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

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

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 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

At the renewal and at the end of the exposure (24 and 48 hours), samples of the old media were taken directly from the test vessels.
Criteria for the analytical Recoveries of the test item should be within ± 20% of the nominal or monitoring (target) initially measured concentration.

Vehicle:

no

Details on test solutions:

Composition of the Culture Medium Elendt M4 according to OECD 202, Annex 3 (2004)
Component Concentration
[mg/L]

CaCl2 x 2 H2O 294
MgSO4 x 7 H2O 123
KCl 5.80
NaHCO3 64.8
Na2SiO3 x 5 H2O 7.47
NaNO3 0.274
KH2PO4 0.143
K2HPO4 0.184
Na2EDTA x 2 H2O 2.50
FeSO4 x 7 H2O 0.996
H3BO3 2.86
MnCl2 x 4 H2O 0.361
LiCl 0.306
SrCl2 x 6 H2O 0.152
RbCl 0.0710
NaBr 0.0160
Na2MoO4 x 2 H2O 0.0615
CuCl x 2 H2O 0.0168
ZnCl2 0.0130
CoCl2 x 6 H2O 0.0100
KI 0.00325
Na2SeO3 0.00219
NH4VO3 0.000575
Thiaminhydrochloride 0.075
Cyanocobalamin 0.0010
Biotin 0.00075
pH 8.2 +/- 0.8

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna STRAUS
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, 16 hours illumination, light intensity of max. 1500 lx.

Culture medium Elendt M4, according to OECD 202, Annex 3 (2004), is used.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

19 – 20 °C

pH:

8.2 +/- 0.8

Dissolved oxygen:

Dissolved oxygen concentration 7.95 - 8.74 [mg/L]

Conductivity:

5.78 - 5.91 [μS/cm]

Details on test conditions:

Test temperature 18 - 22 °C, constant within ± 1 °C.
Illumination (target) Diffuse light, light intensity of max. 1500 lx.
Photoperiod (target) 16/8 hours light/dark cycle.
Feeding The daphnids were not fed during the study.

Reference substance (positive control):

yes

Remarks:

Reference item: potassium dichromate

Key result

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

3.56 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

2.64 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

5.27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

4.52 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

EC50 1.78 mg/L
95% Confidence limits 1.56 - 1.91 mg/L
Valid range 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000)

 Measured Concentrations of the Test Item during the Definitive Test

Sampling date	Fresh media, 0 hours		Old media, 24 hours		Fresh media, 24 hours		Old media, 48 hours		Geometric mean measured test item concentration [mg/L]
Nominal test item concentrations [mg/L]	PU-2018-821
	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%
25.8	26.7	104	24.2	94	27.2	106	24.1	93	25.5
11.7	11.7	100	10.8	92	12.0	102	10.4	89	11.2
5.33	5.27	99	5.21	98	5.77	108	4.87	91	5.27
2.42	2.32	96	2.15	89	2.69	111	1.99	82	2.27
1.10	0.989	90	0.803	73	1.09	99	0.818	74	0.918
0.501	0.410	82	0.269	54	0.473	94	0.331	66	0.363
Control	< LOQ		< LOQ		< LOQ		< LOQ

Meas. conc.   = measured concentration of the test item, mean value of all injections, dilution factors taken into account

LOQ              = limit of quantification (200 µg test item/L)

%                   = percentage of the nominal concentration of the test item

 

Validity criteria fulfilled:

yes

Conclusions:

LC50(48 h) = 4.52 mg/L

Based on the geometric mean measured concentrations of the test item PU-2018-821, the 48 hours-
EC50 for Daphnia magna was 4.52 mg/L (95% confidence limits: 3.53 – 5.28 mg/L).

Executive summary:

The Daphnia magna acute immobilisation test was performed as a limit test.

LC50(48 h) = 4.52 mg/L

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the test item

PU-2018-821 (batch number: 2298695) were determined at the test facility according to OECD 202

(2004) from 2019-06-25 to 2019-06-27.

The study was conducted under semi-static conditions over a period of 48 hours with six

concentrations of the test item (nominal: 0.501 to 25.8 mg/L). They were prepared from a stock

solution of 25.8 mg test item/L in a geometric series with a separation factor of 2.2 between the test

concentrations. For details of the preparation, see section 4.2.

Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each

concentration level and the control.

The concentrations of the test item PU-2018-821 were analytically verified via LC-MS/MS in the fresh

media at the start of the exposure and at the renewal of the test solutions (0 and 24 hours) as well

as in the 24-hours old media at the renewal and at the end of the test (24 and 48 hours) in all

concentration levels and the control. Details of the analytical method are presented in section 14.

The measured concentrations of the test item PU-2018-821 in the fresh media were in the range of

82 to 111% of the nominal values at the start of the exposure (0 hours) and at the renewal (24 hours).

The measured concentrations of the test item in the old media at the renewal and at the end of the

test (24 and 48 hours) were in the range of 54 to 98% of the nominal values. The analytical results

are presented in Table 5. The geometric mean measured concentrations of the test item

PU-2018-821 were 0.363 – 0.918 – 2.27 – 5.27 – 11.2 – 25.5 mg/L.

Since the measured concentrations did not stay within ± 20% of the initially measured concentrations

during the exposure, the ECx-values are based on the geometric mean measured concentrations of

the test item PU-2018-821. For details, see Table 1.

The validity criteria of the test guideline were fulfilled.

Description of key information

LC50(48 h) = 4.52 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

4.52 mg/L