Method for detecting related substances of clemastine fumarate raw material medicine

Abstract

The invention relates to the technical field of medicine analysis, in particular to a method for detecting related substances of a clemastine fumarate raw material medicine. According to the method, high performance liquid chromatography is adopted for detection, a chromatographic column with octadecylsilane chemically bonded silica as a filler is selected, 0.02 mol / L dipotassium phosphate buffer solution (the pH value is adjusted to 8.1 with phosphoric acid) is used as a mobile phase A, acetonitrile is used as a mobile phase B, and gradient elution is performed. By adopting the method disclosed by the invention, the related substances in the clomastine fumarate can be effectively detected, and the method has the advantages of strong specificity, high sensitivity and the like, has good detection capability on specific impurities in the clomastine fumarate, and can effectively guarantee the quality of the clomastine fumarate.

Description

Technical Field

[0001] The invention belongs to the field of drug analysis, and particularly relates to a method for detecting related substances of clemastine fumarate raw material medicine. Background Art

[0002] Clmastine fumarate is an H1 receptor antagonist that selectively acts on H1 receptors, preventing histamine from acting on target cells and inhibiting capillary permeability. By inhibiting the release of histamine, it has an anti-allergic effect and has sedative, antipruritic, antihistamine, anti-allergic, and antipyretic effects. Common dosage forms of clemastine fumarate include clemastine fumarate tablets and clemastine fumarate injection.

[0003] The chemical name of clemastine fumarate is [ R -( R *, R *)]-1-methyl-2-[2-[1-(4-chlorophenyl)-1-phenylethoxy]ethyl]pyrrolidine ( E )-2-butenedioate, the structural formula is as follows:

[0004] Currently, pharmacopoeias in various countries include methods for detecting related substances in clemastine fumarate API. However, these methods are unable to effectively separate the process impurities and degradation impurities in this product, necessitating the development of a method for detecting related substances in clemastine fumarate API. The detection method provided by the present invention can effectively separate the process impurities and degradation impurities in clemastine fumarate API, and has good method specificity, high sensitivity, and simple operation. Summary of the Invention

[0005] The present invention aims to provide a method for analyzing related substances in a clemastine fumarate bulk drug. The method can effectively detect and separate impurities in the clemastine fumarate bulk drug, and is used for quality control of the clemastine fumarate bulk drug.

[0006] The objectives of the present invention are achieved through the following technical solutions.

[0007] A method for detecting related substances in a clemastine fumarate raw material drug comprises the following steps.

[0008] (1) Prepare the solution.

[0009] Diluent: water-acetonitrile (40:60).

[0010] Test solution: Take an appropriate amount of clemastine fumarate bulk drug, dissolve it in solvent and dilute it to make the test solution.

[0011] Control solution: Accurately measure an appropriate amount of the test solution and quantitatively dilute it with solvent to prepare a control solution.

[0012] Sensitivity solution: Accurately measure an appropriate amount of control solution and quantitatively dilute it with solvent to prepare the sensitivity solution.

[0013] Fumaric acid localization solution: Take an appropriate amount of fumaric acid reference substance, dissolve it in solvent and dilute it to make fumaric acid localization solution.

[0014] System suitability solution: Take appropriate amount of clemastine fumarate reference substance and each impurity reference substance, dissolve and dilute with solvent to prepare system suitability solution.

[0015] (2) The chromatographic conditions are as follows.

[0016] Chromatographic column: Octadecylsilane bonded silica gel is used as filler.

[0017] Mobile phase A: potassium hydrogen phosphate buffer.

[0018] Mobile phase B: acetonitrile.

[0019] Detection wavelength: 220~230nm.

[0020] Flow rate: 0.8~1.0ml / min.

[0021] Column temperature: 40~50℃.

[0022] Injection volume: 80~100µl.

[0023] The concentration of the test solution in step (1) is 0.2 mg / ml; the concentration of the control solution is 0.2 μg / ml; the concentration of the sensitivity solution is 0.04 μg / ml; the concentration of the fumaric acid solution is 0.05 mg / ml; the concentration of the main component of the system suitability solution is 0.2 mg / ml, and the impurity concentration is 2 μg / ml.

[0024] The mobile phase A in step (2) is 0.02 mol / L potassium hydrogen phosphate buffer, and the pH value is adjusted to 8.1±0.2 with phosphoric acid.

[0025] The column temperature in step (2) is 45°C.

[0026] The flow rate in step (2) is 0.9 ml / min.

[0027] The wavelength in step (2) is 225 nm.

