High performance liquid chromatography analysis method of montelukast sodium salt

Abstract

The invention discloses a high performance liquid chromatography analysis method of montelukast sodium salt. According to the method, a reverse chromatographic column and an ultraviolet detector are adopted, a phosphate buffer solution-acetonitrile with the pH value of 4.0 is used as a mobile phase, and gradient elution is performed. The method realizes simultaneous quantitative detection of six known impurities and unknown impurities in montelukast sodium salt, and can effectively control the content of the known impurities through a principal component self-contrast method with correction factors, the separation degree between impurity peaks and the separation degree between a main peak and an adjacent impurity peak are greater than 1.5, and the purity factor of the main peak is 1000. The method disclosed by the invention is simple in operation, good in repeatability and strong in specificity, can truly reflect the quality of the montelukast sodium salt serving as a chemical raw material medicine, is beneficial to quality control of downstream products, and has a relatively great positive progress effect and a relatively great practical application value.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical chemical industry, in particular to a high performance liquid chromatography analysis method of montelukast sodium salt. Background Art

[0002] Montelukast sodium salt (CAS: 151767-02-1), chemical name is [R-(E)]-1-ff[1-[3-[2-[7-chloro-2-quinolinyl)vinyl]phenyl-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]thio]methyl]cyclopropaneacetic acid sodium, structural formula is

[0003]

[0004] Montelukast sodium is an oral leukotriene receptor antagonist that can specifically inhibit cysteinyl leukotriene (CysLT1) receptors in the airways, thereby improving airway inflammation and effectively controlling asthma symptoms.

[0005] Montelukast sodium is a non-hormonal anti-inflammatory drug, suitable for the prevention and long-term treatment of asthma in adults and children over 1 year old and the treatment of allergic rhinitis, with three different specifications. According to the 2008 edition of my country's "Guidelines for the Prevention and Treatment of Bronchial Asthma", leukotriene modifiers are the only long-term control drugs that can be used alone except inhaled hormones. They can be used as an alternative treatment for mild asthma and a combined treatment for moderate to severe asthma. Therefore, establishing a detection method for related substances in montelukast sodium salt to fully detect the impurities in montelukast sodium salt is of great significance for effectively controlling the quality of montelukast sodium salt and downstream products. Summary of the invention

[0006] The present invention provides a high performance liquid chromatography analysis method for montelukast sodium salt, comprising the following steps:

[0007] (1) Preparation of the test solution: Take an appropriate amount of montelukast sodium raw material, weigh it accurately, and dissolve it in a diluent (90% methanol) to dilute it into a solution with a montelukast concentration of 1 mg / ml to obtain the test solution.

[0008] (2) Preparation of system suitability solution: Accurately pipette appropriate amounts of impurities B, C, D, E, and F, add diluent (90% methanol) to dissolve and dilute to make solutions of impurities B, C, D, E, and F of 0.03 mg / ml, 0.02 mg / ml, 0.015 mg / ml, 0.015 mg / ml, and 0.015 mg / ml, respectively. Irradiate under natural light for 20 min to generate impurity G, thus obtaining the system suitability solution.

[0009] (3) Preparation of control solution: Take the test solution prepared in step (1) and dilute it 200 times with diluent (90% methanol) to prepare a control solution.

[0010] (4) Chromatographic detection: Take 10 μl of the test solution, system suitability solution and control solution respectively, and record the chromatogram after chromatographic detection.

