4,4'-[(6-chloro-1,3,5-triazine-2,4-diyl)diimino]bis[5-hydroxy-6-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonic] acid, sodium salt

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 October 1992 to 06 November 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Test concentrations: 0/220/ 500 mg/L

Vehicle:

no

Details on test solutions:

Water for dilution was added to the test substance, the mixture was homogenized using an Ultra-Turrax and Ultrasonic Bath and poured into the chamber under stirring with a glass rod.

The test batches were present as dark red solutions. No segregation or formation of precipitates could be observed.

In order to determine the substance concentration, water samples (approx. 20 mL) were taken from the middle of the test chamber using a screw-neck glass bottle at study start, and after 48 and 96 hours.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: zebrafish (Brachydanio rerio) (HAMILTON-BUCHANAN)
Origin: Central Toxicology (Hoechst AG)
Date of hatching: December 15, 1991
Delivery date: May 14, 1992
The fish were kept for 14 days in water for dilution before the start of the study under the following conditions:
Temperature: 22 ± 1 °C
Oxygen content: > 80 % of the saturation value
Duration of light period: 12 hours daily
Density: < 1 g fish / L water
Feeding: twice daily ad libitum
Food: Tetra Min, Tetra Werke, Melle

Prior to study start the body length of 10 representative fish from each batch was measured. These animals were not used during the study.

Part of study Batch No. Date n Body length (cm)
Variation range Mean s
1 16/91/H15/L2 23-Oct-92 10 3.4 – 3.9 3.6 ± 0.21
2 1/92/H1/L1 06-Nov-93 10 2.6 – 3.2 3.0 ± 0.17
Total sample 20 2.6 – 3.9 3.3 ± 0.35

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No post exposure observation

Hardness:

No data

Test temperature:

21.0 - 22.3 °C

pH:

7.0 - 7.8

Dissolved oxygen:

6.1 - 10.1 mg/L

Nominal and measured concentrations:

0, 220 and 500 mg/L

Details on test conditions:

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 liters 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 in a range of 8.3 - 8.5.

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 Z® 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 06.00 a.m. to 06.00 p.m. The light intensity directly over the chambers was approximately 700 lux.

The test chambers were not aerated during the course of the study.

After the test concentration had been prepared and water parameters recorded, 10 fish were assigned at random to each test and control chamber. The fish received feed for the entire study period. Inspection of the fish took place after 2 - 4, 24, 48, 72 and 96 hours and involved recording the mortality and visible alterations in appearance and behaviour. Dead fish were removed from the chambers. Fish were considered dead when there was a lack of opercular movement and no response to slight mechanil stimulus. The water parameters were measured and recorded at the same dates.

Reference substance (positive control):

not required

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 500 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 500 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

220 mg/L

Conc. based on:

test mat.

Basis for effect:

behaviour

Details on results:

Recording of symptoms was disturbed by the dark red discoloration of the test water. The 220 mg/L group showed no effect in comparison with the control. The fish of the 500 mg/L group showed changes in behaviour and swimming behaviour.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

No data

Sublethal observations / clinical signs:

0 mg/L (Control groups)
Hours after study start	Hours -----------------------------------à
	2-4	24	48	72	96
Lethality (absolute)	0/20	0/20	0/20	0/20	0/20
No changes in appearance and behaviour	a	a	a	a	a
220 mg/L
Hours after study start	Hours ----------------------------------à
	2-4	24	48	72	96
Lethality (absolute)	0/10	0/10	0/10	0/10	0/10
No changes in appearance and behaviour	a	a	a	a	s
Recording of symptoms was disturbed	x	x	x	x	s
500 mg/L
Hours after study start	Hours ----------------------------------à
	2-4	24	48	72	96
Lethality (absolute)	0/10	0/10	0/10	0/10	1/10
No changes in appearance and behaviour	a	a	a	a	a
Recording of symptoms was disturbed	x	x	x	x	x

s = some fish

a = all fish

Validity criteria fulfilled:

yes

Conclusions:

In this 96-hour acute toxicity study of Reaktiv-Rot F-67787, only one fish died at 500 mg/L . No mortality occurred in the 220 mg/L group. The LC50 after 48 and 96 hours therefore was > 500 mg/L.

Executive summary:

The acute toxicity to fish with Reaktiv-Rot F-67787 was conducted according to EU-Guideline C.1. Acute Toxicity in Fish Directive 84/449/EEC and theOECD-Guideline for Testing of Chemicals, 203 Fish Acute Toxicity Test. Study conducted in compliance with GLP. 

Testing of lethal effects of Reaktiv-Rot F-67787 for 96 hours in Zebra fish (Brachydanio rerio) according to the method, only one fish died at 500 mg/L . No mortality occurred in the 220 mg/L group. The LC50 after 48 and 96 hours therefore was > 500 mg/L.

 

The test batches were present as dark red solutions.

 

In order to determine the substance concentration, water samples were taken from the middle of the test chamber at study start, and after 48 and 96 hours. All mean values were above 80 % of the nominal concentration.

Description of key information

LC50 (96h) > 500 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Remarks:

(96h)

Effect concentration:

500 mg/L

Additional information

The acute toxicity to fish of Reaktiv-Rot F-67787 was conducted according to EU-Guideline C.1. Acute Toxicity in Fish Directive 84/449/EEC and the OECD-Guideline for Testing of Chemicals, 203 Fish Acute Toxicity Test. The study was conducted in compliance with GLP. 

In order to determine the substance concentration, water samples were taken from the middle of the test chamber at study start, and after 48 and 96 hours. All mean values were above 80% of the nominal concentration.

Testing of lethal effects of Reaktiv-Rot F-67787 for 96 hours in Zebra fish (Brachydanio rerio) up to 500 mg/L, only one fish died at 500 mg/L . No mortality occurred in the 220 mg/L group. The LC50 after 48 and 96 hours therefore was > 500 mg/L.