[0028] The wavelength in step (2) is 225 nm.

[0029] The elution method in step (2) is gradient elution.

[0030] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0 55 45 10 55 45 15 35 65 50 35 65 51 55 45 60 55 45

[0031] The method for detecting related substances in the clemastine fumarate raw material drug provided by the present invention has the advantages of strong specificity, high sensitivity, and the like, and can effectively detect the related substances of clemastine fumarate, thereby effectively ensuring the quality of clemastine fumarate. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is the solution chromatogram of the system suitability of Example 1.

[0033] Figure 2 This is a chromatogram of the test solution of Example 1.

[0034] Figure 3 This is the chromatogram of the test solution of Example 2.

[0035] Figure 4 The chromatogram is compared with that in Example 3.

[0036] Figure 5 The chromatogram is compared with that in Example 4. DETAILED DESCRIPTION

[0037] The present invention will be further described in detail below with reference to examples, but the embodiments of the present invention are not limited thereto.

[0038] Example 1

[0039] Detection of related substances in clemastine fumarate raw material.

[0040] (1) Chromatographic conditions.

[0041] Chromatographic column: Waters XBridge C18, 4.6 mm × 250 mm, 5 μm.

[0042] Detection wavelength: 225nm.

[0043] Flow rate: 0.9ml / min.

[0044] Column temperature: 45℃.

[0045] Injection volume: 90µl.

[0046] Mobile phase A: 0.02 mol / L potassium dihydrogen phosphate buffer (pH adjusted to 8.1 with phosphoric acid).

[0047] Mobile phase B: acetonitrile.

[0048] Elution method: gradient elution.

[0049] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0 55 45 10 55 45 15 35 65 50 35 65 51 55 45 60 55 45

[0050] (2) Solution preparation.

[0051] Diluent: water-acetonitrile (40:60).

[0052] Test solution: Take an appropriate amount of the product, dissolve it in solvent and dilute it to make a solution containing approximately 0.2 mg per 1 ml.

[0053] Control solution: Accurately measure an appropriate amount of the test solution and quantitatively dilute it with solvent to make a solution containing approximately 0.2 μg per 1 ml.

[0054] Sensitivity solution: Accurately measure an appropriate amount of control solution and quantitatively dilute it with solvent to make a solution containing approximately 0.04 μg per 1 ml.

[0055] Fumaric acid reference solution: Take an appropriate amount of fumaric acid reference substance, accurately weigh it, dissolve it in solvent and quantitatively dilute it to make a solution containing approximately 0.05 mg per 1 ml.

[0056] Take appropriate amounts of impurity STE0038-02, impurity STE0058, impurity STE0061, impurity STE0072, impurity STE0085 and impurity STE0087 reference substances, accurately weigh them, dissolve them in acetonitrile and quantitatively dilute them to prepare solutions containing approximately 0.2 mg per 1 ml as stock solutions of each impurity; take appropriate amounts of clemastine fumarate, accurately weigh them, add appropriate amounts of solvent to dissolve them, then accurately measure appropriate amounts of the above impurity stock solutions, dilute them with solvent to prepare mixed solutions containing approximately 0.2 mg of clemastine fumarate and 2 μg of each impurity per 1 ml, and shake well.

[0057] Determination method.

[0058] Accurately measure the test solution and control solution, inject them into the liquid chromatograph respectively, and record the chromatogram; calculate according to the principal component self-control method with the addition of correction factor.

[0059] Test results: System suitability diagram is attached Figure 1 ; See the attached sample solution Figure 2 .

[0060] Element Positioning solution System suitability solution System suitability solution Element Retention time (min) Retention time (min) Separation degree from adjacent impurities STE0087 8.674 8.672 / STE0061 11.929 11.932 6.2 STE0072 17.652 17.662 15.4 STE0058 19.646 19.647 8.2 STE0038-02 21.294 21.394 6.2 STE0085 24.986 25.224 9.6 Clemastine 26.996 27.002 3.9

[0061] The results showed that the blank solution did not interfere with the detection of related substances of this product, the minimum separation between impurity peaks and between impurities and the main peak was 3.9, and the separation was good; in the sensitivity solution chromatogram, the signal-to-noise ratio of the main component peak height was 19.1>10, which met the requirements; only the known impurity STE0085 was detected in the test solution, with a content of 0.04%, and the maximum unknown single impurity was 0.02%, which met the regulations.

[0062] Example 2

[0063] Detection of related substances in clemastine fumarate raw material (influencing factor 60℃30 days).

