Disodium 6-(4,6-dichloro-1,3,5-triazin-2-ylamino)-1-hydroxy-2-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-3-sulfonate

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:

From Sep. 08, 1997 to Nov. 07, 1997

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)

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:

Daily analysis of the only test concentration plus a control samples
- Sampling method: 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 h.
- Sample preparation: The samples were diluted with mobile phase

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method of administration: Direct weighing
- 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.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Strain: HAMILTON BUCHANAN
- Source: Hoechst Marion Roussel Deutschland GmbH, Global Preclinical Development - Drug Safety
- Age at study initiation (mean and range, SD): ~ 3 mo
- Length at study initiation (length definition, mean, range and SD): 2.6 - 2.9 cm

ACCLIMATION
- Acclimation period: 14 d
- Acclimation conditions (same as test or not): Yes
- Keeping water: Reconstituted water, composition according to ISO/DIS 7346/1
- Food type: Tetra Min, Tetra Werke, Melle (Germany)
- Frequency: twice daily ad libitum

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No

Hardness:

2.1 - 2.3 mmol Ca2+ Mg2 / L

Test temperature:

21.7-22.1 °C

pH:

7.6-7.9

Dissolved oxygen:

Oxygen content: 7.2-9.3 mg/L

Salinity:

No data

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured: 103 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass chambers(length 20 cm, width 15 cm, height 20 cm) and stood in a water bath made of Hostalit Z® with a Plexiglas viewing panel
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: 4 L test medium
- Aeration: Not aerated during the course of the study
- Type of flow-through (e.g. peristaltic or proportional diluter): Static
- Method of administration: Direct weighing


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water, composition according to ISO/DIS 7346/1

OTHER TEST CONDITIONS
- Photoperiod: 12 h light/12 h dark
- Light intensity: 700 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality, visible effects on appearance and behaviour

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 100 mg/L

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: -

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 103 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: -

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: -

Details on results:

- Behavioural abnormalities: Showed changes in behavior and swimming behavior during the first 48 h of exposure. From 72 h on only changes in swimming behavior was observed
- Other biological observations: The fish showed changes in respiration rate up to 72 h of exposure
- Mortality: No lethality occurred in the 100 mg/L-group and in the control group

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

No data

Sublethal observations / clinical signs:

Table 1. Results-Lethality values

	after 24 h	after 48 h	after 72 h	after 96 h
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

The 100 mg/L group exhibited symptoms but no lethality 

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the 96 h LC50 value of the test substance was determined to be ≥100 mg/L.

Executive summary:

A study was performed to assess the acute toxicity of test substance to Zebra fish (Brachydanio rerio) over 96 h under static conditions. The study was conducted in accordance with OECD Guideline 203 and EU Method C.1. in compliance with GLP.

Groups of seven fish of the recommended size were exposed to a limit test concentration of nominally 100 mg/L of test substance dissolved in synthetic fresh water. Analysis was performed for the test concentration plus a control on daily basis.  Observations were made on the lethality and visible changes in appearance and behavior of fish after 3, 6, 24, 48, 72 and 96 h of exposure. The water parameters were measured and recorded before study start and after 0, 24, 48, 72 and 96 h.

The tested concentration was present as clear red solution. The test batch was homogeneous. No particulate matter was observed. The substance concentration was determined in the 100 mg/L test batch. The values determined were in a range of ± 10 % of the theoretical value.

 The LC50 h values at 24, 48, 72 and 96 h was determined to be >100 mg/L. The 100 mg/L group exhibited symptoms but no lethality. The fish showed changes in behavior, swimming behavior and respiration rate during the first 48 h of exposure. From 72 h on only changes in swimming behavior and respiration rate were observed

Under the test conditions, the 96 h LC50 value of the test substance was determined to be >100 mg/L.

Description of key information

The 96 h LC50 value of the test substance was determined to be >100 mg/L.

Additional information

A study was performed to assess the acute toxicity of test substance to Zebra fish (Brachydanio rerio) over 96 h under static conditions. The study was conducted in accordance with OECD Guideline 203 and EU Method C.1.

Groups of seven fish of the recommended size were exposed to a limit test concentration of nominally 100 mg/L of test substance dissolved in synthetic fresh water. Analysis was performed for the test concentration plus a control on daily basis.  Observations were made on the lethality and visible changes in appearance and behavior of fish after 3, 6, 24, 48, 72 and 96 h of exposure. The water parameters were measured and recorded before study start and after 0, 24, 48, 72 and 96 h.

The tested concentration was present as clear red solution. The test batch was homogeneous. No particulate matter was observed. The substance concentration was determined in the 100 mg/L test batch. The values determined were in a range of ± 10 % of the theoretical value.

 The LC50 h values at 24, 48, 72 and 100 was determined to be >100 mg/L. The 100 mg/L group exhibited symptoms but no lethality. The fish showed changes in behavior, swimming behavior and respiration rate during the first 48 h of exposure. From 72 h on only changes in swimming behavior and respiration rate were observed

The 96 h LC50 value of the test substance was determined to be >100 mg/L (Dr. Zok S, 1997).