[0011] The high performance liquid chromatography analysis method for montelukast sodium salt as described above is characterized in that the impurity B in step (2) is [1-[[[(1R)-1-[3-[(E)-2-(7-chloroquinoline-2-yl)vinyl]phenyl]-3-[2-(1-methylvinyl)phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid, the impurity C is [1-[[[1-[3-[(E)-2-(7-chloroquinoline-2-yl)vinyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]sulfinyl]methyl]cyclopropyl]acetic acid, and the impurity D is [1-[[[(1R)-1-[3-[(1R)-1-[[[1-(carboxymethyl)cyclopropyl]methyl]mercapto]-2-(7-chloroquinoline-2-yl)ethyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl )phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid, the impurity E is [1-[[[(1R)-1-[3-[(1S)-1-[[[1-(carboxymethyl)cyclopropyl]methyl]mercapto]-2-(7-chloroquinolin-2-yl)ethyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid, the impurity F is [1-[[[(1R )-3-(2-acetylphenyl)-1-[3-[(E)-2-(7-chloroquinoline-2-yl)vinyl]phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid, and the impurity G is [1-[[[(1R)-1-[3-[(Z)-2-(7-chloroquinoline-2-yl)vinyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid.

[0012] The high performance liquid chromatography analysis method of montelukast sodium salt as described above is characterized in that the chromatographic test conditions in step (4) are: using a reverse C18 column, the column temperature is set at 25-35°C, a low concentration phosphate buffer-acetonitrile with pH 4.0 as the mobile phase, the flow rate is 1.0ml / min-1.5ml / min, and the detection wavelength is 230nm-240nm. The gradient elution is as follows,

[0013] Time (min) Mobile phase A% (phosphate buffer) Mobile phase B% (acetonitrile) 0 40 60 3 40 60 34 15 85 36 15 85 37 40 60 40 40 60

[0014] The high performance liquid chromatography analysis method of montelukast sodium salt as described above is characterized in that the concentration of the phosphate buffer is 15mM to 25mM.

[0015] The high performance liquid chromatography analysis method of montelukast sodium salt as described above is characterized in that the phosphate is a mixture of one or more of sodium dihydrogen phosphate and potassium dihydrogen phosphate.

[0016] The high performance liquid chromatography analysis method of montelukast sodium salt as described above is characterized in that the pH adjuster of the low concentration phosphate buffer at pH 4.0 is phosphoric acid.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows: the method of the present invention can effectively control the related substances in the montelukast sodium raw material, the separation between each impurity peak and between the main peak and the adjacent impurity peak is greater than 1.5, and the main peak purity factor is 1000. The method of the present invention can analyze the degradation impurities of montelukast sodium salt in various complex environments, and the method has strong specificity. The method of the present invention can quantitatively analyze each known impurity and accurately control the content of each known impurity. The method of the present invention is conducive to achieving quality control of montelukast sodium downstream products, and has a large positive progressive effect and practical application value. DETAILED DESCRIPTION

[0018] The embodiments described below are exemplary and are only used to explain the present invention, but should not be construed as limiting the present invention.

[0019] A high performance liquid chromatography analysis method for montelukast sodium salt comprises the following steps:

[0020] (1) Preparation of the test solution: Take an appropriate amount of montelukast sodium raw material, weigh it accurately, and dissolve it in a diluent (90% methanol) to dilute it into a solution with a montelukast concentration of 1 mg / ml to obtain the test solution.

[0021] (2) Preparation of system suitability solution: Accurately pipette appropriate amounts of impurities B, C, D, E, and F, add diluent (90% methanol) to dissolve and dilute to make solutions of impurities B, C, D, E, and F of 0.03 mg / ml, 0.02 mg / ml, 0.015 mg / ml, 0.015 mg / ml, and 0.015 mg / ml, respectively. Irradiate under natural light for 20 min to generate impurity G, thus obtaining the system suitability solution.

[0022] (3) Preparation of control solution: Take the test solution prepared in step (1) and dilute it 200 times with diluent (90% methanol) to prepare a control solution.

