Glycerol, propoxylated, esters with acrylic acid

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-05-04 to 2010-05-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All concentration levels and the control were analytically verified in the fresh media (0 h) and in the old media (48 h) by LC-MS/MS.
- Sampling method: For the old media, separate replicates without test organisms were prepared and incubated under test conditions. The method was validated according to SANCO 3029/99 rev.4 (2000-07-11).
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until start of analysis if necessary.

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 and shaken with 20 rpm for 24 h at room temperature.
- 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 (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

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water at test start: 263 (mg CaCO3/L)

Test temperature:

20.1°C

pH:

Water Quality Parameters at Test Start (0 h, New Media)
(measured in one replicate per concentration and control)

Nominal
test item
concentration
mg/L] pH-Value


100 7.85
50.0 7.80
25.0 7.81
12.5 7.79
6.25 7.84
Control 7.93

Water Quality Parameters at Test End (48 h, Old Media)
(measured in all replicates)

Nominal
test item
concentration
[mg/L] pH-Values
Replicates
1 2 3 4
100 7.71 7.68 7.72 7.58
50.0 7.68 7.66 7.69 7.71
25.0 7.68 7.69 7.68 7.64
12.5 7.62 7.70 7.69 7.69
6.25 7.64 7.69 7.68 7.62
Control 7.65 7.64 7.65 7.66

Dissolved oxygen:

Water Quality Parameters at Test Start (0 h, New Media)
(measured in one replicate per concentration and control)

Nominal
test item
concentration
mg/L] O2-Concentration

[mg/L]
100 8.81
50.0 8.83
25.0 8.84
12.5 8.88
6.25 8.97
Control 8.90

Water Quality Parameters at Test End (48 h, Old Media)
(measured in all replicates)

Nominal
test item
concentration
[mg/L] O2 -Concentration [mg/L]
Replicates
1 2 3 4
100 8.32 8.30 8.43 8.31
50.0 8.36 8.34 8.30 8.33
25.0 8.37 8.35 8.50 8.34
12.5 8.28 8.39 8.33 8.30
6.25 8.34 8.33 8.41 8.03
Control 8.37 8.36 8.37 8.40

Salinity:

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): One application at test start, 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: Acc. to Directive 92/69/EEC Method C.2, Annex 1
- Conductivity: Dilution water at test start: 640 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 assessed in all treatment levels after 24 h and 48 h. An animal was considered to be immobile, if it was not able to swim during a 15 second observation period following gentle agitation of the test vessel.

TEST CONCENTRATIONS

- Range finding study
Immobilization Rates of the Preliminary Test in [%]
(n = 20, divided into 2 replicates with 10 daphnids each)
Nominal
test item
concentration
[mg/L] IMMOBILIZATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 80 60 70 100 80 90
10 0 0 0 0 0 0
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

91.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: (72.1-116)

Details on results:

- Behavioural abnormalities: None
- Mortality of control: 0%
- Other adverse effects control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: 1.70 (CI 1.51 - 1.97) mg/L

Reported statistics and error estimates:

The EC10- and the EC50-values were calculated by sigmoidal dose -response regression. The confidence intervals for the EC50-values were calculated from the best-fit values, the standard error and the t-distribution with the software GraphPad prism.

Percentage of Daphnids Incapable of Swimming after 24 and 48 h of Exposure

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

Nominal Test Item Concentration	IMMOBILIZATION [%]
	24 h					48 h
	Replicates					Replicates
[mg/L]	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
100	0	0	0	20	5	80	40	20	80	55
50.0	0	0	0	0	0	0	40	40	0	20
25.0	0	0	0	0	0	0	0	0	0	0
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

Validity criteria fulfilled:

yes

Conclusions:

In this study the registered substance (test item) was found to be toxic to Daphnia magna after 48 h at concentrations of 50.0 mg/L (LOEC48 h) and higher. The NOEC after 48 h was 25.0 mg/L. The EC50 (48 h) was determined to be 91.4 mg/L (CI: 72.1 - 116 mg/L). All effect values are given based on the nominal concentrations of the test item.

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS) the effects of the test item (batch number: S700140014) were determined according to OECD 202 (2004) from 2010-05-04 to 2010-05-07 at Dr.U.Noack-Laboratorien, Käthe-Paulus-Straße 1, D-31157 Sarstedt, Germany.

The study was conducted under static conditions with 5 concentration levels of the test item ranging from 6.25 to 100 mg/L set up in a geometrical series with a dilution factor of 2 over a period of 48 hours. 20 test organisms were exposed to each test concentration and the control.
All concentration levels and the control were analysed via LC-MS/MS after 0 h and 48 h.
The recoveries in the fresh (0 h) and old media (48 h) were in the range of 88 to 103 % of the nominal values, demonstrating that concentrations of Photomer4094 Fwere successfully maintained for the duration of the test. All effect values are given based on the nominal concentrations of the test item (Table 1).
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.

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

              (based on the nominal concentrations of the test item Photomer4094 F)

	Test Duration [h]	Nominal Concentration of the Test Item [mg/L]	Confidence Interval p = 95% [mg/L]
EC10	24	> 100	---
	48	38.0	---
EC50	24	> 100	--
	48	91.4	72.1 – 116
EC100	24	> 100	---
	48	> 100	---
NOEC	48	25.0	---
LOEC	48	50.0	---

Description of key information

Short term toxicity to daphnia - OECD 202 – EC50: 91.4 mg/l, 48h - (PARAD consortium, NOACK, 2010, M. Noack)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

91.4 mg/L

Additional information

Short term toxicity to daphnia - OECD 202 – EC50: 91.4 mg/l, 48h - (PARAD consortium, NOACK, 2010, M. Noack)