D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C18-C18 (unsaturated) acyl] derivs.

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-11-19 to 2013-12-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study, no deficiencies

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
5 test item concentrations in a geometric series with a separation factor of 101/4, prepared by diluting the stock solution of 7.10 mg/L with dilution water, were tested as follows: 0.710 - 1.26 - 2.25 - 3.99 - 7.10 mg/L. The test concentrations were selected based on the results of preliminary tests under static conditions.


- Sampling method:
All concentration levels and the control were analytically verified by LC-MS/MS at the start (0 and 24 h) and at the end of both exposure intervals (24 and 48 h). At the start of the exposure intervals (0 and 24 h), sampling was carried out immediately after preparation of the test concentrations.
At the end of the exposure intervals (24 and 48 h), samples were taken directly from the test vessels. Sorption to the walls of the test vessel was checked as specified below. For this purpose, additional replicates without daphnids were prepared and incubated under test conditions for quantification of sorption to the glassware. The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11).
Recoveries of the test item should be within ± 20 % of the nominal or initially measured concentrations.
The effect levels given are based on the nominal concentrations of the test item C18/C18 unsatd.-Glucamide and the active components Glucamides, because the test item is a UVCB substance with an active content of 80.4 % (w/w) Glucamides.

The sorption of the test item on glass was quantified at the concentration levels of 1.26 – 2.25 – 3.99 mg/L (double determinations) at the end of both exposure intervals (24 and 48 h). Two additional replicates per concentration level without daphnids were prepared. After sampling for the test medium analysis (aqueous phase), the glassware was emptied and an appropriate amount of a suitable solvent was added. The concentration of the test item in this solution was measured and the adsorbed test item amount was calculated from this concentration. Before this procedure, the test vessels were carefully rinsed with demineralised water to remove the remaining test media.


- Sample storage conditions before analysis: All original samples were stored at room temperature until preparation. Prepared samples were stored in an autosampler at room temperature until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution (7.10 mg/L of the test item were weighed out) was freshly prepared with dilution water prior to each exposure interval (0 and 24 h). The stock solution was treated with ultrasound for 30 minutes at 40 °C. 5 test item concentrations in a geometric series with a separation factor of 101/4, prepared by diluting the stock solution of 7.10 mg/L with dilution water, were tested as follows: 0.710 - 1.26 - 2.25 - 3.99 - 7.10 mg/L.




- Dilution water : ISO Test water, according to OECD 202, Annex 3, composition see below

- Controls: Dilution water without test item tested under the same conditions as the test goups.

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: 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, light intensity of max. 20 µExm-2 x s-1;
Culture feeding: at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus with an algae cell density of > 10^6 cells/mL.
Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L

- Feeding during test: No feeding

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

Component Concentration [mg/L]
KCl 5.76
NaHCO3 64.8
CaCl2  2 H2O 294
MgSO4  7 H2O 123

pH-value: 7.8  0.2

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water at test start: 262 (mg CaCO3/L)
Dilution water after 24 hours: 247 (mg CaCO3/L)

Test temperature:

At test start: 19.8°C (Water)
After 24 h: 20.1°C

pH:

Please refer to "Any other information on materials and methods"

Dissolved oxygen:

Please refer to "Any other information on materials and methods"

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, 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 water renewal)
- 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: 633 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 at 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) :
-Immobilisation was determined in all groups after 24 and 48 h. An animal was considered to be immobile, if it was not able to swim in the water phase within 15 seconds after gentle agitation of the test vessel.

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
loading of
the test item
[mL/L] IMMOBILISATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 70 100 85 100 100 100
10 0 10 5 20 60 40
1.00 0 0 0 0 0 0
Control 0 0 0 0 0 0


MV = mean value

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CI (0.9-1.1 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: 2.1 (CI: 1.8 - 2.4) mg/L

Reported statistics and error estimates:

The EC100-values after 24 and 48 h, the NOEC and LOEC after 48 h were deduced directly from the observation data, without mathematical calculation.

The EC10- and the EC50-values were calculated by sigmoidal dose-response regression. The 95 % confidence limits were calculated from the best-fit values,
the standard error and the t-distribution with the software GraphPad prism5.
The EC50-value for the reference item was calculated accordingly. The presented NOEC, LOEC and EC-values were rounded for presentation to one decimal place.

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

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

Nominal concentrations of the		IMMOBILISATION [%]
test item     [mg/L]	active components Glucamides [mg/L]	24 h					48 h
		Replicates					Replicates
		1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
7.10	5.71	60	60	40	60	55	100	100	100	100	100
3.99	3.21	20	20	80	60	45	100	100	100	100	100
2.25	1.81	60	60	60	20	50	100	100	100	100	100
1.26	1.01	0	40	40	0	20	80	80	100	100	90
0.710	0.571	0	0	0	0	0	0	0	20	20	10
Control		0	0	0	0	0	0	0	0	0	0

MV  = mean value

All test item concentrations were visually clear throughout the exposure period.

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item C18/C18 unsatd.-Glucamide the 48 h-EC50 for Daphnia magna was 1.0 mg/L (95 % confidence limits: 0.9 – 1.1 mg/L). Based on the nominal concentrations of the active components Glucamides the 48 h-EC50 for Daphnia magna was 0.8 mg/L (95 % confidence limits: 0.7 – 0.9 mg/L).
The NOEC after 48 h was 0.7 mg test item/L, which corresponds to 0.6 mg Glucamides/L.
The LOEC after 48 h was 1.3 mg test item/L, which corresponds to 1.0 mg Glucamides/L.

