Amines, N-C12–14 (even numbered)-alkyltrimethylenedi-, reaction products with chloroacetic acid and diethylenetriamine, N-C12–14 (even numbered)-alkyl derivs.

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:

2017-12-04 to 2017-12-06

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)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

All concentration levels and the control were analytically verified via LC-MS/MS in the fresh media at the start of exposure and at renewal of the test solutions (0 and 24 hours) and in the 24-hours old media at renewal and at the end of the exposure (24 and 48 hours).
At the start of the exposure and at renewal (0 and 24 hours), samples were taken after preparation of the test item concentrations and analyzed. At renewal and at the end of the exposure (24 and 48 hours), samples of the 24-hours old media were taken directly from the test vessels.
All original samples were stored at room temperature before 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)
A stock solution (100 mg/L of the test item was weighed out) was freshly prepared with demineralized water prior to the start of the exposure (at 0 hours) and prior to the renewal of the test solutions (at 24 hours). The stock solution was mixed thoroughly by manual agitation until the test item is completely dissolved.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: STRAUS (clone 5)
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany
- Feeding during test: The daphnids were not fed during the study.

ACCLIMATION
- Acclimation period: at least 2 h
- Acclimation conditions (same as test or not): same

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

26 - 32 mg CaCO3/L

Test temperature:

20.3 - 20.5°C

pH:

7.28 - 7.49

Dissolved oxygen:

8.30 - 8.94 mg/L

Conductivity:

147 - 184 µS/cm

Nominal and measured concentrations:

0.313- 0.625 - 1.25 - 2.50 - 5.00 mg/L, corresponding to the nominal dry content concentrations: 0.0833 - 0.166 - 0.333 - 0.665 - 1.33 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with watch glasses; 20 mL fill volume
- Renewal rate of test solution (frequency/flow rate): daily renewal
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- se Any other information on materials and methods

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 1500 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobilization was determined in all groups after 24 and 48 hours.

RANGE-FINDING STUDY
- Test concentrations: 0.0500 – 0.500 and 5.00 mg/L in natural river water
- Results used to determine the conditions for the definitive study: 100 % immobilization after 24 hours at 5 mg/L, 0% immobilization at lower concentrations

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.162 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 95% c.i.

Remarks:

0.0844 – 0.319 mg/L

Details on results:

- Mortality of control: no mortality on control

Results with reference substance (positive control):

- Results with reference substance valid?
- Relevant effect levels: 1.64 mg/L (95% c.i. 1.00 – 4.00 mg/L)
- Valid range: 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000); clone 5
0.6 - 2.1 mg/L, acc. to OECD 202 (2004); clone A

Reported statistics and error estimates:

The EC100-values (after 24 and 48 hours) were empirically derived from the observation data (lowest concentration level resulting in 100% immobilization). All effect concentrations (EC10 / 50 / 100) given are based on the nominal dry content concentrations of test item, since the test item is an UVCB substance. The concentration-effect relationships of the test item after 24 and 48 hours are shown graphically.
The EC10- and the EC50-values (after 24 and 48 hours) were calculated by sigmoidal dose-response regression with the software GraphPad Prism5. The respective 95% confidence limits were calculated from the standard error and the t-distribution.

Validity criteria:

In the control group, no daphnids were immobilized or showed any signs of disease or stress, e.g. discoloration or unusual behavior such as trapping on the surface of the water, during the 48-hour test period (required: not more than 10% of the daphnids in the control).

 

The dissolved O2 concentration in the 24-hours old media at the end of the exposure intervals was ≥8.30 mg/L (required: ≥ 3 mg/L in the 24-hours old media) in all test vessels of all tested concentration levels and the control.

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

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

Nominal dry content concentration of test item [mg/L]	IMMOBILIZATION [%]
	24 hours					48 hours
	Replicates					Replicates
	1	2	3	4	Mean	1	2	3	4	Mean
1.33	100	100	100	100	100	100% mortality after 24 hours
0.665	100	100	100	100	100
0.333	0	0	0	20	5	100	100	100	100	100
0.166	0	0	0	0	0	40	100	60	80	70
0.0833	0	0	0	0	0	0	0	20	0	5
Control	0	0	0	0	0	0	0	0	0	0

Absolute Numbers of immobile Daphnids after 24 and 48 h of Exposure in the Definitive Test

