N-[3-(dimethylamino)propyl]-N,N',N'-trimethylpropane-1,3-diamine

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 2018-January 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

02 October 2012

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Test item name: PU-2018-788
Product name: POLYCAT 77 – PU-2018-788
Chemical name: N-[3-(Dimethylamino)propyl]-N,N',N'-Trimethylpropane-1,3-diamine
Molecular formula: C11H27N3
CAS number: 3855-32-1
Batch/Lot number: 1944471
Active substance: 100 % (theoretically)
Purity: 97 %
Carbon content: 65.6 % (calculated)
Stability: Stable in aqueous solution
Water solubility: Soluble (ca. 190 g/L at 25 °C)
Volatility: Vapour pressure 5.453 hPa at 21 °C
Appearance: Liquid at RT
Colour: Colourless
Storage conditions: Room temperature
Expiration date: 26 March 2019
Certificate of analysis: 05 July 2018
Origin: Evonik Nutrition & Care GmbH
Identification: Appearance: Liquid; Colour: Colourless; Batch: 1944471

Analytical monitoring:

yes

Details on sampling:

TOC analysis of the test solutions was performed to determine concentrations of the test item during exposure.
The samples were taken from the additional vessels without test organisms in order to not disturb the TOC analysis. The sampling was conducted according to the following specification:
• After 0 d exposure, the additional vessels of NC, A, B, C, D and E were sampled; 2 samples perreplicate: 4 samples of 6 mL per treatment group.
• After 7 d and 14 d:
• Before renewal of test solutions, the additional vessels of NC, A, B, C, D and E were sampled; 2 samples per replicate: 4 samples of 6 mL per treatment group.
• After renewal of test solutions, the additional vessels of NC, A, B, C, D and E were sampled; 2 samples per replicate: 4 samples of 6 mL per treatment group.
• After 21 d exposure, the additional vessels of NC, A, B, C, D and E are sampled; 2 samples per replicate: 4 samples of 6 mL per treatment group. Of these 4 samples per treatment and sampling time, 2 were analysed directly after sampling and
2 were stored as retain samples in the freezer (≤ -18 °C) until finalisation of the study.

Vehicle:

not specified

Details on test solutions:

The test was performed with Elendt M4 medium according to OECD 202 (2004)
After addition of the stock solutions, except the vitamin stock solution S23, the medium was aerated for 2 – 3 h to achieve oxygen saturation and the pH (7.2 – 8.6), oxygen concentration (7.8 – 8.2 mg/L) and conductivity (683 – 733 µS/cm) was determined. Solution S23 was added directly before use of the medium.

Test organisms (species):

Daphnia magna

Details on test organisms:

The test organism Daphnia magna Straus originates from the clone breeding of the Federal Environment Agency, department IV 2.4, Berlin, and has been cultivated at Hydrotox GmbH since October 2012. 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. The test organisms used were 0.25 – 23.5 h old at the start of the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Remarks on exposure duration:

Daphnids, less than 24 h old, were exposed to different concentrations of the test item in Elendt M4 medium for 21 days.

Post exposure observation period:

Not specified

Hardness:

Not specified

Test temperature:

The temperature during the exposure was 20.6 – 21.8 °C (required: 18 – 22 °C, constant within ± 1 °C).

pH:

The pH was 7.4 – 8.1 in the control and 7.5 – 8.8 in the test item treatments (recommended: 6 - 9)

Dissolved oxygen:

The dissolved oxygen concentration was 8.2 – 9.1 mg/L in the control and 8.2 – 10.0 mg/L in the test item treatment (required: ≥ 3 mg/L).

Salinity:

NA

Conductivity:

Not specified

Nominal and measured concentrations:

As the measured geometric mean test item concentrations are within ± 20 % of the nominal concentrations, according to OECD 211 (2012), all results are given in relation to the nominal test item concentrations.

Details on test conditions:

The test vessels are kept in an incubator (specified temperature 18 – 22 °C, constant within ± 1 °C per day) with 16 h light : 8 h dark (≤ 20 µE m-2 s-1). The pH and the O2-concentrations were measured at the start (0 d) of the exposure, at each renewal of the test solutions (3×/week) and at the end of the exposure (21 d). The temperature was measured continuously.

