1,3,6-Naphthalenetrisulfonic acid, 7-[2-[2-[(aminocarbonyl)amino]-4-[[4-chloro-6-[[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]phenyl]diazenyl]-, sodium salt

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 February 1998 to 13 February 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted to recent EU test guidance in compliance with GLP and reported with a GLP certificate. This study is read across to the Structural Analogue 02, which is considered to be structurally equivalent to the substance to be registered, in that only the halogen ion differs structurally (fluorine instead of chlorine). The Structural Analogue 02 also contains a form where hydroxyl group replaces the chlorine. Neither of these differing analogues are considered to adversely affect the toxicity or intrinsic effects overall between the molecules. Details of structure are detailed below under "illustrations”.

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

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
None

Analytical monitoring:

no

Details on sampling:

- Concentrations:0/100mg/l

Vehicle:

no

Details on test solutions:

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

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 organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Source: Hoechst Marion Roussel Deutschland GmbH
- Length at study initiation (length definition, mean, range and SD): cm, 2.8, 2.5-3.1, ±2
- Feeding during test: No

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions (same as test or not): same as test
- Feeding frequency: twice daily, ad libitum
- Health during acclimation (any mortality observed): No

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:

21.4-21.7C

pH:

7.4-7.9

Dissolved oxygen:

6.9-9.8mg/l

Salinity:

Not applicable - freshwater

Nominal and measured concentrations:

100 mg/l (nominal)

Details on test conditions:

The study was conducted in a static system. The test chambers, which were calibrated to 4 liters, were made of glass (length 20 cm, width 15 cm, height 20 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 concentrations had been prepared and water parameters recorded, 7 fish were assigned to each test and control chamber. The fish received no feed for the entire study period. Inspection of the fish took place after 3, 6, 24, 48, 72 and 96 hours and involved recording the lethality and visible changes in appearance and behavior. Dead fish were removed from the chambers. Fish were considered dead when there was a lack of opercular movement and no response to slight mechanical stimulus. The water parameters were measured and recorded before study start and after 0, 24, 48, 72 and 96 hours.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

In the control and in the 100 mg/L concentration group no lethality was observed. In the concentration group the fish showed changes in behavior and respiration rate. The effects were reversible within 96 hours of exposure.

In this 96-hours acute toxicity study in zebra fish (Danio rerio) no lethality occurred in the 100 mg/L group and the control group.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not reported.

Sublethal observations / clinical signs:

	after 24 hours	after 48 hours	after 72 hours	after 96 hours
LC0 (mg/L)	100	100	100	100
LC50 (mg/L)	> 100	> 100	> 100	> 100
LC100 (mg/L)	not determined	not determined	not determined	not determined

Validity criteria fulfilled:

yes

Conclusions:

In the control and in the 100 mg/L concentration group no lethality was observed. In the concentration group the fish showed changes in behavior and respiration rate. The effects were reversible within 96 hours of exposure.

In this 96-hours acute toxicity study in zebra fish (Danio rerio) no lethality occurred in the 100 mg/L group and the control group.

Under the conditions of this test the LC50 of the substance after 96 hours was > 100 mg/L.

Executive summary:

The study was conducted to EU test guidance 92/69/EEC part C1 and OECD test guideline 203 in compliance with GLP.

LC50 > 100mg/l

The substance is not considered to pose a hazard to fish.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October to November 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Qualifier:

according to guideline

Guideline:

other: Letale Wirkung beirn Zebrabärbling Brachydanio rerio (LC0, LC50, LC100; 48 - 96 Stunden) Empfehlung des UBA zur Durchführung der Prüfung nach §4 Abs.1 Nr. 6 der Verordnung über Anmeldeunterlagen und Prüfnachweise nach dem ChemG Texte 16/84, UBA

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

In order to determine the substance concentration, water samples were taken from the middle of the test chamber with 100 mL neck glass bottle at study start, and after 48 and 96 hours.

