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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
4 February 1992 to 3 December 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
OECD Guideline No. 202, Part 1 and EEC Directive 67/548 Annex 5, C2 as published in 84/449/EEC
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): SP 491 Peroxidase
- Substance type: UVCB
- Physical state: Dark brown crystaline solid
- Lot/batch No.: PPX 3806
- Expiration date of the lot/batch: 04 March 1994
- Storage condition of test material: In the refrigerator at 4 degrees of Celcius

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
no
Details on test solutions:
- Controls: Laboratory tap water, dechlorinated by passage through activated carbon, softened by reverse osmosis and aged by recirculation though a gravel filter bed. Total hardness 150 - 200 mg/I as CaCO3.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus
- Source: Laboratory culture originating from a strain supplied by IRChA, France.
- Age at study initiation: Gravid adults were isolated 24 hours prior to initiation of the test. Young daphnids produced overnight were used for testing.
- Feeding during test: No
- Food type: Cultures were fed daily with a mixture of fry fish food (Liquifry®) and a suspension of mixed algae (predominantly Scenedesmus or Selenastrum spp.).

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
150-200 mg/L CaCO3
Test temperature:
21°C
pH:
7.6-8
Dissolved oxygen:
7.7-8.6 mg O2/L
Conductivity:
1000 pS/cm
Nominal and measured concentrations:
5.6, 10, 18, 32, 56, 100, 180, 320 and 560 mg/L (nominal concentrations)
Details on test conditions:
TEST SYSTEM
- Test vessels: Glass jars each containing 200 ml test solution.
- Experimental design: 9 test concentrations plus 1 control each in duplicate. 20 animals per test concentration.
- Aeration: None.
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water, dechlorinated by passage through activated carbon, softened by reverse osmosis and aged by recirculation though a gravel filter bed. Total hardness 150 - 200 mg/I as CaCO3.
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
Method of initiation: Daphnia were placed in the test solutions after addition of the test substance.
Adjustment of pH: No
Loading: 20 mL test solution per organism.
Photoperiod: 16 h light: 8 h dark.
Medium renewal: None.
Duration of exposure: 48 hours.
Criterion of effect: Daphnia were considered to be immobilised if they were unable to swim for approximately 15 seconds after gentle agitation.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): The numbers of mobile, immobile and floating daphnia were counted approximately 24 and 48h after the start of the study.
TEST CONCENTRATIONS
- 5.6, 10, 18, 32, 56, 100, 180, 320 and 560 mg/L (1.6, 2.8, 5.1, 16, 28.6, 91.4, 160 mg/L active enzyme protein).
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
95 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
26.2 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 560 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 154.6 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
32 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
8.8 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
>= 560 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
>= 154.6 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Details on results:
Details on results
- Mortality of control: 0
- Other adverse effects control: no
- Abnormal responses: no

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Analysis of the immobility of Daphnia magna exposed to peroxidase to concentrations up to 560 mg/L for 48 hours gave an EC50 value of 95 mg/L, equivalent to 26.2 mg/L active enzyme protein.
Executive summary:

The acute toxicity of Peroxidase to Daphnia magna was assessed under static exposure conditions.

The study was conducted in accordance with OECD Guideline No. 202, Part 1 and EEC Directive 67/548 Annex 5, C2 as published in 84/449/EEC. A group of twenty Daphnia 24 hours old was exposed for 48 hours to Peroxidase in concentrations up to 560 mg/L. Dechlorinated tap water was used as test water, having a total hardness of 150 -200 mg/L.

Observations of the Daphnia in each control and test vessel were made after 24 and 48 hours, which produced these findings:

24-hour EC50 value: >560 mg/L (>154.6 mg/L active enzyme protein)

48-hour EC50 value:  95 mg/L (26.2 mg/L active enzyme protein)

24-hour NOEC value: >560 mg/L (>154.6 mg/L active enzyme protein)

48-hour NOEC value:  32 mg/L (8.8 mg/L active enzyme protein)