Registration Dossier

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:
2015-12-07 to 2015-12-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.3 (Algal Inhibition test)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): JNJ-150098-AAC (T001310)
- Molecular formula (if other than submission substance): C6H11NO2.HCl
- Molecular weight (if other than submission substance): 165.62
- Physical state: white powder
- Analytical purity: 97.4 % (base titration), 98.7 % (acid titration)
- Impurities (identity and concentrations): water 0.1%
- Purity test date: 2014-11-25
- Lot/batch No: I14KB4913
- Expiration date of the lot/batch: 2020-11-23 (retest date)
- Storage condition of test material: room temperature
- Stability under storage conditions: stable
- Other: solubility in water: 320 g/L
Specific details on test material used for the study:
Batch I14KB4913
Purity/Composition 97.4%
Test item storage conditions At room temperature
Stable under storage conditions until 23 November 2020 (retest date)

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Sampling method: samples were taken before the start of the test and after 24h and 72 hours from each test medium and from the control. At the end of the exposure period, the replicates with algae were pooled at each concentration before sampling.
- Sample storage conditions before analysis: stored in a freezer until analysis.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: preparation of test solutions started by dissolving test item in medium with a concentration of 100 mg/L. No special treatment other than careful mixing was necessary to achieve complete dissolving. This solution was used as the highest test concentration after adjusting the pH from 5.2 to 7.7 using a 0.1 M NaOH solution (Merck, Germany). Lower test concentrations were prepared by subsequent dilutions of the highest test concentration in test medium
- Controls: yes

Test organisms

Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Strain: NIVA CHL 1
- Source (laboratory, culture collection): In-house laboratory culture
- Method of cultivation: Algae stock cultures were started by inoculating growth medium with algal cells from a pure culture on agar. The suspensions were continuously aerated and exposed to light in a climate room at a temperature of 21-24°C.

ACCLIMATION
- Acclimation period: 3 days
- Culturing media and conditions (same as test or not): same as the test

Study design

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

Test conditions

Hardness:
24 mg CaCO3/L
Test temperature:
21-23°C
pH:
7.8-8.0
Dissolved oxygen:
not reported
Salinity:
not applicable
Nominal and measured concentrations:
nominal test concentrations : 100 mg/L
measured test concentration t= 0 h: 86.1 - 89.1 mg/L
measured test concentration t= 24h: 88.4 - 89.6 mg/L
measured test concentration t= 72 h: 82.1 - 88.6 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 mL all-glass flasks
- Type (delete if not applicable): capped vessels
- Material, size, headspace, fill volume: 100 mL all-glass flasks filled with 50 mL test solution
- Initial cells density: 10,000 cells/mL
- Control end cells density: 118.5 x 10,000 cells/mL
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: M2 according to the OECD 201 guideline, formulated using Milli-RO water
- Culture medium different from test medium: no
- Intervals of water quality measurement: temperature= continuously, pH at the beginning in all concentrations and the end of the test in the control and the limit concentration.

OTHER TEST CONDITIONS
- Sterile test conditions: no information
- Adjustment of pH: yes in the highest test concentration from pH 5.2 to 7.7 using a 0.1 M NaOH solution before subsequent dilutions.
- Photoperiod: continuous illumination
- Light intensity and quality: within the range of 78 to 90 µE.m-2.s-1 using TLD-lamps

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter] At the beginning, cells were counted using a microscope and a counting chamber. thereafter, cell densities were determined by spectrophotometric measurement of samples at 680 nm using a spectrophotometer with immersion probe.
- effect calculated parameters: specific growth rate and yield

TEST CONCENTRATIONS
- Spacing factor for test concentrations: none (limit test)
- Range finding study
- Test concentrations: 0.10, 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes. Concentrations in excess of nominal 100 mg/L have not been tested in accordance with EU Commission Directive 92/69/EE.
Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Details on results:
- Exponential growth in the control (for algal test): yes
- Any stimulation of growth found in any treatment: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: clear test solution
- EC50 (growth rate and yield) > 100 mg T001310 /L.
- EC10 (growth rate and yield) > 100 mg T001310 / L.
- LOEC (growth rate and yield) > 100 mg T001310 / L.
Results with reference substance (positive control):
- Results with reference substance valid? yes
- EC50: 72-h EC50 (growth rate) = 1.5 mg/L (95% CI 1.4 to 1.6 mg/L)
Reported statistics and error estimates:
NOEC and EC values were determined using a two-sample t-test procedure (alpha=0.05, one-sided, smaller)

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
A 72-h growth inhibition test with the unicellular green alga Pseudokirchneriella subcapitata was performed with the test substance T001310 according to the OECD guideline 201 (GLP conditions).
The test substance clearly had no inhibitory effect on the growth (biomass and growth rate) of Pseudokirchneriella subcapitata at the tested concentrations up to and including the nominal test concentration of 100 mg/L. The 72h-ErC50 was beyond the range tested, i.e. exceeding an analytically confirmed nominal concentration of 100 mg/L. The results of the test can be considered reliable without restrictions.