Reaction mass of 5,8,11,14-Tetraoxaoctadecane and 5,8,11,14,17-Pentaoxaheneicosane

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:

2012-06-06 to 2012-06-08

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

Test material information

Constituent 1

Reference substance name:

5,8,11,14,17-pentaoxahenicosane; 5,8,11,14-tetraoxaoctadecane

EC Number:

940-417-3

Molecular formula:

C14H30O4 C16H34O5

IUPAC Name:

5,8,11,14,17-pentaoxahenicosane; 5,8,11,14-tetraoxaoctadecane

Details on test material:

- Name of test material (as cited in study report): Polyglycol BB 300
- Physical state: Liquid, colourless
- Analytical purity: 99.6 % (a/a)
- Purity test date: 2011-06-28
- Lot/batch No.: DEG4155254
- Expiration date of the lot/batch: 2013-06-24
- 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: 12.5 – 25.0 – 50.0 – 100 – 200 mg/L
- Sampling method: The test item Polyglycol BB 300 was analytically verified by GC-MS in the fresh media (0 h) and old media (48 h) of all concentration levels and the control.
Sampling from the freshly prepared media was carried out immediately after homogenisation. Samples of the old media were taken from additional replicates, prepared with test media, but without daphnids. These replicates were incubated under test conditions until sampling. Recoveries of the test item should be within ± 20% of the nominal concentrations.
- Sample storage conditions before analysis:All original samples were stored at 6 ± 2 °C until sample preparation, if necessary. 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: The stock solution (200 mg/L test item was weighed out) was freshly prepared with dilution water and agitated.
- Eluate: Dilution water
- Differential loading: 12.5 – 25.0 – 50.0 – 100 – 200 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: 253 (mg CaCO3/L)

Test temperature:

20.8 °C (Water)

pH:

Water Quality Parameters at 0 h (Fresh Media)
(measured in one additional replicate per concentration level and control)
Polyglycol BB 300
Nominal
Test Item Concentration
[mg/L] pH-Value


200 7.80
100 7.66
50.0 7.75
25.0 7.77
12.5 7.73
Control 7.81

Water Quality Parameters after 48 h (Old Media)
(measured in all replicates)
Polyglycol BB 300
Nominal
Test Item Concentration
[mg/L] pH-Values
Replicates
1 2 3 4
200 7.70 7.70 7.69 7.69
100 7.66 7.67 7.69 7.69
50.0 7.69 7.68 7.67 7.66
25.0 7.72 7.71 7.70 7.69
12.5 7.67 7.69 7.69 7.70
Control 7.66 7.64 7.64 7.65

Dissolved oxygen:

Water Quality Parameters at 0 h (Fresh Media)
(measured in one additional replicate per concentration level and control)
Polyglycol BB 300
Nominal
Test Item Concentration
[mg/L] Dissolved O2-Concentration

[mg/L]
200 8.76
100 8.77
50.0 8.73
25.0 8.74
12.5 8.71
Control 8.89

Water Quality Parameters after 48 h (Old Media)
(measured in all replicates)
Polyglycol BB 300
Nominal
Test Item Concentration
[mg/L] Dissolved O2 -Concentration [mg/L]
Replicates
1 2 3 4
200 8.33 8.41 8.39 8.44
100 8.17 8.29 8.31 8.23
50.0 8.17 8.10 8.18 8.10
25.0 8.30 8.19 8.12 8.10
12.5 8.27 8.26 8.29 8.30
Control 8.26 8.16 8.22 8.35

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:654 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 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
300 80 80 80 100 100 100
100 40 20 30 100 100 100
30 10 10 10 0 0 0
10 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.:

65.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CI 58.6-73.8 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: 1.76 (CI 1.50 - 2.01) mg/L

Reported statistics and error estimates:

EC10- 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 possible. The concentration effect relationships are shown 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)

Polyglycol BB 300	IMMOBILISATION [%]
Nominal Test Item Concentration [mg/L]	24 h					48 h
	Replicates					Replicates
	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
200	80	60	60	60	65	100	100	100	100	100
100	40	40	20	60	40	80	80	60	100	80
50.0	0	0	0	0	0	40	20	40	20	30
25.0	0	0	0	0	0	0	0	0	0	0
12.5	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 Polyglycol BB 300 the 48-h EC50 for Daphnia magna was estimated to be 65.4 mg/L (95 % confidence limits: 58.6 – 73.8 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 immobilisation test with Daphnia magna (STRAUS), the effects of the test item Polyglycol BB 300 (batch number: DEG4155254) were determined according to OECD 202 (2004) from 2012-06-06 to 2012-06-11, with the definitive exposure phase from 2012-06-06 to 2012-06-08,  at Dr.U.Noack-Laboratorien, Käthe-Paulus-Str.1, 31157 Sarstedt, Germany .

The study was conducted in sealed glass flasks under static conditions over a period of 48 h with 5concentration levels of the test item Polyglycol BB 300 in the range of 12.5 to 200 mg/L 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 concentrations of the test item Polyglycol BB 300 were analytically verified by GC-MS in the fresh media (0 h) and in the old media (48 h) of all concentration levels and the control. The measured concentrations of the test item Polyglycol BB 300 in the fresh media (0 h) were in the range of 91 to 105 % of the nominal values. The measured concentrations of the test item in the old media (48 h) were in the range of 91 to 119 % of the nominal values, indicating that the concentrations of the test item Polyglycol BB 300 were successfully maintained for the duration of the test.

The endpoints were assessed based on the nominal concentrations of the test item Polyglycol BB 300 (Table 1).

The water quality parameters (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.

Table 1:    EC₁₀-, EC₅₀- (with Confidence Interval), EC₁₀₀-Values, NOEC and LOEC

                   (based on the nominal concentrations of the test item Polyglycol BB 300)

Endpoints (with 95 % confidence limits)	Test Duration [h]	Polyglycol BB 300
		Nominal Concentration of the Test Item [mg/L]
EC10	24	54.4 (CI: 43.5 – 67.6)
	48	34.5 (CI: 27.0 – 42.1)
EC50	24	132    (CI: 111 - 156)
	48	65.4 (CI: 58.6 – 73.8)
EC100	24	> 200
	48	200
NOEC	48	25.0
LOEC	48	50.0