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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14th March 2001 to 16th March 2001
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: For the stock solution, 100 mg/L test material was dissolved in dilution water one day prior to application, stirred for 24 hours and filtered before use.
- Controls: Test medium and potassium dichromate.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain: STRAUS
- Source: Institut fur Wasser-, Boden-und Lufthygiene des Bundes-gesundheitsamtes, Corrensplatz, D-14195 Berlin
- Age at study initiation (mean and range, SD): <24 hours
- Method of breeding: In 2-3 L glass vessels approximately 1.8 L medium at 21°C in an incubator, 16 hours illumination.

ACCLIMATION
- Acclimation period: Daphnids were acclimatised for 2 hours under test conditions.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
256 CaCO3 mg/L
Test temperature:
21 ± 1°C
pH:
7.54-7.83 (0 h); 7.30-7.55 (48 h)
Dissolved oxygen:
7.86-8.05 mg/L (0 h); 7.42-8.01 mg/L (48 h)
Nominal and measured concentrations:
Measured concentrations: 0.36, 0.60. 1.3, 3.2 and 5.3 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (5 cm ID x 8 cm), 50 mL
- Fill volume: 20 mL
- No. of organisms per vessel: 5 in the vessels with the test substance and control, 10 in the vessels containing the reference material
- No. of vessels per concentration (replicates): 4 vessels in each concentration with the test substance and the control, 2 for each of the concentrations of the reference substance

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: According to EEC 92/69 L383A C.2

OTHER TEST CONDITIONS
- Photoperiod: 16/8 hour light/dark cycle
- Light intensity: Diffuse light, illumination range 1.5-5 µmol/m^-2.s^-1

EFFECT PARAMETERS MEASURED
Immobilisation

TEST CONCENTRATIONS
- Range finding study: A range finding study was performed, the percentage immobility was determined in all test item dilutions of the stock solution, ranging from 1:1 to 1:100. Based on the results of the preliminary test, a definitive test was performed with 5 dilutions of a stock solution ranging from 1:1 to 1:16 with a dilution factor of 2 to enable the determination of 0 and 100 % immobilisation after 24 and 48 hours.
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Duration:
24 h
Dose descriptor:
EC10
Effect conc.:
3.3 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
3.7 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 3.6-3.9 mg/L
Duration:
24 h
Dose descriptor:
EC100
Effect conc.:
5.3 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
2.1 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
2.7 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 2.5-2.9 mg/L
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
5.3 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Immobilisation of control: No immobilisation was observed in the control
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The test substance dilutions were clearly dissolved after filtration of the stock solution with 0.45 µm filter in all tested concentration levels throughout exposure.
Results with reference substance (positive control):
- Results with reference substance valid? The results with the reference substance were valid
- Mortality: The percentage immobility for the reference substance was determined after 24 hours. The EC100 value was determined directly from the test results. The EC10 and EC50 values, with 95% confidence intervals, were determined in a probability network by interpolation according to standard procedures. The EC50 value of the reference substance after 24 hours was within the prescribed concentration range of 1.0-2.5 mg/L of the validity criteria.
- EC50: 1.9 mg/L (CI 1.7-2.1 mg/L)
- EC10: 1.2 mg/L
- EC100: 5.8 mg/L

Table 1: Immobilisation rates of the range finding test

Test dilution*

24 h

48 h

Replicate 1

Replicate 2

Mean

Replicate 1

Replicate 2

Mean

1:1

70

50

60

100

100

100

1:10

0

0

0

0

0

0

1:100

0

0

0

0

0

0

Control

0

0

0

0

0

0

 *stock solution 100 mg/L filtered

Table 2: Percentage of Daphnia immobilised after 24 and 48 hours of exposure

Real test concentration (mg/L)

Immobilsation (%)

24 h

48 h

1

2

3

4

Mean

1

2

3

4

Mean

5.3

100

100

100

100

100

100

100

100

100

100

3.2

0

0

0

20

5

100

60

60

60

70

1.3

0

0

0

0

0

0

0

0

0

0

0.60

0

0

0

40

10

0

0

0

40

10

0.38

0

0

0

0

0

0

0

0

0

0

Control

0

0

0

0

0

0

0

0

0

0

 

Table 3: DOC- Analysis of new media (0 h)

Test material dilution*

DOC (mg/L)

Real test concentration (mg/L)***

Measured

Corrected**

1:1

5.45

3.73

5.3

1:2

3.94

2.22

3.2

1:4

2.63

0.91

1.3

1:8

2.14

0.42

0.60

1:16

1.97

0.25

0.36

Control

1.72

-

-

*stock solution 100 mg/L filtered

**corrected against control

***concentration of test substance based on ThTOC (0.7 mg C/mg)

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the study, the acute daphnia immobilisation 48 hour EC50 of hexenyl-3-cis salicylate was determined to be 2.7 mg/L based on the measured test substance concentration
Executive summary:

The acute immobilisation of (Z)-3-henxenyl salicylate to Daphnia magna Straus was determined according to standardised guidelines OECD 202 and EU Method C.2. The study was conducted under static conditions for 48 hours. 20 Daphnia were added to each test concentration and the control. The test material was clearly dissolved after filtration of the saturated stock solution in all test concentration levels throughout exposure. The real test concentrations were calculated based on DOC analysis to be 0.36, 0.60, 1.3, 3.2 and 5.3 mg/L.

The EC10 and EC50 values were calculated by probit analysis to be:

   Test duration (h)  Concentration (mg/L)  Confidence interval (p = 95%)
 EC10  24  3.3  -
   48  2.1  -
 EC50  24  3.7  3.6 - 3.9
   48  2.7  2.5 - 2.9
 EC100  24  5.3  
   48  5.3  

The EC50 of the reference substance potassium dichromate was calculated to be 1.9 mg/L which was in the prescribed concentration range of 1.0 to 2.5 mg/L. The validity criteria of the test guideline were fulfilled.

Description of key information

Acute Daphnia 48 hour EC50 2.7 mg/L; study conducted in accordance with OECD 202, EU C.2; Noack (2001)

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
2.7 mg/L

Additional information

The acute immobilisation of (Z)-3-henxenyl salicylate to Daphnia magna Straus was determined according to standardised guidelines OECD 202 and EU Method C.2. The study was conducted under static conditions for 48 hours. 20 daphnia were added to each test concentration and the control. The test material was clearly dissolved after filtration of the saturated stock solution in all test concentration levels throughout exposure. The real test concentrations were calculated based on DOC analysis to be 0.36, 0.60, 1.3, 3.2 and 5.3 mg/L.

The EC10 and EC50 values were calculated by probit analysis to be:

   Test duration (h)  Concentration (mg/L)  Confidence interval (p = 95%)
 EC10  24  3.3  -
   48  2.1  -
 EC50  24  3.7  3.6 - 3.9
   48  2.7  2.5 - 2.9
 EC100  24  5.3  
   48  5.3  

The EC50 of the reference substance potassium dichromate was calculated to be 1.9 mg/L which was in the prescribed concentration range of 1.0 to 2.5 mg/L. The validity criteria of the test guideline were fulfilled.