3-morpholinopropylamine

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 20, 2017 - January 24, 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

440/2008

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: OECD No. 23

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss GLP Monitoring Authorities, Date of decission 2015-11-16

Test material

Test material information

Constituent 1

Reference substance name:

3-morpholinopropylamine

EC Number:

204-590-2

EC Name:

3-morpholinopropylamine

Cas Number:

123-00-2

Molecular formula:

C7H16N2O

IUPAC Name:

3-morpholin-4-ylpropan-1-amine

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
Test Item Name: N-morpholinopropylamine
Batch Number: 6H706
CAS-No.: 123-00-2
Purity: 99.7 %
Molecular Formula: C7H16N2O
Appearance: Colorless, clear liquid, substantially free of foreign matter
Relative Density: 0.99
Expiration Date: August 11, 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
Storage Conditions: At room temperature, at 20 ± 5 °C. Keep container tightly closed in a dry and well-ventilated place.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampled concentration: 100 mg/L
- Sampling method: duplicate samples were taken 100mg/L and the control at the start and at the end of the test (after 48 hours). For the 48-hour stability samples, the contents of the respective replicates were combined prior to sampling.
- Sample storage conditions before analysis: all samples were frozen (at -20 ± 5 °C)
- Analysed samples: The concentration of N-morpholinopropylamine were analyzed in one of the duplicate test medium samples from the control and the nominal concentrations of 100 mg/L from both sampling times (0 and 48 hours).

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test medium of the nominal concentration of 100 mg/L was prepared by dissolving 101 µL of the test item completely in 1000 mL of test water. This volume is equivalent to a loading rate of 100 mg/L, considering the relative density of the test item of 0.99. The test item was dissolved by using ultrasonic treatment for 15 minutes and intense stirring for 15 minutes at room temperature. During this stirring period the pH was adjusted from pH 9.5 to 7.5 with 1 M hydrochloric acid solution.
- Eluate: No eluation was necessary, since only the single concnetration of 100mg/L was tested. The test medium was prepared just before the introduction of the daphnids (i.e., start of the test).
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): After the dissolving procedure the test concentration of 100 mg/L was a clear solution.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Daphnia magna Straus
- Source: Daphnia Collection of the University of Basel/Switzerland in 2015
- Age of parental stock: 3-4 Weeks. At the start of the test, the organisms used in the test were 6 to 24 hours old and were not first brood progeny.
- Feeding during test: no


ACCLIMATION
- Acclimation period: no acclimatisation was necessary, since the bred of the daphnia was under identical conditions to those of the test.
- Type and amount of food during the breeding: Algal suspension of the green algae Desmodesmus subspicatus supplied by the Collection of Algal Cultures (SAG, Institute for Plant Physiology, University of Göttingen, 37073 Göttingen / Germany) and cultivated at IES Ltd Laboratories under standardized conditions or a mixture of this algal suspension and a
commercial fish diet (Tetra Min® Hauptfutter, supplied by TETRA-GmbH, 49324 Melle / Germany).
- Feeding frequency: three times a week
- Health during acclimation (any mortality observed): The daphnia were healthy, no mortality was observed.

Study design

Test type:

static

Water media type:

other: ISO Test water (Medium according to the guideline)

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

2.5 mmol/L = 250 mg/L as CaCO3

Test temperature:

21

pH:

at the start of the test at 100 mg/L and the control: 7.5
at the end of the test at 100 mg/L: 8.0 and the control: 7.9

Dissolved oxygen:

at the start of the test at 100 mg/L: 8.2 mg/L oxygen and the control: 8.3 mg/L oxygen
at the end of the test at 100 mg/L: 8.0 mg/L oxygen and the control: 8.1 mg/L oxygen

Salinity:

n.d.

Conductivity:

n.d.