Executive summary:

In the acute immobilisation test withDaphnia magna (STRAUS), the effects of the test item C18/C18 unsatd.-Glucamide were determined according to OECD 202 (2004) from 2013-11-19 to 2013-12-19, with the definitive exposure phase from 2013-11-26 to 2013-11-28, at Dr.U.Noack-Laboratorien, Käthe-Paulus-Str.1, 31157 Sarstedt, Germany.

The study was conducted under semi-static conditions over a period of 48 h with 5 concentrations of the test item C18/C18 unsatd.-Glucamide in the range of 0.710 to 7.10 mg/L in a geometric series with a separation factor of 10^1/4. Twenty daphnids were exposed to each concentration leveland the control.

The test item is an UVCB substance (substances of Unknown or Variable composition, Complex reaction products or Biological materials) and is poorly soluble in water at room temperature.

The concentrations of the main component C18 unsatd.-Glucamide were analytically verified via LC-MS/MS at the start of the exposure intervals (0 and 24 h) and at the end of the exposure intervals (24 and 48 h) in all concentration levels and the control.

The measured concentrations of the main component C18 unsatd.-Glucamide at the start of the exposure intervals (0 and 24 h) were in the range of 99 to 108 % of the nominal values. At the end of the exposure intervals (24 and 48 h), the measured concentrations of the main component C18 unsatd.-Glucamide were in the range of 58 to 73 % of the nominal values.

Adsorption to the glass walls of the test vessels was determined exemplarily at the concentration levels of 1.26 – 2.25 – 3.99 mg/L (double determinations) in separate replicates without daphnids. Adsorption to the glass walls was in the range of 6 – 21 % of the nominal concentrations of the main component C18 unsatd.-Glucamide.

The effect levels given are based on the nominal concentrations of the test item C18/C18 unsatd.-Glucamideand the active components Glucamides.

The water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 and 24 h) and at the end of the exposure intervals (24 and 48 h), were within the acceptable limits. The validity criteria of the test guideline were fulfilled.

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

(based on the nominal concentrations of the test item C18/C18 unsatd.-Glucamide and the active components Glucamides)

Effect levels	Test duration [h]	Nominal concentrations of the test item C18/C18 unsatd.-Glucamide [mg/L]	Nominal concentrations of the active components Glucamides (80.4 % (w/w)) [mg/L]
EC10 (with 95 % confidence limits)	24	1.2 (< 0.7 – 1.3)	1.0 (< 0.6 – 1.1)
	48	0.7 (< 0.7 – 0.8)	0.6 (< 0.6 – 0.7)
EC50 (with 95 % confidence limits)	24	2.4 (1.3 – > 7.1)	1.9 (1.1 – > 5.7)
	48	1.0 (0.9 – 1.1)	0.8 ( 0.7 – 0.9)
EC100	24	> 7.1	> 5.7
	48	2.3	1.8
NOEC	48	0.7	0.6
LOEC	48	1.3	1.0

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-06-08 to 2016-06-10

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

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Determination of the test item
All concentration levels and the control were analytically verified via
LC-MS/MS at the start (0 and 24 hours) and at the end of the exposure intervals (24 and 48 hours). The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000). Details of the analytical method are presented in section 14.


Sampling for the analytical monitoring
At the start of the exposure intervals (0 and 24 hours), sampling was
carried out after preparation of the test concentrations.
At the end of the exposure intervals (24 and 48 hours), samples were taken directly from the test vessels.

Range (target)
Recoveries of the test item should be within ± 20% of the nominal or initially measured concentrations.

Determination of losses due adsorption onto glassware
The adsorption of the test item on glass was quantified in all
concentration levels at the end of the exposure intervals (24 and 48 hours). One additional replicate per concentration level without daphnids was prepared. After sampling for the test medium analysis (aqueous phase), the glassware was emptied and an appropriate amount of a suitable solvent (acetonitrile containing 0.2 % formic acid) was added. The concentration of the test item in this solution was measured and the adsorbed test item amount was calculated from this concentration. Before this procedure, the test vessels were carefully rinsed with demineralized water to remove the remaining test media. The samples, which were analyzed, are listed in Table 4.

Schedule of Specific Test Item Analysis
Sampling Date
[h] Samples analyzed:

0 and 24
(fresh media) aqueous phase without daphnids:
all concentration levels and the control
24 and 48
(old media) aqueous phase with daphnids:
all concentration levels and the control
aqueous phase and glass surface without daphnids:
all concentration levels

Vehicle:

no

Details on test solutions:

Stock solution
The stock solution (7.10 mg/L of the test item were weighed out) was freshly prepared with dilution water (Table 3) before the start of each exposure interval (at 0 and 24 hours).

Treatment
The stock solution was treated with ultrasound for 30 minutes at 40 °C and stirred thereafter with a magnetic stirrer at approximately 1100 rpm for 15 minutes at room temperature.

Test concentrations
5 test item concentrations in a geometric series with a separation factor of 101/4, prepared by dilution of the stock solution 7.10 mg/L with dilution water,
were tested as follows:
0.710 - 1.26 - 2.25 - 3.99 - 7.10 mg/L
The test concentrations were selected based on the results of a non-GLP preliminary range finding test. For results, see section 7.1.