[0023] (4) Chromatographic detection: Take 10 μl of the test solution, system suitability solution and control solution, respectively, and record the chromatogram after chromatographic detection. In the embodiment provided in this application, the conditions for chromatographic detection are: using a reverse C18 column, the column temperature is set at 25-35°C, pH 4.0 low concentration phosphate buffer-acetonitrile is used as the mobile phase, the flow rate is 1.0 ml / min-1.5 ml / min, and the detection wavelength is 230 nm-240 nm. The gradient elution is as follows,

[0024] Time (min) Mobile phase A% (phosphate buffer) Mobile phase B% (acetonitrile) 0 40 60 3 40 60 34 15 85 36 15 85 37 40 60 40 40 60

[0025] In a feasible embodiment, the concentration of the phosphate buffer is 15mM to 25mM, the phosphate is a mixture of one or more of sodium dihydrogen phosphate and potassium dihydrogen phosphate, and the pH adjuster of the low-concentration phosphate buffer with a pH of 4.0 is phosphoric acid.

[0026] Several embodiments are given below to further illustrate the content of the present invention. Those skilled in the art will appreciate that more chromatographic conditions can be designed according to the following embodiments, all of which fall within the protection scope of the present invention.

[0027] Example 1

[0028] (1) Sample configuration:

[0029] Blank solution: methanol: water = 9:1.

[0030] Dilution: Same as blank solution.

[0031] Impurity B reference substance stock solution: Take about 15.13 mg of impurity B reference substance, accurately weigh it, place it in a 50 ml brown volumetric flask, add 1 ml DMSO to dissolve the sample, and then dilute it to the scale with diluent. (0.3 mg / ml)

[0032] Impurity C reference substance stock solution: Take about 10.01 mg of impurity C reference substance, accurately weigh it, place it in a 50 ml brown volumetric flask, and dissolve it with diluent to the scale. (0.2 mg / ml)

[0033] Impurity D reference substance stock solution: Take about 15.30 mg of impurity D reference substance, accurately weigh it, place it in a 100 ml brown volumetric flask, and dissolve it with diluent to the scale. (0.15 mg / ml)

[0034] Impurity E reference substance stock solution: Take about 14.99 mg of impurity E reference substance, accurately weigh it, place it in a 100 ml brown volumetric flask, and dissolve and dilute it to the scale with diluent. (0.15 mg / ml)

[0035] Impurity F reference substance stock solution: Take about 16.08 mg of impurity F reference substance, accurately weigh it, place it in a 100 ml brown volumetric flask, and dissolve and dilute it to the scale with diluent. (0.15 mg / ml)

[0036] Impurity G reference substance stock solution: Take about 15.06 mg of the impurity G reference substance, accurately weigh it, place it in a 100 ml brown volumetric flask, and dissolve and dilute it to the scale with diluent. (0.15 mg / ml)

[0037] Impurity B reference solution: Accurately measure 1 ml of impurity B reference stock solution, place in a 10 ml brown volumetric bottle, add diluent to dissolve and dilute to scale, shake well. (0.03 mg / ml)

[0038] Impurity C reference solution: Accurately measure 1 ml of impurity C reference stock solution, place in a 10 ml brown volumetric bottle, add diluent to dissolve and dilute to scale, shake well. (0.02 mg / ml)

[0039] Impurity D reference solution: Accurately measure 2 ml of impurity D reference stock solution, place in a 10 ml brown volumetric bottle, add diluent to dissolve and dilute to scale, shake well. (0.03 mg / ml)

[0040] Impurity E reference solution: Accurately measure 2 ml of impurity E reference stock solution, place in a 10 ml brown volumetric bottle, add diluent to dissolve and dilute to scale, shake well. (0.03 mg / ml)

[0041] Impurity F reference solution: Accurately measure 2 ml of impurity F reference stock solution, place in a 10 ml brown volumetric bottle, add diluent to dissolve and dilute to scale, shake well. (0.03 mg / ml)

[0042] Impurity G reference solution: Accurately measure 2 ml of impurity G reference stock solution, place in a 10 ml brown volumetric bottle, add diluent to dissolve and dilute to scale, shake well. (0.03 mg / ml)