Nominal and measured concentrations:

nominal concentrations: 100 mg/L
measured concentration (0 and 48h): 104 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Beakers
- Type (delete if not applicable): Open (The test vessels were coverd with glass plates to reduce the loss of water by evaporation and to avoid the entry of dust into the solutions.)
- Material, size, headspace, fill volume: glas, 100 mL, open, 50 mL
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): no, static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates):4
- No. of vessels per control (replicates):4
- No. of vessels per vehicle control (replicates):no
- Biomass loading rate: 1 Daphnia / 10 mL test media

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO Medium Ingredients / Properties [mmol/L] / Concentration [mg/L]
CaCl2 × 2H2O / 2.0 / 294
MgSO4 × 7H2O / 0.5 / 123
NaHCO3 / 0.75 / 65
KCl / 0.075 / 5.8
- Alkalinity: 0.8 mmol/L
- Water Hardness: 2.5 mmol/L = 250 mg/L as CaCO3

OTHER TEST CONDITIONS
- Adjustment of pH: 7.5 with 0.1M hydrochloric acid
- Photoperiod:16-hour light to 8-hour dark cycle with a 30-minute transition period
- Light intensity:15 and 17 µE m-2 s-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobility of the daphnids and other non-lethal effects (e.g. abnormal behavior or appearance) after 24 and 48 hours of exposure.

VEHICLE CONTROL PERFORMED: no, not needed

RANGE-FINDING STUDY with pH adjustment: The pH was adjusted in the 100 mg/L stock solution from pH 9.5 to pH 7.5
- Test concentrations: 1.0 / 10 / 100 mg/L
- Results used to determine the conditions for the definitive study:
48-hour immobilization: 1.0 mg/L / 0%; 10 mg/L / 0%; 100 mg/L / 0%

Reference substance (positive control):

yes

Remarks:

Potassium dichromate / Separate reference studies under GLP twice a year.

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: no abnormalities in the control and the concentration 100 mg/L.
- Mortality of control: no
- Other adverse effects control: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: 0.19-3.0 mg/L
- Dose-response test: Yes
- ECx: 24-hour EC50: 1.3 mg/L

Reported statistics and error estimates:

No statistics needed.
The 24-hour and 48-hour NOEC, EC0, EC50 and EC100 were determined directly from the raw data.

Any other information on results incl. tables

Analytical results

Sampling Day/ Sample Age	Nominal Concentration of Test Item cnom	Measured Concentration  of Test Item x	Sample Preparation Factor F	Determined Concentration of Test Item c	% of Nominal Concentration
[d/h]	[mg/L]	[mg/L]		[mg/L]	[%]
0/0	Control	n.d.	1	< LOQ	n.a.
(fresh)	100	104	1	104	104
2/48	Control	n.d.	1	< LOQ	n.a.
(aged)	100	104	1	104	104

n.d.        = no test item detected

n.a.        = not applicable

LOQ:     19.8 mg/L

Biological Results: Effect of N-morpholinopropylamine on the Mobility of Daphnia magna

 

Nominal Test Item Concentration	No. of Daphnids Tested	Immobilized Daphnids after 24 Hours	Immobilized Daphnids after 24 Hours	Immobilized Daphnids after 48 Hours	Immobilized Daphnids after 48 Hours
[mg/L]		No.	[%]	No.	[%]
Control	20	0	0	0	0
100	20	0	0	0	0

 

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In a valid, reliable and conclusive study according to OECD 202 (2004), the effect of the test item N-morpholinopropylamine on the mobility of young daphnids of the species Daphnia magna after 48 hours was determined. The test item N-morpholinopropylamine had no acute toxic effects on Daphnia magna in a static 48-hour test up to the nominal concentration of 100 mg/L under the conditions of the test.

Executive summary:

The acute toxicity of the test item N-morpholinopropylamine on Daphnia magna was determined in a 48‑hour static test according to the OECD Guideline for Testing of Chemicals, No. 202 (2004) and the Commission Regulation (EC) No. 440/2008, Part C.2.

A limit test was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effect on the test organisms up to and including a nominal concentration of 100 mg/L. Thus, the only nominal concentration tested was 100 mg/L. A control group was tested in parallel.

The measured concentration of the test item N-morpholinopropylamine in the test medium of the single test concentration of 100 mg/L was 104 % of nominal value at the start and the end of the test.

 Thus, the correct dosage of the test item N-morpholinopropylamine was confirmed. The test item was stable in the test medium over the test period of 48 hours. Therefore, the endpoint values were related on the nominal concentration of 100 mg/L.

The biological test results (based on nominal concentration) were as follows:

24-hour EC₅₀ > 100 mg/L

24-hour EC₀ ≥ 100 mg/L

24-hour EC₁₀₀ > 100 mg/L

48-hour EC₅₀ > 100 mg/L

48-hour EC₀ and  48-hour NOEC: ≥ 100 mg/L

48-hour EC₁₀₀ > 100 mg/L