D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-16 (even numbered) and C18 unsaturated acyl] derivs.

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:

2014-05-14 to 2014-05-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline study performed under GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

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:

D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-16 (even numbered) and C18 unsaturated acyl] derivs.

EC Number:

940-422-0

Cas Number:

1591783-13-9

Molecular formula:

C15H31NO6 (C8 derivative) C17H35NO6 (C10 derivative) C19H39NO6 (C12 derivative) C21H43NO6 (C14 derivative) C23H47NO6 (C16 derivative) C25H51NO6 (C18 derivative) C25H49NO6 (C18 unsatd. derivative)

IUPAC Name:

D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-16 (even numbered) and C18 unsaturated acyl] derivs.

Test material form:

other: waxy

Details on test material:

- Molecular formula: C19H39NO6 (C12-Glucamide) and C21H43NO6 (C14-Glucamide)
- Molecular weight: 377.52 – 405.57 g/mol for C12- and C14-Glucamide
- Substance type: UVCB
- Physical state: waxy solid
- Stability under test conditions: Stable (in water and light at room temperature)
- Storage condition of test material: Room temperature, protected from light, in the tightly closed original container

- Other:
- Solubility (in water at RT): Slightly soluble
- pH value (in aqueous solution): Ca. 8-10 at concentration of 10.000 [mg/L]
- Melting point: 70 – 71 °C
- Boiling point: No boiling point, substance decomposes before boiling
- Vapour pressure: < 0.1 Pa

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

All concentration levels and the control were analytically verified the test item by LC-MS/MS at the start (0 h) and at the end of the exposure (48 h).
At the start of the exposure (0 ), sampling was carried out from the freshly prepared test concentrations.
At the end of the exposure (48 h), samples were taken directly from the test vessels.
The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11).

Test solutions

Vehicle:

no

Details on test solutions:

The stock solution (40.0 mg/L) was treated with ultrasound for 30 minutes at 40 °C. Test concentrations were prepared by diluting the stock solution of with dilution water.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Clone 5
- Source:
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
Breeder: DR.U.NOACK-LABORATORIEN, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
- Age at study initiation (mean and range, SD): 2 to 24 h old daphnids from a healthy stock were used for the study
- Method of breeding:
Culture: In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 +- 2°C, in an incubator, 16 h illumination; light intensity
of max. 20 µE x m-2 x s-1
Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
Cultur 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 > 106 cells/mL
- 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 conditions.
- Health during acclimation (any mortality observed): No mortality observed.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

263 mgCaCO3/L

Test temperature:

18 - 22°C, constant within ± 1°C

pH:

pH at the Start of the Exposure (0 h)
(measured in one additional replicate per concentration level and control)

Nominal test item
concentration
[mg/L] pH-value
40.0 6.93
20.0 6.84
10.0 6.81
5.00 6.79
2.50 6.74
Control 7.61

pH at the End of the Exposure (48 h)
(measured in all replicates)

Nominal test item
concentration pH-values
[mg/L] Replicates
1 2 3 4
40.0 7.47 7.48 7.50 7.51
20.0 7.45 7.45 7.47 7.47
10.0 7.36 7.39 7.41 7.42
5.00 7.28 7.30 7.32 7.32
2.50 7.19 7.21 7.25 7.25
Control 7.19 7.14 7.16 7.16

Dissolved oxygen:

O2-concentration at the Start of the Exposure (0 h)
(measured in one additional replicate per concentration level and control)

Nominal test item
concentration
[mg/L] O2-concentration [mg/L]
40.0 8.19
20.0 8.51
10.0 8.63
5.00 8.71
2.50 8.75
Control 8.70

O2-concentration at the End of the Exposure (48 h)
(measured in all replicates)

Nominal test item
concentration O2-concentration [mg/L]
[mg/L] Replicates
1 2 3 4
40.0 8.14 8.03 8.25 8.20
20.0 8.14 8.20 8.19 8.06
10.0 8.37 8.44 8.59 8.37
5.00 8.61 8.59 8.65 8.45
2.50 8.75 8.71 8.74 8.72
Control 8.86 8.64 8.69 8.71

Nominal and measured concentrations:

