CN107976501B - A kind of detection method of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethylamine - Google Patents

Abstract

The invention discloses a method for detecting 1- (3-ethoxy-4-methoxyphenyl) -2- (methylsulfonyl) ethylamine, which adopts a liquid chromatograph to detect and an area normalization method to carry out quantitative analysis, and has the following chromatographic conditions: a chromatographic column: a phenyl column; sample introduction amount: 5-10 mul; flow rate: 0.8-1.2 ml/min; column temperature: 20-30 ℃; detection wavelength: 200-250 nm; mobile phase: mobile phase a 0.05% aqueous trifluoroacetic acid solution to mobile phase B0.05% aqueous trifluoroacetic acid (acetonitrile: methanol: 700:300) solution in a volume ratio of (10-95): (5-90); diluting liquid: acetonitrile to water in a volume ratio of (0.5-1.5) to 1; a detector: an ultraviolet detector; the detection method is simple and rapid, can realize accurate determination of the 1- (3-ethoxy-4-methoxyphenyl) -2- (methylsulfonyl) ethylamine, and has high sensitivity.

Description

A kind of detection of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethylamine test method

technical field

The invention belongs to the technical field of analytical chemistry, in particular to a method for detecting 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine.

Background technique

Apremilast is an oral medication for the treatment of active psoriatic arthritis with the chemical name (S)-2-[1-(3-ethoxy-4-methoxy)phenyl-2- Methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione, CAS No. 608141-41-9. The drug, approved by the FDA in March 2014 under the trade name Otezla (Apremilast), is the first and only FDA-approved oral, selective phosphodiesterase 4 (PDE4) for the treatment of plaque psoriasis ) inhibitor.

1-(3-Ethoxy-4-methoxy)phenyl-2-methanesulfonylethylamine is a key intermediate in the preparation of apremilast, and its purity and impurities directly affect the purity and impurities of apremilast content, thereby affecting the efficacy of the drug.

At present, there are no relevant literature and reports on the purity detection method of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine. For the quality control of ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine, it is necessary to develop its detection method.

SUMMARY OF THE INVENTION

In view of the problems existing in the prior art, the purpose of the present invention is to provide a method for detecting 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine.

The structural formula of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine (AMTO1) is as follows:

3-Ethoxy-4-methoxybenzonitrile (AMT00) is a Intermediate, its structural formula is as follows:

In order to achieve the above object, the present invention adopts the following technical solutions:

A method for detecting 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine, comprising the following steps:

(1) Weigh out 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine and 3-ethoxy-4-methoxybenzonitrile respectively In the same volumetric flask, dissolve and dilute to volume with diluent to prepare a system adaptability solution; 2-(Methylsulfonyl)ethylamine and 20-30mg of 3-ethoxy-4-methoxybenzonitrile were placed in the same 50ml volumetric flask, dissolved with diluent and fixed to volume to prepare a system Adaptation solution; wherein the concentration of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine is 0.4-0.6 mg/ml, 3-ethoxy- The concentration of 4-methoxybenzonitrile is 0.4-0.6 mg/ml;

(2) Precisely weigh the test sample, dissolve it with the diluent and dilute to volume, and prepare the test sample solution; The solution is dissolved and constant to volume, and the solution of the test product with a concentration of 0.3-1 mg/ml is prepared;

(3) Precisely measure the diluent, inject it into the liquid chromatograph, use the mobile phase to carry out gradient elution, and record the chromatogram without interference;

(4) Precisely measure the system adaptability solution, inject it into the liquid chromatograph, use the mobile phase to carry out gradient elution, and record the chromatogram, the theoretical plate number is 1-(3-ethoxy-4-methoxyphenyl) )-2-(methylsulfonyl)ethylamine should not be less than 2000, and the resolution between the main peak and the AMT00 peak should not be less than 1.5;

(5) Precisely measure the test solution, inject it into a liquid chromatograph, use the mobile phase to carry out gradient elution, and record the chromatogram, deduct the blank test chromatographic peak in the chromatogram of the test solution, and use the area normalization method. Calculate product purity and impurity content separately;

