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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:
2013-01-15 to 2013-01-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study, no deficiencies

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Pentapropylensuccinic anhydride, reaction products with ethanolamine, sodium and triethanolamine salts
- Physical state: Brown, viscous liquid
- Analytical purity: 100 % (w/w)
- Purity test date: 2011-11-15
- Lot/batch No.: ESD0010887
- Expiration date of the lot/batch: 2013-12-13
- Stability under test conditions: Not applicable
- Storage condition of test material: Room temperature, protected from light, in original container

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: 1.25 - 2.50 - 5.00 - 10.0 - 20.0 - 40.0 mg/L (dilution factor 2).
- Sampling method:
All test concentrations and the control were analytically verified by LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) as specified below. Sampling at the start of the exposure (0 h) was carried out immediately before application.
At the end of the exposure (48 h), samples were taken from additional replicates, prepared with test media, but without daphnids. These replicates were incubated under test conditions until sampling. The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11).
- Sample storage conditions before analysis: All original samples were stored at room temperature until sample preparation. Prepared samples were stored at room temperature until analysis.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution (40.0 mg/L of the test item were weighed out) was freshly prepared with dilution water. Dispersion treatment was agitation
- Eluate: Dilution water
- Differential loading: 1.25 - 2.50 - 5.00 - 10.0 - 20.0 - 40.0 mg/L (dilution factor 2).
- Controls: Dilution water without test item tested under the same conditions as the test goups.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 µExm-2 x s-1
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

DILUTION WATER:
Dilution water ISO test water, according to OECD 202, Annex 3:
Component Concentration [mg/L]
KCl 5.76
NaHCO3 64.8
CaCl2  2 H2O 294
MgSO4  7 H2O 123
pH-value: 7.8  0.2




Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
No

Test conditions

Hardness:
Dilution water at test start: 240 (mg CaCO3/L)
Test temperature:
19.9 °C (Water)
pH:
Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate per concentration level and control)
Nominal concentration
of the test item
[mg/L] pH-Value

40.0 7.88
20.0 7.80
10.0 7.79
5.00 7.75
2.50 7.73
1.25 7.71
Control 7.86

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Nominal concentration
of the test item
[mg/L] pH-Values
Replicates
1 2 3 4
40.0 7.60 7.61 7.63 7.61
20.0 7.68 7.66 7.69 7.69
10.0 7.72 7.71 7.70 7.75
5.00 7.70 7.71 7.73 7.71
2.50 7.67 7.66 7.69 7.70
1.25 7.77 7.75 7.71 7.66
Control 8.02 8.00 7.93 7.89
Dissolved oxygen:
Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate per concentration level and control)
Nominal concentration
of the test item
[mg/L] Dissolved O2-Concentration

[mg/L]
40.0 8.68
20.0 8.72
10.0 8.71
5.00 8.73
2.50 8.68
1.25 8.57
Control 8.66

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Nominal concentration
of the test item
[mg/L] Dissolved O2 -Concentration [mg/L]
Replicates
1 2 3 4
40.0 7.23 7.47 7.58 7.50
20.0 7.89 7.83 7.88 7.79
10.0 8.27 8.05 7.96 7.99
5.00 8.06 8.23 8.27 8.21
2.50 7.98 8.27 7.94 8.21
1.25 8.42 7.85 8.11 7.60
Control 8.44 8.39 8.41 8.43
Salinity:
Not measured, freshwater
Nominal and measured concentrations:
please refer to "Any other information on materials and methods"
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: see above
- Conductivity: Dilution water at test start:628 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start and test end


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility was determined in all groups after 24 h and 48 h.

TEST CONCENTRATIONS
- Range finding study:
Immobilisation Rates in the non GLP Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)
Nominal
Test Item
Concentration
[mg/L] IMMOBILISATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 100 100 100 100 100 100
10.0 80 90 85 70 50 60
1.00 0 0 0 0 0 0
0.100 0 0 0 0 0 0
Control 0 0 0 0 0 0
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrations
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
11.8 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CI 10.2-13.8 mg/L
Details on results:
- Mortality of control: 0 %
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- EC50/LC50: 1.69 (CI 1.41 - 2.12) mg/L
Reported statistics and error estimates:
EC100-values and the NOEC and LOEC after 48 h were estimated directly from the observation data, without mathematical calculation.

