Disodium 6-(4,6-dichloro-1,3,5-triazin-2-ylamino)-1-hydroxy-2-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-3-sulfonate

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:

From Sep. 03, 1997 to Nov. 07, 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Water samples were taken at study start and study end from all tested concentrations.
- Sample preparation: The samples were diluted with mobile phase

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method of administration: Test substance was weighed into a graduated flask, filled up with test water and stirred for about 5 min with a magnetic stirrer. The amounts corresponding to the test concentrations were pipetted into a graduated flask, made up to a total volume of 200 mL with test water, and then transferred to the test chambers

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Clone 5 (IRChA, France)
- Source: Stock culture of Hoechst Marion Roussel Deutschland GmbH, Global Preclinical Development - Drug Safety, 65926 Frankfurt am Main.
- Age at study initiation (mean and range, SD): 0-24 h
- Method of breeding: 100 mL beakers (height 90 mm, diameter 50 mm), filled with 70 mL test water (according to Elendt, M4) served as culture vessels. One daphnid was used per culture vessel. The animals were used for breeding up to an age of 42 d. The reproduction rate and the state of health of the animals were monitored daily apart from weekends.
- Food type: monocellular green algae (Scenedesmus subspicatus).
- Frequency: Three times a week

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

None

Post exposure observation period:

No

Hardness:

No data

Test temperature:

20.5-21.2 °C

pH:

7.8-8.3

Dissolved oxygen:

8.4-9.0 mg/L

Salinity:

No data

Nominal and measured concentrations:

Nominal: 0, 22, 50, 100, 220, 500 mg/L
Measured (mean): 0, 19.4, 48, 86, 193, 479 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Beakers
- Material, size, headspace, fill volume: 250 mL beakers (height 110 mm, diameter 70 mm, water level about 75 mm), each filled with 200 mL
test water
- Aeration: Aerated until saturated with oxygen
- Renewal rate of test solution: No
- No. of organisms per vessel: 10 daphnids/vessel
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Keeping water: Reconstituted fresh water
- Method of administration: Direct weighing

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Demineralized water with a conductance of < 1 µS /cm was used for preparing the artificial fresh water (according to Elendt)


OTHER TEST CONDITIONS
- Photoperiod: 12 h light/12 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Number of immobile daphnids after 24 h and 48 h

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 117.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 87.88-158.9

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: -

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

362 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 292-468.2

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: -

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: -

Details on results:

- Other biological observations: No
- Mortality of control: No
- No immobility was observed in control group

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

ECSO values were determined after 24 and 48 h by means of the probit analysis (after the method according to LINDER und WEBER, confidence limit according to FIELLER).

Table 1. Immobility-Results

Concentration mg/L	Immobility after 24 h		Immobility after 48 h
	Absolute	%	Absolute	%
22	0/20	0	0/20	0
50	0/20	0	3/20	15
100	0/20	0	14/20	70
220	3/20	15	10/20	50
500	15/20	75	20/20	100

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the 48 h EC50 value of the test substance was determined to be 117.4 mg/L.

Executive summary:

A study was performed to assess the acute toxicity of test substance to Daphnia magna (water flea) over 48 h under static conditions. The study was conducted in accordance with OECD Guideline 202 and EU Method C.2. in compliance with GLP.

Daphnids were exposed to the concentrations tested were 22, 50, 100, 220 and 500 mg/L and a negative control (0 mg/L). The substance concentration was determined in all tested groups at the test start and after 48 h of exposure. Observations were made on the immobilisation rate after 24 and 48 h of exposure.

All tested concentrations were present as clear orange solutions. The values determined were in a range of ± 20 % of the theoretical value.

The 24 and 48 h EC50 values were determined to be 362.8 and 117.4 mg/L respectively. In the control group no immobility was observed. Immobility was observed at 50, 100, 220 and 500 mg/L. The 22 mg/L group showed no effect in comparison to the negative control.

 

Under the test conditions, the 48 h EC50 value of the test substance was determined to be 117.4 mg/L.

Description of key information

The 48 h EC50 value of the test substance was determined to be 117.4 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

117.4 mg/L

Additional information

A study was performed to assess the acute toxicity of test substance to Daphnia magna (water flea) over 48 h under static conditions. The study was conducted in accordance with OECD Guideline 202 and EU Method C.2.

 

Daphnids were exposed to the concentrations tested were 22, 50, 100, 220 and 500 mg/L and a negative control (0 mg/L). Analysis was performed for the test concentration plus a control at the test start and after 48 h of exposure. Observations were made on the immobilisation rate, after 24 and 48 h of exposure.

All tested concentrations were present as clear orange solutions. The substance concentration was determined in all tested groups. The values determined were in a range of ± 20 % of the theoretical value.

The 24 and 48 h EC50 values were determined to be 362.8 and 117.4 mg/L respectively. In the control group no immobility was observed. Immobility was observed at 50, 100, 220 and 500 mg/L. The 22 mg/L group showed no effect in comparison to the negative control.

 

The 48 h EC50 value of the test substance was determined to be 117.4 mg/L (Dr. Zok S, 1997)