N,N,4-trimethylpiperazine-1-ethylamine

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-06-27 to 2017-10-07

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: PFW160141
- Expiration date of the lot/batch: 2018-12-30
- Purity test date: 2016-07-19 (certificate of analysis release date)
- Purity: 99.5%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under test conditions: no data
- Solubility and stability of the test substance in the solvent/vehicle: no data

Analytical monitoring:

yes

Details on sampling:

- Sampling method:
* Range-Finding Test: A sample of each test concentration was taken for chemical analysis at 0 and 48 hours in order to determine the stability of the test item under test conditions.
* Definitive Test: The concentration and stability of the test item in the test preparations were verified by chemical analysis at 0 and 48 hours.
- Sample storage conditions before analysis: All samples were stored frozen prior to analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
* Range-Finding Test: The test concentrations to be used in the definitive test were determined by a preliminary range-finding test.
A nominal amount of test item (50 mg) was dissolved in test water and the volume adjusted to 500 mL to give the 100 mg/L test concentration from which a series of dilutions was made to give further test concentrations of 0.10, 1.0 and 10 mg/L. Each prepared concentration was inverted several times to ensure adequate mixing and homogeneity.
* Definitive Test: Based on the results of the range-finding test the following test concentrations were assigned to the definitive test: 10, 18, 32, 56 and 100 mg/L.
A nominal amount of test item (200 mg) was dissolved in test water and the volume adjusted to 2 liters to give the 100 mg/L test concentration which was pH adjusted to bring it within the range outlined in the General Study Plan of between 6 and 9. A series of dilutions was then made from this stock solution to give further test concentrations of 10, 18, 32 and 56 mg/L.
Each prepared concentration was inverted several times to ensure adequate mixing and homogeneity.
- Controls: yes
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): All solutions were clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water-flea
- Source: in-house laboratory culture with a known history
- Age at study initiation (mean and range, SD): < 24 h (from parental daphnids of more than 2 weeks old, maximum age 4 weeks)
- Method of breeding: Culture conditions ensured that reproduction was by parthenogenesis.
- Feeding during cultivation: mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension, daily
- Feeding during test: no

ACCLIMATION
- Acclimation period: not relevant

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

not reported

Test temperature:

22-23°C

pH:

At t=0h: 8.0-8.1
At t=48h: 7.6-8.1

Dissolved oxygen:

At t=0h: 8.6-8.8
At t=48h: 8.5-8.7

Salinity:

not relevant

Nominal and measured concentrations:

Range finder:
Nominal concentrations: 0.10, 1.0, 10 and 100 mg/L (10 and 100 mg/L analysed)
Measured concentrations (mg/L) at t=0h: 15.0, 115
Measured concentrations (mg/L) at t=48h: 14.4, 114

Final test:
Nominal concentrations: 10, 18, 32, 56 and 100 mg/L
Measured concentrations (mg/L) at t=0h: Measured concentrations (mg/L) at t=48h:

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 150 mL, glass beaker filled with 100 mL
- Aeration: not during the test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Test medium: adjusted ISO medium

OTHER TEST CONDITIONS
- Adjustment of pH: yes
- Photoperiod: 16h light daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Immobility was defined as those organisms which are not able to swim within 15 seconds after gentle agitation of the test container and was monitored after 24 and 48 hours of continuous treatment with the test substance.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0.10, 1.0, 10 and 100 mg/L (setup: combined range finder/limit test)
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

66 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: none

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: no
- EC50/LC50: 48-h EC50 = 1.2 mg/L. The sensitivity of the daphnids was within the range determined with the historical data collected at Charles River Laboratories Den Bosch.

Reported statistics and error estimates:

The EC50 values and associated confidence limits at 48 hours and the slope of the response curve and its standard error were calculated by Probit analysis using Linear Maximum-Likelihood regression. The Lowest Observed Effect Concentration and the No Observed Effect Concentration at 24 and 48 hours were calculated using the Fisher’s Exact Binomial Test with Bonferroni correction. All results were calculated using the ToxRat Professional computer software package (TOXRAT).

