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Ecotoxicological information

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:
2018-08-29 to 2019-05-28
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
according to guideline
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
adopted April 13, 2004
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
- Lot/batch No.of test material:
- Expiration date of the lot/batch:
- Aggregate state at room temperature: Liquid
- Color: Brown-Orange
- Storage conditions at Test Facility: 4°C +/- 4°C
Analytical monitoring:
Details on sampling:
Test Concentrations:: 10, 4.5, 2.1, 0.9 and 0.4 mg test item/L (spacing factor 2) nominal loading rate and a control

- Duplicate samples from the freshly prepared test media of all test concentrations and the control were taken at the start of the test
- For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test period, duplicate samples from the test media of all test concentrations and the control were collected at the end of the test (after 48 hours) by pouring together the contents of the test beakers of each treatment
- All samples were diluted by a factor of 2 with acetonitrile
- Additional samples of the control and the dilution solvent were taken at each sampling test start and test end without any sample treatment
- One sample from the freshly prepared stock solution and duplicate samples from the freshly prepared test media of all test concentrations and the controls were taken at the start of the test

Details on test solutions:
- supersaturated stock suspensions of the test item was prepared separately by suspending test item in test water at concentrations of 10, 4.5, 2.1 and 0.9 mg test item/L
- suspensions were stirred separately for approximately 24 hours to dissolve as much test item as possible
- test solutions were extracted from below the water surface. An adequate volume of the water accommodated fraction of 0.9 mg test item/L was mixed into test water to obtain the desired dilution of nominal 0.4 mg test item/L.
- The test media were prepared just before introduction of the daphnids (= start of the test) and test medium renewal on Day 1
Test organisms (species):
Daphnia magna
Details on test organisms:
- Common name: Daphnia magna
- Strain/clone: Straus
- Age at study initiation (mean and range, SD): From 0 to 0 hours old
- Source: taken from ibacon's in house laboratory culture.
- Feeding during test : no

- Acclimation period: Was not necessary, since the test was performed in the same medium as the culturing
- Acclimation conditions (same as test or not): The daphnids were bred in the laboratories of ibacon under similar temperature and light conditions as used in the test. The cultivation of the parental daphnids was performed in Elendt M4 medium (see 6.5). The test organisms were not first brood progeny.
- Type and amount of food: The daphnids in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the laboratories of ibacon.
- Feeding frequency: on all working days
- Health during acclimation (any mortality observed): no abnormalities observed

For the evaluation of the quality of the Daphnia clone and the consistency of the experimental conditions, the reference item potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions
Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
2.5 mmol/L (= 250 mg/L) as CaCO3
Test temperature:
19.6 to 19.9 °C in the freshly prepared media;
19.2 to 20.7°C in the aged test media
7.8 to 7.9 in the freshly prepared media;
7.9 in the aged test media; and thus the pH-value did not vary by more than 1.5 units
Dissolved oxygen:
8.9 to 9.5 mg/L in the freshly prepared media;
8.6 to 9.0 mg/L in the aged test media
Nominal and measured concentrations:
10, 4.5, 2.1, 0.9 and 0.4 mg test item/L (nominal values)
Details on test conditions:
- Test vessel: Glass beakers of 100 mL volume containing approximately 60 mL of test medium
- Type: closed (test vessels were covered with a lid to reduce the loss of water due to evaporation and to avoid the entry of dust into the solutions)
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 5
- No. of vessels per vehicle control (replicates): 4

Reconstituted Water (Elendt "M4").

- Water Hardness: 2.5 mmol/L (= 250 mg/L) as CaCO3
- Alkalinity: 0.9 mmol/L


- Photoperiod: 16 h light : 8 h dark
- Light intensity: The light intensity was 0 to 0 lux (measured once during the test)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mobility


- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
Key result
48 h
Dose descriptor:
Effect conc.:
0.705 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
48 h
Dose descriptor:
Effect conc.:
0.397 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
48 h
Dose descriptor:
Effect conc.:
0.4 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Details on results:
- Behavioural abnormalities: not reported
- Observations on body length and weight: not reported
- Other biological observations: not reported
- Mortality of control: none
- Other adverse effects control: not reported
- Abnormal responses: not reported
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: not reported
- Effect concentrations exceeding solubility of substance in test medium: no
Reported statistics and error estimates:
The 24-hour and 48-hour EC50, EC20 and EC10 and the 95 % confidence limits were calculated by weibull analysis.
The NOEC and LOEC after 24 and 48 hours were calculated by Step-down Cochran Armitage test procedure.
The software used to perform the statistical analysis was ToxRat Professional, Version 3.2.1, ToxRat Solutions GmbH.

After 48 hours of exposure no immobilisation of the test animals was observed in the control and the lowest test item concentration of 0.4 mg test item/L. At the concentration of 0.9 mg test item/L, 18 animals were immobile and 20 animals were immobile at the concentrations of 4.5 and 10 mg test item/L.

Table 1. Summary of biological results

   % of immobilised daphnids after 24 hours   % of immobilised daphnids after 48 hours

 EC50 [mg test item/L]

95 % CI [mg test item/L]


1.014 - 7.483


0.605 - 0.821

EC20 [mg test item/L] 

95 % CI [mg test item/L]


0.222 - 5.738


0.401 - 0.621

EC10 [mg test item/L] 

95 % CI [mg test item/L]


0.068 - 5.710 


0.300 - 0.525

NOEC [mg test item/L] 



 LOEC [mg test item/L] 



Validity criteria fulfilled:
For the conducted semi-static acute immobilization test with Daphnia magna, applying nominal concentrations of 0.4 to 10 mg/L jasmine absolute, effect concentrations were determined on basis of measured concentrations:
- 48 h EC50: 0.705 mg/L
- 48h LOEC: 0.9 mg/L
- 48h NOEC: 0.4 mg/L
Executive summary:

The 48-hr-acute toxicity of jasmine absolute to Daphnia magna (Straus) was studied under semi-static conditions. Daphnids were exposed to control and test chemical at nominal concentrations of 10, 4.5, 2.1, 0.9 and 0.4 mg test item/L for 48 hr. Immobilisation and sublethal effects were observed daily.  The 48-hour EC50 was 0.705 mg/L. The 48-hr NOEC based on immobilization was 0.4 mg/L

Based on the results of this study, jasmine absolute would be classified as toxic to D. magna in accordance with the EU CLP Regulation. 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.


Description of key information

A 48-h semi-static acute toxicity study with Daphnia magna was conducted according to the OECD guideline 202 and in accordance with the principles of GLP. The derived 48 h EC50 was 0.705 mg/l, based on the nominal concentrations. The 48-hr NOEC based on immobilization was 0.4 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
0.705 mg/L

Additional information