Hexahydrocyclopenta[c]pyrrol-2(1H)-amine

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

15 Feb. 2010 to 10 Jun. 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

yes

Principles of method if other than guideline:

Deviations :
• The temperature for the main and reference study lay in a range of 21 — 25 °C and therefore higher than stated in the guideline. Because the controls showed a normal exponential growth, this can be stated as uncritical.
• In OECD Guideline 201 and EU Method C.3, different compositions of stock solution II are stated. The study was conducted in accordance with OECD Guideline 201 , therefore, the composition of stock solution II must be stated as deviation from EU Method C.3.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Hexahydrocyclopenta[c]pyrrol-2(1H)-amine

EC Number:

259-201-9

EC Name:

Hexahydrocyclopenta[c]pyrrol-2(1H)-amine

Cas Number:

54528-00-6

Molecular formula:

C7H14N2

IUPAC Name:

octahydrocyclopenta[c]pyrrol-2-amine

Test material form:

liquid

Remarks:

Colourless

Specific details on test material used for the study:

A solution stock solution containing 125 mg test item /L (nominal concentration) in deionised water was prepared. The concentrations to be tested were prepared by diluting of this stock solution with deionised water.

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

Stock Solution I:
NH4Cl 1500 mg
MgC12*6H20 1200 mg
CaCl2*2H20 1800 mg
MgS04*7H20 1500 mg
KH2P04 160 mg
H2O deionised ad 1000 mL

Stock Solution II:
FeCl3*6H20 64 mg
NaEDTA*2H20 100 mg
H2O deionised ad 1000 mL

Stock Solution III:
H3B03 185 mg
MnCl2*4H20 415 mg
ZnCl2 3 mg
CoCl2*6H20 1.5 mg
CuCl2*2H20 0.01 mg
Na2MoO4*2H20 7 mg
H2O deionised ad 1000 mL

Stock Solution IV:
NaHCO3 50 g
H2O deionised ad 1000 mL

Pre-culture-medium
Stock Solution I 10.0mL
Stock Solution II 1.0mL
Stock Solution III 1.0mL
Stock Solution IV 1.0mL
H2O deionised ad 1000mL

Nutrient-medium (10-fold concentrated)
Stock Solution I 100mL
Stock Solution II 10mL
Stock Solution III 10mL
Stock Solution IV 10mL
H2O deionised ad 1000mL

Test organisms

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

The culture of Desmodesmus subspicatus was obtained in Dec. 2009 by MBM Sciencebridge GmbH (Institut für Pflanzenpohysiologie of Universität Göttingen). The algae are kept as stock culture on solid agar at 8 C. From an aliquot of this stock culture the pre-culture was prepared.

Study design

Test type:

static

Water media type:

freshwater

Total exposure duration:

72 h

Test conditions

Test temperature:

Temperature range was 21-25°C.

pH:

The pH values range was 7.3 to 7.7.

Dissolved oxygen:

Not specified

Salinity:

Not specified

Conductivity:

Not specified

Nominal and measured concentrations:

Nominal : 0 ; 1 ; 10 & 100 mg/L

Reference substance (positive control):

yes

Remarks:

Potassium dichromate K2Cr207 (CAS No. 7778-50-9) was used as positive control.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

< 1 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.:

< 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: AUC or Area Under Curve according to EU Method C.3

Remarks:

means the integral of the biomass

Effect concentrations 3

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

< 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Yield (according to OECD Guideline 201)

Remarks:

It is defined as the biomass at the end of the exposure period minus the biomass at the start of the exposure period.

Effect concentrations 4

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 5

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: AUC or Area Under Curve according to EU Method C.3

Remarks:

means the integral of the biomass

Effect concentrations 6

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Yield (according to OECD Guideline 201)

Remarks:

It is defined as the biomass at the end of the exposure period minus the biomass at the start of the exposure period.

Effect concentrations 7

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

ca. 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 8

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 1 - < 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: AUC or Area Under Curve according to EU Method C.3

Remarks:

means the integral of the biomass

Effect concentrations 9

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

< 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Yield (according to OECD Guideline 201)

Remarks:

It is defined as the biomass at the end of the exposure period minus the biomass at the start of the exposure period.

Effect concentrations 10

Duration:

72 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 11

Duration:

72 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: AUC or Area Under Curve according to EU Method C.3

Remarks:

means the integral of the biomass

Effect concentrations 12

Duration:

72 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Yield (according to OECD Guideline 201)

Remarks:

It is defined as the biomass at the end of the exposure period minus the biomass at the start of the exposure period.

Results with reference substance (positive control):

NOEC 72 h = 0.04m/L (95 IC = not deteminable)
LOEC 72 h = 0.063m/L (95 IC = not deteminable)
72h ErC50 = 1.8mg/L (95 IC = 1.4 - 2.3mg/L)
72h EbC50 = 0.55m/L (95 IC = 0.48 - 0.62 mg/L)
72h EyC50 = 0.35mg/L (95 IC = 0.30 - 0.4 mg/L)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

All validity criteria were met.
The EC50s of potassium dichromate were determined in a current reference test. For the estimation of the EC50s of the positive control, the fits showed sufficient statistical correspondence of the data with the dose-response-equation, The values were within the normal range of the laboratory.
In the lowest concentrated treatment (1 mg/L), significant inhibition was observed. Therefore no NOEC can be stated.
In the main study, replicate 2 of the control and in the reference study, replicate 3 of the control showed abnormally large absorptions. Upon inspection of the replicates, a contamination was discovered. Therefore, these replicates were excluded from the evaluation.
The result of the test can be considered valid.

Executive summary:

The study was performed using three concentrations containing 1, 10 and 100 mg/L. The treatments were used to incubate the unicellular freshwater green alga Desmodesmus subspicatus for a period of 72 hours. The cell concentration of each replicate was determined by measuring the absorption of the cuvettes at 440 nm every 24 hours with a spectral photometer. With these measured values, the number of cells was calculated (linear correlation between cell concentration and absorption given). Then the growth rate µ, the  area under the growth curve (AUC) and the Yield were determined.

The EC50s of potassium dichromate were tested in a current reference test. The values lay within the normal range of the laboratory.

The following results for the test item was determined:

Endpoint	NOEC	LOEC	EC50
Growth Rate	<1mg/L	1mg/L	approx. 10 mg/L
AUC	<1mg/L	1mg/L	1 -10 mg/L
Yield	<1mg/L	1mg/L	<1mg/L