Catalase

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:

4 February 1992 to 3 December 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Catalase belongs to the same sub-subclass as peroxidase and due to the similarity of the two enzymes, read-across can be applied.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

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

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of peroxidase (EC no. 232-668-6, CAS no. 9001-05-2, EC name Peroxidase, Enzyme class no. 1.11.1.7)

Molecular formula:

Not applicable, see remarks.

IUPAC Name:

Active enzyme protein of peroxidase (EC no. 232-668-6, CAS no. 9001-05-2, EC name Peroxidase, Enzyme class no. 1.11.1.7)

Constituent 2

Reference substance name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available

IUPAC Name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 3

Reference substance name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 4

Reference substance name:

Carbohydrates constituent of enzyme deriving from the fermentation or extraction process.

Molecular formula:

Not available. See remarks.

IUPAC Name:

Carbohydrates constituent of enzyme deriving from the fermentation or extraction process.

Constituent 5

Reference substance name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

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 concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

95 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

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

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 560 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 154.6 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Effect concentrations 5

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 6

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

8.8 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Effect concentrations 7

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

>= 560 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 8

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)