Reaction products of 7-amino-4-hydroxynaphthalene-2-sulfonic acid reacted with 2-((1,3,5-triazin-2-yl)amino)ethan-1-ol, coupled with diazotised 3-amino-4-hydroxybenzenesulfonic acid, subsequently coupled with diazotised 2-amino-5-methoxybenzenesulfonic acid, chelated with copper, sodium salt

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 1991-04-17 to 1991-05-15

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

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0.32, 0.56, 1.0, 1.8, 3.2, 5.6, 10, 18 and 32 mg/L
- Sampling method: The test sample solutions were diluted using aged dechlorinated water to a theoretical nominal concentration of 0.32 mg/L or 3.2 mg/L. The samples were measured after 0 and 48 hours.
- Sample storage conditions before analysis: 21 °C

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Direct dispersion in water. A stock solution of 1.00 g test material per litre of diluent was prepared from which serial dilutions were made to give the test series.
- 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
- Stage and instar at study initiation: Daphnia magna (Straus) were tested in the 1st instar
- Source: Laboratory culture originating from a strain supplied by I.R.CH.A., France
- Method of breeding: Culture conditions ensure that reproduction is by parthenogenesis
- Feeding during test: no

ACCLIMATION
- Acclimation period: 24 hours
- Acclimation conditions (same as test or not): At 21°C in polypropylene vessels containing two litres of dechlorinated and aged tap water.
- Type and amount of food:
- Feeding frequency: Cultures were fed daily with a suspension of mixed algae (predominantly Chlorella spp.)
- Health during acclimation (any mortality observed): no

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

50 mg/L CaCO3

Test temperature:

21 °C

pH:

7.1 - 7.2

Dissolved oxygen:

8.0 - 8.3 mg/L

Nominal and measured concentrations:

- Nominal concentrations: 0.32, 0.56, 1.0, 1.8, 3.2, 5.6, 10, 18 and 32 mg/L
- Measured concentrations:
At the start of the test (0 h): 0.325, 1.18, 3.17, 10.6 and 35.3 mg/L corresponding to 101.6, 118.0, 99.1, 106.0, 110.3 % of the nominal concentrations.
At the end of the test (48 h), after storage at ambient temperature in light: 0.348, 1.28, 3.29, 10.7 and 34.6 mg/L corresponding to 107.1, 108.5, 103.6, 101.3 and 97.9 % of the nominal concentrations.
At the end of the test (48 h), after storage at ambient temperature in dark: 0.358, 1.27, 3.31, 10.7 and 34.2 mg/L corresponding to 110.2, 107.0, 104.3, 101.3 and 96.7 % of the nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass jars
- Type: closed, covered with foil to reduce evaporation.
- Volume of solution: 200 mL
- Aeration: None
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Biomass loading rate: Loading: 20 mL test solution per organism.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated (with sodium thiosulphate) and aged laboratory tap water.
- Hardness: Total hardness approximately 50 mg/L as CaCO3.
- Culture medium different from test medium: No
- Intervals of water quality measurement: pH, temperature and oxygen were measured each 24 hours.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: Photoperiod: 16h light: 8h dark

EFFECT PARAMETERS MEASURED: Mobility of daphnids after 0, 24 and 48 hours

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
No range finding test was performed

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 2.6 - 3.8 mg/L

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

2.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 1.9 - 2.7 mg/L

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.56 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: Not observed
- Observations on body length and weight: Not observed
- Other biological observations: Not observed
- Mortality of control: No
- Other adverse effects control: Not observed
- Immobilisation of control: No
- Abnormal responses: Not observed
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

After 48 hours, the EC50 was determined to be 2.3 mg/L

Results with reference substance (positive control):

No test with reference substance was performed.

