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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-compliant guideline study available as unpublished report, no restrictions, fully adequate for assessment

Data source

Reference Type:
study report
Report Date:

Materials and methods

Test guidelineopen allclose all
according to
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
according to
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
yes (incl. certificate)
BASF AG Experimental Toxicology and Ecology

Test material

Test material form:
other: liquid
Details on test material:
- Name of test substance: 1,2,3.4-Tetrahydroisoquinoline
- Purity: 87.3area- %
- Physical state / appearance: Liquid / redwish-yellow
- Homogeneity: homogeneous
- Water solubility: 20 g/L
- Storage conditions: The test substance was stored at ambient temperature
- Storage stability: stability of test substance under storage conditions over test period guaranteed by sponsor
- Test substance number: 05/0797-1
- Batch identification: LJ 32741/75
- Date of production: 10 Oct 2005

Sampling and analysis

Analytical monitoring:
Details on sampling:
At the start of the exposure (0 h) samples from vessels without daphnids and at the end of the exposure (48 h) samples from vessels with daphnids were analysed. Retained samples were stored in the freezer.

Test solutions

Details on test solutions:
The test solutions were prepared by serial dilution of a stock solution. Preparation of the stock solution: 100 mg test substance was weighed and made up with 1000 mL M4 medium. The mixture was stirred (magnetic stir plate) for about 20 minutes at 20 ± 2 °C. By dilution of this stock solution with M4 medium the test concentrations were prepared. The highest concentration (100 mg/L) is identical with the stock solution.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
- Test species: Daphnia magna STRAUS;
- Origin: Clone of Daphnia magna STRAUS 1820 used was supplied by the Institut National de Recherche Chimique Appliquée, France, in 1978. From this date on, clone was cultured and bred continuously in the Laboratory of Ecotoxicology of the Experimental Toxicology and Ecology of BASF SE in Ludwigshafen;

- Photoperiod: 16 h light, 8 h dark;
- Temperature: 20 ± 1°C;
- Diet: unicellular algae Desmodesmus subspicatus;
- Feeding: daily;
- Renewal of culture water: twice weekly;
- Culture medium: Elendt M4;

Study design

Test type:
Water media type:
Limit test:
Total exposure duration:
48 h

Test conditions

2.40 mmol/L
Test temperature:
20.3 - 20.7 °C;
8.1 - 8.5
Dissolved oxygen:
8.4 - 8.9 mg/L
Nominal and measured concentrations:
- Nominal concentrations: 0, 6.25, 12.5, 25, 50 and 100 mg/L
- Measured concentrations (at t=0): n.d., 6.26, 11.5, 22.4, 47.6 and 101.9 mg/L
- Measured concentrations (at t=48h): n.d., 5.74, 11.8, 23.1, 46.6 and 101.0 mg/L
Details on test conditions:
- A synthetic fresh water (Elendt M4) was used as media for culture and test purposes. For the composition of this M4 medium see OECD 202.
- Properties of this medium:
- Total hardness: 2.20 - 3.20 mmol/L;
- Acid capacity up to pH 4.3: 0.8 - 1.00 mmol/L;
- Molar ratio Ca:Mg: about 4:1;
- pH-value: 7.5 - 8.5;
- Conductivity: 550 - 650 µS/cm;
- Prior to use the M4 medium was aerated for approximately 24 hours until saturation with oxygen was reached.

The exposure was carried out in a static system with no aeration.
- Test temperature: 20.3 – 20.7 °C
- Test vessel: Test tubes (glass) with flat bottom(nominal volume 20 mL)
- Test volume: 10 mL
- Loading (animals/mL): 0.5
- Number of animals/vessel: 5
- Total number of animals/ concentration: 20
- Number of replicates per concentration: 4 replicates with test animals (5 animals per replicate) 1 additional replicate (replicate 5, without animals) per test concentration and control for measurement of the pH and oxygen content at the beginn of the exposure. This replicate was discarded after measurement. 1 replicate without animals for analytical control analyses
- Age of the animals at the start of the exposure: > 2 h < 24 h (at least 3rd brood progeny)
- Age of parent animals: 2 - 4 weeks
- Illumination: Artificial light, type warm white (OSRAM L58 W31) 16 hours day : 8 hours night
- Test parameter: Immobilisation
- Feeding: None

- Each test vessel was visually checked for immobilised daphnids after 0, 24 and 48 hours. In addition any abnormal behaviour or appearance was documented.
- Determination of the chemical and physical parameter of the test medium prior to use: Total hardness, acid capacity, pH value and conductivity of the water after aeration and new preparation.
- Measurement of the pH and oxygen content at the start of the exposure (in an additional replicate) and at the end of the exposure in replicate 1 of all test concentrations and the control.
- Measurement of temperature continuously during the whole period in a separate vessel filled with water proximal to the test vessels.
Reference substance (positive control):
Potassium dichromate

Results and discussion

Effect concentrations
48 h
Dose descriptor:
Effect conc.:
25.9 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: 95% C.L.: 6.0-1304 mg/L
Details on results:
After 48 hours no immobilisation was noted at the lowest test concentration of 6.25 mg/L. At test concentrations of 12.5 and 25 mg/L immobilisation was observed in 17/20 and 15/20 daphnids, respectively. At the highest test concentrations of 50 and 100 mg/L all animals were immobile. No additional adverse effects or abnormal behaviour were observed in any of the test treatments.
Results with reference substance (positive control):
The EC50(24h) of the control substance potassium dichromate was 1.26 mg/L. This result is within the range of 0.6 – 1.7 mg/L (ISO 6341) and indicates that the culture of Daphnia magna used in this study is responding normally to toxic stress.
Reported statistics and error estimates:
For the statistical calculation / evaluation of the EC50 the probit method based on Finney moving average method with SAS (Version 8.2) software was used.

Any other information on results incl. tables


- None of the control daphnids were immobilized or showed any other signs of disease or stress.

- The O2-concentration was > 3 mg/L in all tested replicates.

In conclusion the test was considered to be valid.

Applicant's summary and conclusion