α,α,α-trifluoro-p-toluidine

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 21 july 1995 to 09 feb 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was performed according to the standardised method and GLP. Sufficient information was available about the test substance and the test conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Cited as Directive 84/449/EEC, C.2 (Acute toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

α,α,α-trifluoro-p-toluidine

EC Number:

207-236-5

EC Name:

α,α,α-trifluoro-p-toluidine

Cas Number:

455-14-1

Molecular formula:

C7H6F3N

IUPAC Name:

4-(trifluoromethyl)aniline

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Not relevant

Test solutions

Vehicle:

no

Details on test solutions:

- Concentrations: Only the stock solution was analysed at T0 (end of  centrifugation), T24H and T48H

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult  test substances)
- Method: Stock solutions were prepared by dissolving 100 mg of substance  in 1000 g of test medium. The flasks were stirred for 24h at 30°C, then  for 24h at 20°C and finally centrifugated at circa 9000 g for 20 min.  This extracted solution was then used as stock solution The tested 
concentrations were prepared by dilution of the stock  solution. These tested concentrations are expressed as % of the extracted  solution 
(Concentration of the extracted solution at T0: 39 mg/l, T24H:  33 mg/l, T48H: 29 mg/l).
- Controls: yes
- Chemical name of vehicle: none 
- Concentration of vehicle in test medium (stock solution and final test  solution): none
- Evidence of undissolved material: no data

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia sp.
- Strain: Daphnia magna strauss 1820 (Cladocera crustacea)
- Source: :  IRCHA-INERIS (Daphnia breeding in Décines Rhône-Poulenc  laboratory)
- Age at study initiation: no data 
- Weight at study initiation : no data 
- Length at study initiation: 560- Method of breeding: no data
- Feeding during test: no data

ACCLIMATION
- Acclimation period: no data
- Acclimation conditions: no data
- Type and amount of food: no data
- Feeding frequency:  no data
- Health during acclimation: no data

QUARANTINE (wild caught)
- Duration: no data
- Health/mortality: no data

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

no data

Test temperature:

19.8 - 20°C

pH:

7.9 - 8.4

Dissolved oxygen:

96 - 99%

Salinity:

no data

Nominal and measured concentrations:

0, 1.2, 1.9, 2.8, 4.4, 6.5, 10,  15, 22, 33 and 50% of the extracted solution

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 ml glass pyrex beaker     
- Type: open     
- Material, size, headspace, fill volume: 100 ml filled with 40g of  solution     
- Aeration: no
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 1 daphnia per 2g of tested soutions

WATER PARAMETERS
- Source/preparation of dilution water: bidistillated water
- Total organic carbon: no data
- Particulate matter: no data
- Metals: no data
- Pesticides: no data
- Chlorine: no data
- Alkalinity: NaHCO3: 0.2 g/l
- Ca/mg ratio: CaCl2,2H2O: 0.297 g/l; MgCl2,6H2O: 0.167 g/l
- other: K2SO4: 0.026 g/l - Conductivity: 1.3 µS/cm (of the bidistillated water)
- Culture medium different from test medium: no data
- Intervals of water quality measurement: no data

TEST MEDIUM PARAMETERS  
- Ca/Mg ratio: CaCl2,2H2O: 0.297 g/l; MgCl2,6H2O: 0.167 g/l
- conductivity: 1.3 µS/cm (of the bidistillated water)
- pH : 7.9 at 20°C
- O2 : 100% saturation

Test conditions: 
- Adjustment of pH: no
- Photoperiod: exposure in the dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : 

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5 - Range finding study     
- Test concentrations: 0.1, 0.35, 1.0, 3.5, 10, 35 and 100% of the  extracted solution     
- Results used to determine the conditions for the definitive study:  % of immobilisation at 48H: 25% at 3.5%, 35% at 10%, 70% at 35% and 100%  at 100% of the extracted solution (repeated twice for the latter result)

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.85 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

behaviour

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

9.1 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

behaviour

Details on results:

The measured effect is the mortality of the daphnids  estimated through their immobilisation
- Behavioural abnormalities: no data
- Observations on body length and weight: no data
- Other biological observations: no data
- Mortality of control: 0%
- Other adverse effects control: no data
- Abnormal responses: no data
- Any observations (e.g. precipitation) that might cause a difference  between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium:  no

Results with reference substance (positive control):

Potassium dichromate EC50-24h = 1.2 mg/L

Reported statistics and error estimates:

- 95% conf. limits: 1.4-3.9 %  (of the extracted solution) mg/L (probit method). In the conditions of  the test, substance concentration decreased by approximately 26% on 48H  exposure period (calculated on the stock solution concentration)

Any other information on results incl. tables

EC50 (48h) PTFMA: 2.6% extraction solution=  0.85 mg/l calculated on the basis of geometric mean of concentrations.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Substance considered as very toxic to aquatic organisms according to directive 67/548/EEC.

Executive summary:

The 24 and 48 hours toxicity of p-TFMA to Daphnia magna was studied under static conditions (RP Industrialisation, 1996). Daphnids were exposed to control and test chemical at concentrations of 0 to 50% of the extracted solution for 24 and 48 hours.

Immobilization effects were observed and interpretade as mortality.

The 48 hours EC 50 was 0.85 mg/L (geometric mean measured concentrations).

Based on the result of this study, p-TFMA should be classified very toxic for daphnia magna in accordance with the EC classification criteria.