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

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-08-07 - 2017-12-06

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 202 (Daphnia sp. Acute Immobilisation Test)

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:

All tested concentration levels (pH-neutralized and un neutralized) and the control were analytically verified via LC-MS/MS in the fresh media at the start of
exposure (0 hours) and in the old media at the end of the exposure (48 hours).
At the start of the exposure (0 hours), samples of the fresh media were taken after preparation of all test item concentrations (pH neutralized and un neutralized) and analyzed. At the end of the exposure (48 hours), samples of the old media were taken from the test vessels.

Test solutions

Vehicle:

no

Details on test solutions:

Stock solution The stock solution (100 mg/L of the test item was weighed out) was freshly prepared with dilution water (see Table 2) prior to the
start of the exposure (at 0 hours). The stock solution was mixed thoroughly by manual agitation.

Test concentrations 5 test item concentrations in a geometric series with a separation factor of 2, prepared by dilution of the stock solution 100 mg/L with
dilution water (see Table 2), were tested as follows:
6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
pH-neutralized The highest concentration level of 100 mg/L was tested pH-neutralized in addition to the un-neutralized concentration level to
concentration distinguish toxic effects of the test item from pH effects. For this purpose, an appropriate aliquot of the highest concentration level
was adjusted to pH 7 ± 0.2 by the addition of 1 M HCl prior to the start of the exposure (0 hours) and tested under the same conditions as the un neutralized concentration.

The test concentrations are based on the results of a non-GLP preliminary range finding test.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system Daphnia magna STRAUS (Clone 5).
Origin Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany

Breeder Noack Laboratorien GmbH, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany

Culture In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20  2 °C, in an incubator, 16 hours illumination,
light intensity of max. 1500 lx

Culture medium Elendt M4, according to OECD 202, Annex 3 (2004), modified to a total hardness of 160 to 180 mg CaCO3/L, is used.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

Elendt M4, according to OECD 202, Annex 3 (2004), modified to a total hardness of 160 to 180 mg CaCO3/L, is used.

Test temperature:

18 - 22 °C, constant within ± 1 °C

Nominal and measured concentrations:

Sampling date 0 hours 48 hours
Nominal
test item concentration
[mg/L] N,N`-bis(3-aminopropyl)piperazine
Meas. conc. Meas. conc.
[mg/L] % [mg/L] %
100 119 119 115 115
50.0 54.8 110 54.5 109
25.0 26.9 108 24.5 98
12.5 10.9 87 11.5 92
6.25 5.11 82 6.13 98
100 pH neutralizd 103 103 115 115
Control < LOQ < LOQ
Meas. conc. = measured concentration of the test item, mean value of two injections, dilution factors taken into account
% = percent of the nominal concentration of the test item
LOQ = limit of quantification of the analytical method (0.03 mg/L of the test item)

Reference substance (positive control):

yes

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

44.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 5

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

47.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 6

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

EC50-Value (with 95% confidence limits) of the Reference Item Potassium dichromate
based on nominal concentrations [mg/L], (0 - 24 hours)
Current Study Valid Range
EC50 2.10 mg/L 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000); clone 5
95% confidence limits 1.92 - 2.43 mg/L 0.6 - 2.1 mg/L, acc. to OECD 202 (2004); clone A

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item N,N`-bis(3-aminopropyl)piperazine, the 48 hours-EC50 for Daphnia magna was 47.9 mg/L (95% confidence limits: 25.9 – 88.8 mg/L).

Executive summary:

In the acute immobilization test withDaphnia magna(STRAUS), the effects of the test itemN,N`-bis(3-aminopropyl)piperazine(batch no.:JH5262736A) were determined at the test facility according to OECD 202 (2004) from 2017-11-28 to 2017-11-30. The study was conducted under static conditions over a period of 48 hours with five nominal concentrations of the test item ranging from6.25 to 100 mg/Lin a geometric series with a separation factor of 2. Additionally, the highest concentration of 100 mg/L was tested as a pH neutralized solution to determine any unspecific effects caused by pH. The test item is a colorless liquid. All test item concentrations were visually clear throughout the exposure period.

Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration leveland the control. The concentrationsof the test item were analytically verified via LC-MS/MS in fresh media at the start of the exposure (0 hours) and in old media at the end of the exposure period (48 hours) in all tested concentration levels and in the control.

The measured concentrations of the test item in fresh media at the start of the exposure (0 hours) were in the range of 82 to 119 % of the nominal values in all test item concentrations. At the end of the exposure period (48 hours), the measured test item concentrations in the old media were in the range of to 92 % 115 of the nominal values of all test concentrations.The measured concentrations of the test item N,N`-bis(3-aminopropyl)piperazine remained stable within ±20% of the nominal concentrations throughout the exposure period, which indicates that the test item concentrations were successfully maintained for the duration of the test.

All effect concentrations (EC_{10 / 50 / 100}) given in the table below are based on the nominal concentrations of the test item N,N`-bis(3 -aminopropyl)piperazine.The pH neutralized test item solution showed the same effects as the non pH neutralized. Therefore, all effects are considered to be specific effects on Daphnia by the test item.

The validity criteria of the test guideline were fulfilled.

 

EC₁₀ -, EC₅₀ - (with 95% Confidence Limits) and EC₁₀₀ -Values

(based on the nominal concentrations of the test item)

N,N`-bis(3-aminopropyl)piperazine
Effect concentrations	Test duration [hours]	nominal concentrations of the test item [mg/L]
EC10	24	>100*
	48	44.1 (<6.25 – 87.2)
EC50	24	>100*
	48	47.9 (25.9 – 88.8)
EC100	24	>100*
	48	100*

               * = Empirically derived, no Confidence Interval given