The operating conditions of the liquid chromatograph are:

Chromatographic column: phenyl column;

Injection volume: 5-10μl;

Flow rate: 0.8-1.2ml/min;

Column temperature: 20-30℃;

Detection wavelength: 200-250nm;

Mobile phase: mobile phase A 0.05% trifluoroacetic acid aqueous solution: mobile phase B 0.05% trifluoroacetic acid (acetonitrile: methanol=700:300) solution, the volume ratio is (10-95): (5-90);

Diluent: acetonitrile: water, its volume ratio is (0.5-1.5): 1;

Detector: UV detector.

Preferably, the chromatographic column is Agilent Zorbax SB phenyl, the size is 4.6*250mm, 5μm.

Preferably, the injection volume is 5 μl.

Preferably, the flow rate is 1 ml/min.

Preferably, the detection wavelength is 230 nm.

Preferably, the column temperature is 25°C.

Preferably, the diluent is formed by mixing acetonitrile and water in a volume ratio of 1:1.

Preferably, the gradient elution conditions are: 0-30min: mobile phase A 95%, mobile phase B 5%; 30-35min: mobile phase A 10%, mobile phase B 90%; 35-40min: mobile phase A 95%, mobile phase B 5%.

The mobile phase in the present invention is trifluoroacetic acid aqueous solution, trifluoroacetic acid methanol-acetonitrile solution, and gradient elution is carried out according to a certain ratio, so that 1-(3-ethoxy-4-methoxyphenyl)-2-( Methylsulfonyl)ethylamine and related impurities can be effectively separated, and adding trifluoroacetic acid to the mobile phase can effectively improve the peak shape.

The beneficial effects of the present invention

(1) The detection method of the present invention is simple and fast, can realize the accurate determination of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine, and has high sensitivity;

(2) The detection method of the present invention enables effective separation of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine and its known impurities, and the degree of separation Not less than 1.5, meeting the standard of complete separation;

(3) The detection method of the present invention provides quality support for the industrial production of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine.

Description of drawings

Fig. 1 is the high performance liquid chromatogram of system adaptability solution in embodiment 1; Chromatographic peak 1 is 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethylamine (AMT01); Chromatographic peak 2 is 3-ethoxy-4-methoxybenzonitrile (AMT00);

Fig. 2 is the high performance liquid chromatogram of test sample 2.

Detailed ways

In order to better explain the present invention, further description will now be made in conjunction with the following specific embodiments, but the present invention is not limited to the specific embodiments.

Example 1

A method for detecting 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine,

1. Instruments and testing conditions

Shimadzu LC-15C high performance liquid chromatograph, the chromatographic column is Agilent ZORBAX SB-phenyl (4.6*250mm, 5μm);

Injection volume: 5μl;

Flow rate: 1ml/min;

Column temperature: 25℃;

Detection wavelength: 230nm;

Mobile phase: mobile phase A 0.05% trifluoroacetic acid aqueous solution, mobile phase B 0.05% trifluoroacetic acid (acetonitrile: methanol = 700: 300) solution, using gradient elution, the gradient elution conditions are: 0-30min: mobile Phase A 95%, mobile phase B 5%; 30-35min: mobile phase A 10%, mobile phase B 90%; 35-40min: mobile phase A 95%, mobile phase B 5%.

Diluent: acetonitrile and water in a volume ratio of 1:1.

Detector: UV detector.

2. Experimental steps

High performance liquid chromatography was used for detection, and the detection steps were as follows:

(1) Precisely weigh 25 mg of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine and 25 mg of 3-ethoxy-4-methoxyphenyl Benzonitrile was placed in the same 50ml volumetric flask, dissolved with diluent and adjusted to volume to prepare a system adaptability solution;

(2) Accurately weigh 25mg of the test product, place it in a 50ml volumetric flask, dissolve it with a diluent and dilute to volume, and prepare a test solution with a concentration of 0.5mg/ml;

(3) Precisely measure 5 μl of the diluent, inject it into a liquid chromatograph, use the mobile phase to carry out gradient elution, and record the chromatogram;

