Tetrasodium 4-amino-5-hydroxy-6-[[2-methoxy-5-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]azo]-3-[[4-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonate

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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

see read-cross justification in section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Validity criteria fulfilled:

yes

Conclusions:

The EC0 is above 100 mg/L

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13. Nov. 1991 to 05. Mar 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 times: begin and end of the study from each test vessel
- Sample size: ca. 10 mL
- Sample storage conditions before analysis: NA

Vehicle:

no

Details on test solutions:

The TS was directly weighed into the test vessels and homeneneously diluted with dilution water adjusted to pH 6 to prevent the rapid disintegration of the TS to the oxy-dye (Oxyfarbstoff = Reactive Black 5-OH) by means of an ultra sonic sound.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Species: Daphnia magna STRAUS
- Source: Hoechst AG Kastengrund
- 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 CSB)
- Frequency: trice per week

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

NA

Hardness:

measured once a week in the dilution water

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

Salinity:

NA

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 (see attachment)
- Source/preparation of dilution water: modified freshwater according to Elendt
- Intervals of water quality measurement: after addition of TS and at study end


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


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility rate

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca 1.8
- Range finding study: yes

Reference substance (positive control):

not required

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

560 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

748 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: geometric mean

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

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):

NA

Reported statistics and error estimates:

not possible

Validity criteria fulfilled:

yes

Conclusions:

The EC50 of Reactive Black 5 in Daphnia magna after 48 hours is determined to be 748 mg/L

Executive summary:

Reactive Black 5 in its bis-ester form, was tested for acute toxic effects on Daphnia magna according to OECD 202 at concentrations of 320, 560 and 1000 mg/L over an exposure time of 48 hours in a semi-static test under slightly acidic test conditions to avoid abiotic degradation of the test substance. No adverse effects were observed during this study at 320 and 560 mg/L. At 1000 mg/L all daphnids were immobile after 48 hours.

The EC0 was 560 mg/L, the EC50 was calculated (as the geometric mean between 560 and 1000 mg/) to be 748 mg/L, the EC100 was 1000 mg/L.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11. Dec to 13. Dec 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Deviations:

no

GLP compliance:

yes

Remarks:

without inspection and statement

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Daphnia magna

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

NA

Hardness:

no data

Test temperature:

no data

pH:

change of pH during study was less than 1

Dissolved oxygen:

> 2 mg/L at study end

Salinity:

NA

Nominal and measured concentrations:

0, 2, 4, 8, 16, 32, 64, 128 mg/L
the test substance was stable over the testing period.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

> 128 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 128 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 128 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

EC50: 0.9 to 2.4 mg/L potassium dichromate

Validity criteria fulfilled:

yes

Conclusions:

The EC50 in Daphnia magna after 48 hours is above 128 mg/L

Executive summary:

Reactive Black 5 -OH was tested for acute toxic effects on Daphnia magna according to OECD 202 at concentrations of 2 to 128 mg/L over an exposure time of 48 hours. No adverse effects were observed during this study. The EC0 and EC50 values are above the highest test concentration of 128 mg/L, tested.

Description of key information

The test substance showed a low acute toxicity to Daphnia magna with an EC50 of 748 mg/L calculated as the geometric mean of the test concentration with and without effect.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

748 mg/L

Additional information

Reactive Black 5-OH, the hydrolysis product of Reactive Black 5, a structural analogue and impurity of the test compound, was tested for acute toxic effects on Daphnia magna according to OECD 202 at concentrations of 2 to 128 mg/L over an exposure time of 48 hours. No adverse effects were observed during this study. The EC0 and EC50 values are above the highest test concentration of 128 mg/L, tested.

Reactive Black 5, a structural analogue and impurity of the test compound, was tested for acute toxic effects on Daphnia magna according to OECD 202 at concentrations of 320, 560 and 1000 mg/L over an exposure time of 48 hours in a semi-static test under slightly acidic test conditions (pH 6) to avoid abiotic degradation of the test substance. No adverse effects were observed during this study at 320 and 560 mg/L. At 1000 mg/L all daphnids were immobile after 48 hours. The EC0 was 560 mg/L, the EC50 was calculated (as the geometric mean between 560 and 1000 mg/) to be 748 mg/L, the EC100 was 1000 mg/L.