4-[3-(1-naphthylamino)propyl]morpholine

Stability in organic solvents and identity of relevant degradation products

Administrative data

Endpoint:

stability in organic solvents and identity of relevant degradation products

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 4, 2016 to September 2, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Principles of method if other than guideline:

Experimental Design:
Stability assessment at concentrations of 0.250, 2.5, 175, 250 mg/mL was based on the measured concentrations of 4-(3-(1-naphthylamino)propyl)morpholine from PG in comparison to initial results.
The stability samples were stored under ambient temperature, exposed to normal lighting conditions, and stirred continuously for the duration of the stability assessment. Stability samples in PG were analyzed on days 2 (initial), 3 (low dose only), 9, 16, and 23 to determine test material concentration.
Stability assessment at concentrations of approximately 0.2 – 25 μg/mL, and 5,000 μg/mL was based on the measured concentrations of 4-(3-(1-naphthylamino)propyl)morpholine from acetonitrile + 0.1% formic acid in comparison to initial results.
Stability samples in acetonitrile + 0.1% formic acid at concentrations of approximately 0.25 – 25 μg/mL were analyzed on days 0 (initial) and 7 to determine test material concentration. The stability sample in acetonitrile +0.1% formic acid at a concentration of approximately 5,000 μg/mL was analyzed on days 0 (initial) and 21 to determine test material concentration.
Solubility of the test material at 250 mg/mL PG and approximately 5,000 μg/mL acetonitrile + 0.1% formic acid stored under ambient temperature, exposed to normal lighting conditions was assessed by visual inspection.
Analytical method validation was performed by fortification of PG with 4-(3-(1- naphthylamino)propyl)morpholine at the concentration range of interest, as per TERC SOP MTH-ACL-2. This included nine fortified (spiked) samples (triplicate analyses at three concentration levels). Samples of control PG were analyzed as well. Appropriately sized aliquots (0.5 – 1 mL) of the samples were analyzed to obtain an accurate determination of test chemical concentration.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-[3-(1-naphthylamino)propyl]morpholine

EC Number:

226-033-2

EC Name:

4-[3-(1-naphthylamino)propyl]morpholine

Cas Number:

5235-82-5

Molecular formula:

C17H22N2O

IUPAC Name:

N-[3-(morpholin-4-yl)propyl]naphthalen-1-amine

Test material form:

solid: granular

Details on test material:

Colour: Dark Brown to Black
GLP Characterized: January 21, 2016

Specific details on test material used for the study:

Test Material Name: 4-(3-(1-naphthylamino)propyl)morpholine
Chemical Name: N-1-Naphthalenyl-4-morpholinepropanamine
Synonyms: None
Lot/Reference/Batch Number: ZA01212016
Purity/Characterization (Method of Analysis and Reference): The purity of the test material was determined to be 94.4% by liquid chromatography with identification by nuclear magnetic resonance spectroscopy and liquid chromatography mass spectrometry (Ferrer, 2016a). Standards were not corrected for purity.
Test Material Stability Under Storage Conditions: 4-(3-(1-Naphthylamino)propyl)morpholine, lot ZA01212016, was determined to be stable for 2 weeks at 54°C which is equivalent to 24 months under ambient storage conditions as tested under USEPA OPPTS Guideline 830.6313 (Ferrer, 2016b).

Results and discussion

Test substance stable:

yes

Transformation products:

not specified

Any other information on results incl. tables

Upon mixing, the 0.250 and 2.5 mg/mL doses contained particles of test materials swirling in the vortex. The 175 and 250 mg/mL doses were opaque with particles of test material. After two days of mixing, the 0.250 and 2.5 mg/mL doses were a light greyish pink solution. No particles of test material were observed. The 175 and 250 mg/mL doses were an opaque grey suspension with no visible particles of test material observed.

Stability of 4-(3-(1-naphthylamino)propyl)morpholine in PG stored at ambient temperature, exposed to light, and stirred continuously was determined. For the test material to be considered stable, 4-(3-(1-naphthylamino)propyl)morpholine would have to be 85% to 115% of the Day 2 (PG) or Day 0 (acetonitrile + 0.1% formic acid) concentration per laboratory standard operating procedures. 4-(3-(1-naphthylamino)propyl)morpholine was found to be stable in PG for 21 days at 0.393, 2.5, 175, and 250 mg/mL concentrations. The 0.393 mg/mL dose was mixed at a target concentration of 0.250 mg/mL. Upon initial analysis, it was found to have been mixed at a higher concentration. After consultation with study personal, it was determined to continue analysis on this dose and assess stability at 0.393 mg/mL, the mean concentration of the initial timepoint.

