4-nitrobenzoic acid

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: Study according to OECD Guideline No. 203

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-nitrobenzoic acid

EC Number:

200-526-2

EC Name:

4-nitrobenzoic acid

Cas Number:

62-23-7

Molecular formula:

C7H5NO4

IUPAC Name:

4-nitrobenzoic acid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Appr. 100 mL samples were taken from the middle of the test vessel at 0, 48 and 96 hours respectively.

Test solutions

Details on test solutions:

PREPARATION OF TEST SOLUTION
0.095 mL Tween 80 /L was used in the final test concentration. After homogenizitation by ultrasound and intensive stirring the mixture was incorporated into the test vessel and stirred for another 2 hours. For the medium and high dose concentration the resulting acidic pH was adjusted with diluted sodium hydroxide solution to the pH of the test system.
Concentration of the test item in the treatment vessel (high concentration only) was determined after 0, 48 and 96 h respectively.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Strain: Brachydanio rerio (Hamilton-Buchanan)
- Source: West Aquarium, D-37431 Bad Lauterberg, Germany
- Age at study initiation: appr. 4 months
- Length at study initiation (mean, range and SD): 3.1 cm, 2,7 - 3.5 cm, SD 0.21
- Feeding during test: No feeding during the test

ACCLIMATION
- Acclimation period: at least 14 days
- Type and amount of food: Tetra Min, ad libitum
- Feeding frequency: 2 times per day

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1°C
- Photoperiod (hrs dark / hrs light): 12/12
- The dissolved oxygen concentration was more than 80 % of the air saturation value

Study design

Test type:

static

Water media type:

freshwater

Total exposure duration:

96 h

Post exposure observation period:

Observations were carried out during exposure at the beginning of the experiment and after 2 - 4, 24, 48, 72 and 96 hours.

Test conditions

Test temperature:

21.5°C - 23.0°C (treatment), 21.2°C - 23.0°C (control)

pH:

6.9 - 8.0 (treatment), 7.4 - 8.4 (control)

Dissolved oxygen:

4.3 - 9.5 mg/L (treatment); 4.6 - 9.5 mg/L (control)

Nominal and measured concentrations:

Nominal concentrations: 50, 100, 220 mg/L
Nominal concentration: 500 mg/L
Measured concentrations: 504 mg/L - 501 mg/L - 500 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass-aquaria 30 cm x 22 cm x 24 cm, 10 L per vessel
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2 (1 with solutizer)
- Biomass loading rate: fish density in the tanks was less than 1 g per litre test solution.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water, quality according ISO 7346/1. The water was saturated with oxygen. pH-value: 8.0 - 8.3.
pH , temperature and oxygen saturation were observed at the beginning, after 2-4, 24, 48, 72 and 96 h in all vessels


OTHER TEST CONDITIONS
- Photoperiod (hrs dark / hrs light): 12/12
- Light intensity: 700 Lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and behavior of the test fish and control fish were observed at 2-4, 24, 48, 72, 96 h.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 500 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality (fish)

Details on results:

There was no mortality in control as well as in treated fish at any observation time point.
At different observation time points at all dose levels and the solvent control as well the following abnormalities were observed: hypoactivity, irregular respirate, swimming posture tail-heavy, projecting opercula, increased and decreased respiration rate, increased fright reaction, swimming at the surface

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In this study the test item did not cause any mortalities to zebrafish after 96 h when tested with different concentrations up to 500 mg/L (nominal) solution using Tween 80 as solutizer.

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) at Hoechst AG, Frankfurt, Germany. A static test with concentrations 50, 100, 220 and 500 mg/L solution, using a solubilizer was conducted. The preparation method has been chosen with regard to the test item properties and in accordance with the relevant OECD guidance document. Duration of the test was 96 h. 10 test organisms were exposed to the test solution respectively and the control. Water quality parameters pH-value, temperature and oxygen-saturation measured after 0, 2 -4, 24, 48, 72 and 96 h were determined to be within the acceptable limits. Test item analysis was carried out at 0, 48 and 96 h. Concentrations were above 80 % of the initial concentration.

The maximum test concentration of 500 mg/L solution of the test item didn´t show any lethality at all observation time points up to 96 hours.