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Short-term toxicity to aquatic invertebrates

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short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
according to guideline
ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
- Melting point: -30°c
- Boiling point: 314°c
- Vapour pressure: < 0.001 hPa@20°c
- Water solubility (under test conditions): 1.02 g/l
- Henry's law constant: 2.17 E-006 atm-m³/mole
- log Pow: 2.10
- pKa:
- Stability in water:
- Stability in light:
- pH dependence on stability:

OTHER PROPERTIES (if relevant for this endpoint)
- Results of test for ready biodegradability: not readily biodegradable
- Other:
Analytical monitoring:
Details on sampling:
- Concentrations: Based on the result of the range-finding test, a range of nominal concentrations 10, 18, 32, 56 and 100 mg/l
- Sampling method: The batch of BBSA tested was a clear, colourless liquid with a purity of 99.8%. In a pre-test BBSA proved to be completely soluble in test medium at 100 mg/l after 10 minutes of stirring followed by a 10-minute treatment with ultrasonic waves. Preparation of test solutions started with a stock solution of 100 mg/l
- Sample storage conditions before analysis:
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: In a pre-test BBSA proved to be completely soluble in test medium at 100 mg/l after 10 min of stirring followed by a 10-min treatment with ultrasonic waves.
- Eluate:
- Differential loading:
- Controls: test medium without test substance or other additives
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): /
- Concentration of vehicle in test medium (stock solution and final test solution): /
- Evidence of undissolved material (e.g. precipitate, surface film, etc):no, they were all clear and colourless
Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnia magna (Crustacea)
- Common name: Daphnia magna
- Strain: crustacea, cladocera
- Source: no data
- Age at study initiation (mean and range, SD): age of < 24h.
- Weight at study initiation (mean and range, SD): /
- Length at study initiation (length definition, mean, range and SD): /
- Valve height at study initiation, for shell deposition study (mean and range, SD): /
- Peripheral shell growth removed prior to test initiation: /
- Method of breeding: start of each batch: with new-born animals, i.e less than 3 days old, by placing about 250 of them into 10l of medium in an all-glass culture vessel.
max. age of the cultures: 4 weeks
renewal of the cultures: after 7d of cultivation half of the medium twice a week.
temp. of the medium: 18-22°c, const. within +- 1°c
feeding: daily, a suspension of fresh water algae.
medium: M7, as prescribed by Dr. Elendt-Schneider.
- Feeding during test
- Food type: no feeding
- Amount:/
- Frequency:/

- Acclimation period:no acclimation
- Acclimation conditions (same as test or not):no acclimation
- Type and amount of food:no acclimation
- Feeding frequency: no acclimation
- Health during acclimation (any mortality observed):no acclimation

QUARANTINE (wild caught)
- Duration: /
- Health/mortality:/
Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
Post exposure observation period:
250 mg/L expressed as CaCo3
Test temperature:
Nominal concentration BBSA (mg/l) start (t=0h) pH End (t=48h) pH
0 7.9 8.1
10 7.9 8.1
18 8.0 8.0
32 8.0 8.0
59 8.0 8.0
100 8.0 8.0
Dissolved oxygen:
Nominal concentration BBSA (mg/l) start (t=0h) O2 End (t=48h) O2
0 8.4 8.5
10 8.5 8.5
18 8.4 8.4
32 8.6 8.4
59 8.7 8.4
100 8.6 8.4
Nominal and measured concentrations:
The 24h EC50 was based on nominal concentrations (95% confidence interval between 51 and 82 mg/l)
The 48h EC50 was based on nominal concentrations (95% confidence interval between 49 and 69 mg/l)
Details on test conditions:
- Test vessel:100 ml, all-glass
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 80 ml medium
- Aeration: no aeration of the test solutions.
- Type of flow-through (e.g. peristaltic or proportional diluter): /
- Renewal rate of test solution (frequency/flow rate):/
- No. of organisms per vessel: 10 per vessel
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates):2
- No. of vessels per vehicle control (replicates):/
- Biomass loading rate:/

- ISO-medium
- Total organic carbon:/
- Particulate matter:/
- Metals:/
- Pesticides:/
- Chlorine:/
- Alkalinity:/
- Ca/mg ratio:/
- Conductivity:/
- Culture medium different from test medium:/
- Intervals of water quality measurement: /

- Adjustment of pH:
- Photoperiod: 16h daily
- Light intensity: 16h photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study:
Reference substance (positive control):
potassium dichromate
48 h
Dose descriptor:
Effect conc.:
56 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
24 h
Dose descriptor:
Effect conc.:
62 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Details on results:
- Behavioural abnormalities: none
- Observations on body length and weight:none
- Other biological observations:/
- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: no
- 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):
Results with reference substance valid?
-Mortality: no (Daphnia-immobilisation)
- EC50/LC50: 24h EC50 and 48h EC50 by K2Cr2O7 was 0.75 mg/l with 95% fiducial limits ranging from 0.56 mg/l to 1.0 mg/l .
- Other:
Reported statistics and error estimates:
Finney, D.j.,1971 Probit analysis, cambridge University Press,Cambridge, UK 3rd edition)
logit-model (cox,D.R.1977 analysis of binary data,Methuen&Co LTD
Validity criteria fulfilled:
Under the conditions of the present study BBSA did not induce acute immobilisation of Daphnia magna at 32 mg/l after 48h of exposure (NOEC).
The 24h-EC50 was 62 mg/l based on nominal conc.
The 48h-EC50 was 56 mg/l based on nominal conc.

Description of key information

The acute toxicity of BBSA for aquatic Invertebrates was determined in an ISO 6341 15 test (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea)). The measured 48hr-EC50 was 56 mg/lL (nominal).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
56 mg/L

Additional information