2.50 - 5.00 - 10.0 - 20.0 - 40.0 mg test item/L (factor 2)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 cm ID x 7 cm H), 50 mL capacity, loosely covered with watch glasses
- Test volume: 20 mL
- Aeration: None
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO Test water, according to OECD 202, Annex 3
Component Concentration [mg/L]
KCl 5.76
NaHCO3 64.8
CaCl2 x 2 H2O 294
MgSO4 x 7 H2O 123

- Water Quality Parameters of the Dilution Water prior to the Start of the Exposure (0 h):
pH-value DissolvedO2-concentration[mg/L] Temperature[°C] Conductivity[µS/cm] Total hardness [mg CaCO3/L]
7.61 8.70 21.8 654 263

- Culture medium different from test medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: test start (0h) and test end (48h)

OTHER TEST CONDITIONS
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 20 µE x m-2 x s-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Immobilisation, 24 h interval

TEST CONCENTRATIONS
- Test concentrations: 2.50 - 5.00 - 10.0 - 20.0 - 40.0 mg test item/L (spacing factor 2 )

- Range finding study:
A non GLP preliminary range finding test was conducted under semi-static conditions with two concentrations of the test item of 1.00 and 10.0 mg/L.
The percentage immobility was determined in all concentrations and control groups after 24 and 48 h of exposure. The biological results are presented in the
following table.

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
10.0 0 0 0 90 30 60
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

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

4.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Remarks on result:

other: < 2.50 - 6.92

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

4.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Remarks on result:

other: < 2.50 - 5.94

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

20.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Remarks on result:

other: 14.6 - 27.9

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

7.29 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Remarks on result:

other: 5.54 - 9.59

Effect concentrations 5

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 40 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 6

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

40 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 7

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

2.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 8

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Details on results:

See below

Results with reference substance (positive control):

The percentage immobility for the reference item was determined after 24 h. The EC50-value with 95 % confidence interval (CI) was calculated by
sigmoidal dose-response regression. The EC50-value for the most recent of the monthly performed reference tests was:
EC50: 1.62 mg/L (CI 1.57 - 1.66 mg/L)
The EC50-value of the reference item potassium dichromate after 24 h is within the prescribed concentration range of 0.6 - 2.4 mg/L of quality
criteria according to AQS P 9/2 (02/2000) for daphnids clone 5 cultured in Elendt M4 medium. The EC50-value of the reference item is also within therecommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.

Any other information on results incl. tables

Biological Data

The percentage immobility, determined in all concentration levels and the control after 24 and 48 h under static conditions, is given in the table below.

 

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
40.0	32.4	100	80	60	60	75	100	100	100	100	100
20.0	16.2	60	40	60	40	50	100	60	100	100	90
10.0	8.10	40	20	0	40	25	100	60	40	100	75
5.00	4.05	0	0	20	40	15	0	0	20	60	20
2.50	2.03	0	0	0	0	0	0	0	0	0	0
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. At the end of the exposure (48 h), white flakes were observed at the bottom of the test vessels of the concentration levels 20.0 and 40.0 mg/L.

Measured Exposure Concentrations during the Definitive Test

The concentrations of the main components C12-Glucamide and C14-Glucamide were analytically verified via LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) in all concentration levels and the control.

The measured concentrations of the component C12-Glucamide at the start of the exposure (0 h) were in the range of 90 to 109 % of the nominal values and 98 to 106 % of the nominal values at the end of the exposure (48 h). The measured concentrations of the component C14-Glucamide at the start of the exposure (0 h) were in the range of 87 to 110 % of the nominal values and 48 to 91 % of the nominal values at the end of the exposure (48 h).The analytical results are presented in the tables below.

