Benzenesulfonic acid, 2-[2-[5-(aminocarbonyl)-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridinyl]diazenyl]-4-[[4-chloro-6-[[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-, sodium salt (1:2)

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 June 1993 to 04 August 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to EU & OECD test guidance in compliance with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

79/831/EEC

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Details on test material:

See below

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Sample concentrations: 0, 560, 1000 mg/l
In order to determine the substance concentration, water samples were taken at study start and study end. The concentration value of the 1000 mg/l-group decreased from 89.5 % (895 mg/l) of the nominal

Test solutions

Vehicle:

no

Details on test solutions:

The test substance was weighed directly into the test chambers, mixed with water for dilution and homogenized with the Ultra-Turrax and ultrasonic bath. The test batches were present as yellow solutions.
No separation or formation of deposits was observed with any of the concentrations tested. In order to determine the substance concentration, water samples were taken at study start and study end. The concentration value of the 1000 mg/l-group decreased from 89.5 % (895 mg/1) of the nominal concentration at study start to 77.5 % (775 mg/1) after 48 hours.
All concentration-values of the 560 mg/l-group were in the range of ± 20 % of the nominal concentration.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna - Clone 5 (IRChA, France)
Origin: Stockculture of the Department of Toxicology (Hoechst AG)
Breeding conditions: 100 ml beakers (height 90 mm, diameter 50 mm), filled with 70 ml test water (treated fresh water, for composition see Appendix page 18) served as culture vessels. One daphnid was used per culture vessel. The animals were used for breeding up to an age of 42 days. The reproduction rate and the state of health of the animals were monitored daily apart from weekends. The animals were fed three times a week with monocellular green algae (Scenedesmus subspicatus).

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation.

Test conditions

Hardness:

Recorded by not reported

Test temperature:

20.4 - 21.5C

pH:

7.5 - 8.6

Dissolved oxygen:

8.5 - 8.8mg/l

Salinity:

Not applicable.

Nominal and measured concentrations:

0, 560, 1000 mg/l (nominal)

Details on test conditions:

The study was conducted in a static system. The test chambers were 250 ml beakers (height 110 mm, diameter 70 mm, water level about 75 mm), each filled with 200 ml test water. The room temperature was about 21⁰C. The room was illuminated by fluorescent tubes in a day/night cycle, with each period lasting 12 hours.

After the test concentrations had been prepared and water parameters recorded, 10 daphnids were used in each test chamber. The daphnids received no feed for the entire study period. Inspection of the daphnids took place after 24 and 48 hours and involved recording the immobility. Immobile daphnids were removed from the chambers. Daphnids were considered immobile when there was a 1 ack of movement 15 sec. after touching the test chamber. The water parameters were measured after adding the substance and at the end of the test.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

In this 48-hour acute toxicity study of Reaktiv-Gelb F66 923 FW in daphnia magna no immobility occurred in the 560 and 1000 mg/l group. The mean of the analytical determined concentration-values in the 1000 mg/1 group was 835.0 mg/l.
The LC50 after 96 hours therefore exceeds 1000 mg/l (nominal concentration) or 835.0 mg/l (mean of the analytical determined concentration), respectively.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

No data

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In this 48-hour acute toxicity study of Reaktiv-Gelb F66 923 FW in daphnia magna no immobility occurred in the 560 and 1000 mg/l group. The mean of the analytical determined concentration-values in the 1000 mg/l group was 835.0 mg/l.

The LC50 after 96 hours therefore exceeds 1000 mg/1 (nominal concentration) or 835.0 mg/1 (mean of the analytical determined concentration), respectively.
The substance is not harmful to Daphnia.

Executive summary:

The acute toxicity of Reaktiv-Gelb F-66 923 FW in Daphnia magna (waterflea) was determined in accordancewith the requirements of EEC-Guideline, EEC Directive 79/831 Annex V, Methods for the Determination of Ecotoxicity 5.1.2 Acute Toxicity for Daphnia as from Feb. 1999 and the OECD-Guideline, OECD-Guideline for testing of chemicals, Section 2 - Effects on Biotic Systems Test Guideline 202 Daphnia sp., 14 -day Reproduction Test (including an Acute Immobilisation Test) Adopted: April 4, 1984. Study conducted in complince with GLP.

 

When testing the immobility effect of Reaktiv-Gelb F-66 923 FW in Daphnia magna (water flea) over a total of 48 hours according to the method described under section 4, no lethality occurred at 1000 mg/l.The concentrations tested were 560, 1000 mg/l and a negative control (0 mg/l).

All tested concentrations were present as yellow solutions.

All test batches were homogeneous. No particulate matter was observed.

The mean of the analytical determined concentration-values in the 1000 mg/l group was 835.0 mg/l.

The EC50 after 48 hours therefore exceeds 1000 mg/l (nominal concentration) or 835.0 mg/l (mean of the analytical determined concentration), respectively.

The concentration value of the 1000 mg/l-group decreased from 89.5 % of the nominal concentration at study start to 77.5% after 48 hours. All concentration-values of the 560 mg/l-group were in the range of + 20 % of the nominal concentration.

 

The test substance is not considered to be harmful to Daphnia.