Tetrasodium 7-[[2-[(aminocarbonyl)amino]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]phenyl]azo]naphthalene-1,3,6-trisulphonate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

key study

Study period:

From October 26, 1989 to January 18, 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The test was conducted by means of Read Across approach. The reliability of the source study report is 1. Further information was attached at section 13

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:

Test concentrations: 0/ 560 / 1000 mg/L

Vehicle:

no

Details on test solutions:

The relevant amount of the test substance was weighed into a graduated flask (200 mL), filled up with test water and stirred for about 5 minutes with a magnetic stirrer. The test batches were present as red solutions. No segregation or formation of precipitates could be observed.In order to determine the test substance concentration, water samples were taken at study end.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (STRAUS)
Origin: Stock culture of Department of Toxicology (Hoechst AG)
Breeding conditions: beakers (height 235 mm, diameter 128 mm), filled with 2L test water (according ISO/DIS 7346/1) served as culture vessels. 20 adult daphnids were used per culture vessel. Animals were transferred to fresh prepared vessels once a week. The room temperature was at 20 ± 1 °C. The breeding room was illuminated by diffuse light in a day/night cycle of 12 hours. The animals were fed with monocellular green algae (Scenedemus subspicatus) and a fine suspension of Tetra min dry food (Tetra Werke, Melle).To collect test animals with an age of at least 6 hours and a maximum of 24 hours, approx. 3 weeks old parent animals were separated in a cristallising dish (3L volume) filled with test medium for 18 hours. After 18 hours the adults were separted from the young daphnids by sieving through sieves of different mesh size. Adults remain on the upper crude sieve while the young daphnids are found on the fine sieve. These animals were used for the study after further 6 hours.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation period

Hardness:

No data

Test temperature:

20.4 - 21.3 °C

pH:

7.7 - 8.3

Dissolved oxygen:

8.6 - 9.5 mg/L

Salinity:

Not applicable.

Nominal and measured concentrations:

0, 560 and 1000 mg/L.

Details on test conditions:

The study was conducted in 2 stages. 20 daphnids, divided into two parallel groups of 10 animals, were used for each concentration.

Water for dilution
Reconstituted water, composition according to ISO/DIS 7346/1 was used as water for dilution.
Preparation was carried out in a unit consisting of two Hostalen®- lined steel vessels with a capacity of 1700 litres each.

The dilution water was prepared as described and was aerated until oxygen saturation. The pH was measured before the use of each study batch. It was at 8.3.

Test conditions
The study was conducted in a static system. As test chambers 300 mL cristallising dishes were used (height 55 mm, diameter 95 mm, water level about 32 mm), each filled with 200 mL test water. The room temperature was about 21 °C. The room was illuminated by fluorescent tubes for 12 hours each day.

Test procedure
After the test concentrations had been prepared and water parameters were recorded, 10 daphnids were used in each test chamber. The daphnids were not fed during the entire study period. Inspection of the daphnids took place after 24 and 48 hours and involved recording the immobility and further possible deviations to the control group. Immobile daphnids were removed from the chambers. Those animals which were not able to swim within 15 seconds after gentle agitation of the test container were considered to be immobile. The water parameters were measured after adding the substance and at the end of the test.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Probit analysis could not be carried out due to the lethality rate. As the factor for the concentration steps was 1.8 no further concentrations were tested. The EC50 range was given.

Under the conditions of this test with Reaktiv-Rot F-66 813 FW the following immobility rates were observed after 24 and 48 hours:

Concentration (mg/l) 24 hours 48 hours
Absolute % Absolute %
0 mg/l 0/40 0 0/40 0
560 mg/l 0/20 0 0/20 0
1000 mg/l 0/20 0 2/20 10


EC0 (mg/L) 1000 560
EC50 (mg/L > 1000 > 1000

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

No data

Immobility – individual values

Concentration (mg/L)	Test-chamber No.	Study-start (date)	Immobility hours after study start
			24	48
0	80a	10-Jan-90	0/10	0/10
0	80b	10-Jan-90	0/10	0/10
1000	36a	10-Jan-90	0/10	0/10
1000	36b	10-Jan-90	0/10	2/10
0	90a	16-Jan-90	0/10	0/10
0	90b	16-Jan-90	0/10	0/10
560	1a	16-Jan-90	0/10	0/10
560	1b	16-Jan-90	0/10	0/10

 

Validity criteria fulfilled:

yes

Conclusions:

EC50 > 1000 mg/L

Executive summary:

Method

The study on acute toxicity to daphnids meets the requirements of EEC Directive 79/831 Annex V Methods for the determination of ecotoxicity 5.1.2 Acute toxicity for daphnia and the OECD Guidelines for Testing of Chemicals, Section 2 - Effects on biotic systems 202 Daphnia sp., 14-day reproduction test (including an acute immobilisation test).

The concentrations tested were 0 / 560 / 1000 mg/L. The test batches were present as red solutions. No segregation or formation of precipitates could be observed.

Results

Testing for acute toxicity in Daphnia magna (water flea) over a total of 48 hours yielded the following immobility values: 

after 24 hours (mg/L)    after 48 hours (mg/L)

EC₀       1000                                     560

EC₅₀       > 1000                                 > 1000

 

Description of key information

EC50  > 1000 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1 000 mg/L