EC10- and EC50-values were calculated by sigmoidal dose-response regression. The confidence intervals were calculated from the best-fit values, the standard error and the t-distribution with the software GraphPad prism5. The concentration effect relationships are presented graphically.

Any other information on results incl. tables

Immobilisation Rates in the Definitive Test after 24 and 48 h of Exposure

                 (n = 20, divided into 4 replicates with 5 daphnids each)

Nominal

Test Item

Concentration

[mg/L]

IMMOBILIZATION [%]

24 h

48 h

Replicates

Replicates

1

2

3

4

MV

1

2

3

4

MV

40.0

80

60

60

80

70

100

100

100

100

100

20.0

40

40

60

60

50

  60

  80

  80

  60

  70

10.0

  0

20

40

40

25

  40

  40

  60

  60

  50

  5.00

  0

  0

  0

  0

  0

    0

    0

    0

    0

    0

  2.50

  0

  0

  0

  0

  0

    0

    0

    0

    0

    0

  1.25

  0

  0

  0

  0

  0

    0

    0

    0

    0

    0

Control

  0

  0

  0

  0

  0

    0

    0

    0

    0

    0

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Based on the nominal concentrations of the test item Pentapropylensuccinic anhydride, reaction products with ethanolamine, sodium and triethanolamine salts, the 48-h EC50 for Daphnia magna was estimated to be 11.8 mg/L (95 % confidence limits: 10.2 – 13.8 mg/L). The NOEC after 48 h was 5.00 mg/L. The LOEC after 48 h was 10.0 mg/L.

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the test item Pentapropylensuccinic anhydride, reaction products with ethanolamine, sodium and triethanolamine salts (batch number: ESD0010887) were determined according to OECD 202 (2004) from 2013-01-15 to 2013-01-18, with the definitive exposure phase from 2013-01-15 to 2013-01-17, at Dr.U.Noack-Laboratorien, Käthe-Paulus-Str.1, 31157 Sarstedt, Germany. The study was conducted under static conditions over a period of 48 h. Sixconcentration levels of the test item Pentapropylensuccinic anhydride, reaction products with ethanolamine, sodium and triethanolamine salts in the range of 1.25 to 40.0 mg/Lwere prepared with dilution water in a geometric series with a factor of 2and tested. Twenty daphnids were exposed to each concentration level and the control. The concentrations of the test item Pentapropylensuccinic anhydride, reaction products with ethanolamine, sodium and triethanolamine salts were analytically verified by LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) of all concentration levels and the control. The measured concentrations of the test item at the start of the exposure (0 h) were in the range of 97 to 102 % of the nominal values. The measured concentrations of the test item at the end of the exposure (48 h) were in the range of 97 to 102 % of the nominal values, indicating that the concentrations of the test item were successfully maintained for the duration of the test. The endpoints were based on the nominal concentrations of the test item Pentapropylensuccinic anhydride, reaction products with ethanolamine, sodium and triethanolamine salts.

The water quality parameters (i.e. pH-values and dissolved oxygen concentrations), measured at the start (0 h) and the end of the exposure (48 h), were within the acceptable limits. The validity criteria of the test guideline were fulfilled.

EC10-, EC50- (with Confidence Interval), EC100-Values

                   (based on the nominal concentrations of the test item)

Endpoints

Test

Duration

[h]

Pentapropylensuccinic anhydride, reaction products with ethanolamine, sodium and triethanolamine salts

Nominal Concenrations
of the Test Item

[mg/L]

95 % confidence limits

[mg/L]

EC10

24

     6.82

4.65 – 9.27

48

     5.02

3.72 – 6.51

EC50

24

   19.1

15.2 – 24.6

48

   11.8

10.2 – 13.8

EC100

24

> 40.0

Not applicable

48

   40.0

Not applicable

NOEC

48

     5.00

Not applicable

LOEC

48

   10.0

Not applicable