Vehicle:

no

Details on test solutions:

The test substance was weighed into a beaker. Water for dilution was added, the mixture was homogenized using an Ultra-Turrax and Ultrasonic Bath and poured into the chamber under stirring with a glass rod.
Test concentrations: 0/100/500/710 and 1000 mg/L

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Species: Zebra fish - Brachydanio rerio (HAMILTON-BUCHANAN)
- Source: West Aquarium, Bad Lauterberg, Germany
- Date of hatching: July 07, 1988 / August 11, 1988
- Delivery date: September 29, 1988 / October 11, 1988 / October 18, 1988
- Start of study: October 17,1988
- Acclimatisation: The fish were kept for 14 days before the start of the study in water for dilution
- Length at study initiation (length definition, mean, range and SD): mean: 2.9±0.21; range: 2.4 - 3.3
- Method of breeding:
- Feeding during test: none
- Food type: NA
- Amount: NA
- Frequency: NA


ACCLIMATION
- Acclimation period: 14 days
- Feeding: ad libitum
- Food type: Tetra Min, Tetra Werke, Melle (Germany)
- Frequency: twice daily
- Temperature: 22 ± 1 °C
- Oxygen content: >= 80 % of the saturation value
- Duration of light period: 12 hours daily
- Population density: :<= 1 g fish/L water
- Health during acclimation (any mortality observed): none

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

-

Hardness:

-

Test temperature:

Test groups: 21 - 23°C
Control: 21.3 - 23°C

pH:

Test groups: 7.3 - 7.8
Control: 7.5 - 8.1

Dissolved oxygen:

Test groups: 7.1 - 9.4 mg/L
Control: 7.1 - 9.4 mg/L

Salinity:

-

Nominal and measured concentrations:

Nominal concentration Measured concentration
(mg/L) mean (mg/L) %
100 101.7 101.7
500 506.2 101.2
710 722.1 101.7
1000 961.4 96.1

Details on 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 with a Plexiglas viewing panel. The temperature of the water bath was regulated by a thermostat to 22 ±1°C. The chambers were illuminated above 12 hours. The light intensity directly over the chambers was
approximately 700 lux. The test chambers were not aerated during the course of the study.

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 Iitres 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 7.9 - 8.1.

After the test concentrations had been prepared and water parameters recorded, 10 fish were assigned to each test and control chamber. The fish received no 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 lethality and visible changes in appearance and behavior. Dead fish were removed from the chambers. Fish were considered dead when there was a lack of opercular movement and no response to slight mechanical stimulus. The water parameters were measured and recorded before study start and after 0, 2 - 4, 24, 48, 72 and 96 hours.

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

842.6 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

710 - 1 000 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

596 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: CI: 525 - 675 mg/L

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

1 000 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

other: mortality and behaviour

Details on results:

Because of the colouration of the test solutions the observation of symptoms was disturbed in the 710 and 1000 mg/L groups, The fish showed changes in behavior, swimming behavior and partly in respiration rate in the 500,710 and 1000 mg/L groups.
Symptoms occurred during the entire exposure time. The 100 mg/L group showed no effect in comparison with the negative control.
Dead fish showed redded opercula, lock-jaw, partly dark or yellow or light coloured integument.

Reported statistics and error estimates:

Probit analysis could not be carried out after 24, 48 and 72 hours because of the lethality rate. As the factor for the concentration steps was 1.41 (2nd root of 10) no further concentrations were tested. After 48 hours the geometric mean of the LC0 and LC100 and after 24 and 72 hours the LC50 range are given.
On the basis of the lethality rate, the LC50 values were determined after 96 hours by means of the probit analysis.

Validity criteria fulfilled:

yes

Conclusions:

The LC50 after 96 hours was 596 mg/L under the conditions of this test

Executive summary:

The acute toxicity of the test substance in zebra fish was determined in a study over 96 hours. The concentrations tested were 100, 500, 710 and 1000 mg/L and a negative control (0 mg/L).

The test batches were present as orange to dark red solutions. No particulate matter was observed. The substance concentration was determined in all test batches. All values determined were in a range of ±20% of the theoretical value.

In this 96-hour acute toxicity study in zebra fish (Brachydanio rerio) over 96 hours the following lethality values were determined:

	24 h	48 h	72 h	96 h
LC0 [mg/L]	1000	710	500	100
LC50 [mg/L]	> 1000	842.6	710 - 1000	596 (CI: 525 - 675)
LC1000 [mg/L]	not determined	1000	1000	1000

Symptoms and lethality were observed at 500, 710 and 1000 mg/L. The 100 mg/L group showed no symptoms and no lethality.

Description of key information

The LC50 for acute fish toxicity was determined to be higher than 100 mg/L after 96 h exposure as determined in close structural analogues.

Additional information

LC50 (96 h): > 100 mg/L test mat.