Reaction mass of Reaction product of 1-chloro-3-{[1-chloro-3-(dodecyloxy)propan-2-yl]oxy}propan-2-ol with methyl diethanolamine and [3-(dodecyloxy)-2-hydroxypropyl]bis(2-hydroxyethyl)methylammonium chloride

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:

2018-03-19 to 2018-03-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Reference 2

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of Reaction product of 1-chloro-3-{[1-chloro-3-(dodecyloxy)propan-2-yl]oxy}propan-2-ol with methyl diethanolamine and [3-(dodecyloxy)-2-hydroxypropyl]bis(2-hydroxyethyl)methylammonium chloride

IUPAC Name:

Reaction mass of Reaction product of 1-chloro-3-{[1-chloro-3-(dodecyloxy)propan-2-yl]oxy}propan-2-ol with methyl diethanolamine and [3-(dodecyloxy)-2-hydroxypropyl]bis(2-hydroxyethyl)methylammonium chloride

Test material form:

other: solid, very viscous yellow paste

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Experiment 1: 1.0 / 2.2 / 4.6 / 10 / 22 mg/L; Experiment 2: 0.1 / 0.22 / 0.46 / 1.0 /2.2 mg/L
- Sampling method: taken at the beginning and at the end of the test
- Sample storage conditions before analysis: storage in a fridge for samples not measured immediately. The mean stability of the test item in Daphnia medium was determined to be 119.4 % after 48h.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Stock solutions containing 22.3 mg/L and 22.5 mg/L test item in dilution water were pre-pared for experiment 1 and experiment 2, respectively. The lower concentrations were prepared by dilution of this stock solution with dilution water.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Berlin
- Source: Umweltbundesamt Berlin
- Food type: unicellular green algae (Desmodesmus subspicatus)
SELECTION OF DAPHNIA
- Experiment 1: 18 hours 40 minutes before the start of the test, the adult animals were separated from the young. 0.75 hours before test start, the adults were caught with the help of a glass tube, and the newborn daphnia, aged between 0 and 17 hours 55 minutes, were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in period, animals which showed no apparent damage were used for the test.
- Experiment 2: 18 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 17 hours, were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in period, animals which showed no apparent damage were used for the test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

250 mg CaCO3/L

Test temperature:

Experiment 1: 20.3 - 21.8°C
Experiment 2: 19.7 - 20.3°C

pH:

Experiment 1: 7.8-8.0
Experiment 2: 7.8-7.9

Dissolved oxygen:

Experiment 1: 8.5-9.2 mg/L
Experiment 2: 8.8-9.1 mg/L

Nominal and measured concentrations:

Experiment 1: 1.0 / 2.2 / 4.6 / 10 / 22 mg/L (nominal); 1.2, 2.7, 5.4, 13.1, 29.2 mg/L (measured)
Experiment 2: 0.1 / 0.22 / 0.46 / 1.0 /2.2 mg/L (nominal); 0.1, 0.2, 0.5, 1.1, 2.5 mg/L (measured)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers, nominal volume 50 mL, tall shape
- Fill volume: 20 ± 5 mL test solution/ dilution water
- Renewal rate of test solution (frequency/flow rate): 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
- pH: 7.8
- Dissolved oxygen concentration: 9.2 mg/L
- Total hardness: 250 mg/L as CaCO3
- Culture medium different from test medium: yes, M4-medium for husbandry

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 hours

EFFECT PARAMETERS MEASURED
- Biological Assessment: after 24 h and 48 h: count the immobilised daphnids

RANGE-FINDING STUDY
- Test concentrations: 1 / 10 / 100 mg/L.
- Results used to determine the conditions for the definitive study: At 1 mg/L 0% toxicity was observed and at 10 and 100 mg/L 100% toxicity was observed.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.795 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

1.1 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

IMMOBILITY
- Experiment 1: All concentrations showed toxicity between 95 and 100 % immobilisation. Trapping on the surface or clotting was not observed. None of the animals was immobilised in the blank control.
- Experiment 2: The two highest concentrations showed toxicity between 90 and 100 % immobilisation. None of the animals was immobilised in the lower concentrations and in the blank control.

