α,α,α-trifluoro-o-toluidine

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:

1988

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:

1988

Report date:

1988

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)

Version / remarks:

84/449/EEC

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

α,α,α-trifluoro-o-toluidine

EC Number:

201-806-7

EC Name:

α,α,α-trifluoro-o-toluidine

Cas Number:

88-17-5

Molecular formula:

C7H6F3N

IUPAC Name:

2-(trifluoromethyl)aniline

Sampling and analysis

Analytical monitoring:

no

Test solutions

Details on test solutions:

PREPARATION OF TEST SOLUTION
The test item was diluted with water. After homogenizitation by ultrasound and intensive stirring the mixture was incorporated into the test vessel. No phase separations were observed at any test concentration.

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: ca. 3 months
- Length at study initiation (mean (n=40), range and SD): 2.9 cm, 2,4 - 3.5 cm, SD 0.29
- 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: twice 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:

21.4°C - 23.0°C (treatment), 21.6°C - 23.0°C (control)

pH:

7.4 - 8.1 (treatment), 7.5 - 8.1 (control)

Dissolved oxygen:

6.3 - 9.4 mg/L (treatment); 7.1 - 9.4 mg/L (control)

Nominal and measured concentrations:

Nominal concentrations: 10, 25, 35, 50 and 100 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): 1
- 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: 7.8 - 8.0.
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

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

> 25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 25 - < 35 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

- There was no mortality in control at any observation time point.
- At concentrations 35 mg/L and higher mortalities were observed after 72 h. In the high dose group animals died before the first observation time. Dead animals were light and dark coloured. One of the dead animals had lockjaw.
- Behavioural abnormalities at concentration levels 10 - 50 mg/L: swimming posture tail-heavy, upside-down and on side; swimming rotation; increased and decreased activity; increased fright reaction; swimming and sitting on the base of the tank; projecting opercula; convulsions, twitching, integument dark coloured

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table 1: test concentrations and results

Concentration	Mortality
[mg/L]	2 – 4 h	24 h	48 h	72 h	96 h
Experiment 1
0	0 /10	0 /10	0 /10	0 /10	0 /10
100	10 / 10	10 / 10	10 / 10	10 / 10	10 / 10
Experiment 2
0	0 /10	0 /10	0 /10	0 /10	0 /10
10	0 /10	0 /10	0 /10	0 /10	0 /10
Experiment 3
0	0 /10	0 /10	0 /10	0 /10	0 /10
25	0 /10	0 /10	0 /10	0 /10	0 /10
35	0 /10	0 /10	0 /10	4 / 10	8 / 10
Experiment 4
0	0 /10	0 /10	0 /10	0 /10	0 /10
50	0 /10	0 /10	5 / 10	9 / 10	10 / 10

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

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. Static test with the concentrations 10, 25, 35, 50 and 100 mg/L solution were 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 and the control respectively. 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.

There were no mortalities at a concentration of 25 mg/L at any observation time point. At all other concentration levels up to 100 % mortalities were observed after 2 -4 h to 72 h.

At all test concentration levels the following signs of toxicity were observed: swimming posture tail-heavy, upside-down and on side; swimming rotation; increased and decreased activity; increased fright reaction; swimming and sitting on the base of the tank; projecting opercula; convulsions, twitching, integument dark coloured.

The median lethal concentration of the test item to zebrafish observed is:

LC₅₀ (96 h): > 25 - < 35 mg/L