Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 12 June 2001 to 14 June 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 the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report Date:
2001

Materials and methods

Test guidelineopen allclose all
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)
Remarks:
24-25th March 1997

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Sponsor's identification: Wollwachsalkohol I Lanolinalkohol
Description: Yellow, type wax
Batch number: Eucerit 6480
Active ingredient: Wool waxalcohol
Storage conditions: 7 ± 2 °C, protected from light and moisture
Composition: Aliphates: 25.5 %
Sterines: 60.6 %
Cholesterine: 31.2 %
CH-5dien-70ne: 8.9 %
Dehydrolanosterine: 5.5 %
Lanosterine: 10.9 %

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations:
Limit concentration: 1 : 1 = 0.13 mg DOC I L

- Sampling method: Not reported
- Sample storage conditions before analysis: Not reported

Test solutions

Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
The solution (1000 mg/L test item was weighed out) was prepared with dilution water one day prior to application and filtered (0.45 µm, single use)
Dispersion treatment: 24 h horizontal shaking

Control:
Dilution water without test item tested under same conditions as the test group.

Solubility of the test item:
The test item was clearly dissolved at the limit concentration.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name:
Water Flea

- Source:
Origin: Institut fOr Wasser-, Boden- und Lufthygiene des Bundesgesundheitsamtes, Corrensplatz, 14195 Berlin, FRG.
Breeder: DR.U.NoACK-LASORATORIUM, 31157 Sarstedt, FRG

- Age at study initiation:
Less than 24 h old

- Feeding during test:
Received no food during exposure

ACCLIMATION
- Acclimation period:
2 h


- Acclimation conditions:
2 h in dilution water


- Type and amount of food:
5 x weekly ad libitum with a mix of Scenedesmus subspicatus and ChIarella vulgaris, with an algae cell density of > 10E6 cells/mL.

- Health during acclimation:
No mortality observed

QUARANTINE (wild caught)

- Duration:
Not applicable

- Health/mortality:
Not applicable

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
Not applicable

Test conditions

Hardness:
Total hardness: 263 mg CaCO3/L.
Test temperature:
20 ± 2 °C constant
Measured using Hygro-thermograph (LUFFT).
pH:
7.64 - 7.82
The pH was measured using a pH-Meter 537 (WTW)
Dissolved oxygen:
8.06 mg/l - 8.61 mg/l
The dissolved oxygen concentration was measured using an Oximeter, Oxi 530 (WTW)
Salinity:

freshwater used.
Nominal and measured concentrations:
Limit Concentration: 0.13 mg/l
Details on test conditions:
TEST SYSTEM
The definitive study was performed as limit test with the saturated solution as specified above under static conditions to enable the determination of immobilization after 24 and 48 hours.

- Test vessel:
Glass beakers (5 cm 10 x 8 cm H), 50 mL volume were used

- Type (delete if not applicable):
closed

- Material, size, headspace, fill volume:
20 mL

- Number of study organisms per concentration and control:
20 animals, divided into 4 parallel samples, each with 5 animals

-Number of animals per test vessel: 5

- Number of parallel samples per concentration: 4


OTHER TEST CONDITIONS
- Adjustment of pH:
Not reported

- Photoperiod:
16/8 h ligh/dark cycle

- Light intensity:
Diffuse light, illumination strength max. 20 µmol.m-2 . S·1 (corresponding to 1200 Ix)



Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrations
Duration:
48 h
Dose descriptor:
other: Limit concentration
Effect conc.:
0.13 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Remarks on result:
other: Not reported
Details on results:
- Behavioural abnormalities: Not recorded
- Other biological observations: None recorded

- Immobilisation of control:
The percentage immobility was determined in the tested limit concentration and the control after 24 hand 48 h. The immobilisation rates in % after 24 hand 48 h of exposure are given in Table 1.

There is no biologically significant effect neither in the tested limit concentration nor in control group.


- Other adverse effects control: No other effects observed.

- Abnormal responses: None recorded

- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
The test item was clearly dissolved at the limit concentration.

Results with reference substance (positive control):
- Results with reference substance valid?
Yes

- Mortality:
No mortalities recorded.

- EC50/LC50:

The percentage immobility for the reference item was determined after 24 h. The EC100 value was determined directly from the test results. The EC10 and EC50 with 95 % confidence interval (CI) were determined by probit analysis according to standard procedures.

EC- values after 24 h of the reference item in mg/L were as follows:

EC10 : 1.2
EC50 : 1.9 (CI 1.7-2.1)
EC100 : 5.8


The ECso - value of reference item potassium dichromate after 24 h is within the prescribed concentration range of 1.0 - 2.5 mg/L of validity criteria according to AQS: DIN Guideline 38412 L 30.
- Other:
None.
Reported statistics and error estimates:
An estimate of the EC50 value at 3 hours was given by inspection of the immobilisation data.
The EC50 value and associated confidence limits at 24 hours were calculated by the trimmed Spearman-Karber method (Hamilton et al 1977) and the EC50 value and associated confidence limits at 48 hours and the slope of the response curve and its standard error were calculated by the maximum-likelihood probit method (Finney 1971) using the ToxCalc computer software package (Toxcalc 1999).
Probit analysis is used when two or more partial responses to exposure are shown.
When only one partial response is shown the trimmed Spearman-Karber method is appropriate.

Any other information on results incl. tables

Table 1. Percentage number of daphnids incapable of swimming after 24 and 48 h of exposure (n = 20)

IMMOBILISATION [%]

Limit

Conc.

[mgDOC/L]

24 h

48 h

Replicates

Replicates

1

2

3

4

MV

1

2

3

4

MV

0.13

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

MV = Mean value

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In the tested limit concentration (0.13 mg DOC/L) there is no biologically significant effect neither in the tested limit concentration nor in control group.
Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS) the effect of the saturated solution as limit concentration (0.13 mg DOC / L) of the test item Lanolin alcohol (Wollwachsalkohol/Lanolinalkohol).

Batch number Eucerit 6480, was determined according to OECD Guideline 202 I/EEC Directive 92/69/EEC Method C.2. The limit test was conducted from 12 -06 -2001 to 14 -06 -2001 at DR.U.NOACK-LABORATORIUM FOR ANGEWANDTE BIOLOGIE D-31157 Sarstedt.

The test item was clearly dissolved at the limit concentration.

The study was conducted under static conditions over a duration of 48 hours. 20 test organisms were exposed to the limit test concentration and control.

Table 1. Percentage number of daphnids incapable of swimming after 24 and 48 h of exposure (n = 20)

IMMOBILISATION [%]

Limit

Conc.

[mgDOC/L]

24 h

48 h

Replicates

Replicates

1

2

3

4

MV

1

2

3

4

MV

0.13

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

MV = Mean value

A reference test is carried out once per month with potassium dichromate as reference item and the reference toxicity determined. The ECso -value of the reference item at 1.9 mg/l after 24 h was within the prescribed concentration range of 1.0 to 2.5 mg/l according to AQS, DIN Guideline 38412 l 30.

Water quality parameters of pH-value and dissolved oxygen concentration measured at 0 and 48 h were determined to be within the acceptable limits.

The validity criteria of the test guideline were fulfilled.

In the tested limit concentration (0.13 mg DOC/L) no biologically significant effect was determined.