N1,N3-diallylpropane-1,3-diamine dihydrochloride

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:

31 March 2020 to 29 June 2020

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)

Version / remarks:

13 April 2004

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 12.50, 25, 50, 100 and 200 mg/L
- Sampling method: After 0 h and 48 h exposure, two replicates of 6 ml each were sampled. Per replicate 1.5 ml from each test vessel were pooled to obtain 6 mL in total. To each sample 6 mL Acetonitrile was added. The samples were filled into glass screw-cap vials and are stored in the freezer (≤ -18 °C).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The stock solution was prepared by adding 200 mg test item to 1000 mL test medium (OECD 202 test medium) and stirring on a magnetic stirrer.
- Controls: negative control

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea, Daphnia magna Strauß
- Age at study initiation: 0.25 - 23.0 h
- Method of breeding: The daphnids are cultured at 20 ± 2 °C with 16 h light : 8 h dark and placed into fresh Elendt M4 medium twice a week. They are kept at approx. 10 animals / 200 mL and fed on workdays with Desmodesmus subspicatus algae corresponding to 0.1 mg C × daphnid-1 × day-1.
- Source: Federal Environment Agency, Department IV 2.4, Berlin (2012)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20.3 - 21.6 °C

pH:

7.3 - 8.0

Dissolved oxygen:

8.4 - 8.5 mg/l

Nominal and measured concentrations:

nominal: 12.50, 25, 50, 100, 200 mg/L (measured concentrations were within 95.6 - 105.5 % of nominal concentrations)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers 50 ml, Schott, Mainz
- Volume of solution: 20
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h light : 8h dark

EFFECT PARAMETERS MEASURED : immobilisation

RANGE-FINDING STUDY
- Test concentrations: 1, 10 and 100 mg/L

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

30.85 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

< 12.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

69.63 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

< 12.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: none reported
- Mortality of control: 0
- Other adverse effects control: none reported
- Immobilisation of control: 0

Results with reference substance (positive control):

Quality assurance takes place at regular intervals by testing the sensitivity of the test organisms to the reference item potassium dichromate (Honeywell Speciality Chemicals Seelze GmgH, Seelze, Germany). The recent quality testing was performed in December 2019 (GLP 1504) with EC50(24h) = 1.95 mg/L, which is in the declared range of 0.6 - 2.1 mg/L (24h) according to OECD 202.

Reported statistics and error estimates:

The effect occurrence after 24 h and 48 h exposure was calculated. With the statistical software ToxRat Professional 3.3.0 (ToxRat Solutions GmbH, Alsdorf, Germany), ECx and LOEC/NOEC were determined.

Table 1: Effect concentrations

Lowest/No Observed) Effect Concentration	Nominal test item concentration (mg/L)
95 % Confidence Limits (CL)	24h	48h
EC50	69.63	30.85
CL	46.08 – 117.9	22.73 – 40.27
EC10	9.84	10.36
CL	2.32 – 18.43	5.07 – 15.32
LOEC	≤12.50	≤12.50
NOEC	<12.50	< 12.50

Table 2: Analytically measured test item concentrations after 0h and 48h exposure

Nominal test item concentration (mg/L)	Sample	Sampling time [h]	Test item concentration (mg/L)	Recovery [%] nominal	Mean recovery
NC	1527-NC1-0h	0	< LOC*	--	--
	1527-NC1-48h	48	< LOC	--
12.5	1527-A1-0h	0	12.2	97.6	96.8
	1527-A1-48h	48	12.0	96.0
25.5	1527-B1-0h	0	25.8	103.2	100.4
	1527-B1-48h	48	24.4	97.6
50.5	1527-C1-0h	0	51.7	103.4	99.5
	1527-C1-48h	48	47.8	95.6
100.0	1527-D1-0h	0	101	101.0	103.5
	1527-D1-48h	48	106	106.0
200.0	1527-E1-0h	0	202	101.0	103.3
	1527-E1-48h	48	211	105.5

* LOC = Limit of Calibration (4 mg/L)

Table 3: Immobility after 24h and 48h exposure

Nominal test item concentration (mg/L)	Daphnia introduced				Immobile daphnia								Immobility [%]
Sampling time	0h				24h				48h				24h	48h
Replicates	1	2	3	4	1	2	3	4	1	2	3	4
NC	5	5	5	5	0	0	0	0	0	0	0	0	0	0
12.5	5	5	5	5	1	1	1	0	1	1	1	1	15	20
25.0	5	5	5	5	1	1	1	0	2	1	1	2	15	30
50	5	5	5	5	3	2	1	4	3	4	3	5	50	75
100	5	5	5	5	2	4	3	4	4	5	4	5	65	90
200	5	5	5	5	4	4	4	2	5	5	5	5	70	100

Validity criteria fulfilled:

yes

Conclusions:

Effects were observed at the lowest test concentration used in the study, therefore, it can be concluded, that N1,N3-diallylpropane-1,3-diamine dihydrochloride already has toxic effects on Daphnia magna at concentrations below 12.50 mg/L.
The 48-hr EC50 is 30.85 mg/L.

Executive summary:

The 48-hr acute toxicity of N1,N3-diallylpropane-1,3-diamine dihydrochloride to Daphnia magna Strauß was studied under static conditions. Test species were exposed to N1,N3-diallylpropane-1,3-diamine dihydrochloride at nominal concentrations of 0, 12.50, 25, 50, 100 and 200 mg/L for 48 hr.  Immobilisation was observed at all concentrations.  The 48-hour EC₅₀ was 30.85 mg/L.  The 48-hr NOEC based on immobilization was < 12.50 mg/L. 

 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age: 0.25 - 23 h

Test Type: static

 

NOEC:  < 12.5 mg/L                       

EC₅₀:  30.85 mg/L                       95% C.I.:  22.73 to 40.27 mg/L

Endpoint(s) Effected: Immobility

Description of key information

In a single key study on the acute toxicity of N1,N3-diallylpropane-1,3-diamine dihydrochloride to Daphnia magna according to OECD guideline 202, an 48-hr EC₅₀ of 30.85 mg/L was reported. The test was conducted under static conditions and fulfilled all criteria for a valid test.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

30.85 mg/L

Additional information

Daphnia magna were exposed to N1,N3-diallylpropane-1,3-diamine dihydrochloride at nominal concentrations of 0, 12.50, 25, 50, 100 and 200 mg/L for 48 hr. Immobilisation was observed at all concentrations. The 48-hr EC₅₀ was 30.85 mg/L. The 48-hr NOEC based on immobilization was < 12.50 mg/L.