ANALYTICAL DETERMINATIONS
- Experiment 1: 123 - 135 % of the nominal concentrations at the beginning of the test and 108 - 131 % of the nominal concentration after 48 hours.
- Experiment 2: 79 - 110 % of the nominal concentrations at the beginning of the test and 76 - 115 % of the nominal concentration after 48 hours.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: 24h-EC50: 1.6 mg/L

Reported statistics and error estimates:

Since immobility in experiment 1 was observed from 95 to 100% in all treatments, no statistical evaluation was conducted. However biological results were given in a range of concentrations or determined as lower orlower as or higher than a certain concentration. The estimation of the biological results from experiment 2 was accomplished using the software ToxRat® Professional, version 3.2.1.

Any other information on results incl. tables

Biological results

Immobility experiment 1:

Nominal Concentration in mg/L	Immobility 24 hours					Immobility 48 hours
	absolute				in %	absolute				in %
Blank control	0	0	0	0	0	0	0	0	0	0
1.0	0	0	0	0	0	5	5	5	4	95
2.2	5	5	5	5	100	5	5	5	5	100
4.6	5	5	5	5	100	5	5	5	5	100
10	5	5	5	5	100	5	5	5	5	100
22	5	5	5	5	100	5	5	5	5	100

 

Immobility experiment 2:

Nominal Concentration in mg/L	Immobility 24 hours					Immobility 48 hours
	absolute				in %	absolute				in %
Blank control	0	0	0	0	0	0	0	0	0	0
0.1	0	0	0	0	0	0	0	0	0	0
0.22	0	0	0	0	0	0	0	0	0	0
0.46	0	0	0	0	0	0	0	0	0	0
1.0	0	0	2	0	10	4	4	5	5	90
2.2	5	5	5	5	100	5	5	5	5	100

Analytical results:

Measured concentrations experiment 1:

Nominal Concentration Test Item	Measured concentrations  t = 0 h	Measured concentrations  t = 48 h	% of Nominal concentration  t = 0 h	% of Nominal concentration  t = 48 h
mg/L	mg/L	mg/L	%	%
Blank control	0.00	0.00	--	--
1	1.23	1.23	123	123
2.2	2.75	2.72	125	123
4.6	5.86	4.99	127	108
10	13.25	12.91	132	129
22	29.71	28.72	135	131

Measured concentrations experiment 2:

Nominal Concentration Test Item	Measured concentrations  t = 0 h	Measured concentrations  t = 48 h	% of Nominal concentration  t = 0 h	% of Nominal concentration  t = 48 h
mg/L	mg/L	mg/L	%	%
Blank control	0.00	0.00	--	--
0.1	0.08	0.08	79	76
0.22	0.22	0.19	101	86
0.46	0.46	0.44	101	95
1.0	1.06	1.08	106	108
2.2	2.42	2.52	110	115

Effect values:

Biological Results Test Item Experiment 1 (based on geometric mean of measured concentrations):

Parameter	Value
24h EC50	1.2 - 2.7 mg/L
48h EC50	< 1.2 mg/L
48h NOEC	< 1.2 mg/L
48h LOEC	≤ 1.2 mg/L

 

Biological results test item experiment 2 (based on geometric mean of measured concentrations):

Parameter	Value	95% confidence interval
24h EC50	1.278 mg/L	n.d.
48h EC50	0.795 mg/L	0.689 - 0.922 mg/L
48h NOEC	0.5 mg/L	--
48h LOEC	1.1 mg/L	--

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (48 h) for immobilisation was determined to be 0.795 mg/L. The corresponding NOEC (48 h) was 0.5 mg/L.

Executive summary:

The short term toxicity of test item to Daphnia magna STRAUS was assessed according to OECD 202 resp. EU C.2 over a period of 48 hours.

Two valid experiments were performed. In the first experiment all concentrations showed toxicity between 95 and 100 % immobilisation. In the second experiment the two highest concentrations showed toxicity between 90 and 100 % immobilisation.

The content of the test item in the test solutions was analytically verified using LC-MS-MS-determination. The determination of the biological results was based on the geometric mean of the measured concentrations.

The EC50 (48 h) for immobilisation was determined to be 0.795 mg/L. The corresponding NOEC (48 h) was 0.5 mg/L.