7-acetamido-4-hydroxy-3-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2-sulphonic acid, sodium salt

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

from 1985-12-18 to 1986-02-13

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP compliant with international guideline; test period 24 h

Qualifier:

according to guideline

Guideline:

other: DIN 38 412, part 11

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

(EEC-Direktive 79/831, Part C)

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 (control) and 250 mg/L
- Sampling method: The test concentrations were measured at start and at the end of the test period
- Sample storage conditions before analysis: Room temperature

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test solutions were performed with the successive dissolution of the stock solution (1 g/L in deionized water) with distilled water.
- Controls: Blank control performed with the test solution without the test substance and positive control performed with potassium dichromate
- Evidence of undissolved material: no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: Hoechst AG, Brüningstr. 64, 65929 Frankfurt, Germany
- Age at study initiation: less then 24 hours
- Method of breeding:at 23 ± 2 °C, 12 hours cycle dark/light
- Food type:algae (Scenedesmus) and TETRA MIN (powder)
- Frequency: once per day

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

24 h

Test temperature:

22.1 - 22.7 °C

pH:

7.1- 7.5

Dissolved oxygen:

8.3 - 8.5 mg/L

Nominal and measured concentrations:

- Nominal concentrations: 500 , 363.3, 250, 176.8, 125.6, 88.4 and 0 (control) mg/L
- Measured concentrations: Test preparation analytics at 250 mg/L did not show any change in extinction at the end of study compared to the extinction at study start.

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL crystallizing dish
- Material, size, headspace, fill volume: 20 mL
- Aeration: no
- No. of organisms per vessel: 20 animals
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- No. of vessels per vehicle control: 0

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod:12 hours cycle dark/light

TEST CONCENTRATIONS
- Range finding study: yes: 500, 100, 10 and 0 mg/L

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

ca. 88.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

ca. 355 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 297-424 mg/L

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

ca. 600 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: no
- Any observations that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no
After 24 h the EC50 was determined to be 355 mg/L

Results with reference substance (positive control):

- Results with reference substance valid? - Yes
- Mortality: 30% at 0.9 mg/L; 100% at 1.9 mg/L

Reported statistics and error estimates:

Calculation of EC50, regression and correlation; conficence interval according to Litchfield-Wilcoxon

Absorbance:

At the start 9.5 at end of test 9.6. A measurable change in the reactive orange concentration during the test could not be found.

RANGE FINDING TEST

product (mg/L)	normal organims	immobilized organisms	% immobilization
500	3	17	85
100	17	3	15
10	20	0	0
0	19	1	5

test item

product (mg/L)	normal organims	immobilized organisms	% immobilization
500	4	16	80
353.6	8	12	60
250	18	2	10
176.8	17	3	15
125.6	19	1	5
88.4	20	0	0
0	36	4	10

Validity criteria fulfilled:

yes

Conclusions:

After 24 h the EC50 was determined to be 355 mg/L.

Executive summary:

The acute toxicity study of the test substance was tested on Daphnia according to DIN 38 412 "Daphnien-Kurzzeittest" (eqaul to OECD 202) for 24 hours.

The EC0 of the test item is 88.4 mg/L

The EC50: 355 mg/L

The EC100: 600 mg/L

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 1997-09-03 to 1997-11-06

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:

- Concentrations: Following nominal concentrations were measured: 0, 22, 50, 100, 220, 500 mg/L
- 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.
- Sample storage conditions before analysis: Room temperature

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- 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.
- Controls: Blank control performed with the test solutions without test substance
- Evidence of undissolved material: no

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 concentrations: 0, 22, 50, 100, 220, 500 mg/L
Measured concentrations (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: Number of immobile daphnids after 24 h and 48 h

Reference substance (positive control):

no

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: -

Key result

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:

EC100

Effect conc.:

500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: -

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:

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

Details on results:

- Other biological observations: No
- Mortality of control: No
- No immobility was observed in control group
After 48 hours the EC50 was determined to be 117.4 mg/L

Results with reference substance (positive control):

No test with reference substance was performed.