Two days of mixing was required before a homogenous mixture was observed. Test material adhesion to the sides of the test vessel at concentrations of 175 and 250 mg/mL was observed throughout the study. Adhered test material was incorporated into the test mixture approximately 1 hour before sampling. It is recommended to incorporate the test material as soon as it is noticed to prevent adhesion or at least 1 hour before sampling if adhesion is observed. The test material was found to be stable in acetonitrile + 0.1% formic acid for 21days at a concentration of approximately 5000 μg/mL. The test material was found to be stable in acetonitrile + 0.1% formic acid for 7days at concentrations of approximately 0.25 to 25 μg/mL. Although percent of initial target for Standard B is greater than ± 15% of initial target, the percent of nominal target is within ± 15%. The average of all standards is within ± 15% for percent of initial and percent of nominal concentrations.

A solubility assessment was performed by dissolving approximately 0.1 g of 4-(3-(1-naphthylamino)propyl)morpholine (Lot # ZA01212016) in approximately 25 mL of acetonitrile + 0.1% formic acid. Solubility was confirmed by visual inspection at a concentration of approximately 5000 μg/mL acetonitrile + 0.1% formic acid.

The HPLC/UV instrumentation exhibited a linear response for 4-(3-(1-naphthylamino)propyl)morpholine over a concentration range of approximately 0.25 -25 μg/mL acetonitrile + 0.1% formic acid.

For the recoveries of 4-(3-(1-naphthylamino)propyl)morpholine in PG at approximate concentrations of 0.25, 2.5, and 250 mg/mL, the average recovery across all concentrations was 96.4% ± 1.83% (n=37).

Applicant's summary and conclusion

Conclusions:

4-(3-(1-naphthylamino)propyl)morpholine mixed in PG was shown to be stable at ambient temperature, exposed to light, stirring continuously, over the course of 21 days at concentrations ranging from 0.393 to 250 mg/mL. All dose levels required 2 days of mixing to form a homogenous mixture. The high doses of 175 and 250 mg/mL required reincorporation of test material due to adhesion of test material to the sides of the test vessel.
4-(3-(1-naphthylamino)propyl)morpholine in acetonitrile + 0.1% formic acid was shown to be stable at ambient temperature, exposed to light, over the course of 7 days at concentrations of approximately 0.25 –25 μg/mL and 21 days at a concentration of approximately 5,000 μg/mL.
Solubility of 4-(3-(1-naphthylamino)propyl)morpholine (Lot # ZA01212016) has been demonstrated at a concentration of approximately 5,000 mg/mL in acetonitrile + 0.1% formic acid.
The analytical method has been validated for use in analyzing and quantifying 4-(3-(1-naphthylamino)propyl)morpholine in PG.

Executive summary:

This study was conducted to determine the stability of 4-(3-(1-naphthylamino)propyl)morpholine in propylene glycol (PG) at ambient temperature, exposed to normal lighting conditions and stirred

continuously. When mixed in PG, 4-(3-(1-naphthylamino)propyl)morpholine was shown to be stable over the course of 21 days at concentrations ranging from 0.393 to 250 mg/mL. At least 2 days of mixing was required to obtain homogenous mixtures. 4-(3-(1-naphthylamino)propyl)morpholine formed a suspension in PG at a concentration of 250 mg/mL.

Samples were prepared by diluting with acetonitrile + 0.1% formic acid. Samples were then sonicated for a minimum of 15 minutes and allowed to equilibrate for a minimum of 30 minutes before being further diluted with acetonitrile + 0.1% formic acid for analysis. Analysis was performed by high performance liquid chromatography with ultraviolet detection.

Standards were shown to be linear using linear regression with 1/x weighting over the concentration range of approximately 0.25 – 25 μg 4-(3-(1-naphthylamino)propyl)morpholine /mL acetonitrile + 0.1% formic acid. Quantitation was performed using external standard calibration.

Recoveries of the test material, fortified into PG at concentrations of approximately 0.25 to 250 mg/mL, ranged from 93.9% ± 1.38% to 98.4% ± 1.23%. Fortified samples were prepared for the purpose of method validation and spike recovery.

The analytical method has been validated for use in analyzing and quantifying 4-(3-(1-naphthylamino)propyl)morpholine in PG.