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of JeffCat TAP to Daphnia magna was determined in a 48 hour static test according to OECD guideline 202. Based on nominal concentrations, the 48-h EC50 was 66 mg/L. The results of the test can be considered reliable without restriction.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2001-11-14 to 2002-03-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Purity: 99.2% (GC)
- Source and lot/batch No.of test material: 180601
- Expiration date of the lot/batch: 2002-10-25 (retest date)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature in the dark
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: at least 96h in water

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preperation of test solutions started with a stock in ISO-medium at 100 mg/L. No special treatment other than magnetic stirring for a short period was necessary to completely dissolve the test substance in the test medium. The test concentrations were prepared by subsequent dillutions of the stock in test medium.
- Controls: yes
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): All solutions were clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water-flea
- Source: in-house laboratory culture with a known history
- Age at study initiation (mean and range, SD): < 24 h (from parental daphnids of more than 2 weeks old, maximum age 4 weeks)
- Method of breeding: Culture conditions ensured that reproduction was by parthenogenesis.
- Feeding during cultivation: fresh water algae, daily
- Feeding during test: no

ACCLIMATION
- Acclimation period: not relevant

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 mg/L expressed as CaCO3

Test temperature:

18.9 to 20.2°C

pH:

At t=0h: 8.0-9.1
At t=48h: 7.9-8.6

Dissolved oxygen:

At t=0h: 9.1-9.2
At t=48h: 9.3-9.5

Salinity:

not relevant

Nominal and measured concentrations:

Range finder:
Nominal concentrations: 0.10, 1.0, 10 and 100 mg/L

Final test:
Nominal concentrations: 10, 18, 32, 56 and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 100 mL, all-glass beaker filled with 80 mL
- Aeration: not during the test
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2 (final test)
- No. of vessels per control (replicates): 2 (final test)
- Test medium: M7 (adjusted ISO-medium)

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

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Immobility was defined as those organisms which are not able to swim within 15 seconds after gentle agitation of the test container and was monitored after 24 and 48 hours of continuous treatment with the test substance.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: x 1.8
- Range finding study
- Test concentrations: 0.10, 1.0, 10 and 100 mg/L (setup: combined range finder/limit test)
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

39 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% C.L.: 29 - 53 mg/L

Details on results:

- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: none

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: no
- EC50/LC50: 48-h EC50 = 0.55 mg/L. The sensitivity of the daphnids was within the range determined with the historical data collected at Notox.

Reported statistics and error estimates:

The EC50-value was calculated at 24 and 48 hours of exposure from the probits of the percentages of affected daphnia and the logarithms of the corresponding test substance concentrations using the maximum likelihood estimation method (Finney, D.J., 1971: probit analysis, Cambridge University Press, Cambridge, U.K., 3rd edition)

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of JeffCat TAP to Daphnia magna was determined in a 48 hour static test according to OECD guideline 202. Based on nominal concentrations, the 48-h EC50 was determined to be 39 mg/L (95% C.L. 29 - 53 mg/L). The results of the test can be considered reliable without restriction.

Description of key information

Two relevant studies were identified to cover this endpoint.

The studies of Migchielsen (2002) and Squance (2018), investigating the acute toxicity of the test substance in Daphnia magna according to OECD guideline 202, were considered as high quality studies for endpoint coverage.

Based on nominal concentrations, the study of Migchielsen (2002) revelated a 48-h EC50 of 39 mg/L. An EC50 of 66 mg/L (based on nominal test concentrations) was found after an exposure of 48 hours in a static test in freshwater (Squance, 2018).

The study of Migchielsen (2002) was considered for the PNEC derivation. Based on the results it can be concluded that the test substance is harmful to aquatic invertebrates.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

39 mg/L