N,N'-bis(3-aminopropyl)piperazine

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:

2017-06-02 - 2017-11-23

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-bis(3-aminopropyl)piperazine

EC Number:

230-589-1

EC Name:

N,N'-bis(3-aminopropyl)piperazine

Cas Number:

7209-38-3

Molecular formula:

C10H24N4

IUPAC Name:

3-[4-(3-aminopropyl)piperazin-1-yl]propan-1-amine

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The concentrations of N,N'-bis(3-aminopropyl)piperazine were determined in fresh media (0 hours) and old media (72 hours) of all tested concentration levels and in the control via LC-MS/MS.

Test solutions

Vehicle:

no

Details on test solutions:

A stock solution with a nominal test item concentration of 100 mg/L was freshly prepared with dilution water. The stock solution was agitated until the test item was completely dissolved.
Out of the stock solution five concentration levels were tested in a geometrical series with a dilution facor of √10: 1.00 - 3.16 - 10.0 - 31.6 - 100 mg/L. The concentrations were based on the result of a preliminary range finding test.

Test organisms

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

Origin: Sammlung von Algenkulturen (SAG), Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, 37073 Göttingen, Germany
Cultivation at test facility: Fresh stocks are prepared every month on Z-agar. Light intensity amounted to 2567-5130 lux for 24 hours per day.
Culture medium: Nutrient medium Z according to Lüttge et al. (1994)

Study design

Test type:

static

Water media type:

freshwater

Remarks:

dilution water according to the test guideline

Limit test:

no

Total exposure duration:

72 h

Test conditions

Hardness:

The test medium had a nominal hardness of 0.24 mmol Ca + Mg /L.

Test temperature:

min.: 21.5 °C, max.: 22.5 °C, mean value: 22 °C

pH:

"Geometric mean measured test item concentration
[mg/L]" pH-value
0 hours 72 hours
98.2 9.97 8.56
28.7 9.24 8.45
6.97 8.34 8.70
2.42 8.02 9.04
1.02 8.01 9.21
Control 8.27 9.33

Nominal and measured concentrations:

Nominal test
item concentration
[mg/L] measures concentrations Geometric mean measured test item concentration
0 hours % 72 hours % [mg/L]
100 94.7 95 102 102 98.2
31.6 28.0 88 29.4 93 28.7
10 8.32 83 5.84 58 6.97
3.16 2.97 94 1.97 62 2.42
1.0 1.08 108 0.966 97 1.02
Control

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

6.97 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Key result

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

28.7 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

Key result

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

8.03 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: 95% CI: 5.54 - 11.3 mg/L

Effect concentrations 4

Key result

Duration:

72 h

Dose descriptor:

EC20

Effect conc.:

13.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: 95% CI: 10.1 - 16.5 mg/L

Effect concentrations 5

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

32.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: 95% CI: 28.3 - 38.0 mg/L

Effect concentrations 6

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

2.42 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

cell number

Effect concentrations 7

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

6.97 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

cell number

Effect concentrations 8

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

2.94 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

cell number

Remarks on result:

other: 95% CI: 2.05 - 4.22 mg/L

Effect concentrations 9

Duration:

72 h

Dose descriptor:

EC20

Effect conc.:

3.91 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

cell number

Remarks on result:

other: 95% CI: 2.71 - 5.43 mg/L

Effect concentrations 10

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

8.73 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

cell number

Remarks on result:

other: 95% CI: 6.30 - 13.1 mg/L

Results with reference substance (positive control):

current study valid range (average ± 3xSD)
growth rate inhibition
ErC50 0.559 0.754 ± 0.518
95% confidence interval 0.538-0.581
yield inhibition
EyC50 0.320 0.398 ± 0.291
95% confidence interval 0.242-0.341

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In this study, N,N'-bis(3-aminopropyl)piperazine was found to inhibit the growth of the freshwater green alga Pseudokirchnerella subcapitata after 72 hours with the following effect values (bases on geometric mean measured test item concentrations):
The EC50-values with 95% confidence intervals for inhibition of growth rate (ErC50) and yield (EyC50 after 72 hours were 32.3 (28.3 - 38.0) mg/L and 8.73 (6.30 - 13.1) mg/L, respectively. The NOEC-values for inhibition of growth rate and yield after 72 hours were 6.97 mg/L and 2.42 mg/L, respectively. All effect levels are bases on the geometric mean measured test item concentrations.

Executive summary:

In this study, N,N'-bis(3-aminopropyl)piperazine was found to inhibit the growth of the freshwater green alga Pseudokirchnerella subcapitata after 72 hours with the following effect values (bases on geometric mean measured test item concentrations):

The EC50-values with 95% confidence intervals for inhibition of growth rate (ErC50) and yield (EyC50 after 72 hours were 32.3 (28.3 - 38.0) mg/L and 8.73 (6.30 - 13.1) mg/L, respectively. The NOEC-values for inhibition of growth rate and yield after 72 hours were 6.97 mg/L and 2.42 mg/L, respectively. All effect levels are bases on the geometric mean measured test item concentrations.