Control
Dilution water without test item incubated under the same conditions as the test groups

Test method
Based on the results of a preliminary range finding test (section 7.1), the study was performed with a semi-static test design with a renewal of the test
solutions after 24 hours, since the measured concentrations did not remain stable over 48 hours under test conditions in the range finding test.

Test duration 48 hours

Test vessels Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with watch glasses

Renewal of the test solutions
The test solutions were renewed after 24 hours. For this purpose, a second set of test vessels was filled with the freshly prepared test solutions and the
daphnids were transferred by pipette (see ‘Application’).

Dilution water
Same as culture medium (see Table 3)

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system
Daphnia magna STRAUS (Clone 5).

Reason for the selection of the test system
Daphnia magna is the preferred species in accordance with the
test guideline and is bred at the test facility.

Origin
Institut für Wasser-, Boden- und Lufthygiene (WaBoLu),
14195 Berlin, Germany

Breeder
Noack Laboratorien GmbH,
Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany

Culture
In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20  2 °C, in an incubator, 16 hours illumination, light intensity of max. 1500 lx.

Culture medium
Elendt M4, according to OECD 202, Annex 3 (2004), modified to a total hardness of 160 to 180 mg CaCO3/L, is used. The composition of the culture medium is presented

Culture feeding
The culture daphnids were fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and
Desmodesmus subspicatus, with an algae cell density of > 106 cells/mL. The algae were cultured at the test facility.

Origin of the food algae
Sammlung von Algenkulturen (SAG),
Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, 37073 Göttingen, Germany

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

total hardness [mg CaCO3/L]
0 hours: 2016-06-08 167
24 hours: 2016-06-09 175

Test temperature:

Dilution water

Temperature [°C]
0 hours: 2016-06-08 20.8
24 hours: 2016-06-09 20.4

pH:

Dilution water
pH-Value

0 hours: 2016-06-08 7.67
24 hours: 2016-06-09 7.70

Dissolved oxygen:

Dilution water
Dissolved
O2 concentration
[mg/L]

0 hours: 2016-06-08 8.67
24 hours: 2016-06-09 8.67

Conductivity:

Dilution water


Conductivity
[µS/cm]

0 hours: 2016-06-08 462
24 hours: 2016-06-09 500

Nominal and measured concentrations:

The concentrations of the main component C18 unsatd.-Glucamide were analytically verified via LC MS/MS at the start of the exposure intervals
(0 and 24 hours) and at the end of the exposure intervals (24 and 48 hours) in all tested concentration levels and in the control. Details of the analytical
method are presented.
The measured concentrations of the main component C18 unsatd.-Glucamide at the start of the exposure intervals (0 and 24 hours) were in the range
of 95 to 105% of the nominal values. At the end of the exposure intervals (24 and 48 hours), the measured concentrations of the main component C18
unsatd.-Glucamide were in the range of 36 to 61% of the nominal values. The analytical results are presented.
Adsorption to the glass walls of the test vessels was determined in separate replicates without daphnids at the end of both exposure intervals
(24 and 48 hours) in all concentration levels.
One additional replicates per concentration level without daphnids were prepared. After sampling for the test medium analysis (aqueous phase), the
glassware was emptied, rinsed carefully with demineralized water and 10 mL acetonitrile containing 0.2% formic acid was added. The concentration of the
test item in this solution was measured and the adsorbed test item amount was calculated from this concentration.
Adsorption to the glass walls was in the range of 11 – 23% of the nominal concentrations of the main component C18 unsatd.-Glucamide.

Details on test conditions:

Test temperature (target) 18 - 22 °C, constant within ± 1 °C

Illumination (target) Diffuse light, light intensity of max. 1500 lx

Photoperiod (target) 16/8 hours light/dark cycle

Feeding The daphnids were not fed during the study.

Reference substance (positive control):

yes

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

1.78 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

0.988 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 7.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

2.11 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 7.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 7.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Valuesbased on Nominal Concentrations

C18/C18unsatd.-Glucamide
Effect levels	Test duration	Toxicity endpoint based on the
	[hours]	nominal concentrations of the test item   [mg/L]		nominal concentrations of the active components (80.0% (w/w)) [mg/L]
EC10 (with 95% confidence limits)	24	1.78	(Cl: 1.25 - 2.16)	1.42	(Cl: 1.00 - 1.73)
	48	0.988	(Cl: 0.855 - 1.13)	0.792	(Cl: 0.685 - 0.907)
EC50 (with 95% confidence limits)	24	> 7.10	(Cl: Not applicable)	> 5.68	(Cl: Not applicable)
	48	2.11	(Cl: 1.93 - 2.31)	1.69	(Cl: 1.54 - 1.85)
EC100	24	> 7.10		> 5.68
	48	> 7.10		> 5.68

 

 

 EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Values based on Measured Concentrations

C18/C18unsatd.-Glucamide
Effect levels	Test duration	Toxicity endpoint based on the
	[hours]	geometric mean measured concentrations of the test item [mg/L]		geometric mean measured concentrations of the active components (80.0% (w/w)) [mg/L]
EC10 (with 95% confidence limits)	24	1.21	(Cl: 0.821 - 1.51)	0.969	(Cl: 0.657 - 1.20)
	48	0.670	(Cl: 0.600 - 0.751)	0.536	(Cl: 0.480 - 0.600)
EC50 (with 95% confidence limits)	24	> 5.58	(Cl: Not applicable)	> 4.47	(Cl: Not applicable)
	48	1.45	(Cl: 1.31 - 1.61)	1.16	(Cl: 1.05 - 1.29)
EC100	24	> 5.58		> 4.47
	48	> 5.58		> 4.47

