Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2019

Materials and methods

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

Test material

Constituent 1
Chemical structure
Reference substance name:
(-)-1-Benzyl-4-(-4-fluor-phenyl)-3-hydroxymethyl-1,2,3,6-tetrahydropyridine-L-dibenzoyl-tartaric salt
EC Number:
606-459-9
Cas Number:
201855-57-4
Molecular formula:
C37H34FNO9
IUPAC Name:
(-)-1-Benzyl-4-(-4-fluor-phenyl)-3-hydroxymethyl-1,2,3,6-tetrahydropyridine-L-dibenzoyl-tartaric salt
Test material form:
solid: crystalline

Sampling and analysis

Analytical monitoring:
yes

Test solutions

Vehicle:
yes
Details on test solutions:
Preparation of stock solution: The preparation of stock and test solutions was based on the preliminary non GLP range finding test [9]. As the Range Finding Test was not performed in compliance with the GLP-Regulations it is excluded from the Statement of Compliance in the final report, but the raw data of this test are archived under the study code of present study. For the stock solution 36.20 mg of test item was measured into 1 000 ml Elendt M4 medium. The suspension was stirred till complete dissolution. After stirring the stock solution was clear and colourless. The concentration of stock solution according to the measured amount was 36.20 mg/L.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Test species: Daphnia magna Straus
Age: < 24 hours neonates
Origin: National Institute of Environmental Health
Date of arriving: 16 May 2013
Culture conditions: The test species is maintained in Elendt M4 medium with culture conditions at 21±1 °C and a light/dark cycle of 16/8 hours with light intensities of less than 1000 Lux. The cultures are fed with centrifugalized green alga (Pseudokirchneriella subcapitata) cells daily at weekdays.
Breeding conditions:
The stock, breeding and test cultures were kept at the same conditions in thermostat (temperature: 21 ± 1 °C; light/dark cycle: 16/8 hour). These conditions ensure that the reproduction was by parthenogenesis.
Stock cultures: The stock cultures contained Daphnia magna of different age. Occasionally the adult animals were separated by sieving to start a new breeding culture.
Breeding cultures: Adult daphnids which were able to give birth to neonates were kept in breeding cultures. The volume of breeding cultures was 1 500 ml and each vessel contained 20-60 adult daphnids. The pH and oxygen concentration of these cultures were measured weekly. The progeny was separated at least weekly from adults by sieving. The first progeny of a new breeding culture was not used for the test.
Neonates: Young daphnids were used in the test were less than 24 hours old and were not first brood progeny.

Study design

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

Test conditions

Test temperature:
The temperature of thermostat was recorded daily and registered continuously by Extech datalogger. The temperature inside the thermostat at the test vessels was measured and registered continuously by Extech SD200 thermometer and datalogger.
pH:
The pH were measured at the beginning and at the end of the test. At the end of the test the pH and the dissolved oxygen were measured in each test vessel.
Dissolved oxygen:
At the beginning of the test dissolved oxygen were measured from the remaining test solutions of the concurrent test vessels at each dilution levels.
Nominal and measured concentrations:
In the test the following nominal concentrations were applied: 36.20; 18.10; 10.86; 6.03; 3.62; 1.81 and 0.91 mg/L.
Details on test conditions:
The test was maintained under static conditions for a period of 48 hours. At the start of the study each test vessels contained 50 ml of test solutions and five young daphnids. In the test four replicates total of 20 animals at each test concentration level were investigated. The test vessels were covered with plastic lids and were kept in thermostat at 21 ± 1 °C with a 16/8 hours light/dark cycle. Each control vessels containing five young daphnids in Elendt M4 medium were
prepared in four replicates and kept under the same static conditions. The number of immobilised and abnormally behaved daphnids was recorded after 24 and 48h. A daphnia is considered to be immobile if it is not able to swim within 15 seconds after a gentle agitation of the test vessel. The daphnids were not fed during the test.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC0
Effect conc.:
2.42 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
other: EC20
Effect conc.:
3.2 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
5.45 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
The No Observed Effect Concentration (NOEC) and Lowest Observed Effect Concentration (LOEC) of the test item were determined with Fisher’s exact binominal test with Bonferroni correction by ToxRat® software. The results were not conclusive.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The EC50 was 5.45 mg/L at 48 h. In this Daphnia sp., Acute Immobilisation Test for (-)-Hydroxymethyl-tetrahydropiridine-L-dibenzoyl-tartarate the results indicated that the test item is hazardous to the aquatic environment and classified into the second acute toxic hazardous category [GHS Acute aquatic hazard Category Acute II. – 48 hr EC50 (for Crustacea) > 1 mg/L but ≤ 10 mg/L].