2,2'-[[3-chloro-4-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]phenyl]imino]bisethyl bis(hydrogen sulphate), potassium sodium salt

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:

2017-08-01 to 2018-01-31

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-[[3-chloro-4-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]phenyl]imino]bisethyl bis(hydrogen sulphate), potassium sodium salt

EC Number:

287-719-5

EC Name:

2,2'-[[3-chloro-4-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]phenyl]imino]bisethyl bis(hydrogen sulphate), potassium sodium salt

Cas Number:

85567-07-3

Molecular formula:

C36H42Cl2KN6NaO28S8

IUPAC Name:

2,2'-[[3-chloro-4-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]phenyl]imino]bisethyl bis(hydrogen sulphate), potassium sodium salt

Test material form:

solid

Details on test material:

Batch: 1283067001

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100, 31.3, 9.77, 3.05 and 0.954 mg/L and control
- Sampling method: taken from all test item concentrations and control at test start and after 24 hours from fresh and aged solutions and after 48 hours from aged solutions. For each sampling also a retain sample was taken.
- Sample storage conditions before analysis: mixed with formic acid (0.5% v/v final concentration) immediately after sampling and stored frozen until analysis

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Stock solution S1 – S3: weight necessary amount of test item in volumetric flask and added test medium up to the benchmark, then homogenised by shaking. Lower test solutions: dilution appropriate solution with test medium
- Test concentration: 100, 31.3, 9.77, 3.05 and 0.954 mg/L and control

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain/clone: Daphnia magna (Straus), clone V
- Source: originally purchased from the Federal Environment Agency in Berlin/Germany
- Age at Test Start: Less than 24 hours
- Breeding Conditions: feed with single cell green algae (Desmodesmus subspicatus) at least three times a week.
- Feeding during test: no

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

232 mg CaCO3/L

Test temperature:

19.6 to 20.2 °C

pH:

7.71-8.01

Dissolved oxygen:

9.0-9.2 mg/L

Nominal and measured concentrations:

Nominal: 100, 31.3, 9.77, 3.05, 0.954 mg/L
Measured: 90.0, 26.9, 8.70, 2.62, 0.687 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass vessel (100 mL)
- Type: covered with glass plate
- Material, size, headspace, fill volume: 100 mL, filled up with ~ 50 mL test solution
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: ~ 10 mL of test solution for each animal

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium
- pH: 7.75
- Dissolved oxygen concentration: 9.2 mg/L
- Total hardness:13°dH (232 mg/L as CaCO3)
- Culture medium different from test medium: no

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

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Biological Assessment: after 24 h and 48 h: count the immobilised daphnids and record abnormal behaviour

RANGE-FINDING STUDY
- Test concentrations: 100, 10.0 and 1.00 mg/L and control
- Results used to determine the conditions for the definitive study: After 48 hours of exposure no immobilisation above the allowed control immobilisation was observed in the control and at test item concentration of 1.00 and 10.0 mg/L. 25 % of the daphnids were immobile at at 100 mg/L.

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.:

> 90 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.:

8.7 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

See details in Any other information on results.

Results with reference substance (positive control):

EC50 (24 h) < 1.00 mg/L, in accordance with the requirements of the OECD guideline 202 and fall within the historical data, the daphnids were suitable for the determination of the toxicological effects of the test item.

Any other information on results incl. tables

Immobilisation in the non GLP range-finding test:

	nominal test concentration (mg/L)
	control	1.00	10.0	100
Group 1	0	0	0	1
Group 2	0	0	0	2
Group 3	0	0	0	1
Group 4	0	0	0	1
sum	0	0	0	3
%	0	0	0	25

Results of the main test, after 48 h exppsure:

	Nominal test item concentration [mg/L]
	Control	0.954	3.05	9.77	31.3	100
	Immobilised daphnids after 48 h
Group 1	0	0	0	0	2	2
Group 2	0	0	0	0	1	1
Group 3	0	0	0	0	1	2
Group 4	0	0	0	0	2	3
Σ	0	0	0	0	6	8
%	0	0	0	0	30	40

EC₅₀and NOEC-values of daphnids exposed to the test item evaluated using nominal and actual concentrations:

	[mg/L] (nominal)		[mg/L] (actual*)
	24 h	48h	24 h	48h
NOEC	9.77	9.77	8.70	8.70
EC50	> 100	> 100	> 90.0	> 90.0
95 % confidence limit of EC50	N/A	N/A	N/A	N/A

*The actual concentrations are calculated as geometric mean.

Analytical results:

Nominal concentration (mg/)	Sampling (h)	Measured concentration (mg/L)	% of nominal	Actual concentration (mg/L)*
Control	0, fresh	n.d.	-	-
	24, aged	n.d.	-
	24, fresh	n.d.	-
	48, aged	n.d.	-
0.954	0, fresh	0.725	76	0.687
	24, aged	0.725	76
	24, fresh	0.673	71
	48, aged	0.62	65
3.05	0, fresh	2.61	86	2.62
	24, aged	2.58	85
	24, fresh	2.69	88
	48, aged	2.58	85
9.77	0, fresh	8.73	89	8.7
	24, aged	8.57	88
	24, fresh	8.88	91
	48, aged	8.83	90
31.3	0, fresh	26.6	85	26.9
	24, aged	26.6	85
	24, fresh	26.9	86
	48, aged	27.7	88
100	0, fresh	87.8	88	90
	24, aged	90.9	91
	24, fresh	89.9	90
	48, aged	93.0	93

*The actual concentrations are calculated as geometric mean.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (48 h) for immobilisation is determined to be > 100 mg/L (nominal) and > 90.0 mg/L (actual). The corresponding NOEC (48 h) was 9.77 mg/L (nominal) and 8.70 mg/L (actual).

Executive summary:

The purpose of this study was to determine the immobilisation effect of the test item on the water flea Daphnia magna according to OECD 202 (2004). A semi-static dose-response test with twenty organisms per test concentration (4 replicates of 5 animals each) were used. The duration of the test was 48 hours. Nominal concentrations of 100, 31.3, 9.77, 3.05 and 0.954 mg/L and control were tested.

Analytical samples taken at 0 hours (initial value) and 24 hours from fresh and aged and after 48 hours from aged test solutions were analysed from control and all test item concentrations. The initial measured content of test item was between 71 and 91 % of nominal and the aged measured content was between 65 and 93 % of nominal. Therefore toxicological endpoints were evaluated using nominal and actual concentrations (based on the geometric mean of measured concentrations) of the test item.

According to the results of the test, the EC50 (48 h) was determined to be > 100 mg/L (nominal) and > 90.0 mg/L (actual). The corresponding NOEC (48 h) was 9.77 mg/L (nominal) and 8.70 mg/L (actual).