[0043] Separation solution: Take 103.64 mg of montelukast sodium raw material, weigh accurately, place in a 100 ml volumetric flask, add appropriate amount of diluent to dissolve, then accurately measure 1 ml of impurity B, C, D, E, F, G reference substance stock solution and place in the same volumetric flask, dilute to the scale with diluent, and shake well. (Impurity B: 0.003 mg / ml; impurity C: 0.002 mg / ml; impurity D: 0.0015 mg / ml; impurity E: 0.0015 mg / ml; impurity F: 0.0015 mg / ml; impurity G: 0.0015 mg / ml; montelukast: 1 mg / ml)

[0044] System suitability solution: Accurately pipette 1 ml of each impurity B, C, D, E, and F stock solution into a 10 ml transparent volumetric flask, mix well, and irradiate under natural light for 20 min.

[0045] Test solution: Take about 103.53 mg of montelukast sodium raw material, weigh accurately, place in a 100 ml brown volumetric flask, dissolve and dilute to the scale with diluent, and shake well. (Montelukast concentration: 1 mg / ml)

[0046] Reference solution: Accurately measure 1 ml of the test solution, place it in a 200 ml volumetric flask, dilute to the mark with diluent, and shake well. (Montelukast concentration: 5 μg / ml)

[0047] Stock solution of raw material: take about 1040.06 mg of montelukast sodium raw material, weigh accurately, put into a 100 ml brown volumetric flask, dissolve and dilute to scale with diluent, shake well, and set aside. (Montelukast: 10 mg / ml)

[0048] API solution (undamaged): Accurately measure 5 ml of montelukast sodium API mother solution, place in a 50 ml brown volumetric flask, dilute to scale with diluent, and shake well. (Montelukast: 1 mg / ml).

[0049] API solution (acid destruction): Accurately measure 5 ml of montelukast sodium API mother solution, place in a 50 ml volumetric flask, add 1 ml of 1 mol / L HCl solution, heat in an 80°C water bath for 2 min, take out, cool, neutralize with 1 ml of 1 mol / L NaOH solution, dilute to the scale with diluent, and shake well.

[0050] Blank solution (acid destruction): Take the diluent and operate in the same manner.

[0051] API solution (alkali destruction): Accurately measure 5 ml of montelukast sodium API mother solution, place in a 50 ml volumetric flask, add 5 ml of 5 mol / L NaOH solution, heat in a 90°C water bath for 2 h, take out, cool, neutralize with 5 ml of 5 mol / L HCl solution, dilute to the scale with diluent, and shake well.

[0052] Blank solution (alkaline destruction): take the diluent and operate in the same manner.

[0053] API solution (oxidative damage): Accurately measure 5 ml of montelukast sodium API mother solution, place in a 50 ml volumetric flask, add 0.1 ml of 3% hydrogen peroxide solution, heat in a 60°C water bath for 15 min, take out, cool, dilute to the scale with a diluent, and shake well.

[0054] Blank solution (oxidative destruction): Take the diluent and operate in the same manner.

[0055] API solution (light-damaged): Accurately measure 5 ml of montelukast sodium API mother solution and place it in a 50 ml volumetric flask. Irradiate it under a light intensity of 4500 LX for 1.5 h, then take it out, dilute it to the scale with a diluent, and shake it well.

[0056] Blank solution (light-destroyed): Take the diluent and operate in the same manner.

[0057] API solution (high temperature destruction): Accurately measure 5 ml of montelukast sodium API mother solution and place it in a 50 ml volumetric flask. Heat in a 90°C water bath for 15 h. Take it out, cool it, dilute it to the mark with a diluent, and shake it well.

[0058] Blank solution (high temperature destruction): Take the diluent and operate in the same way.

