80 to obtain the final test solutions.

The concentration ofthe test solutions were analyzed by HPLC at 0,48 and 96 hours. The concentration of canrenone in the stock solution was determined by TOC only. It was 15.2 g/L.

No mortality but abnormal behavior was observed in the dilutions 1 :2.5, 1:5 and 1: 10 of canrenone. Fish at the highest concentration had to be killed after 24 hours in a moribund status for humane reasons.

The observations showed that, although not mortality occurred up to a eoneentration of 7.7 mgIL (1:2.5 dilution), slight effeets were seen at 1.5 mg/L (1: 10 dilution) and severe effects from 3.2 mg/L (1:5 dilution) upwards. The highest concentration without effects was 0.8 mg/L (dilution 1 :20).