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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline Study
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Chemical name: Amines, coco alkyl

Sampling and analysis

Analytical monitoring:
yes

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Dispersion treatment: Stock solution was 5 mg/L. 6 μL/L test item were applied to 3 L natural river water. This solution was treated for 30 minutes with ultrasound at 40 °C. After a cooling period of 10 minutes the test item concentrations were prepared out of this stock solution.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain: Daphnia magna STRAUS (Clone 5)
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu); Breeder DR.U.NOACK-LABORATORIEN, D-31157 Sarstedt, Germany
- Age at study initiation: 2 - 24 h old
- Feeding during test: The daphnids were not fed during the study.

HOUSING:
- Conditions: In 2-3 L glass vessels with approximately 1.8 L culture medium, at 21 °C (temperatures of 20 - 25 °C are tolerated), in an
incubator, 16 h illumination, illumination strength max. 20 μE⋅m-2 ⋅s-1.
- Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.
- Type and amount of food: Feeding ad libitum with a mix of Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 106 cells/mL.
- Feeding frequency: 5x weekly



ACCLIMATION
- Acclimation period: At least 2 h in dilution water

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
131.4 mg CO3/L
Test temperature:
19.6 °C
pH:
7.6 - 7.8
Dissolved oxygen:
7.89 - 9.25 mg/L
Nominal and measured concentrations:
Nominal concentrations: 5.00 - 2.00 - 0.80 - 0.32 - 0.123 - 0.051 mg/L (factor 2.5)
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, fill volume: Glass beakers (5 cm ID x 8 cm H), 50 mL; Volume of the study medium is 20 mL
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4



TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: A natural occurring river water (River Böhme) was used as test medium.
Test water parameter:
pH: 7.68
Conductivity [μS/cm]: 451
DOC [mg C/L]: 6.3
TOC [mg C/L]: 7.1
Ammonium-N [mg N/L]: < 1.94
Nitrate-N [mg N/L]: 3.9
o-Phosphate-P [mg P/L]: 0.029
Total Phosphate mg P/L]: 0.080
Suspended Matter [mg/L]: 16.9
Total Hardness [mg CO3/L]: 131.4
- Culture medium different from test medium: yes
- Intervals of water quality measurement: Prior to test start (0 h) pH-value, dissolved oxygen concentration, temperature, conductivity and total hardness of the dilution water were measured. At the beginning of the test water parameters (pH value, oxygen concentration) were measured in one additional replicate per concentration and control. After 48 h the water parameters in old media were measured in all replicates per concentration and control. The room temperature was recorded throughout the test with a thermohygrograph.


OTHER TEST CONDITIONS
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, illumination range max. 20 μE⋅m-2 ⋅ s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The percentage immobility was determined in all test and control groups after 24 h and 48 h.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
0.23 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.32 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
0.8 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility

Any other information on results incl. tables

Table 1: Percentage of Daphnids incapable of swimming after 24 and 48 h of exposure with

Amines, coco alkyl

Nominal conc.(mg/l)

IMMOBILISATION [%]

 

24 h

48 h

 

Replicates

Replicates

 

1

2

3

4

MW

1

2

3

4

MW

5.00

100

100

100

100

100

100

100

100

100

100

2.00

100

100

100

100

100

100

100

100

100

100

0.80

80

60

80

80

75

100

100

100

100

100

0.32

0

0

0

20

5

60

40

40

40

45

0.128

0

0

0

0

0

0

0

0

0

0

0.051

0

0

0

0

0

0

0

0

20

5

Control

0

0

0

0

0

0

0

0

0

0

Table 2: Concentrations of Amines, coco alkyl

Nominal test item conc.

