Pyridinium, 3-[(diethylamino)carbonyl]-1-(3-sulfopropyl)-, inner salt

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:

18.11.2014 to 21.11.2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Version / remarks:

(2006/20111)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Version / remarks:

(2008/2009)

GLP compliance:

yes (incl. QA statement)

Remarks:

LAUS GmbH, Auf der Schafweide 20, D-67489 Kirrweiler (Germany)

Test material

Test material information

Constituent 1

Reference substance name:

3-(diethylcarbamoyl)-1-(3-sulfonatopropyl)pyridin-1-ium

EC Number:

807-112-6

Cas Number:

1393708-17-2

Molecular formula:

C13H20N2O4S

IUPAC Name:

3-(diethylcarbamoyl)-1-(3-sulfonatopropyl)pyridin-1-ium

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Pyridiniumsulfonate
- Physical state: white powder
- Analytical purity: 99.8% (HPLC)
- Impurities (identity and concentrations):
- Purity test date:
- Lot/batch No.: CHZT000001
- Expiration date of the lot/batch: 2/2016
- Stability under test conditions: stable
- Storage condition of test material: test material stored in closed vessel dark and dry at room temperature (min. -5 - max. +40°C)

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The content of the test item in the test vessels was determined at the beginning and at the end of the test via HPLC/UV

Test solutions

Vehicle:

no

Details on test solutions:

A stock solution containing 100.5 mg/L test item in nutrient medium was prepared. The lower test concentrations were prepared by serial dilution of the stock solution with nutient medium.

Test organisms

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

TEST ORGANISM
- Common name: green alga (Desmodesmus subspicatus)
- Strain: SAG Strain Number: 86.81
- Source (laboratory, culture collection): The culture of Desmodesmus subspicatus was obtained in February 2014 by MBM Sciencebridge GmbH (Institut für Pflanzenphysiologie of Universität Göttingen).
- Age of inoculum (at test initiation):
- Method of cultivation: The algae were kept as stock culture on solid agar at 7°C. From the stock culture, a permanent culture was prepared. From an aliquot of the permanent culture, the pre-culture was prepared.

ACCLIMATION
- Acclimation period: 96 h
- Culturing media and conditions (same as test or not): no: nutrient medium was 10-fold concentrated in comparison with the pre-culture medium
- Any deformed or abnormal cells observed: no

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test conditions

Test temperature:

21.3-23.2°C

pH:

at time 0: 7.7
after 72 h: 7.6-7.9

Nominal and measured concentrations:

nominal concentrations: 0 (control), 10, 18, 32, 56, 100 mg/L
measured concentrations: not detected (control), 9.94-9.96, 17.86-18.18, 32.09-32.16, 56.01-56.64, and 99.61-100.86 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass flasks
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass/65 mL/-/45 mL
- Aeration: not reported
- Type of flow-through (e.g. peristaltic or proportional diluter): test conducted under static conditions
- Renewal rate of test solution (frequency/flow rate): test conducted under static conditions
- Initial cells density: ca. 2 x 10E3 cells/mL
- Control end cells density: 183619 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
- No. of vessels per vehicle control (replicates): no vehicle used

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to guideline
- Culture medium different from test medium: nutrient medium was 10-fold concentrated in comparison with the pre-culture medium
- Intervals of water quality measurement: At test start and test end the pH value in treatments and control was measured

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: continuous lighting
- Light intensity and quality: 5800 lux during the test


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : growth, yield
- Determination of cell concentrations: light absorption

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 1.7
- Justification for using less concentrations than requested by guideline: -
- Range finding study: not reported

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

NOEC

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:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

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 4

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: yield

Effect concentrations 5

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: yield

Effect concentrations 6

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: yield

Results with reference substance (positive control):

The EC50 of potassium dichromate were tested in a separate reference test under GLP conditions. The measured values laid within the normal range (72 h ErC50=0.99 mg/L; 72 h EyC50=0.55 mg/L; EU C.3: 0.60-1.03 mg/L).

Reported statistics and error estimates:

t-test

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In a guideline study, the 72 h NOEC and EC50 values based on growth rate and yield were determined to be >=100 mg/L and >100 mg/L, respectively.

Executive summary:

The toxicity of pyridiniumsulfonate towards the unicellular green alga Desmodesmus subspicatus was investigated in a study conducted according to OECD Guideline 201 (Alga, Growth Inhibition Test; 2006/2011) and EU Method C.3 (Algal Inhibition Test; 2008/2009). Cultures of Desmodesmus subspicatus were exposed to 5 pyridiniumsulfonate concentrations ranging from 10 to 100 mg/L (nominal). At the beginning and at the end of the test, the content of the test substance was analysed via HPLC/UV. The measured concentrations laid in a range from 99 -101% of the nominal concentration. The 72 h ErC50 and 72 h EyC50 were determined to be >100 mg/L (nominal), respectively. The 72 h NOEC based on growth rate and yield was determined to be >=100 mg/L (nominal).

The study is considered as reliable without restriction and fulfills the requirements of the guideline.

Results Synopsis
Test organism: Desmodesmus subspicatus
Test Type: static
72 h ErC50/EyC50: >100 mg/L (nominal)

72 h NOEC: >=100 mg/L (growth rate, yield; nominal)

Endpoint(s) Effected: growth rate, yield (biomass)