Measured Concentrations of the ComponentC12-Glucamideof theTest ItemGlucamide CCduring the Definitive Test

Sampling date		2014-05-14 0 h Start of the exposure interval		2014-05-16 48 h End of the exposure interval
Nominalconcentrationof the		C12-Glucamide, Component of the Test Item Glucamide CC
test item [mg/L]	component C12-Glucamide [mg/L]	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%
40.0	15.8	17.3	109	16.8	106*
20.0	7.92	7.96	101	8.08	102*
10.0	3.96	3.73	94	4.12	104
5.00	1.98	1.79	90	2.07	105
2.50	0.990	0.907	92	0.974	98*
Control		< LOQ		< LOQ

Meas. conc.    = measured concentration of C12-Glucamide of the test item Glucamide CC, mean value of 2 injections,
  dilution factor taken into account

%                     = percent of the nominal concentration of the test item

LOQ                = limit of quantification (30.0 µg/L of the test item, corresponding to 12.0 µg/L C12-Glucamide)

*                       = reanalysed on 2014-05-16, mean value of 3 replicates

Measured Concentrations of the ComponentC14-Glucamideof theTest ItemGlucamide CCduring the Definitive Test

Sampling date		2014-05-14 0 h Start of the exposure interval		2014-05-16 48 h End of the exposure interval
Nominalconcentrationof the		C14-Glucamide, Component of the Test Item Glucamide CC
test item [mg/L]	component C14-Glucamide [mg/L]	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%
40.0	5.44	5.97	110	2.61	48*
20.0	2.72	2.62	96	1.84	68*
10.0	1.36	1.19	88	1.18	87
5.00	0.680	0.593	87	0.617	91
2.50	0.340	0.305	90	0.277	81**
Control		< LOQ		< LOQ

Meas. conc.    = measured concentration of C14 -Glucamide of the test item Glucamide CC, mean value of 2 injections,
  dilution factor taken into account

%                     = percent of the nominal concentration of the test item

LOQ                = limit of quantification (30.0 µg/L of the test item corresponding to 4.12 µg/L C12-Glucamide)

*                       = reanalysed on 2014-05-16, mean value of 3 replicates

**         = reanalysed on 2014-05-16, mean value of 2 replicates

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item Glucamide CC the 48 h-EC50 for Daphnia magna was 7.29 mg/L
(95 % confidence limits: 5.54 – 9.59 mg/L).
Based on the nominal concentrations of the active components Glucamides the 48 h-EC50 for Daphnia magna was 5.91 mg/L
(95 % confidence limits: 4.49 – 7.77 mg/L).
The NOEC after 48 h was 2.50 mg test item/L, which corresponds to 2.03 mg Glucamides/L.
The LOEC after 48 h was 5.00 mg test item/L, which corresponds to 4.05 mg Glucamides/L.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item Glucamide CC (batch number:EK13/177) were determined according to OECD 202 (2004) from 2014-05-14 to 2014-05-16, 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 concentrations of the test item Glucamide CC in the range of 2.50 to 40.0 mg/Lin a geometric series with a separation factor of 2. 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 slightly soluble in water at room temperature.

The concentrations of the main components C12 -Glucamide and C14 -Glucamide were analytically verified via LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) in all concentration levels and the control.

The measured concentrations of the component C12 -Glucamide at the start of the exposure (0 h) were in the range of 90 to 109 % of the nominal values and 98 to 106 % of the nominal values at the end of the exposure (48 h). The measured concentrations of the component C14 -Glucamide at the start of the exposure (0 h) were in the range of 87 to 110 % of the nominal values and 48 to 91 % of the nominal values at the end of the exposure (48 h).

Nevertheless the effect levels are based on the nominal concentrations of the test item Glucamide CC and the active components Glucamides and presented in the table below.

The water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 h) and at the end of the exposure interval (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 itemGlucamide CCand the active components Glucamides)

Effect levels	Test duration [h]	Nominal concentrations of the test item Glucamide CC [mg/L]	Nominal concentrations of the active components Glucamides (81.0 % (w/w) [mg/L]
EC10 (with 95 % confidence limits)	24	4.40 (< 2.50 – 6.92)	3.57 (< 2.03 – 5.60)
	48	4.05 (< 2.50 – 5.94)	3.28 (< 2.03 – 4.81)
EC50 (with 95 % confidence limits)	24	20.9 (14.6 – 27.9)	16.9 (11.8 – 22.6)
	48	7.29 (5.54 – 9.59)	5.91 ( 4.49 – 7.77)
EC100	24	> 40.0	> 32.4
	48	40.0	32.4
NOEC	48	2.50	2.03
LOEC	48	5.00	4.05