Registration Dossier

Ecotoxicological information

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
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Constituent
Test material form:
liquid
Specific details on test material used for the study:
Test Item: Baccartol Crude
Batch Number: VE00364716
Purity: Not applicable - complex mixture
Appearance: yellow viscous liquid
Expiry date: 2017-04-02
Recommended storage: Keep container tightly closed in a dry and well-ventilated place at ambient temperature (10 - 30 °C) protected from light.

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
At the start of the exposure and at renewal (0 and 24 hours), samples were taken from the freshly prepared WAFs and analyzed.
At renewal and at the end of the exposure (24 and 48 hours), samples of the 24-hour old media were taken directly from the test vessels and analyzed. Samples were analyzed immediately following sampling.

Test solutions

Vehicle:
no
Details on test solutions:
Five Water Accommodated Fractions (WAF) of the test item were individually prepared at loading levels of 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L in a geometric series with a separation factor of 2. For each loading level, an appropriate volume of the test item (relative density: 0.8905 (D20/4) was taken into account) was placed by a positive displacement pipette onto the water surface.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Test system: Daphnia magna STRAUS (Clone 5)
Reason for the selection of the test system: Daphnia magna is the preferred species in accordance with the test guidelines and is bred at the test facility.
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany
Breeder: Noack Laboratorien GmbH, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
Culture: In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 ± 2 °C, in an incubator, 16 hours illumination, light intensity of max. 1500 lx
Culture medium: Elendt M4, according to OECD 202, Annex 3 (2004), modified to a total hardness of 160 to 180 mg CaC03/L, is used.
Culture feeding: The culture daphnids are fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus, with an algae cell density of > 106cells/mL. The algae are cultured at the test facility.
Origin of the food algae: Sammlung von Algenkulturen (SAG), Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, 37073 Göttingen, Germany

Study design

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

Test conditions

Test temperature:
18-22 °C, constant within ± 1 °C
Nominal and measured concentrations:
Range finding (nominal): 1, 10 and 100 mg/L
Definitive test (nominal): 6.25 - 12.5 - 25.0 - 50.0 -100 mg/L in a geometric series with a separation factor of 2.
Definitive test (geonetric mean measured): 3.10, 4.34, 11.1, 32.7 and 69.6 mg/L
Details on test conditions:
Test method: The study was performed with a semi-static test design, since the measured concentrations of the test item were not within the required range of ± 20% of the nominal values during the non-GLP preliminary range finding test. Due to the volatility of the test item, the study was performed in a closed system without headspace, to reduce contact with air and losses of the test item by evaporation.
Test vessels / volume: Sealed clear glass flasks (conical shoulder bottles with a narrow neck, 3.7 (ID) x 8 (H) cm) with glass stoppers were used. A test volume of approximately 60 mL was provided in each test vessel, when filled up to the top with the test solutions without any headspace.
Number of daphnids and replicates: 20 daphnids, divided into 4 replicates, each with 5 daphnids, were used per WAF and control.
Age of the daphnids at the start of the exposure: Less than 24 hours old daphnids from a healthy stock were used for the study. Juvenile daphnids were removed from the culture vessels at the latest 24 hours before the start of the exposure and discarded. The juveniles born within the following period of max. 24 hours preceding the exposure were used for the test. No first brood progeny was used for the test.
Acclimatization: Acclimatization of the daphnids was not necessary, because the dilution water was equivalent to the culture medium.
Application: The test vessels were filled up to the top with the test solutions. The daphnids were inserted with a small amount of dilution water (start of the exposure) or test solution (water renewal) by pipette. Thereafter, the test vessels were closed immediately with glass stoppers. There was no headspace in the test vessels.
Illumination (target): Diffuse light, light intensity of max. 1500 lx
Photoperiod (target): 16/8 hours light/dark cycle
Feeding: The daphnids were not fed during the study.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate p.a. (Sigma)

Results and discussion

Effect concentrations
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
11.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: (95% confidence limits: 10.8 - 12.7 mg/L)
Results with reference substance (positive control):
The percentage of immobility for the reference item potassium dichromate (Sigma-Aldrich, batch number MKBV090ÛV, purity 99.0%, CAS RN 7778-50-9) was determined after 24 hours from 2017-02-16 to 2017-02-17. EC50 = 2.00 mg/L (95% CL 1.80 -2.28 mg/L)

Any other information on results incl. tables

Validity Criteria

The study was performed according to OECD Guideline 202 (2004). The validity criteria were fulfilled:

• In the control group, no daphnids were immobilized or showed any signs of disease or stress, e.g. discoloration or unusual behavior such as trapping on the surface of the water, during the 48-hour test period (required: not more than 10% of the daphnids in the control).

• The dissolved O2 concentration in the 24-hour old media at the end of the exposure intervals was 2 6.48 mg/L (required: 2 3 mg/L at the test end of the exposure intervals) in the test vessels of all tested WAFs and the control.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Based on the nominal loadings of the test item, the 48 hour-EL50 for Daphnia magna was 11.9 mg/L (95% confidence limits: 10.8 - 12.7 mg/L).