Sodium m-[[4-[[p-[bis(2-hydroxyethyl)amino]phenyl]azo]-1-naphthyl]azo]benzenesulphonate

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Nov - Dec 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

31 July 1992

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Sodium m-[[4-[[p-[bis(2-hydroxyethyl)amino]phenyl]azo]-1-naphthyl]azo]benzenesulphonate

EC Number:

266-867-4

EC Name:

Sodium m-[[4-[[p-[bis(2-hydroxyethyl)amino]phenyl]azo]-1-naphthyl]azo]benzenesulphonate

Cas Number:

67674-28-6

Molecular formula:

C26H25N5O5S.Na

IUPAC Name:

sodium m-[[4-[[p-[bis(2-hydroxyethyl)amino]phenyl]azo]-1-naphthyl]azo]benzenesulphonate

Test material form:

solid

Details on test material:

Acid Red 299

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 10, 29, 83 mg/L
- Sampling method: the samples were taken from the middle of the water body using pippetes

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
stock solution: The test item (1000mg) were dissolved in 250mL distilled water.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish (Danio rerio)
- Source: ASCANIA-AQUARIUM, Aschersleben, Germany
- Age at study initiation: juvenile fish
- Length at study initiation (mean): 33.7mm
- Mortality of test population: <5%

ACCLIMATION
- Acclimation period: minimum 7 days
- Acclimation conditions: same as test
- Type and amount of food during acclimation: standard food daily until 2 days before commencement of test

FEEDING DURING TEST
- Food type: standard food (pestled trout fish food)
- Frequency: daily until 2 days before commencement of test

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

c(Ca++ and Mg++) = 2.5mmol/L

Test temperature:

22.7-23.1°C

pH:

pH Control: 8.0
pH Test: 8.0 - 8.2

Dissolved oxygen:

88-95%

Nominal and measured concentrations:

nominal: 0, 10, 17, 29, 49, 83 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: aquarium
- fill volume: 4L
- Aeration: yes
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 1

OTHER TEST CONDITIONS
- Adjustment of pH: no

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

29 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

49 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

37.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: LC50 as geometric mean between LC0 and LC100

Details on results:

- Behavioural abnormalities:
- Observations: animals of test group no: 1 a light pink coloring was observed
- Other biological observations: animals of test group no: 2 and 3 were slightly damaged and swam during the first hour on the water surface and reacted oversensitive to external stimuli. At the end of the exposure a reddish coloring on the animales was observed.
- Due to the intense coloring of the test solution no observation could be made in respect of test group 4 and 5.
- At the end of the exposure period a dark, flaky precipitation was observed on the bottom of the test vessels (increased with higher concentrations). These is probably caused of the excrements of the fish.
- Mortality of control: no
- Other adverse effects control: none observed

Any other information on results incl. tables

Sublethal observations / clinical signs:

	Exposure Time
Conc. (mg/L)	24 (hours)	48 (hours)	72 (hours)	96 (hours)
LC0	29	29	29	29
LC100	49	49	49	49

Mortality

		Live animals after:
Test Group No.	Conc. (mg/L)	24 h	48 h	72 h	96 h	Mortality Rate after 96 h (%)
0 (Control)	0	7	7	7	7	0
1	10	7	7	7	7	0
2	17	7	7	7	7	0
3	29	7	7	7	7	0
4	49	0	-	-	-	100
5	83	0	-	-	-	100

pH values during exposure

Test Group		pH values during exposure (hours)
No.	Conc. (mg/L)	2	24	48	72	96
0	0	8,0	8,0	8,0	8,0	8,0
1	10	8,0	8,0	8,0	8,0	8,0
2	17	8,0	8,0	8,0	8,0	8,0
3	29	8,1	8,0	8,0	8,1	8,1
4	49	8,1	8,1	-	-	-
5	83	8,2	8,1	-	-	-

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 96-h LC50 to Danio rerio was determined to be > 29 and < 49mg/L. The calculated LC50 (as geometric mean value between LC0 and LC100) is 37.7 mg/L. The substance is acutely harmful to fish.