[0059] (2) Detection: Take 10 μl of each sample solution and inject it, and record the chromatogram respectively. The chromatographic column is Kromasil C18 4.6 nm × 150 nm, 3.5 μm. The chromatographic conditions are 20 mM sodium dihydrogen phosphate (pH adjusted to 4.0 with phosphoric acid) as mobile phase A and acetonitrile as mobile phase B. The detection wavelength is 234 nm. The flow rate is 1.2 ml / min. The column temperature is 30 °C. The elution gradient is as follows:

[0060] Time (min) Mobile phase A% (phosphate buffer) Mobile phase B% (acetonitrile) 0 40 60 3 40 60 34 15 85 36 15 85 37 40 60 40 40 60

[0061] Table 1 Separation solution results

[0062] Impurity C Impurity E Impurity D Impurity G Montelukast Impurity F Impurity B Retention time (min) 7.725 11.537 12.083 14.886 19.037 19.988 32.264 Separation / 17.0 2.2 10.6 14.6 3.2 38.2

[0063] Table 2 Specificity test results

[0064]

[0065] The blank solution has no interference at the main peak and the peak positions of each impurity. Under each forced degradation test, the separation between the main peak and the adjacent impurity peak is greater than 1.5, and the main peak purity factor is 1000. Under each forced degradation test, the degradation amount of the main peak and the growth amount of impurities should be conserved. Under each forced degradation test, the ratio of the sum of the main peak and impurity peak areas of the test sample to the sum of the main peak and impurity peak areas of the undegraded test sample is between 98.0% and 102.0%.

[0066] Example 2

[0067] A high performance liquid chromatography analysis method for montelukast sodium salt comprises the following steps:

[0068] (1) Preparation of the test solution: Take an appropriate amount of montelukast sodium raw material, weigh it accurately, and dissolve it in a diluent (90% methanol) to dilute it into a solution with a montelukast concentration of 1 mg / ml to obtain the test solution.

[0069] (2) Preparation of system suitability solution: Accurately pipette appropriate amounts of impurities B, C, D, E, and F, add diluent (90% methanol) to dissolve and dilute to make solutions of impurities B, C, D, E, and F of 0.03 mg / ml, 0.02 mg / ml, 0.015 mg / ml, 0.015 mg / ml, and 0.015 mg / ml, respectively. Irradiate under natural light for 20 min to generate impurity G, thus obtaining the system suitability solution.

[0070] (3) Preparation of control solution: Take the test solution prepared in step (1) and dilute it 200 times with diluent (90% methanol) to prepare a control solution.

[0071] (4) Chromatographic detection: Take 10 μl of each sample solution and inject it, and record the chromatogram respectively. The chromatographic column is Kromasil C18 4.6 nm × 150 nm, 3.5 μm. The chromatographic conditions are 20 mM sodium dihydrogen phosphate (pH adjusted to 4.0 with phosphoric acid) as mobile phase A and acetonitrile as mobile phase B. The detection wavelength is 234 nm. The flow rate is 1.2 ml / min. The column temperature is 25 °C. The elution gradient is as follows:

[0072] Time (min) Mobile phase A% (phosphate buffer) Mobile phase B% (acetonitrile) 0 40 60 3 40 60 34 15 85 36 15 85 37 40 60 40 40 60

[0073] Example 3

[0074] A high performance liquid chromatography analysis method for montelukast sodium salt comprises the following steps:

[0075] (4) Preparation of the test solution: Take an appropriate amount of montelukast sodium raw material, weigh it accurately, and dissolve it in a diluent (90% methanol) to dilute it into a solution with a montelukast concentration of 1 mg / ml to obtain the test solution.

[0076] (5) Preparation of system suitability solution: Accurately pipette appropriate amounts of impurities B, C, D, E, and F, add diluent (90% methanol) to dissolve and dilute to make solutions of 0.03 mg / ml, 0.02 mg / ml, 0.015 mg / ml, 0.015 mg / ml, and 0.015 mg / ml of impurities B, C, D, E, and F, respectively. Irradiate under natural light for 20 min to generate impurity G, thus obtaining the system suitability solution.

[0077] (6) Preparation of control solution: Take the test solution prepared in step (1) and dilute it 200 times with diluent (90% methanol) to prepare a control solution.

