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

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 Nov. 2004 to 26 Nov. 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 (control), 25 and 100 mg/L sampled at test start and after 72 h of exposure
- Sampling method: chemical analysis of incoulated replicates
- Sample storage conditions before analysis: room temperature, analysed following day (storage < 24 h)

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test substance was stirred in test medium for about 10 min at 20 ± 2°C. Test solutions prepared by dilution of stock solution.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none used
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The stock solution was clear.

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

TEST ORGANISM
- Common name: green algae
- Strain: SAG 86.81
- Source (laboratory, culture collection): SAG (collection of algal cultures, Göttingen, Germany)
- Age of inoculum (at test initiation): seed culture: 4 days ( 23±2 °C; final cell density: 277E4 cells/mL); pre-culture: 3 days ( 23±2 °C; final cell density: 61E4 cells/mL)
- Method of cultivation: algae kept in liquid culture in the Laboratory of Experimental Toxicology and Ecology (BASF, Ludwigshafen, Germany)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test temperature:

23±2 °C

pH:

7.5 to 8.3

Nominal and measured concentrations:

Nominal (mean measured) concentrations: control, 12.5, 25 (22.5), 50 and 100 (100) mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed with gas permeable silicone sponge caps
- Material, size, headspace, fill volume: Erlenmeyer flasks, 250 mL nominal volume, fill volume 100 mL
- Initial cells density: 10000 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3

GROWTH MEDIUM
- Standard medium used: yes

OTHER TEST CONDITIONS
- Adjustment of pH: yes
- Photoperiod: permanent illumination
- Light intensity and quality: artificial light, universal white (OSRAM L 25), about 60 to 120 µE/(m2*s) at 400 to 700 nm

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- test parameter: in-vivo chlorophyll-a-fluorescence (pulsed excitation with light flashes having a wavelength of 435 nm)
- Determination of cell concentrations: fluorimeter after 0, 24, 48, 72 h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

Reference substance (positive control):

no

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: verified by analytical measurement

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

93.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: verified by analytical measurement

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: verified by analytical measurement

Duration:

72 h

Dose descriptor:

EC90

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: verified by analytical measurement

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: verified by analytical measurement

effect values based on biomass:
EbC10 (72 h): 76.3 mg/L
EbC50 (72 h): >100 mg/L
EbC90 (72 h): >100 mg/L
NOEbC (72 h): >/= 100 mg/L
LOEbC (72 h): > 100 mg/L

 

pH values:

Concentration [mg/L]	uninoculated		inoculated
	0 h	72 h	72 h
Control	8.0	8.3	8.3
12.5	7.9	8.0	8.2
25	7.9	8.0	7.5
50	7.9	8.1	7.6
100	7.9	7.8	7.8

 

Concentration control analysis:

Nominal concentration	Measured concentration
	0 h	72 h	mean
(mg/L)	(mg/L)	(mg/L)	(mg/L)	(% of nominal)
0 (control)	< 10*	< 10*	-	-
25	23*	22*	22.5	90
100	101**	99**	100	100

*: based on two measurements

**: based on three measurements

 

Test results: Calculation of biomass and growth rate and their inhibition after 72 h

Nominal conc.	Biomass after 72 h		Growth rate after 72 h
(mg/L)	(rel. units)	inhibition (%)	(rel. units)	inhibition (%)
0 (control)	178	0.0	0.0590	0.0
12.5	206	-15.5	0.0607	-2.8
25	195	-9.4	0.0582	1.4
50	193	-8.6	0.0580	1.7
100	139	22.0	0.0526	10.9

 

Test Results: Cell density after O h, 24 h, 48 h and 72 h: 

In this test, surrogates for biomass, such as cell counts, fluorescence, etc. are typically measured and the use of the term ―biomass thus refers to these surrogate measures as well.

The measured fluorescence was corrected by the fluorescence of the test substance and converted into cell densities (cells x 10000/ml) by the following correlation:  cell density (cells x 10000/ml) = fluorescence (relative units) / slope (m)  M = 12,38  

linear correlation was obtained from a calibration curve (cell density versus fluorescence;         

 

		concentration
time (h)	flask No.	0 (/control)	100 mg/l	50 mg/l	25 mg/l	12,5 mg/l
0	1	3,1	3,5	3,3	3,2	3,1
	2	3,0	3,4	3,4	3,3	3,0
	3	3,2	3,2	3,2	3,2	3,0
	mean	3,1	3,4	3,3	3,2	3,0
24	1	12,1	13,7	14,5	13,7	13,7
	2	12,9	12,1	12,9	12,9	13,7
	3	12,9	8,9	13,7	13,7	13,7
	mean	12,7	11,6	13,7	13,5	13,7
48	1	68,7	70,3	83,2	80,8	75,9
	2	61,4	67,0	80,8	80,8	83,2
	3	66,2	33,1	78,4	85,6	80,8
	mean	65,4	56,8	80,8	82,4	80,0
72	1	216,5	214,1	228,6	190,6	227,8
	2	222,1	169,6	212,4	217,3	232,6
	3	207,6	90,5	201,9	234,2	256,9
	mean	215,4	158,1	214,3	214,1	239,1

mean = mean value

 

Test Results: Calculation of biomass and inhibition of biomass after 72 h:

		concentration
parameter	flask No.	0 (/control)	100 mg/l	50 mg/l	25 mg/l	12,5 mg/l
biomass [rel]	1	181	182	204	182	196
	2	178	155	191	194	206
	3	175	79	185	208	215
	mean	178	139	193	195	206
inhibition[%]		0	22	-8,6	-9,4	-15,5

mean = mean value

 

Description of key information

With high probability acutely not harmful to algae.

Key value for chemical safety assessment

EC10 or NOEC for freshwater algae:

50 mg/L

Additional information

There is one reliable study available regarding the toxic effect of N,N'-bis(3 -aminopropyl)ethylenediamine (CAS 10563 -26 -5). The study was performed according to OECD 201 under GLP conditions. The test species was Desmodesmus subspicatus. The test concentrations were analytically verified (90 to 100% mean recovery). The 72 -h NOErC was determined to be 50 mg/L, the 72 -h ErC50 was > 100 mg/L (BASF AG, 2005; report no. 60E0402/043218).