[0064] The chromatographic conditions were the same as in Example 1.

[0065] The solution preparation was the same as in Example 1.

[0066] The results showed that: only the known impurity STE0085 was detected in the test solution of clemastine fumarate raw material (affecting factor 60℃30 days), with a content of 0.05%, and the maximum unknown single impurity was 0.03%, which was in compliance with the regulations; the test solution is attached Figure 3 .

[0067] Example 3

[0068] Comparative Examples 1-3: The chromatographic conditions of each comparative example are basically the same as those in Example 1, except that the pH value of the mobile phase is different; the solution preparation is the same as that in Example 1.

[0069] The pH value of the mobile phase in Comparative Example 1 was 8.1.

[0070] The pH value of the mobile phase in Comparative Example 2 was 7.9.

[0071] The pH value of the mobile phase in Comparative Example 3 was 8.3.

[0072] By the attached Figure 4 It can be seen that the pH value of the mobile phase has a great influence on the peak elution of impurities STE0038-02, impurity STE0085 and clemastine. When the pH value of the mobile phase is 7.9, the separation degree of impurity STE0085 and clemastine becomes larger; when the pH value of the mobile phase is 8.3, the separation degree of impurity STE0085 and clemastine becomes smaller; therefore, the pH value of the mobile phase is controlled in the range of 7.9~8.3.

[0073] Example 4

[0074] Comparative Examples 1-3: The chromatographic conditions of each comparative example are basically the same as those of Example 1, except that the column temperature is different; the solution preparation is the same as that of Example 1.

[0075] The column temperature of Comparative Example 1 was 45°C.

[0076] The column temperature of Comparative Example 2 was 43°C.

[0077] The column temperature of Comparative Example 3 was 47°C.

[0078] By the attached Figure 5 It can be seen that the column temperature has a great influence on the peak elution of impurities STE0085 and clemastine. When the column temperature is 43°C, the separation degree of impurities STE0085 and clemastine becomes larger; when the column temperature is 47°C, the separation degree of impurities STE0085 and clemastine becomes smaller. At the same time, considering the separation of other known impurities and unknown impurities, the column temperature is controlled in the range of 43°C~47°C.

Claims

1. A method for determining related substances in clemastine fumarate raw material, characterized in that: The following steps are included: (1) Prepare the solution as follows: Diluent: water-acetonitrile (40:60) Test solution: Take an appropriate amount of clemastine fumarate bulk drug, dissolve it in solvent and dilute it to make the test solution; Control solution: Accurately measure an appropriate amount of the test solution and dilute it with a solvent to prepare a control solution; Sensitivity solution: Accurately measure an appropriate amount of control solution and quantitatively dilute it with solvent to prepare sensitivity solution; Fumaric acid localization solution: Take an appropriate amount of fumaric acid reference substance, dissolve it in solvent and dilute it to make fumaric acid localization solution; System suitability solution: Take appropriate amount of clemastine fumarate reference substance and each impurity reference substance, dissolve and dilute with solvent to prepare system suitability solution.

2. (2) Chromatographic conditions are as follows: Chromatographic column: octadecylsilane bonded silica gel as filler; Mobile phase A: dipotassium hydrogen phosphate buffer; Mobile phase B: acetonitrile; Detection wavelength: 220~230nm; Flow rate: 0.8~1.0ml / min; Column temperature: 40~50℃; Injection volume: 80~100µl.

3. The method according to claim 1, characterized in that The concentration of the test solution in step (1) is 0.2 mg / ml; the concentration of the control solution is 0.2 μg / ml; the concentration of the sensitivity solution is 0.04 μg / ml; the concentration of the fumaric acid solution is 0.05 mg / ml; the concentration of the main component of the system suitability solution is 0.2 mg / ml, and the impurity concentration is 2 μg / ml.

4. The method according to claim 1, characterized in that The mobile phase A in step (2) is 0.02 mol / L potassium hydrogen phosphate buffer, and the pH value is adjusted to 8.1±0.2 with phosphoric acid.

5. The method according to claim 1, characterized in that The column temperature in step (2) is 45°C.

6. The method according to claim 1, characterized in that The flow rate in step (2) is 0.9 ml / min.

7. The method according to claim 1, characterized in that The wavelength in step (2) is 225 nm.

8. The method according to claim 1, characterized in that The chromatographic column in step (2) is Waters XBridge C18, 4.6 mm × 250 mm, 5 μm.

9. The method according to claim 1, characterized in that The elution method in step (2) is gradient elution. 10.

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