Registration Dossier

Diss Factsheets

Ecotoxicological information

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25th April 2006 - 27th April 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report date:
2006

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EU Method C.3 (Algal Inhibition test)
Version / remarks:
29 December 1992
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Version / remarks:
Adopted 7 June 1984
GLP compliance:
yes

Test material

Constituent 1
Test material form:
solid: particulate/powder
Details on test material:
Light pink tinted powder
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: test substance (ref Y12511/002/001, bottle no. 287981)
- Purity of the lot/batch::99.8%
- Purity test date: 08 May 2006

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations:
- Sampling method: The concentrations of UL125 in the test solutions were measured at 0 and 72 hours using HPLC.

Samples for analysis were taken at 0 hours from the excess test solutions and at 72 hours from the remaining blank solutions.
- Sample storage conditions before analysis: The samples were analyzed on the day of receipt

Test solutions

Vehicle:
yes
Remarks:
Dimethylformamide
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The study was run with a culture medium control and a solvent control together with nominal UL125 concentrations of 1.0, 1.8, 3.2, 5.6, 10, 18 and 32 mg/l.

A 5.0 ml volume of primary stock concentrate of UL125, nominally 320000 mg/l, was prepared by dissolving 1.600 g of UL125 in dimethylformamide (DMF). A 5.0 ml volume of secondary stock, nominally 56000 mg/l, was prepared by the dilution of an aliquot of the primary stock in DMF. The test solutions were prepared from these stock solutions by the addition of appropriate volumes to 1000 ml of culture medium and stirring thoroughly. A solvent control was similarly prepared by the addition of 100 µl of DMF to 1000 ml of culture medium, to give a solvent concentration of 100 µl/l (0.010% v/v). Equalising volumes of FMG were aded, where applicable, to each of the test exposure solutions to give a final DMF concentration of 100 µl/l (0.010% v/v) in each exposure vessel. All addition were made sub-surface, gradually, by microlitre syringe, into solutions vioroously stirred by magnetic follower. All test solutions were subjected to continued stirring >= 100 minutes plus approximately 2 minutes ultrasonication.
- Controls: Medium and solvent
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Dimethylformamide
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): 100µl/l
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): The resultant solutions were all clear and colourless, but at the nominal 3.2 mg/l test concentration and above, particles were visible in stirred suspension, increasing in density with nominal concentration: numrous particles, some large, were observed in the nominal 32 mg/l concentration. The control consisted of culture medium only.

Test organisms

Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Common name: green algae
- Strain: Pseudokirchneriella subcapitata
- Source (laboratory, culture collection): ATCC 22662
- Method of cultivation: liquid cultures

ACCLIMATION
- Acclimation period: sufficient to ensure exponential growth
- Culturing media and conditions (same as test or not): same as test
- Any deformed or abnormal cells observed: none

Study design

Test type:
static
Water media type:
other: Culture medium described by Miller et al
Remarks:
Miller W E, Greene J C and Shiroyama T (1978). Selenastrum capricornutum Printz. Algal Assay Bottle Test: Experimental Design, Application and Data Interpretation Protocol. EPA-600/9-78-018, Corvallis, OR
Limit test:
no
Total exposure duration:
72 h
Remarks on exposure duration:
Standard
Post exposure observation period:
not applicable

Test conditions

Hardness:
not specified
Test temperature:
24 +/- 2 degrees centigrade
pH:
7.3 - 10.2
Dissolved oxygen:
n/a
Salinity:
n/a
Nominal and measured concentrations:
the measured concentrations at the start of the test ranged from 72 to 100% of the nominal values. The measured concentrations in the blanks after 72 hours ranged from 69 to 98% of the nominal values. the overall artimetic means of the measured concentrations ranged from 72% to 100% of the nominal values. The percentrage loss in the measured concentraions over the test period ranged from 2 to 6%.
On the basis of the analytical data the arthimetic means of the measured UL125 concentrations were adopted for the calculation and reporting of results.
Details on test conditions:
TEST SYSTEM
- Test vessel:250ml borosilicate flasks closed with polyurethane foam bungs and filled to 100ml with test solution.
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
- No. of vessels per vehicle control (replicates): 6


GROWTH MEDIUM
- Standard medium used:

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: continuous illumination
- Light intensity and quality: measured in terms of quantum response and was 102.0 µ Einsteins / m2 s. Also 8060 lux by cosine receptor
- Temperature: 24 ± 2 °C
- Rotation: Orbital shaking at 160 rpm in a Gallenkamp type INR-401 orbital incubator


EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Determination of cell concentrations: cell densities determined using a Coulter® Multisizer Particle Counter model Z1. Triplicate determinations were made for each sample.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable – limit test
- Test concentrations: control, solvent control (DMF) and nominal concentrations of 1.0, 1.8, 3.2, 5.6, 10, 18 and 32 mg/l

Results and discussion

Effect concentrationsopen allclose all
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
13.4 mg/L
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 23 mg/L
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
3.1 mg/L
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
3.1 mg/L
Basis for effect:
biomass

Applicant's summary and conclusion

Validity criteria fulfilled:
yes

Categories Display