 

Measured Exposure Concentrations of the Main Component Unsatd. C18 fraction of theTest ItemC18/C18 unsatd.-Glucamide, the Test Item and the Active Components Glucamides (80.0% (w/w))during the Definitive Test

C18/C18 unsatd.-Glucamide Nominal test item concentration [mg/L]				7.10	3.99	2.25	1.26	0.710	Control
Sum of Glucamides (80.0% (w/w)) Nominal concentration of the active components [mg/L]				5.68	3.19	1.80	1.01	0.568
C18 unsatd.-Glucamide Nominal concentration of the main component [mg/L]				5.31	2.98	1.68	0.942	0.531
Sampling date		Start of analysis		Measured concentrations [mg/L]
2016-06-08 Start of the exposure interval, 0 hours		2016-06-08 ^1)	Test item	7.18	3.88	2.15	1.23	0.721	< LOQ
			Main component	5.37	2.90	1.61	0.922	0.539
			%	101	97	95	98	102
2016-06-09 End of the exposure interval, 24 hours		2016-06-09	Test item	4.22	2.08	1.19	0.665	0.422	< LOQ
			Main component	3.15	1.56	0.888	0.497	0.316
			%	59	52	53	53	59
2016-06-09 Start of the exposure interval, 24 hours		2016-06-09	Test item	7.46	4.21	2.30	1.28	0.742	< LOQ
			Main component	5.58	3.15	1.72	0.956	0.555
			%	105	105	102	101	105
2016-06-10 End of the exposure interval, 48 hours		2016-06-10	Test item	4.31	2.41	1.07	0.454	0.349	< LOQ
			Main component	3.22	1.81	0.797	0.339	0.261
			%	61	61	47	36	49
Geometric mean measured concentrations of the	test item C18/C18 unsatd.-Glucamide [mg/L]			5.58	3.01	1.58	0.830	0.530	< LOQ
	active components Glucamides (80.0% (w/w)) [mg/L]			4.47	2.41	1.26	0.664	0.424

Meas. conc.    = measured concentrationofC18 unsatd.-Glucamide(maincomponent) of the test item

                          C18/C18 unsatd.-Glucamide, mean value of 2 injections, dilution factors taken into account

%                     = percent of the nominal concentrationofC18 unsatd.-Glucamide(maincomponent) of the test item

LOQ                = limit of quantification of theanalytical method(2.24 µg/L of the main component)

1)                     = reanalyzed on 2016-06-15

 

 

 

   Measured Concentrations of the Main Component Unsatd. C18 fractionof the Test Item in Aqueous Phase and at Glass Walls of Test Vessels without Daphnids after 24 hours

C18 unsatd.-Glucamide, main component ofthe test itemC18/C18 unsatd.-Glucamide
Nominal concentration of the		End of the 1^stexposure interval (24 hours)				End of the 2^ndexposure interval (48 hours)
test item C18/C18 unsatd.- Glucamide [mg/L]	main component C18 unsatd.- Glucamide [mg/L]	Aqueous Phase 20 mL		Adsorbed to Glass		Aqueous Phase 20 mL		Adsorbed to Glass
		Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%
7.10	5.31	4.35	82	0.892	17	3.52	66	0.627	12
3.99	2.98	1.94	65	0.499	17	2.07	69	0.332	11
2.25	1.68	1.00	60	0.307	18	1.12	67	0.188	11
1.26	0.942	0.554	59	0.160	17	0.432	46	0.217	23
0.710	0.531	0.311	59	0.0647	12	0.330	62	0.0590	11

Meas. conc.    = measured concentrationofC18 unsatd.-Glucamide(maincomponent) of the test item

                          C18/C18 unsatd.-Glucamide, mean value of 2 injections, dilution factor taken into account

%                     = percent of the nominal concentrationofC18 unsatd.-Glucamide(maincomponent) of the test item

Conclusions:

Based on the nominal concentrations of the test item C18/C18 unsatd.-Glucamide, the 48 hour EC50 for Daphnia magna was 2.11 mg/L (95% confidence limits: 1.93 – 2.31 mg/L).
Based on the nominal concentrations of the active components Glucamides (80.0% (w/w)), the 48 hour EC50 for Daphnia magna was 1.69 mg/L (95% confidence limits: 1.54 – 1.85 mg/L).
Based on the geometric mean measured concentrations of the test item C18/C18 unsatd.-Glucamide, the 48 hour EC50 for Daphnia magna was 1.45 mg/L (95% confidence limits: 1.31 – 1.61 mg/L).
Based on the geometric mean measured concentrations of the active components Glucamides (80.0% (w/w)), the 48 hour EC50 for Daphnia magna was 1.16 mg/L (95% confidence limits: 1.05 – 1.29 mg/L).

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the test itemC18/C18 unsatd.-Glucamide were determined according to OECD 202 (2004)

from2016-06-08 to 2016-06-20, with the definitive exposure phase from 2016-06-08 to 2016‑06-10,at the test facility.

The study was conducted under semi-static conditions over a period of 48 hours with five concentrations of the test itemC18/C18 unsatd.-Glucamideranging from0.710 to 7.10 mg/Lprepared in Elendt M4 medium (as specified inTable3)in a geometric series with a separation factor of 10^1/4.

The test item is an UVCB substance and is poorly soluble in water at room temperature (cmc: 0.0071±0.0032 g/L).

