(4-chlorophenyl)hydrazine

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- TS: CPH, BAS 500 F - Stufe 100
- Purity: 98.6 area-%

- Batch No.: 29898/133
- Water solubility: about 100 mg/L (data from sponsor)

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

A stock solution in aqua bidest. (containing 0.1998 g test compound/2l) was prepared. For homogenization the stock solution was incubated in an ultra sonic bath for about 10 minutes. Seperately for each concentration 5, 10, 22, 50 and 100 ml from the stock solution were added to 10 L test water using an ultra-turrax stirrer. The fish were introduced into the test solutions directly after preparation to test the combined effects of the parent compound and its degradation products.

Test organisms

Test organisms (species):

other: Danio rerio (Zebra fish)

Details on test organisms:

- Test fish: Zebra fish (Brachydanio rerio HAM. and BUCH.)
- Supplier: Zierfisch-Center Kloeckner, Ludwigshafen/Rhein, Germany Body length: 3.3 (2.9 - 3.6) cm Body weight: 0.34 (0.24 - 0.53) g

Housing and adaption
- Culture conditions: oil-free aerated and charcoal filtered tap water, flow-through system;
- Water temperature: 21 - 22 °C
- Duration of housing: about 2 weeks
- Duration of adaptation to test water and test temperature: 3 days
- Medical treatment: none
- Diet: "TETRA MIN" standard feed for aquarium fish (TETRA - WERKE, MELLE, Germany), ad libitum; and on workdays generally additionally live brine shrimp (artemia).

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Test temperature:

22 °C

pH:

8.1 - 8.4

Dissolved oxygen:

6.1 - 8.6 mg/L

Nominal and measured concentrations:

Nominal test concentrations: 0, 0.05, 0.1, 0.22, 0.5, 1.0 mg/L (based on the results of a range-finding test). In addition, a control was tested in parallel. The analytically detected concentrations of the parent compound at the end of the exposure was below the detection limit. Therefore all mentioned LC values are based on nominal concentrations.

Details on test conditions:

- Test water: Dilution water: Municipal water of the city of Frankenthal, not chlorinated and passed through a charcoal filter, aerated.

- Dilution water chemistry:
Total hardness : about 2 .5 mmol/l (about 250 mg/l CaC03)
Acid capacity : about 5.5 mmol/l
Ca content : about 90 mg/l
Mg content : about 10 mg/l
pH : about 8.0 - 8.6

- Volume of water: 10 L
- No of animals per test concentration and control: 10
- Loading rate (g fish/l test water): 0.3
- Test vessels: All glass aquaria (30 x 22 x 24 cm)
- Photoperiod: 16 : 8 hours day-night regime
- Aeration: none
- Withdrawal of food: 1 day before and during exposure.

- Observations:
Mortality after 1, 4, 24, 48, 72, and 96 h
Symptoms after 1, 4, 24 and 48 h
Measurements: pH and oxygen content after 1, 24, 48, 72 and 96 h

- Statistical evaluation of data:
The median lethal concentration (LC50) was calculated using Probit Analysis (Finney D.J., Probit Analysis, Cambr. Univ. Press, 3rd Edition, 1971)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

0.22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Any other information on results incl. tables

Sublethal observations / clinical signs:

LC 50 (1h): > 1 mg/l
LC 50 (4h): about 0.78 mg/l
LC 50 (24h): about 0.15 mg/l
LC 50 (48h): about 0.15 mg/l
LC 50 (72h): about 0.15 mg/l
LC 50 (96h): about 0.15 mg/l


Cumulative mortalitiy

Nominal concentration (mg/l) 1h 4h 24h 48h 72h 96h
Control        0 0 0 0 0 0
0.05        0 0 0 0 0 0
0.10        0 0 0 0 0 0
0.22        0 0 10 10 10 10
0.50        0 0 10 10 10 10
1.0        0 9 10 10 10 10


Symptoms of intoxication

Nominal concentration (mg/l) 1h 4h 24h 48h 72h 96h
Control        - - - - - -
0.05        - - - - - -
0.10        - - - - - -
0.22        - - - - - -
0.50        - - - - - -
1.0        - T - - - -

key:
-: No symptoms/all fish dead
T: Tumbling


Nominal oxygen content(mg/l):
concentration(mg/l) 0h 24h 48h 72h 96h
0(Control)        8.4 8.2 8.1 8.1 8.1
0.05        8.4 8.1 8.1 8.1 8.1
0.10        8.4 8.2 8.1 8.1 8.1
0.22        8.3 8.2
0.50        8.3 8.2
1.0        8.3 8.2


pH:
concentration(mg/l) 0h 24h 48h 72h 96h
0(Control)   8.4 6.8 6.5 6.3 6.1
0.05        8.5 6.8 6.6 6.3 6.2
0.10        8.6 6.8 6.6 6.3 6.1
0.22        8.5 7.3
0.50        8.6 7.7
1.0        8.6 7.9