N,N'-[1,4-phenylenebis(methylene)]bis{N,N-dimethyl-3-[(prop-2-enoyl)amino]propan-1-aminium} dichloride

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:

27 Feb - 01 Mar 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Rheinland-Pfalz, Mainz, Germany

Test material

Test material information

Constituent 1

Reference substance name:

{[4-({dimethyl[3-(prop-2-enamido)propyl]azaniumyl}methyl)phenyl]methyl}dimethyl[3-(prop-2-enamido)propyl]azanium dichloride

EC Number:

812-548-5

Cas Number:

1621424-03-0

Molecular formula:

C24H40Cl2N4O2

IUPAC Name:

{[4-({dimethyl[3-(prop-2-enamido)propyl]azaniumyl}methyl)phenyl]methyl}dimethyl[3-(prop-2-enamido)propyl]azanium dichloride

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All test concentrations (0 h, 48 h)

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution containing 100.0 mg/L test item in dilution water was prepared. The lower concentrations were prepared by dilution of this stock solution with dilution water.
- Differential loading: no
- Controls: dilution water only

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: Umweltbundesamt Berlin, in-house breeding since 27. September 2007
- Age at study initiation: 0 - 18 h

Method of breeding:
- Similar to the method described in the OECD guideline
- Vessels: preserving glasses (nominal volume: 2 L)
- Medium: M4-Medium (recipe of ELENDT)
- Food: green algae (Desmodesmus subspicatus)
- Medium renewal: twice a week
- Photo period: 16/8 hours (using neon tubes)
- Temperature 20 ± 2 °C

- Feeding during test : no

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS: 19 hours before the start of the test, the adult animals were separated from the young. 1 hour before test start, the adults were caught with the help of a glass tube, and the newborn daphnia, aged between 0 and 18 hours, were sieved from the medium and immediately placed into a beaker containing dilution water.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

19.8 - 21.2°C

pH:

7.7 - 7.8

Dissolved oxygen:

8.4 - 9.0 mg/L

Nominal and measured concentrations:

Nominal concentrations: 0 (control), 4.6, 10, 22, 46 and 100 mg/L
Measured concentrations initial (t-=0) / test end (t-72h) mg/L: 4.30/4.57, 9.86/10.95, 22.63/24.90, 47.99/53.59, 104.59/114.57 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Beakers, nominal volume 50 mL
- Material: glass, fill volume: 20 ± 5 mL
- Aeration: no
- 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
- Culture medium different from test medium: yes, switching from M4-medium (husbandry) to dilution water (test) has been shown not to cause any detrimental effects for test daphnia.
- Intervals of water quality measurement: pH, the concentration of dissolved oxygen (0 , 48 h)

OTHER TEST CONDITIONS
- Adjustment of pH: no

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilised daphnia (0 , 24, 48 h)

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7)

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 5%

Results with reference substance (positive control):

EC50 (24 h): 1.9 mg/L. The 24h-EC50 value is within the requested range for the 24h-EC50 between 0.6 - 2.1 mg/L of potassium dichromate K2Cr2O7 as specified in the OECD guideline.

Reported statistics and error estimates:

Estimation of the biological results was accomplished using the software ToxRat® Professional, version 3.2.1.

Any other information on results incl. tables

Table 1: Measured test item concentrations

Nominal Concentration Test Item	Measured Concentration Test Item t = 0 h	Measured Concentration Test Item t = 48 h	% of Nominal concentration t = 0 h	% of Nominal concentration t = 48 h
mg/L	mg/L	mg/L	%	%
Blank control	< LOQ	< LOQ	--	--
4.6	4.30	4.57	94	99
10	9.86	10.95	99	109
22	22.63	24.90	103	113
46	47.99	53.59	104	116
100	104.59	114.57	105	115

LOQ (Limit of quantification) = 4 mg/L

Table 2: Immobility

Nominal Concentration in mg/L	Immobility 24 hours					Immobility 48 hours
	absolute				in %	absolute				in %
Blank control	0	0	0	0	0	0	1	0	0	5
4.6	0	0	0	0	0	0	0	0	0	0
10	0	0	0	0	0	0	0	0	0	0
22	0	0	0	0	0	0	0	0	0	0
46	0	0	0	0	0	0	0	2	0	10
100	0	0	2	0	10	1	1	2	0	20

Table 3: Validity criteria for OECD 202

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	5%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	8.4 - 9.0 mg/L	yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For further details, refer to "Any other information on results incl. tables".