The highest concentration level 7.10 mg/L was slightly turbid (Tyndall effects observed) and foamed during the preparation. All other test item concentrations were visually clear throughout the exposure period.

Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration leveland the control.

The concentrations of the main component C18 unsatd.-Glucamide were analytically verified via LC‑MS/MS at the start of the exposure intervals (0 and 24 hours) and at the end of the exposure intervals

(24 and 48 hours) in all tested concentration levels and in the control. Details of the analytical method are presented in section14.

The measured concentrations of the main componentC18 unsatd.-Glucamideat the start of the exposure intervals (0 and 24 hours) were in the range of 95 to 105% of the nominal values. At the end of the exposure intervals (24 and 48 hours), the measured concentrations of the main componentC18 unsatd.-Glucamidewere in the range of 36 to 61% of the nominal values.Theanalytical results are presented .

The geometric mean measured concentrations of the test item were: 0.530 - 0.830 - 1.58 - 3.01 - 5.58 mg/L. The geometric mean measured concentrations of the

active components (80.0% (w/w) were: 0.424 - 0.664 - 1.26 - 2.41 - 4.47 mg/L.

Adsorption to the glass walls of the test vessels was determined in all concentration in separate replicates without daphnids. Adsorption to the glass walls was in the range of 11 – 23% of the nominal concentrations.

The effect levels given are based on the nominal concentrations as well as the geometric mean measured concentrations of the test itemC18/C18 unsatd.-Glucamideand the active components Glucamides

The validity criteria of the test guideline were fulfilled.

 

 

 EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Valuesbased on Nominal Concentrations

C18/C18unsatd.-Glucamide
Effect levels	Test duration	Toxicity endpoint based on the
	[hours]	nominal concentrations of the test item   [mg/L]		nominal concentrations of the active components (80.0% (w/w)) [mg/L]
EC10 (with 95% confidence limits)	24	1.78	(Cl: 1.25 - 2.16)	1.42	(Cl: 1.00 - 1.73)
	48	0.988	(Cl: 0.855 - 1.13)	0.792	(Cl: 0.685 - 0.907)
EC50 (with 95% confidence limits)	24	> 7.10	(Cl: Not applicable)	> 5.68	(Cl: Not applicable)
	48	2.11	(Cl: 1.93 - 2.31)	1.69	(Cl: 1.54 - 1.85)
EC100	24	> 7.10		> 5.68
	48	> 7.10		> 5.68

 

 

EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Values based on Measured Concentrations

C18/C18unsatd.-Glucamide
Effect levels	Test duration	Toxicity endpoint based on the
	[hours]	geometric mean measured concentrations of the test item [mg/L]		geometric mean measured concentrations of the active components (80.0% (w/w)) [mg/L]
EC10 (with 95% confidence limits)	24	1.21	(Cl: 0.821 - 1.51)	0.969	(Cl: 0.657 - 1.20)
	48	0.670	(Cl: 0.600 - 0.751)	0.536	(Cl: 0.480 - 0.600)
EC50 (with 95% confidence limits)	24	> 5.58	(Cl: Not applicable)	> 4.47	(Cl: Not applicable)
	48	1.45	(Cl: 1.31 - 1.61)	1.16	(Cl: 1.05 - 1.29)
EC100	24	> 5.58		> 4.47
	48	> 5.58		> 4.47

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-06-08 to 2016-06-15

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)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Determination of the test item
All concentration levels and the control were analytically verified via
LC-MS/MS at the start (0 and 24 hours) and at the end of the exposure intervals (24 and 48 hours). The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000). Details of the analytical method are presented


Sampling for the analytical monitoring
At the start of the exposure intervals (0 and 24 hours), sampling was
carried out after preparation of the test concentrations.
At the end of the exposure intervals (24 and 48 hours), samples were taken directly from the test vessels.

Range (target)
Recoveries of the test item should be within ± 20% of the nominal or initially measured concentrations.

Determination of losses due adsorption onto glassware
The adsorption of the test item on glass was quantified in all
concentration levels at the end of the exposure intervals (24 and 48 hours). One additional replicate per concentration level without daphnids was prepared. After sampling for the test medium analysis (aqueous phase), the glassware was emptied and an appropriate amount of a suitable solvent was added. The concentration of the test item in this solution was measured and the adsorbed test item amount was calculated from this concentration. Before this procedure, the test vessels were carefully rinsed with demineralised water to remove the remaining test media.

Schedule of Specific Test Item Analysis
Sampling Date
[h] Samples analysed:

0 and 24
(fresh media) aqueous phase without daphnids:
all concentration levels and the control
24 and 48
(old media) aqueous phase with daphnids:
all concentration levels and the control
aqueous phase and glass surface without daphnids:
all concentration levels

Vehicle:

no

Details on test solutions:

Stock solution
The stock solution (7.10 mg/L of the test item were weighed out) was freshly prepared with ISO test water (acc. to OECD 202, Annex 3 as specified below) before the start of each exposure interval (at 0 and 24 hours).

Treatment
The stock solution was treated with ultrasound for 30 minutes at 40 °C and stirred thereafter with a magnetic stirrer at approximately 1100 rpm for 15 minutes at room temperature.

Test concentrations
5 test item concentrations in a geometric series with a separation factor of 101/4, prepared by dilution of the stock solution 7.10 mg/L with ISO test water (acc. to OECD 202, Annex 3 as specified below), were tested as follows:
0.710 - 1.26 - 2.25 - 3.99 - 7.10 mg/L
The test concentrations were selected based on the results of a non-GLP preliminary range finding test. For results, see section 7.1.

