4-Piperidinecarboxylic acid, hydrochloride (1:1)

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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-piperidinecarboxylic acid hydrochloride

EC Number:

611-901-9

Cas Number:

5984-56-5

Molecular formula:

C6H11NO2.HCl

IUPAC Name:

4-piperidinecarboxylic acid hydrochloride

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study reports): JNJ-150098-AAC (T001310)
- Physical state: solid
- Appearance white powder

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):

Raphidocelis subcapitata (previous names: Pseudokirchneriella 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 concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

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.