[mg/L]

 

Meas. conc. [mg/L]after 0 h

Meas. conc. [mg/L]after 48 h

5.00

5.9

0.02

2.00

2.3

< detection limit

0.80

0.9

< detection limit

0.32

0.38

< detection limit

0.128

0.19

< detection limit

0.051

0.06

0.03

Control

< detection limit

< detection limit

Applicant's summary and conclusion

Conclusions:
The total amount of test item recovered from the the stock solution and glassware (aqueous phase and adsorption to glass) was higher than expected. Therefore a correction was applied.
The measured initial concentrations in the test replicates (0 h) showed a good accordance with the geometrical series of dilution. After a 48 h exposure all concentrations (except the lowest one) had dropped below the limit of quantification (10 μg/L).
The adsorption of the test item to glassware used during the test was quantified at the two concentrations which framed the EC50 (0-48 h). As expected, Amines, coco alkyl showed only a limited adsorption to glassware. The amount recovered from the glassware did not exceed 3.6 % of the applied amount. These results indicate that adsorption to glassware contributes only little to the observed decrease of the test item during the test. Under test conditions with natural river water, adsorption to glass can be considered negligible.
Other effects must have caused the observed decrease of concentration in the aqueous phases. Adsorption to humic acids, suspended matter or daphnia surfaces are the most likely mechanisms.
Executive summary:

The Acute Immobilisation of the test item Amines, coco alkyl (batch number: S000358) to Daphnia magna (STRAUS) was determined according to OECD Guideline 202 / 92/69/EC Method C.2 from 2006-03-01 to 2006-03-03 at DR.U.NOACK-LABORATORIEN, D-31157 Sarstedt, Germany. The study was conducted with 6 nominal concentrations ranging from 0.051 to 5.00 mg/L in a geometric series with a factor of 2.5 under static conditions over a duration of 48 h. 20 test organisms were exposed to each test concentration and control. The water quality parameters pH-value and dissolved oxygen concentration were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled. All concentration levels and the control groups were analytically verified via LC-MS/MS after 0 h (new media) and 48 h (old media). The measured initial concentration levels were in good accordance, whereas test concentration after 48 h exposure were below the limit of quanitification (= 10 μg/L). Adsorption to the test vessels after 48 h of exposure, determined from the dosage levels framing the LC50, was found to be negligible (for details of the analytical method and results please refer to chapter 9).

The test item has a low water solubility and sorbs to organic and inorganic materials by different mechanisms. The sorption processes are mostly non-linear, means are concentration dependent. Due to these properties the test item is difficult to test in synthetic water (e.g. sorption to the test organism and walls of the test vessel) and results from such tests depend from the test settings applied. Using natural river water which contains particulate as well as dissolved organic carbon to which the test item can sorb partially reduces the difficulties encountered in tests with synthetic water e.g. preventing that the test item settles onto surfaces. The sorbed fraction of the test item is difficult to extract from the test system which normally leads to low analytical recoveries. Due to the short exposure period these low recoveries cannot be associated to biodegradation. This means the test substance is present in the test system and therefore available for exposure (dissolved in water and sorbed also called bulk). Due to the properties of the test item nominal concentrations have to be used instead of measured ones (see Table 1). This so called Bulk Approach is described by ECETOC (2003). Sorption of the test item to the glass ware of the test system was monitored and found to be small.

   Test Duration [h]   Nominal Test Item Concentration [mg/L]   Confidence Interval p = 95% [mg/L] 
 EC10   24   0.369   --- 
 48   0.236   --- 
 EC50   24   0.613   0.516 – 0.729 
 48   0.323   0.296 – 0.353 
 EC100   24   2.00   --- 
 48   0.80   --- 

The study was performed according to OECD Guideline 202 (2004) and Directive 92/69/EC

Method C.2 (1992). The validity criteria were fulfilled:

• In the control group no Daphnia were immobilised or trapped on the surface of the water.

• The EC50 -value after 24 h of the reference item was in the range of 1.0 to 2.5 mg/L.

• The dissolved O2 concentration was > 3 mg/L.

• The pH-value did not vary by more than 1.5 unit.

• The recovery rates of the test item were > 80 % at test begin and < LOQ (10 μg/L) at test

end. Due to the properties of the test item all effect levels are given based on nominal test

item concentrations.