Control
Dilution water without test item incubated under the same conditions as the test groups

Test method
Based on the results of a preliminary range finding test, the study was performed with a semi-static test design with a renewal of the test solutions after 24 hours, since the measured concentrations did not remain stable over 48 hours under test conditions in the range finding test.

Test duration
48 hours

Test vessels
Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with watch glasses

Renewal of the test solutions
The test solutions were renewed after 24 hours. For this purpose, a second set of test vessels was filled with the freshly prepared test solutions and the daphnids were transferred by pipette (see ‘Application’).

Dilution water ISO Test water, according to OECD 202, Annex 3
Component Concentration [mg/L]
KCl 5.75
NaHCO3 64.8
CaCl2  2 H2O 294
MgSO4  7 H2O 123

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system
Daphnia magna STRAUS (Clone 5).

Reason for the selection of the test system
Daphnia magna is the preferred species in accordance with the
test guideline and is bred at the test facility.

Origin
Institut für Wasser-, Boden- und Lufthygiene (WaBoLu),
14195 Berlin, Germany

Breeder
Noack Laboratorien GmbH,
Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany

Culture
In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20  2 °C, in an incubator, 16 hours illumination, light intensity of max. 1500 lx.

Culture medium
Elendt M4, according to OECD 202, Annex 3 (2004), modified to a total hardness of 160 to 180 mg CaCO3/L, is used. The composition of the culture medium is presented

Culture feeding
The culture daphnids were fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus, with an algae cell density of > 106 cells/mL. The algae were cultured at the test facility.

Origin of the food algae
Sammlung von Algenkulturen (SAG),
Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, 37073 Göttingen, Germany

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Total hardness [mg CaCO3/L]
0 hours 2016-06-08 265
24 hours 2016-06-09 262

Test temperature:

Temperatur [°C]
0 hours 2016-06-08 20.4
24 hours 2016-06-09 20.2

pH:

Nominal
test item
concentration
[mg/L] 0 hours 24 hours
pH-value Dissolved
O2 concentration [mg/L] pH-value Dissolved
O2 concentration [mg/L]
7.10 7.43 7.64 8.30 7.11
3.99 7.44 8.00 8.31 7.91
2.25 7.37 7.88 8.26 8.59
1.26 7.30 8.31 8.06 8.64
0.710 7.22 8.01 7.98 7.56
Control 7.69 8.75 7.74 8.86

Nominal
test item
concentration
[mg/L] pH-values Dissolved O2 concentration [mg/L]
Replicates Replicates
1 2 3 4 1 2 3 4
7.10 7.82 7.83 7.82 7.82 8.24 8.32 8.19 8.13
3.99 7.73 7.82 7.81 7.82 7.83 7.36 8.74 8.46
2.25 7.80 7.70 7.83 7.81 8.18 8.24 7.99 8.44
1.26 7.81 7.82 7.82 7.78 8.60 7.57 8.12 7.75
0.710 7.81 7.82 7.80 7.83 8.75 8.36 8.01 7.83
Control 7.79 7.82 7.80 7.80 6.65 7.75 8.59 8.50

 
Table 12: Water Quality Parameters at the End of the Exposure Interval (48 hours)
(measured in all replicates containing daphnids)
Nominal
test item
concentration
[mg/L] pH-values Dissolved O2 concentration [mg/L]
Replicates Replicates
1 2 3 4 1 2 3 4
7.10 7.77 n.d. 7.77 7.77 8.25 n.d. 8.10 8.24
3.99 7.75 7.75 7.76 7.75 7.86 8.18 8.22 7.71
2.25 7.72 7.73 7.74 7.75 7.57 7.15 8.22 8.14
1.26 7.68 7.69 7.71 7.70 7.30 8.15 8.32 7.50
0.710 7.52 7.56 7.59 7.66 8.26 8.35 8.04 8.32
Control 7.03 7.37 7.44 7.47 7.20 7.99 8.22 8.33
n.d. = not determined, due to 100% mortality after 24 hours

Dissolved oxygen:

Nominal
test item
concentration
[mg/L] 0 hours 24 hours
pH-value Dissolved
O2 concentration [mg/L] pH-value Dissolved
O2 concentration [mg/L]
7.10 7.43 7.64 8.30 7.11
3.99 7.44 8.00 8.31 7.91
2.25 7.37 7.88 8.26 8.59
1.26 7.30 8.31 8.06 8.64
0.710 7.22 8.01 7.98 7.56
Control 7.69 8.75 7.74 8.86

Nominal
test item
concentration
[mg/L] pH-values Dissolved O2 concentration [mg/L]
Replicates Replicates
1 2 3 4 1 2 3 4
7.10 7.82 7.83 7.82 7.82 8.24 8.32 8.19 8.13
3.99 7.73 7.82 7.81 7.82 7.83 7.36 8.74 8.46
2.25 7.80 7.70 7.83 7.81 8.18 8.24 7.99 8.44
1.26 7.81 7.82 7.82 7.78 8.60 7.57 8.12 7.75
0.710 7.81 7.82 7.80 7.83 8.75 8.36 8.01 7.83
Control 7.79 7.82 7.80 7.80 6.65 7.75 8.59 8.50

