Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to an appropriate EU test method and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Qualifier:
according to
Guideline:
ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))
Qualifier:
according to
Guideline:
other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Identification
EPYRRON-OPL

Molecular formula
C23H35NO2
Molecular weight
357.53
CAS Number
159325-45-8
Description
Cream powder with lumps (determined at WIL Research Europe B.V.)
Batch
DGJ524K1NAT
Purity/Composition
85-95%
Test substance storage
At room temperature in the dark
Stability under storage conditions
Stable
Expiry date
15 January 2014 (allocated by WIL Research Europe B.V., 1 year after receipt of the test substance)

Sampling and analysis

Analytical monitoring:
yes

Test solutions

Vehicle:
yes
Details on test solutions:
The standard test procedures required generation of test solutions, which should contain completely dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The testing of concentrations that disturb the test system should be prevented (e.g. film of the test substance on the water surface). No correction was made for the purity/composition of the test substance.
The batch of EPYRRON-OPL tested was a cream powder with lumps with a purity of 85-95% and the test substance was not completely soluble in test medium at the loading rates initially prepared.
In the combined limit/range-finding test preparation of test solutions started with individual loading rates of 1.0, 10 and 100 mg/l. In the final test preparation of test solutions started with a loading rate of 100 mg/l. These mixtures were treated for 15 minutes with ultrasonic waves and subsequently a 2 day-period of magnetic stirring was applied to ensure maximum dissolution in the test medium. Obtained solutions were filtered through a 0.45 μm membrane filter (Whatman; RC55) to remove the undissolved fraction of test substance. In the final test the lower test concentrations were prepared by subsequent dilutions of the filtrate in test medium. The final test solutions for both experiments were all clear and colourless.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST SYSTEM

Species Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions.
Source In-house laboratory culture with a known history.
Reason for selection This system has been selected as an internationally accepted invertebrate species.
Validity of batch Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
Characteristics For the test selection of young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.

BREEDING

Start of each batch With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.
Maximum age of the cultures 4 weeks
Renewal of the cultures After 7 days of cultivation half of the medium twice a week.
Temperature of medium 18-22°C
Feeding Daily, a suspension of fresh water algae.
Medium M7, as prescribed by Dr. Elendt-Schneider
(Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33)

Study design

Test type:
static
Water media type:
freshwater
Total exposure duration:
48 h

Test conditions

Hardness:
180 mg/l expressed as CaCO3 and the pH: 7.7 ± 0.3.
Test temperature:
18.9 and 19.5°C
pH:
7.8-8.1
Dissolved oxygen:
8.6-9.1
Nominal and measured concentrations:
Actual concentrations at the start were 3.1, 5.7 and 11 mg/l for solutions containing 32, 56 and 100% of the filtrate, respectively. These concentrations decreased to 48-50% of initial at the end of the test. Consequently, average exposure concentrations were calculated to correspond with 2.2, 4.1 and 7.4 mg/l for the solutions containing 32, 56 and 100% of the filtrate, respectively.
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 ml, all-glass

- Material, size, headspace, fill volume: 80 ml
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): static
- Renewal rate of test solution (frequency/flow rate): none
- No. of organisms per vessel:5
- No. of vessels per concentration (replicates):4
- No. of vessels per control (replicates):4
- No. of vessels per vehicle control (replicates):0
- Biomass loading rate:

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:M7, as prescribed by Dr. Elendt-Schneider
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Conductivity:
- Culture medium different from test medium:
- Intervals of water quality measurement:

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod:
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- 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):
no

Results and discussion

Effect concentrations
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
5.1 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: (4.4 - 6.1)

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The 48h-EC50 of Epyrron was 5.1 mg/l based on average exposure concentrations (95% confidence interval between 4.4 and 6.1 mg/l), and by read acros the 48h-EC50 of Epyrrol is considered to be similar.
Executive summary:

The 48h-EC50 of Epyrron was 5.1 mg/l based on average exposure concentrations (95% confidence interval between 4.4 and 6.1 mg/l), and by read acros the 48h-EC50 of Epyrrol is considered to be similar.