[0078] (4) Chromatographic detection: Take 10 μl of each sample solution and inject it, and record the chromatogram respectively. The chromatographic column is Kromasil C18 4.6 nm × 150 nm, 3.5 μm. The chromatographic conditions are 20 mM sodium dihydrogen phosphate (pH adjusted to 4.0 with phosphoric acid) as mobile phase A and acetonitrile as mobile phase B. The detection wavelength is 234 nm. The flow rate is 1.1 ml / min. The column temperature is 30°C. The elution gradient is as follows:

[0079]

[0080]

[0081] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A high performance liquid chromatography analysis method for montelukast sodium salt, characterized in that: The following steps are involved: (1) Preparation of test solution: Take an appropriate amount of montelukast sodium raw material, weigh it accurately, and dissolve it in a diluent (90% methanol) to dilute it into a solution with a montelukast concentration of 1 mg / ml to obtain the test solution. (2) Preparation of system suitability solution: Accurately pipette appropriate amounts of impurities B, C, D, E, and F, add diluent (90% methanol) to dissolve and dilute to make solutions of impurities B, C, D, E, and F of 0.03 mg / ml, 0.02 mg / ml, 0.015 mg / ml, 0.015 mg / ml, and 0.015 mg / ml, respectively. Irradiate under natural light for 20 min to generate impurity G, thus obtaining the system suitability solution. (3) Preparation of control solution: Take the test solution prepared in step (1) and dilute it 200 times with diluent (90% methanol) to prepare a control solution. (4) Chromatographic detection: Take 10 μl of the test solution, system suitability solution and control solution respectively, and record the chromatogram after chromatographic detection.

2. The high performance liquid chromatography analysis method of montelukast sodium salt according to claim 1, characterized in that: In step (2), the impurity B is [1-[[[(1R)-1-[3-[(E)-2-(7-chloroquinoline-2-yl)vinyl]phenyl]-3-[2-(1-methylvinyl)phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid, the impurity C is [1-[[[1-[3-[(E)-2-(7-chloroquinoline-2-yl)vinyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]sulfinyl]methyl]cyclopropyl]acetic acid, and the impurity D is [1-[[[(1R)-1-[3-[(1R)-1-[[[1-(carboxymethyl)cyclopropyl]methyl]mercapto]-2-(7-chloroquinoline-2-yl)ethyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]mercapto]methyl]cyclopropyl The impurity E is [1-[[[(1R)-1-[3-[(1S)-1-[[[1-(carboxymethyl)cyclopropyl]methyl]mercapto]-2-(7-chloroquinolin-2-yl)ethyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid, the impurity F is [1-[[[(1R)-3-(2-acetylphenyl)-1-[3-[(E)-2-(7-chloroquinolin-2-yl)vinyl]phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid, and the impurity G is [1-[[[(1R)-1-[3-[(Z)-2-(7-chloroquinolin-2-yl)vinyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]mercapto]methyl]cyclopropyl]acetic acid.

3. The high performance liquid chromatography analysis method of montelukast sodium salt according to claim 1, characterized in that: The chromatographic test conditions in step (4) are: using a reverse C18 column, the column temperature is set at 25-35°C, a low concentration phosphate buffer solution at pH 4.0-acetonitrile is used as the mobile phase, the flow rate is 1.0 ml / min-1.5 ml / min, and the detection wavelength is 230 nm-240 nm. Gradient elution.

4. The high performance liquid chromatography analysis method of montelukast sodium salt according to claim 3, characterized in that: The concentration of the phosphate buffer is 15 mM to 25 mM.

5. The high performance liquid chromatography analysis method of montelukast sodium salt according to claim 3, characterized in that: The phosphate is a mixture of one or more of sodium dihydrogen phosphate and potassium dihydrogen phosphate.

6. The high performance liquid chromatography analysis method of montelukast sodium salt according to claim 3, characterized in that: The pH regulator of the low-concentration phosphate buffer with a pH of 4.0 is phosphoric acid.

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