2-(4-(diethylaminopropylcarbamoyl)phenylazo)-3-oxo-N-(2,3-dihydro-2-oxobenzimidazol-5-yl)butyramide

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:

Aug - Oct 1989

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

adopted 4. Apr. 1984

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-(4-(diethylaminopropylcarbamoyl)phenylazo)-3-oxo-N-(2,3-dihydro-2-oxobenzimidazol-5-yl)butyramide

EC Number:

404-910-2

EC Name:

2-(4-(diethylaminopropylcarbamoyl)phenylazo)-3-oxo-N-(2,3-dihydro-2-oxobenzimidazol-5-yl)butyramide

Cas Number:

164578-14-7

Molecular formula:

C25H31N7O4

IUPAC Name:

N-[3-(diethylamino)propyl]-4-[(1E)-2-{2-oxo-1-[(2-oxo-2,3-dihydro-1H-1,3-benzodiazol-5-yl)carbamoyl]propyl}diazen-1-yl]benzamide

Test material form:

solid: bulk

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
10, 22 and 50 mg/L

- Sampling method:
Samples for analysis were taken (from the vessel middle) at 0 and after 24 h of the fresh preparation.

Test solutions

Vehicle:

yes

Remarks:

0.095 mL Tween 80/L water

Details on test solutions:

water was prepared according to ISO/DIS 7346/1

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
0.095 mL Tween 80 were added to the test item and filled up with water. After homgenisation by sonification and Ultra-Turrax the resulting solution filled in the test bassins.
Water: dilution water according to ISO/DIS 7346/1
Afterwards the solution was stirred for 2 h receiving a yellow solution with slight precipitation. Since the test item did not dissolve completely in water the test solution was prepared and changed every 24 h.
- Controls:
solvent control: 0.095 mL Tween 80/L

- Chemical name of vehicle (organic solvent, emulsifier or dispersant):

Tween 80
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)):
0.095 mL/L

- Evidence of undissolved material (e.g. precipitate, surface film, etc.):
yes, visible precipitation

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Danio rerio
- Strain: HAMILTON-BUCHANAN
- Source: West Aquarium, Bad Lauterberg, Germany
- Age: approx. 10 weeks
- Length at study initiation (length definition, mean, range and SD): 3 +/- 0.15 cm

ACCLIMATION
- Acclimation period: at least 14 days
- Acclimation conditions (same as test or not): yes
- Type and amount of food during acclimation: Tetra Mein (Tetra Werke, Melle, Gemany) twice daily, ad libitum
- Feeding frequency during acclimation: twice daily


FEEDING DURING TEST
no feedind during exposure period

Holding:
in glas vessels (30 x 22 x 24 cm)filled with 10 L solution/water, 22 +/- 1°C, 12 light (700 Lux), test solution was changed every 24 h

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

2 days

Test conditions

Hardness:

according to ISO/DIS 7346/1

Test temperature:

test groups: 21 - 23°C
controls: 21 - 23°C

pH:

test groups: 7.6 -8.3
controls: 7.5 - 8.3

Dissolved oxygen:

test groups: 7.0 -9.3 mg/L
controls: 7.1 - 9.4 mg/L

Salinity:

according to ISO/DIS 7346/1

Conductivity:

according to ISO/DIS 7346/1

Nominal and measured concentrations:

Nominal concentrations:
10, 22, 50 mg/L
Measured concentrations:
10.9 - 10.1 mg/L, 20.8- 19.9 mg/L at 0 and 24 h of freshly prepared test solution, respectively

Details on test conditions:

TEST SYSTEM
- Test vessel: glas vessel (30 x 22 x 24 cm, length x width x heigth), filled with 10 L volume
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): every 24 h, manually
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 3


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

OTHER TEST CONDITIONS
- Adjustment of pH: not needed
- Photoperiod: 12 h
- Light intensity: 700 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: concentrations applied based on result of pre test
- Range finding study : performed

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

LC0

Effect conc.:

22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

33 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 4

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

>= 22 - <= 50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 5

Key result

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 6

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities:
at 10 mg/L and above symptoms like reduced activity, increased breathing frequency, chges in the shape of swimming were observed
due to preciptiation no observation could be made at 50 mg/L

- Other biological observations: fish, that died, were pale or yellow colored; substance preciptation on body surface and reddened gills were reported

- Mortality of control:
- Other adverse effects control: no mortality observed

- Effect concentrations exceeding solubility of substance in test medium: (at least slight) precipitation of test item was observed at all test concentrations

Results with reference substance (positive control):

n.a.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the observed mortality the LC50 was set at 22-55 mg/L (96 h) and 33 mg/L (48 h).

Executive summary:

The acute toxicity of the test item to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1984).

A semi-static limit test with daily renewal of the test media was conducted.

0.095 mL Tween 80 were added to the test item and filled up with water. After homgenisation by sonification and Ultra-Turrax the resulting solution filled in the test bassins.

The dilution water was prepared according to ISO/DIS 7346/1. Afterwards the solution was stirred for 2 h receiving a yellow solution with slight precipitation. Since the test item did not dissolve completely in water the test solution was prepared and changed every 24

Duration of the test was 96 h. 7 test organisms were exposed to 10, 22, 50 mg/L test item and the control.

Water quality parameters pH-value, temperature and oxygen-saturation measured were determined to be within the acceptable limits.