Reference substance (positive control):

not specified

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

10.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

2.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

The test is valid according to OECD Test Guideline 211 (02 October 2012).
The measured geometric mean TOC concentrations in the treatment groups C, D and E were 85.9 – 106.7 % of the nominal concentrations.

Reported statistics and error estimates:

The effect concentrations after 21 d exposure were calculated with the statistical software ToxRat Professional 3.2.1
As the measured geometric mean test item concentrations are within ± 20 % of the nominal concentrations, according to OECD 211 (2012), all results are given in relation to the nominal test item concentrations.

Validity criteria fulfilled:

yes

Conclusions:

In a semi-static Daphnia magna Reproduction Test according to OECD 211, the 21day EC50 for reproduction was determined as 10.3 mg/L. The NOEC for reproduction was determined as 2.2 mg/L.

Executive summary:

The purpose of this study was to investigate the chronical effects of the test item on Daphnia magna. Daphnids, less than 24 h old, were exposed to different concentrations of the test item in Elendt M4 medium for 21 days. A semi-static test design was applied (renewal of test solutions 3×/week) with an accompanying TOC analysis. The vitality and reproduction of the daphnids was assessed on each workday, the length of the parent animals was measured at the end of the exposure. In the present study, five nominal concentrations of 1.0 – 23.4 mg/L test item were used

(based on a Daphnia magna acute toxicity test with an EC50 (48 h) of 35.4 mg/L.

The test was performed with Elendt M4 medium according to OECD 202 (2004).

Daphnids were 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. The test organisms used were 0.25 – 23.5 h old at the start of the test.

The stock solution was prepared by adding 46.5 – 47.0 mg test item to 1000 mL test medium and shaking for 10 min using an overhead shaker. Ten replicates, with one daphnid each, were prepared for each test item treatment and the negative control (NC; test medium). The test item

concentrations were prepared by diluting the stock solution with test medium.

Every 2 – 3 d (Friday, Monday and Wednesday), the daphnids were transferred into test vessels with new test solutions.

The test vessels are kept in an incubator (specified temperature 18 – 22 °C, constant within ± 1 °C per day) with 16 h light : 8 h dark (≤ 20 µE m-2 s-1). The pH and the O2-concentrations were measured at the start (0 d) of the exposure, at each renewal of the test solutions (3×/week) and at

the end of the exposure (21 d). The temperature was measured continuously.

On every workday, the following parameters were documented:

• Vitality of parent animals

• Number of living offspring, dead offspring and aborted eggs

• Number of male offspring and resting eggs

At the end of the test, the total length of the parent animals (without apical spine) was measured

The effect occurrence was calculated on the basis of the biological parameters. With the statistical software ToxRat Professional 3.2.1 (ToxRat Solutions GmbH, Alsdorf, Germany), ECx and LOEC/NOEC were determined.

TOC analysis of the test solutions was performed to determine concentrations of the test item during exposure. The Non-Purgable-OC-method (NPOC; sparging with oxygen) was applied with a total carbon analyser (Shimadzu TOC-L with autosampler ASI-L) using a non-dispersive infrared

(NDIR) detector. The TOC attributable to the test substance in the test item solutions was calculated by subtraction of the mean value of the control (NC) measurements.

The test item concentrations in the treatment groups A and B were too low for a reliable TOC quantification with the available equipment. The measured geometric mean TOC concentrations in the treatment groups C, D and E were 85.9 – 106.7 % of the nominal concentrations (Table 6). As the measured geometric mean TOC concentrations are within ± 20 %

of the nominal concentrations, according to OECD 211 (2012) and OECD 23 (2000), all results are given in relation to the nominal test item concentrations.

No mortalitues were observed except in teh highest concentration (23.4 mg/L) where 100% mortality was observed.

During the exposure, no dead offspring, aborted eggs and male offspring was observed.

The test is valid according to OECD Test Guideline 211 (02 October 2012)

Description of key information

 In a semi-static Daphnia magna Reproduction Test according to OECD 211, the 21day EC50 for reproduction was determined as 10.3 mg/L. The NOEC for reproduction was determined as 2.2 mg/L. 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

2.2 mg/L