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 fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 April 1993 to 23 April 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.1 (Acute Toxicity for Fish)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity 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:

Test concentrations: 0 / 100 / 220 mg/l
In order to determine the substance concentration, water samples were taken from the middle of the test chamber by pipetting approx. 20 ml into a screw-neck glass bottle at study start, and after 48 and 96 hours.

Test solutions

Vehicle:

no

Details on test solutions:

The test substance was weighed in a beaker, water for dilution was added and the mixture homogenized using an Ultra-Turrax and Ultrasonic Bath and poured into the chamber under stirring with a glass rod. All tested concentrations were present as yellow solutions.
At study start the pH-value of the control group and the concentration groups (100 mg/l and 220 mg/l) were adjusted to pH 7.
All test batches were homogeneous. No particulate matter was observed.
In order to determine the substance concentration, water samples were taken from the middle of the test chamber by pipetting approx. 20 ml into a screw-neck glass bottle at study start, and after 48 and 96 hours. The following values were determined during the study:
Nominal - Measured
(mg/l) x (mg/l) %
220 193.3 87.9

All analytical values were in a range of ± 20 % of the nominal concentration.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: zebra fish Brachydanio rerio (HAMILTON-BUCHANAN)
Breeding station: Central Toxicology (Hoechst AG)
Date of hatching: 03/01/1992
Delivery date: 10/27/1992

The fish were kept for at least 14 days before the start of the study in water for dilution under the following conditions:

Temperature: 22 ± 1⁰C
Oxygen content: > 80 % of the saturation value
Duration of light period: 12 hours daily
Population density: < 1 g fish/L water
Feeding: twice daily ad libitum
Feed: Tetra Min, Tetra Werke', Helle (FRG]

The body length of 7 representative fish from each batch was measured.

Body length (cm)
Part of study Batch No. Date n Variation range x s
1 2/92/H2/L2 04/09/93 7 3.1 – 3.4 3.3 0.11
2 2/92/H2/L2 04/23/93 7 3.2 – 3.5 3.3 0.12
Total sample 14 3.1 – 3.5 3.3 0.12

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No post observation period

Test conditions

Hardness:

No data

Test temperature:

21.0C - 22.8C

pH:

7.0 - 7.7

Dissolved oxygen:

6.5 - 9.5 mg/l

Salinity:

Not applicable.

Nominal and measured concentrations:

100 and 220 mg/l

Details on test conditions:

Water for dilution
Reconstituted water, composition according to IS0/D1S 7346/1 was used as water for dilution.
Preparation was carried out in a unit consisting of two Hostalen14- lined steel vessels with a capacity of 1700 litres each.
The dilution water was prepared as described was aerated until oxygen saturation. The pH was measured before the use of each study batch and was 8.2-8.3.

Test conditions
The study was conducted in a static system. The test chambers, which were calibrated to 10 liters, were made of glass (length 30 cm, width 22 cm, height 24 cm) and stood in a water bath made of Hostalit ZR with a Plexiglas viewing panel. The temperature of the water bath was regulated by a thermostat to 22 ± 1 °C. The chambers were illuminated from above from 6.00 a.m. to 6.00 p.m. The test chambers were not aerated during the course of the study.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 220 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The fish showed changes in appearance and behaviour and swimming behaviour. No lethality occurred in the 220 mg/l group.
The LC50 after 96 hours therefore exceeds 220 mg/1.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

All analytical values were in a range of ± 20 % of the nominal concentration.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In this 96-hour acute toxicity study of Reaktiv-Gelb F-66 923 FW in zebra fish (Brachydanio rerio) no lethality occurred in the 220 mg/l group.
The LC50 after 96 hours therefore exceeds 220 mg/l.

Executive summary:

The acute toxicity of Reaktiv-GelbF-66923 FW in fish was determined compliance with the requirements of EEC-Guideline79/831/EWG Appendix V Methods for the determination of ecotoxicity C.1. Acute toxicity on fish and the OECD-Guideline for testing of chemicals,203 Fish, Acute Toxicity Test.Study performed in compliance with the Principles of Good Laboratory Practice (GLP).

 When testing the lethal effect of Reaktiv-Gelb F-66 923 FW in zebra fish (Brachydanio rerio) over a total of 96 hours according to the method described under section 4, no lethality occurred at 220 mg/1. The LC50 after 96 hours therefore exceeds 220 mg/1.

At study start the pH-values of the control group and the concentration groups (100 mg/1 and 220 mg/1) were adjusted to pH 7.

 

All tested concentrations were present as yellow solutions. All test batches were homogeneous. No particulate matter was observed.

The substance concentration was determined in the 220 mg/1 group. All values determined were in a range of + 20 % of the theoretical value.

 

The substance is not considered as harmful to fish.