Table III: Exposure of Daphnia magna for 48 hours

Concentration (mg/L)	0 hours			24 hours	48 hours
	pH	mgO2/L	T°C	T°C	pH	mgO2/L	T °C
Control R1- R2	7.1- 7.1	8.6 - 8.6	21.0-21.0	21.0-21.0	7.2-7.1	8.2-8.1	21.0-21.0
0.32 R1 R2	7.1-7.1	8.6 - 8.6	21.0 -21.0	21.0- 21.0	7.2-7.2	8.0-8.2	21.0-21.0
0.56 R1 R2	7.1-7.1	8.6 - 8.6	21.0-21.0	21.0-21.0	7.2-7.2	8.2-8.1	21.0-21.0
1.0 R1 R2	7.1-7.1	8.6 - 8.6	21.0-21.0	21.0-21.0	7.2-7.2	8.2-8.0	21.0-21.0
3.2 R1 R2	7.1-7.1	8.6 - 8.6	21.0-21.0	21.0-21.0	7.2-7.2	8.2-8.1	21.0-21.0
5.6 R1 R2	7.2-7.2	8.6 - 8.6	21.0-21.0	21.0-21.0	7.2-7.2	8.2-8.2	21.0-21.0
10 R1 R2	7.2-7.2	8.6 - 8.6	21.0-21.0	21.0-21.0	7.3-7.3	8.2-8.3	21.0-21.0
18 R1 R2	7.3-7.3	8.6 -8.6	21.0-21.0	21.0-21.0	7.3-7.3	8.1-8.0	21.0-21.0
32 R1 R2	7.3-7.3	8.6 -8.6	21.0-21.0	21.0-21.0	7.3-7.3	8.1-8.0	21.0-21.0

Medium renewal: None.

Duration of exposure: 48 hours.

Criteria of effect: Daphnia were considered to be immobilised if they were unable to swim for approximately 15 seconds after gentle agitation.

Verification of test concentrations (spectro-photometrically) and Validation of method of analysis were done.

 

Results:

Table IV : Cummulative Immobilsation Data for Daphnia magna exposed for 48 hours

 

R1 – R2:Replicates

 

	Cumulative immobilised Daphnia (initial population: 10 per replicate)
	24 hours				48 hours
Concentration (mg/l)	R1	R2	total	%	R1	R2	total	%
Control	0	0	0	0	0	0	0	0
0,32	0	0	0	0	0	0	0	0
0,56	0	0	0	0	0	0	0	0
1,0	0	0	0	0	0	1	1	5
1,8	1	2	3	15	3	2	5	25
3,2	4	6	10	50	7	8	15	75
5,6	9	8	17	85	10	10	20	100
10	10	10	20	100	10	10	20	100
18	10	10	20	100	10	10	20	100
32	10	10	20	100	10	10	20	100

 

 

Table V: Analysis of the imobility data by the method of Thompson (1947) gave the following results:

Time (h)	EC50 mg/L	95% confidence limits (mg/l)
24	3.2	2.6 – 3.8
48	2.3	1.9 – 2.7

No observed Effect Concentration (NOEC) at 24h: 1.0 mg/L

NOEC at 48 h: 0.56 mg/L

The No Observed Effect Concentration showed that the measured test concentrations were near nominal. It was therefore considered justifiable to calculate the EC50 values in terms of the nominal test concentrations only.

Validity criteria fulfilled:

yes

Conclusions:

No observed Effect Concentration (NOEC) at 24h: 1.0 mg/L
NOEC at 48 h: 0.56 mg/L
The 48h-EC50 was determined to be 2.3 mg/L

Executive summary:

The acute Daphnia toxicity of Direct Red 83:1 was tested in a static system within 24 and 48 h respectively in a GLP laboratory according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

The values of the abiotic parameters (oxygen, pH and temperature) were determined from the total volume of the parallel solutions.

The validity criteria were fulfilled. The analytical value indicated a very good recovery therefore the following results are considered as valid:

Study results

after 24 hours	After 48 hours
95% Confidence limits (g/L) ^2.6 -3.8	1.9 to 2.7
EC50:	2.3 mg/L >= 99.1 mg/L (test substance; including inorganic components)

No Observed Effect Concentration (NOEC) at 24h: 1.0 mg/L

NOEC at 48 h: 0.56 mg/L

The No Observed Effect Concentration (NOEC) is based upon zero immobilisation at these concentrations. Were near nominal. It was therefore considered justifiable to calculate the EC50 values in terms of the nominal test concentrations only.

 

 

Description of key information

A short-term toxicity test to aquatic invertebrates was performed according to OECD 202. After 96 h the EC50 was determined to be 2.3 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

2.3 mg/L