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

Nominal dry content concentration of test item [mg/L]	Number of immobile Daphnids / Total number of Daphnids
	24 h					48 h
	Replicates					Replicates
	1	2	3	4	MV	1	2	3	4	MV
1.33	5 / 5	5 / 5	5 / 5	5 / 5	20 / 20	5 / 5	5 / 5	5 / 5	5 / 5	20 / 20
0.665	5 / 5	5 / 5	5 / 5	5 / 5	20 / 20	5 / 5	5 / 5	5 / 5	5 / 5	20 / 20
0.333	0 / 5	0 / 5	0 / 5	1 / 5	1 / 20	5 / 5	5 / 5	5 / 5	5 / 5	20 / 20
0.166	0 / 5	0 / 5	0 / 5	0 / 5	0 / 20	2 / 5	5 / 5	3 / 5	4 / 5	14 / 20
0.0833	0 / 5	0 / 5	0 / 5	0 / 5	0 / 20	0 / 5	0 / 5	1 / 5	0 / 5	1 / 20
Control	0 / 5	0 / 5	0 / 5	0 / 5	0 / 20	0 / 5	0 / 5	0 / 5	0 / 5	0 / 20

 

 

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal dry content concentrations of the test item, the 48 h EC50 for Daphnia magna was 0.162 mg/L (95% confidence limits: 0.0844 – 0.319 mg/L).

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS), the effects of Reaction product of lauryl-PDA/lauryl-DETA with chloroacetic acid (26.6% a.i.) were determined according to OECD TG 202 (2004) and EU Method C.2 (2008).

The study was conducted under semi-static conditions over a period of 48 hours with natural river water with five nominal concentrations (separation factor of 2): 0.313 – 0.625 – 1.25 – 2.50 – 5.00 mg of the test item, corresponding to the nominal dry content concentrations: 0.0833 – 0.166 – 0.333 – 0.665 – 1.33 mg/L. A stock solution with a nominal concentration of 100 mg/L of the test item was freshly prepared with demineralized water before the start of the exposure period and at renewal (0 and 24 hours) and was used as the stock solution for the preparation of all tested concentration levels. The tested concentration levels were visually clear throughout the exposure period.

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

The exposure concentrations of the test item were analytically verified via LC‑MS/MS in fresh media of all concentration levels and the control at the start of the exposure and at renewal (0 and 24 hours) and in 24-hours aged media at renewal and at the end of the test (24 and 48 hours).

The measured concentrations of the test item in fresh media at the start of the exposure and at renewal (0 and 24 hours) were in the range of 100 to 114% of the nominal values and 59 to 85% in aged media at renewal and at the end of the test (24 and 48 hours).

Since the test item is an UVCB substance all effect concentrations are based on the nominal dry content concentrations of the test item. The validity criteria of the test guidelines were fulfilled.

The 48 h EC50 was 0.162 mg/L (95% confidence limits: 0.0844 – 0.319 mg/L)

 

Result synopsis

Test Organism Age: max. 24 h old daphnids

Test Type: Static Renewal

EC50: 0.162 mg/L (95% confidence limits: 0.0844 – 0.319 mg/L)

Endpoint(s) Effected:  mobility

Description of key information

48 h EC50 = 0.162 mg/L (95% confidence limits: 0.0844 – 0.319 mg/L)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

162 µg/L

Additional information

In the acute immobilization test with Daphnia magna (STRAUS), the effects of Reaction product of lauryl-PDA/lauryl-DETA with chloroacetic acid (26.6% a.i.) were determined according to OECD TG 202 (2004) and EU Method C.2 (2008).

The study was conducted under semi-static conditions over a period of 48 hours with natural river water with five nominal concentrations (separation factor of2): 0.313 – 0.625 – 1.25 – 2.50 – 5.00 mg of the test item, corresponding to the nominal dry content concentrations: 0.0833 – 0.166 – 0.333 – 0.665 – 1.33 mg/L. A stock solution with a nominal concentration of100 mg/Lof the test item was freshly prepared with demineralized water before the start of the exposure period and at renewal (0 and 24 hours) and was used as the stock solution for the preparation of all tested concentration levels. The tested concentration levels were visually clear throughout the exposure period.

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

The exposure concentrations of the test item were analytically verified via LC‑MS/MS in fresh media of all concentration levels and the control at the start of the exposure and at renewal (0 and 24 hours) and in 24-hours aged media at renewal and at the end of the test (24 and 48 hours).

The measured concentrations of the test item in fresh media at the start of the exposure and at renewal (0 and 24 hours) were in the range of 100 to 114% of the nominal values and 59 to 85% in aged media at renewal and at the end of the test (24 and 48 hours).

Since the test item is an UVCB substance all effect concentrations are based on the nominal dry content concentrations of the test item.The validity criteria of the test guidelines were fulfilled.

The 48 h EC50 was 0.162 mg/L (95% confidence limits: 0.0844 – 0.319 mg/L)

The 48 h EC50 of the source substance DOPA-Glycinate in Daphnia magna was 33.30 µg a.i./L (95% CL 26.05–42.57 µg a.i./L).