Reported statistics and error estimates:

EC5O 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.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

from 1991-11-13 to 1992-03-18

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: UBA-Recommendation §4 Abs 1 Nr 6

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 320, 560, 1000 mg/L
- Sampling method: the test concentrations were measured at the start and at the end of the study from each test vessel
- Sample size: ca. 10 mL
- Sample storage conditions before analysis: Room temperature

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The Test substance was directly weighed into the test vessels and homeneneously diluted with dilution water adjusted to pH 6 to prevent the disintegration of the TS to the oxy-dye (Oxyfarbstoff) by means of an ultra sonic sound.
- Controls: Blank control performed with the test solution without the test substance
- Evidence of undissolved material: no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Species: Daphnia magna STRAUS
- Source: Hoechst AG Kastengrund, Germany
- Method of breeding: in a 100 mL beaker glass (90 mm x 50 mm) filled with 70 mL dilution water (modified freshwater according to Elendt). One Daphnia was inocculated into each vessel. Animals were used for breeding until 42 days of age. Reproduction rate and health status was recorded once on weekdays.
- Food type: Scenedesmus subspicatus
- Amount: depending on age (0.3 to 1.6 mg/L COD)
- Frequency: trice per week

ACCLIMATION
- Acclimation conditions: same as test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

NA

Test temperature:

control: 21 to 21.9 °C
test groups: 20.9 to 21.9 °C

pH:

dilution water adjusted to pH 6
measured values:
control: 6 to 7.1
test groups: 5.9 to 7.2

Dissolved oxygen:

control: 8.5 to 9.1 mg/L
test groups: 8.3 to 8.9 mg/L

Nominal and measured concentrations:

Nominal: 0, 320, 560, 1000 mg/L
Measured:
Control: nothing detected
320 mg/L: 0 h: Day1: 323 mg/L; Day 2: 295 mg/L
24 h: Day 1: 300 mg/L Day 2: 273 mg/L
560 mg/L: 0 h: Day 1: 537 mg/L Day 2: 610 mg/L
24 h: Day 1: 511 mg/L Day 2: 521 mg/L
1000 mg/L: 0 h: Day 1: 1035 mg/L Day 2: 864 mg/L
24 h: Day 1: 963 mg/L Day 2: 1027 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: 250 mL beeker glass
- Material, size, headspace, fill volume: glass, 110 mm x 70 mm, filled with 200 mL test water
- Aeration: dilution water was saturated with oxygen prior to study start
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): NA

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: modified freshwater according to Elendt
- Intervals of water quality measurement: after addition of the test substance and at the end of the study (48 hours)

OTHER TEST CONDITIONS
- Adjustment of pH: 6
- Photoperiod: 12/12
- Light intensity: illumination with fluorescent lamp

EFFECT PARAMETERS MEASURED: Immobility rate after 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca 1.8
- Range finding study: yes (10 - 500 mg/L)

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

560 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

748 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

1 000 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Details on results:

- Mortality of control: no
- Effect concentrations exceeding solubility of substance in test medium: no
- Any observations: In a test vessel of the 320 mg/L group, a 100% immobilisation rate was observed. Because in the second test vessel of this group and in the 560 mg/L groups no immobility appeared, this effect was not considered in the result.

Results with reference substance (positive control):

No test with reference substance was performed.

Table 1. Immobility-Results

Concentration mg/L	Immobility after 24 h		Immobility after 48 h
	Absolute	%	Absolute	%
0	0/20	0	0/20	0
320	8/20	40	10/20	50
560	0/20	0	0/20	0
1000	17/20	85	20/20	100

Validity criteria fulfilled:

yes

Conclusions:

The EC50 of the test substance (SA03) in Daphnia magna after 48 hours is determined to be 748 mg/L

Description of key information

 The 48h-EC50 for the read across substance (structural analogue 02) was determined according to OECD 202 to be 117.4 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

117.4 mg/L

Additional information

An acute toxicity study with the structural analogue 01 as lithium salt was carried-out with Daphnia Magna according to DIN 38 412 "Daphnien-Kurzzeittest" (equal to OECD 202) for 24 hours.

Daphnids were exposed to the test substance at concentrations of 88.4, 125.6, 176.8, 250, 353.6 and 500 mg/L and a negative control (0 mg/L). Observations were made on the immobilisation rate after 24 h of exposure.

 Table 1. Immobility-Results

Concentration mg/L	Immobility after 24 h
	Absolute	%
0	4/40	10
88.4	0/20	0
125.6	1/20	5
176.8	3/20	15
250	2/20	10
353.6	8/20	60
500	16/20	80

The EC0 of the test item was 88.4 mg/L

The EC50: 355 mg/L

The EC100 :600 mg/L

With another structural analogue (SA02), a study was performed to assess the acute toxicity of the 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 test substance at concentrations of 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.

 

Table 2. 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

 

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