Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated

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:

2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

All concentration levels and the control were analytically verified via LC-MS at the start (0 hours) of the exposure with algae. The mean recoveries of the three lowest concentration levels were

Vehicle:

no

Details on test solutions:

Composition of Dilution Water (OECD TG 201 medium)
Component Concentration
[mg/L]
NH4Cl 15
MgCl2 x 6 H2O 12
CaCl2 x 2 H2O 18
MgSO4 x 7 H2O 15
KH2PO4 1.6
FeCl3 x 6 H2O 0.064
Na2EDTA x 2 H2O 0.1
H3BO3 0.185
MnCl2 x 4 H2O 0.415
ZnCl2 3 x 10-3
Na2MoO4 x 2 H2O 7 x 10-3
CoCl2 x 6 H2O 1.5 x 10-3
CuCl2 x 2 H2O 1 x 10-5
NaHCO3 50
pH 8.1 +/- 0.2
This medium has a nominal hardness of 0.24 mmol Ca+Mg/L.

Test organisms (species):

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

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Hardness:

nominal hardness of 0.24 mmol Ca+Mg/L.

Test temperature:

Nominal range: 21 - 24 °C, controlled at ± 2 °C

pH:

pH 8.1 +/- 0.2

Conductivity:

139 [μS/cm]

Nominal and measured concentrations:

Nominal test item concentrations [μg/L]
100
31.6
10.0
3.16
1.00

Details on test conditions:

Light intensity [lux]
mean value: 5671

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

3.16 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

10 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

27.1 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

EC20

Effect conc.:

43.7 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

71.3 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Results with reference substance (positive control):

ErC50 0.701 [mg/L]
95% confidence interval 0.660 – 0.750 [mg/L]
Valid range 0.752 ± 0.534 [mg/L]

NOEC, LOEC and EC_x-values and 95% Confidence Intervals ofPU-2018-821(0 - 72 hours)

based on the nominal test item concentration [µg/L]

	Inhibition of Growth Rate [µg/L]
NOEC	3.16
LOEC	10.0
ErC10	27.1 (10.6 – 93.4)
ErC20	43.7 (34.0 – 94.8)
ErC50	71.3 (42.6 – 97.1)
	Inhibition of Yield [µg/L]
NOEC	< 1.00
LOEC	1.00
EyC10	< 1.00
EyC20	< 1.00
EyC50	29.9 (20.0 – 41.1)

 

Validity criteria fulfilled:

yes

Remarks:

The study meets the validity criteria of the guideline.

Conclusions:

EC10 growth rate = 0,027 mg/L

In this study, PU-2018-821 was found to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours with the following effect values (based on the nominal test item concentration): The EC50-values for inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were 71.3 (42.6 – 97.1) μg/L and 29.9 (20.0 – 41.1) μg/L, respectively. The NOEC-values for inhibition of growth rate and yield after 72 hours were 3.16 and < 1.00 μg/L

Executive summary:

EC10 growth rate = 0,027 mg/L

The toxicity of PU-2018-821 (Batch no.: 2298695) to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined according to the principles of OECD 201 and Council Regulation (EC) No. 266/2016/Method C.3 from 2019-07-01 to 2019-07-09 at the test facility. The aim of the study was the determination of the effects on growth rate and yield over a period of 72 hours.

The study was conducted under static conditions with an initial cell density of 5416 cells/mL. A stock solution with a nominal concentration of 100 mg test item/L was prepared with dilution water

(see Table 2). An appropriate amount of the test item was pipetted onto the surface of the dilution water. For this purpose, the density was taken into account. The dispersion was agitated until the

solution was visually clear. The dispersion was checked via laser beam (Tyndall effect) for undissolved test item. The Tyndall effect was negative. The dispersion was diluted to receive a stock solution of 1.00 mg test item/L. Five test item concentrations were tested in a geometrical series with a dilution factor of √10: 1.00 - 3.16 - 10.0 - 31.6 - 100 μg/L. Three replicates were tested for each test item concentration and six replicates for the control. The environmental conditions were within the acceptable limits.

The test media were visually clear throughout the test period. The concentrations of the test item PU-2018-821 in all concentration levels and the control were analytically verified by LC-MS at the

start and at the end of exposure.

The measured concentrations of the test item PU-2018-821 in the fresh media were in the range < LOQ to 101% of the nominal values at the start of the exposure (0 hours) and between < LOQ

and 98% of the nominal values at the end of the exposure (72 hours).

As the lower three test concentrations could not be determined analytically and the two higher concentrations were acceptably stable, all effect values given are based on the nominal test item

concentrations of PU-2018-821.

Description of key information

EC10 growth rate = 0,027 mg/L

Key value for chemical safety assessment

EC10 or NOEC for freshwater algae:

0.027 mg/L