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:
2012-10-31 to 2012-11-02
Reliability:
1 (reliable without restriction)

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
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): Sorbitan monocaprylate
- Physical state: Viscous liquid, brown, clear
- Analytical purity: 99.7 % (w/w)
- Purity test date: 2011-09-21
- Lot/batch No.: DEG4149186
- Expiration date of the lot/batch: 2013-07-06
- Stability under test conditions: Stable in water and light at room temperature
- Storage condition of test material: Room temperature, protected from light, in the original container

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
- Sampling method: All tested concentration levels of the test item and the control were analytically verified by analysis of dissolved organic carbon (DOC, according to DIN EN 1484) at the start of the exposure (fresh media, 0 h) and at the end of the exposure (old media, 48 h).
For the old media, an additional replicate of each concentration level and the control was prepared with test media but without daphnids and incubated under test conditions until sampling.
- Sample storage conditions before analysis: None, direct analysis

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution of 100 mg/L was freshly prepared with dilution water and agitated.
- Eluate: Dilution water
- Differential loading: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
- Controls: Dilution water without test item tested under the same conditions as the test groups.

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: 239 (mg CaCO3/L)
Test temperature:
19.9 °C
pH:
Water Quality Parameters at Test Start (0 h)
(measured in one additional replicate per concentration level and control)
Nominal Concentration
of the Test Item
[mg/L] pH-Value


100 8.13
50.0 8.13
25.0 8.09
12.5 8.12
6.25 8.03
Control 7.98

Water Quality Parameters at the Test End (48 h)
(measured in all replicates)
Nominal Concentration
of the Test Item
[mg/L] pH-Values
Replicates
1 2 3 4
100 7.67 7.66 7.67 7.64
50.0 7.71 7.73 7.73 7.71
25.0 7.80 7.75 7.74 7.73
12.5 7.85 7.84 7.84 7.84
6.25 7.86 7.86 7.84 7.88
Control 8.18 7.94 7.86 7.88
Dissolved oxygen:
Water Quality Parameters at Test Start (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]
100 9.26
50.0 9.22
25.0 9.17
12.5 9.06
6.25 9.26
Control 8.93

Water Quality Parameters at the Test End (48 h)
(measured in all replicates)
Nominal Concentration
of the Test Item
[mg/L] Dissolved O2 –Concentration [mg/L]
Replicates
1 2 3 4
100 6.84 6.99 6.96 7.16
50.0 7.06 7.01 7.06 7.20
25.0 7.21 6.87 7.03 7.04
12.5 8.11 8.02 7.88 7.94
6.25 8.14 8.06 8.06 8.10
Control 8.17 7.97 8.15 8.12
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:632 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:
Immobilization Rates in the non GLP Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)
Sorbitanmonocaprylate
Nominal Concentration
of the Test Item
[mg/L] IMMOBILIZATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 50 60 55 100 80 90
10 0 0 0 0 0 0
1 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 concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
40.1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CI (37.1-42.3 mg/L)
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
28.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CI (25.2-32.5 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: 1.92 (CI 1.74 - 2.10) mg/L
Reported statistics and error estimates:
EC10-value after 48 h and the 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 wherever feasible. The concentration-effect relationships are graphically presented.
The EC50-value for the reference item and its confidence limits were calculated accordingly.

Any other information on results incl. tables

Immobilization Rates in the Definitive Test after 24 and 48 hours of Exposure

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

Sorbitanmonocaprylate

Nominal Concentration

of the Test Item

[mg/L]

IMMOBILIZATION [%]

24 h

48 h

Replicates

Replicates

1.

2.

3.

4.

MV

1.

2.

3.

4.

MV

100

80

100

80

60

80

100

100

100

100

100

  50.0

60

  40

80

60

60

  80

  80

  80

  80

  80

  25.0

  0

    0

  0

  0

  0

    0

  20

    0

    0

    5

  12.5

  0

    0

  0

  0

  0

    0

    0

    0

    0

    0

    6.25

  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 Sorbitanmonocaprylate, the 48-h EC50 for Daphnia magna was calculated to be 40.1 mg/L (95 % confidence limits: 37.1 – 42.3 mg/L). The NOEC after 48 h was 25.0 mg/L. The LOEC after 48 h was 50.0 mg/L.
Executive summary:

In the acute immobilization test to Daphnia magna (STRAUS), the effects of the test item Sorbitanmonocaprylate (batch number: DEG4149186) were determined according to OECD 202 (2004) from 2012-10-31 to 2012-11-02 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 with 5 concentration levels of the test item Sorbitanmonocaprylate. The nominal concentrations in the range of 6.25 to 100 mg/L were prepared with dilution water in a geometric series with a dilution factor of 2. Twenty daphnids were exposed to each concentration level and the control. The biological results are presented in Table 3.

The tested concentration levels of the test item of 6.25 to 100 mg/L and the control were analytically verified by analysis of dissolved organic carbon (DOC, according to DIN EN 1484) at the start of the exposure (fresh media, 0 h) and at the end of the exposure (old media, 48 h).

The measured concentrations of dissolved organic carbon in the fresh media (0 h) were in the range of 4.82 to 54.4 mg C/L in the concentration levels of 6.25 to 100 mg/L, and 0.803 mg C/L in the control. In the old media (48 h), the measured concentrations of dissolved organic carbon were in the range of 5.67 to 60.8 mg C/L in the concentration levels of 6.25 to 100 mg/L, and 1.47 mg C/L in the control.

All effect values given are based on the nominal concentrations of the test item Sorbitanmonocaprylate (Table 1).

Water quality parameters (pH-value and dissolved oxygen concentration), measured in the fresh (0 h) and old media (48 h), were within acceptable limits. The validity criteria of the test guideline were fulfilled.

Table 1:    EC10-, EC50-, EC100 - Values, NOEC and LOEC

                   (based on the nominal concentrations of the test item)

Endpoints

Test

Duration

[h]

Sorbitanmonocaprylate

Nominal Concentration

of the Test Item

[mg/L]

95 % confidence limits

[mg/L]

EC10

24

25.0 – 50.0

Not applicable

48

     28.2

25.2 – 32.5

EC50

24

     49.2

Not applicable

48

     40.1

37.1 – 42.3

EC100

24

> 100

Not applicable

48

   100

Not applicable

NOEC

48

     25.0

Not applicable

LOEC

48

     50.0

Not applicable