(4) Precisely measure 5 μl of the system adaptability solution, inject it into the liquid chromatograph, use the mobile phase to carry out gradient elution, and record the chromatogram. The number of theoretical plates is 1-(3-ethoxy-4-methoxybenzene) base)-2-(methylsulfonyl)ethylamine should not be less than 2000, and the resolution between the main peak and the AMT00 peak should not be less than 1.5;

(5) Precisely measure 5 μl of the test solution, inject it into a liquid chromatograph, use the mobile phase to carry out gradient elution, and record the chromatogram. The blank test chromatographic peak is deducted from the chromatogram of the test solution, and normalized by area. The product purity and impurity content were calculated separately.

Example 2

Take three batches of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethylamine products produced by the same production specification instruction, which are respectively test samples 1-3, according to the implementation The detection method of Example 1 is detected, and the area normalization method is used to calculate the purity and impurities thereof, and the detection results are shown in the following table 1-3:

Table 1 is the test result of test sample 1

Table 2 is the test result of test product 2

Table 3 is the test result of test sample 3

The detection method of the invention can simply, accurately, quickly, efficiently and reliably detect the purity and impurity content of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine , has high sensitivity, is easy to operate, and can achieve complete separation, thereby providing the basis for research and development and quality detection for the study of such compounds.

The above descriptions are only specific embodiments of the present invention, and are not intended to limit the scope of the present invention. All equivalent transformations made by the present invention, or directly or indirectly applied in other related technical fields, are similarly included in the present invention. within the scope of patent protection.

Claims (9)

1. a detection method of 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethylamine, is characterized in that, comprises the following steps: (1) Weigh out 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethylamine and 3-ethoxy-4-methoxybenzonitrile respectively In the same volumetric flask, dissolve and dilute with diluent to prepare a system adaptability solution; (2) Precisely weigh the test sample, dissolve it with diluent and make up to volume, and prepare a test sample solution; (3) inject the described system adaptability solution, need testing solution and blank sample into liquid chromatograph respectively, adopt mobile phase to carry out gradient elution, and record chromatographic peak; Wherein, blank sample is diluent; The operating conditions of the liquid chromatograph are: Chromatographic column: phenyl column; Injection volume: 5-10μl; Flow rate: 0.8-1.2ml/min; Column temperature: 20-30℃; Detection wavelength: 200-250nm; Mobile phase: mobile phase A 0.05% trifluoroacetic acid aqueous solution: mobile phase B 0.05% trifluoroacetic acid solution, its volume ratio is (10-95): (5-90); in the trifluoroacetic acid solution described in mobile phase B The solvent for dissolving trifluoroacetic acid is a mixed solution of acetonitrile and methanol, wherein the volume ratio of acetonitrile: methanol is 700:300; Diluent: acetonitrile: water, its volume ratio is (0.5-1.5): 1; Detector: UV detector; The gradient elution conditions are: 0-30min: mobile phase A 95%, mobile phase B 5%; 30-35min: mobile phase A 10%, mobile phase B 90%; 35-40min: mobile phase A 95%, mobile phase B Phase B 5%. 2. The detection method according to claim 1, wherein in the system adaptability solution, 1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl ) ethylamine at a concentration of 0.4-0.6 mg/ml and 3-ethoxy-4-methoxybenzonitrile at a concentration of 0.4-0.6 mg/ml. 3. The detection method according to claim 1, wherein the concentration of the test substance is 0.3-1 mg/ml. 4. The detection method according to claim 1, wherein the chromatographic column is Agilent Zorbax SBphenyl, and the size is 4.6*250mm, 5μm. 5 . The detection method according to claim 1 , wherein the injection volume is 5 μl. 6 . 6. The detection method according to claim 1, wherein the flow rate is 1 ml/min. 7. The detection method according to claim 1, wherein the detection wavelength is 230 nm. 8 . The detection method according to claim 1 , wherein the column temperature is 25° C. 9 . 9. detection method according to claim 1, is characterized in that, described diluent is acetonitrile and water are mixed by volume ratio of 1:1.

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