 
Table 12: Water Quality Parameters at the End of the Exposure Interval (48 hours)
(measured in all replicates containing daphnids)
Nominal
test item
concentration
[mg/L] pH-values Dissolved O2 concentration [mg/L]
Replicates Replicates
1 2 3 4 1 2 3 4
7.10 7.77 n.d. 7.77 7.77 8.25 n.d. 8.10 8.24
3.99 7.75 7.75 7.76 7.75 7.86 8.18 8.22 7.71
2.25 7.72 7.73 7.74 7.75 7.57 7.15 8.22 8.14
1.26 7.68 7.69 7.71 7.70 7.30 8.15 8.32 7.50
0.710 7.52 7.56 7.59 7.66 8.26 8.35 8.04 8.32
Control 7.03 7.37 7.44 7.47 7.20 7.99 8.22 8.33
n.d. = not determined, due to 100% mortality after 24 hours

Conductivity:

Conductivity (µS/cm)
0 hours 2016-06-08 652
24 hours 2016-06-09 667

Nominal and measured concentrations:

The geometric mean measured concentrations of the test item were: 0.500 - 0.837 - 1.51 - 2.83 - 4.68 mg/L. The geometric mean measured concentrations of the active components (80.0% (w/w) were: 0.400 - 0.670 - 1.21 - 2.26 - 3.74 mg/L.
Adsorption to the glass walls of the test vessels was determined in separate replicates without daphnids at the end of both exposure intervals (24 and 48 hours) in all concentration levels.
One additional replicates per concentration level without daphnids were prepared. After sampling for the test medium analysis (aqueous phase), the glassware was emptied, rinsed carefully with demineralized water and 10 mL acetonitrile containing 0.2% formic acid was added.. The concentration of the test item in this solution was measured and the adsorbed test item amount was calculated from this concentration.
Adsorption to the glass walls was in the range of 7 – 20% of the nominal concentrations. For details, see Table 9.

Details on test conditions:

and replicates Number of daphnids
20 daphnids, divided into 4 replicates, each with 5 daphnids, were
used per concentration level and control.

Age of the daphnids at the start of the exposure
were removed from the culture vessels at the latest 22 hours before the start of the exposure and discarded. The juveniles born within this period of max. 22 hours preceding the exposure were used for the test after an acclimatization phase of at least 2 hours in the dilution water. No first brood progeny was used for the test.

Acclimatization
At least 2 hours in dilution water

Application
20 g test solution per replicate was weighed into each test vessel. This corresponds to 20 mL per test vessel. The daphnids were inserted with a small amount of dilution water (start of the exposure) or test solution (water renewal) by a pipette.

Test temperature (target)
18 - 22 °C, constant within ± 1 °C

Illumination (target)
Diffuse light, light intensity of max. 1500 lx

Photoperiod (target)
16/8 hours light/dark cycle

Feeding
The daphnids were not fed during the study.

Reference substance (positive control):

yes

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

3.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

0.494 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

6.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

2.83 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 7.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

7.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Values based on Nominal Concentrations

C18/C18unsatd.-Glucamide
Effect levels	Test duration	Toxicity endpoint based on the
	[hours]	nominal concentrations of the test item   [mg/L]		nominal concentrations of the active components (80.0% (w/w)) [mg/L]
EC10 (with 95% confidence limits)	24	3.60	(Cl: 2.60 - 4.70)	2.88	(Cl: 2.08 - 3.76)
	48	0.494	(Cl: < 0.710 - 1.78)	0.412	(Cl: < 0.568 - 1.43)
EC50 (with 95% confidence limits)	24	6.10	(Cl: 5.24 - 7.01)	4.88	(Cl: 4.19 - 5.61)
	48	2.83	(Cl: 2.40 - 3.31)	2.26	(Cl: 1.92 - 2.65)
EC100	24	> 7.10		> 5.68
	48	7.10		5.68

 

 

 EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Values based on Measured Concentrations

C18/C18unsatd.-Glucamide
Effect levels	Test duration	Toxicity endpoint based on the
	[hours]	geometric mean measured concentrations of the test item [mg/L]		geometric mean measured concentrations of the active components (80.0% (w/w)) [mg/L]
EC10 (with 95% confidence limits)	24	2.57	(Cl: 1.91 - 3.26)	2.05	(Cl: 1.53 - 2.61)
	48	0.539	(Cl: < 0.500 - 1.16)	0.426	(Cl: < 0.400 - 0.930)
EC50 (with 95% confidence limits)	24	4.10	(Cl: 3.59 - 4.63)	3.28	(Cl: 2.87 - 3.70)
	48	1.94	(Cl: 1.62 - 2.31)	1.56	(Cl: 1.30 - 1.85)
EC100	24	> 4.68		> 3.74
	48	4.68		3.74

 

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item C18/C18 unsatd.-Glucamide, the 48 hour EC50 for Daphnia magna was 2.83 mg/L (95% confidence limits: 2.40 – 3.31 mg/L).
Based on the nominal concentrations of the active components Glucamides (80.0% (w/w)), the 48 hour EC50 for Daphnia magna was 2.26 mg/L (95% confidence limits: 1.92 – 2.65 mg/L).
Based on the geometric mean measured concentrations of the test item C18/C18 unsatd.-Glucamide, the 48 hour EC50 for Daphnia magna was 1.94 mg/L (95% confidence limits: 1.62 – 2.31 mg/L).
Based on the geometric mean measured concentrations of the active components Glucamides (80.0% (w/w)), the 48 hour EC50 for Daphnia magna was 1.56 mg/L (95% confidence limits: 1.30 – 1.85 mg/L).

