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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Remarks:
Daphnia magna
Type of information:
read-across based on grouping of substances (category approach)
Remarks:
The alpha amylase study was used as a key study for read-across given the sensitivity of this enzyme towards aquatic organisms and the similarity to beta-amylase.
Adequacy of study:
key study
Study period:
1992-08-17 to 1993-06-11
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
The alpha amylase study was used as a key study for read-across given the sensitivity of this enzyme towards aquatic organisms.
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test substance was dissolved in test water to give an initial stock solution of 8790 mg enzyme concentrate dry matter/L. Serial dilutions of this stock solution were prepared with test water to give the desired series of exposure levels.
- Controls: test water
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia
- Strain: Daphnia magna Straus
- Source: Institute National de Recherche Chimique Appliqué (IRChA), France
- Age at study initiation (mean and range, SD): less than 24 hours
- Feeding during test: no



Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
171 mg CaCO3/L
Test temperature:
21 °C
pH:
7.7 - 8.1
Dissolved oxygen:
4.8 - 8.6 mg O2/L
Nominal and measured concentrations:
Nominal: 281.3, 492.2, 879.0, 1582.2, 2812.8, 4922.4 mg enzyme concentrate dry matter/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass jar
- Type (delete if not applicable): loosely covered with aluminium foil
- Material, fill volume: glass, 200 mL
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water filtered, dechlorinated and softened by passage through an Elga® water purification system. The water passes through a high pressure sand filter to remove colloids and particulate matter and a high grade activated carbon filter to move chlorine and any organic contaminants. A proportion of the supply then passes through a water softener before final reverse osmosis treatment to produce a highly purified water supply. The two grades of dechlorinated water are then remixed in the ratio of approximately 1:1 to give a supply with the desired water hardness. Batches of softened water were "aged" with aeration for at least 12 hours prior to use by recirculating through a gravel filter bed. Chlorine levels remained below detection limit at all times.
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light: 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
Reference substance (positive control):
no
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
1 758 mg/L
Nominal / measured:
nominal
Conc. based on:
other: enzyme concentrate dry matter
Basis for effect:
mobility
Remarks on result:
other: 95% CI: 1494-2022 mg enzyme concentrate dry matter/L
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
879 mg/L
Nominal / measured:
nominal
Conc. based on:
other: enzyme concentrate dry matter
Basis for effect:
mobility
Details on results:
- Mortality of control: 0
Reported statistics and error estimates:
EC50 values and 95% confidence limits were calculated according to the method of Thompson and Weil (Thompson, W.R. & Weil, C.S., 1952, Biometrics 8; 51-54).
The NOEC (no-observed-effect concentration) is the highest concentration at and below which the incidence of immobilisation is equal or less than 10%.
Validity criteria fulfilled:
yes
Conclusions:
1,4 -alpha-glucan branching enzyme was not tested with daphnids, however, read-across was performed with alpha-amylase.
The 48-hr EC50 value for alpha-amylase with Daphnia magna was 1758 enzyme concentrate dry matter/L (equivant to to 212 mg active enzyme protein/L). NOEC was 879 mg enzyme concentrate dry matter/L.
Executive summary:

1,4 -alpha-glucan branching enzyme was not tested, but a read-across study was used, where the acute toxicity of alpha-amylase to Daphnia magna was assessed.

The study was conducted in accordance with OECD Guideline for Testing of Chemicals No.202, and in compliance with GLP.

Groups of 20, 1st instar Daphnia (less than 24 hours old) were exposed for 48 hours to six concentrations of the test substance dissolved in water. The incidence of immobilisation was recorded for each test and control group at 24 and 48 hours and the following values determined:

EC50 (24h): 4307 mg enzyme concentrate dry matter/L.

EC50(48h): 1758 mg enzyme concentrate dry matter/L (equivalent to 212 mg active enzyme protein/L).

NOEC (48h): 879 enzyme concentrate dry matter/L.

Description of key information

The 48-hr EC50 value for alpha-amylase with Daphnia magna was 1758 enzyme concentrate dry matter/L (equivalent to 212 mg active enzyme protein/L). NOEC was 879 mg enzyme concentrate dry matter/L.


Due to the similarity between enzymes, similar results are expected for beta-amylase.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Dose descriptor:
EC50
Remarks:
Enzyme concentrate dry matter
Effect concentration:
1 758 mg/L

Additional information