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the test itemC18/C18 unsatd.-Glucamide were determined according to OECD 202 (2004)

from 2016-06-08 to 2016-06-15, with the definitive exposure phase from 2016-06-08 to 2016‑06-10,at the test facility.

The study was conducted under semi-static conditions over a period of 48 hours with five concentrations of the test itemC18/C18 unsatd.-Glucamideranging from0.710 to 7.10 mg/Lprepared in ISO test water (acc. to OECD 202, Annex 3) in a geometric series with a separation factor of10^1/4.

The test item is an UVCB substance and is poorly soluble in water at room temperature (cmc: 0.0071±0.0032 g/L).

The highest concentration level 7.10 mg/L was slightly turbid (Tyndall effects observed) and foamed during the preparation. All other test item concentrations were visually clear throughout the exposure period.

Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration leveland the control.

The concentrations of the main component C18 unsatd.-Glucamide were analytically verified via LC‑MS/MS at the start of the exposure intervals (0 and 24 hours) and at the end of the exposure intervals

(24 and 48 hours) in all tested concentration levels and in the control. Details of the analytical method are presented in section14.

The measured concentrations of the main componentC18 unsatd.-Glucamideat the start of the exposure intervals (0 and 24 hours) were in the range of 89 to 99% of the nominal values. At the end of the exposure intervals (24 and 48 hours), the measured concentrations of the main componentC18 unsatd.-Glucamidewere in the range of 41 to 54% of the nominal values.Theanalytical results are presented.

The geometric mean measured concentrations of the test item were: 0.500 - 0.837 - 1.51 - 2.83 - 4.68 mg/L. The geometric mean measured concentrations of the active components (80.0% (w/w) were:

0.400 - 0.670 - 1.21 - 2.26 - 3.74 mg/L.

Adsorption to the glass walls of the test vessels was determined in all concentration in separate replicates without daphnids. Adsorption to the glass walls was in the range of 7 – 20% of the nominal concentrations.

The effect levels given are based on the nominal concentrations as well as the geometric mean measured concentrations of the test itemC18/C18 unsatd.-Glucamideand the active components Glucamides

The validity criteria of the test guideline were fulfilled.

 

: EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Valuesbased on Nominal Concentrations

C18/C18unsatd.-Glucamide
Effect levels	Test duration	Toxicity endpoint based on the
	[hours]	nominal concentrations of the test item   [mg/L]		nominal concentrations of the active components (80.0% (w/w)) [mg/L]
EC10 (with 95% confidence limits)	24	3.60	(Cl: 2.60 - 4.70)	2.88	(Cl: 2.08 - 3.76)
	48	0.494	(Cl: < 0.710 - 1.78)	0.412	(Cl: < 0.568 - 1.43)
EC50 (with 95% confidence limits)	24	6.10	(Cl: 5.24 - 7.01)	4.88	(Cl: 4.19 - 5.61)
	48	2.83	(Cl: 2.40 - 3.31)	2.26	(Cl: 1.92 - 2.65)
EC100	24	> 7.10		> 5.68
	48	7.10		5.68

 

 

  EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Values based on Measured Concentrations

C18/C18unsatd.-Glucamide
Effect levels	Test duration	Toxicity endpoint based on the
	[hours]	geometric mean measured concentrations of the test item [mg/L]		geometric mean measured concentrations of the active components (80.0% (w/w)) [mg/L]
EC10 (with 95% confidence limits)	24	2.57	(Cl: 1.91 - 3.26)	2.05	(Cl: 1.53 - 2.61)
	48	0.539	(Cl: < 0.500 - 1.16)	0.426	(Cl: < 0.400 - 0.930)
EC50 (with 95% confidence limits)	24	4.10	(Cl: 3.59 - 4.63)	3.28	(Cl: 2.87 - 3.70)
	48	1.94	(Cl: 1.62 - 2.31)	1.56	(Cl: 1.30 - 1.85)
EC100	24	> 4.68		> 3.74
	48	4.68		3.74

 

Description of key information

In three Acute immobilization Tests with Daphnia magna, the effectsof the test item C18/C18 unsatd.-Glucamide were determined according to OECD 202 (2004) and are rated according to Klimisch with reliability 1. All studies were conducted under semi-static conditions over a period of 48 hours. Therefore it is justified to apply a weight of evidence approach (WoE) and to calculate the geometric mean of the three results. According to the REACH Guidance R.7.8.5 it is also justified to harmonise by geometric mean when there is more than one set of data with the same reliability rating is available and the results are not more than one order of magnitude apart.

 

Geometric mean calculation

 

	Report	Test item (100 %)		a.i. (80 %)
		EC50 nominal (mg/L)	EC50 measured mg/L)	EC50 nominal (mg/L)	EC50 measured mg/L)
1st Study (ISO Medium)	131014CS/DAI15740	1	0.78	0.8	0.63
2nd Study (ISO Medium)	160518CS/DAI15740	2.83	1.94	2.26	1.56
3rd Study (Elendt Medium)	160517CS/DAI15740	2.11	1.45	1.69	1.16
Geometric mean		1.81	1.30	1.45	1.04

 

It has been shown that all calculated EC₅₀values are above 1 mg/L. The EC₅₀derived as the geometric mean of all three studies and based on the measured concentrations of the active ingredient was determined to be 1.04 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1.3 mg/L