Analysis method for measuring content of dihydroergoline mesylate bulk drug

Abstract

The invention relates to the technical field of drug analysis, and particularly discloses an analysis method for measuring the content of a dihydroergoline mesylate bulk drug, which comprises the following steps: step S1, adding the dihydroergoline mesylate bulk drug into anhydrous acetonitrile to obtain a stock solution to be measured, precisely measuring 1mL of the stock solution, adding a cosolvent, uniformly shaking, and fixing the volume to obtain a solution to be measured; s2, taking a dihydroergoline mesylate raw material medicine, adding the dihydroergoline mesylate raw material medicine into anhydrous acetonitrile to obtain a control stock solution, precisely measuring 1mL of the control stock solution, adding a cosolvent, uniformly shaking, and fixing the volume to obtain a control solution; s3, measuring the content of the dihydroergoline mesylate raw material medicine by using a high performance liquid chromatography; according to the present invention, the cosolvent contains the 1-butyl-3-methylimidazole tosilate, the mobile phase of the high performance liquid chromatography contains the elution reinforcing agent, and through the reasonable ratio of the components, the solubility of the dihydroergoline mesylate can be improved, and the accuracy of the high performance liquid chromatography analysis can be improved.

Description

Technical Field

[0001] The present application relates to the technical field of drug analysis, and more specifically, to an analytical method for determining the content of dihydroergotamine mesylate raw material drug. Background Art

[0002] Ergotamine mesylate is a calcium ion antagonist, which is suitable for the treatment of age-related mental degeneration, Alzheimer's disease, cerebrovascular accident, peripheral vascular disease, and subjective vascular symptoms caused by arterial hypertension.

[0003] In the prior art, triethylamine is often used as the mobile phase to analyze the above-mentioned raw materials. However, triethylamine is easily adsorbed on the silica gel filler and is not easy to be eluted. In addition, its continuous use has a great influence on the column efficiency of the chromatographic column, thereby affecting the accurate determination of the experimental results.

[0004] In addition, when preparing the dihydroergotamine mesylate control solution, dilute hydrochloric acid solution is used as a medium for direct dissolution, which requires long-term shaking and has the problem of inability to completely dissolve.

[0005] Based on the above statements, the present application provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug. Summary of the invention

[0006] In order to solve the problems in the prior art of using high performance liquid chromatography to analyze the content of dihydroergot mesylate, the mobile phase is not easy to elute, which affects the accuracy of the experiment, and there is a problem that dilute hydrochloric acid cannot completely dissolve the dihydroergot mesylate raw material drug, the present application provides an analytical method for determining the content of dihydroergot mesylate raw material drug.

[0007] An analytical method for measuring the content of dihydroergotamine mesylate raw material drug comprises the following steps: Step S1, taking dihydroergotamine mesylate raw material drug and adding it to anhydrous acetonitrile to obtain a stock solution to be tested, accurately measuring 1 mL, adding a cosolvent, shaking well, and fixing the volume to obtain a solution to be tested; Step S2, taking dihydroergotamine mesylate reference substance and adding it to anhydrous acetonitrile to obtain a reference stock solution, accurately measuring 1 mL, adding a cosolvent, shaking well, and fixing the volume to obtain a reference solution; Step S3, using high performance liquid chromatography to determine the content of the dihydroergotamine mesylate bulk drug.

[0008] Preferably, in step S1, the co-solvent is prepared by mixing triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.1-1.5:100:1.2-1.6.

[0009] Preferably, in step S1, the concentration of the stock solution to be tested is 80-100 µg / mL, and the concentration of the solution to be tested is 5-7 µg / mL.

[0010] Preferably, in step S2, the concentration of the control stock solution is 80-100 µg / mL, and the concentration of the control solution is 5-7 µg / mL.

[0011] Preferably, in step S3, the specific process of the high performance liquid chromatography is: after the test solution and the control solution are centrifuged, 20 μL is accurately measured, injected into the high performance liquid chromatography, and the chromatogram is recorded.

[0012] Preferably, the centrifugal treatment is performed at a relative centrifugal force of 15000-20000 g and a centrifugal time of 5-10 min.

[0013] Preferably, the HPLC conditions are: Chromatographic column: octadecylsilane bonded silica gel as filler; Mobile phase: includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is an elution enhancer, and the volume ratio of phase A, phase B and phase C is 60-80:40-45:1.8-2.2.

[0014] Preferably, the elution enhancer is prepared by the following steps: 1-aminopropyl-3-vinylimidazole bistrifluoromethanesulfonyl imide salt and platinum catalyst are added to isopropanol, heated to 30-40°C, stirred evenly, adjusted to pH 8-9, and then dripped with a mixed solution of 1,1,3,3-tetramethyldisiloxane and isopropanol, and the dripping is controlled to be completed within 10 minutes. After the dripping is completed, the temperature is raised to reflux, and the stirring reaction is continued for 6-8 hours. After the reaction is completed, it is cooled to room temperature, rotary evaporated, washed, and dried to obtain an elution enhancer, wherein 1-aminopropyl-3-vinylimidazole bistrifluoromethanesulfonyl The mass ratio of the imide salt, the platinum catalyst, the isopropanol and the mixed liquid a is 6-10:2-3:55-65:30-40. In the mixed liquid a, the mass ratio of 1,1,3,3-tetramethyldisiloxane and the isopropanol is 26-30:1-1.6. In the above reaction process, the isopropanol is used as a solvent, and the unsaturated double bonds on the 1-aminopropyl-3-vinylimidazole bistrifluoromethanesulfonyl imide salt and the active hydrogen on the 1,1,3,3-tetramethyldisiloxane undergo a silylation reaction to obtain an elution enhancer.

[0015] Preferably, the platinum catalyst is an isopropanol solution of chloroplatinic acid with a mass fraction of 1%.

[0016] In summary, the present application has the following beneficial effects: the analytical method for the content of the dihydroergotamine mesylate raw material drug used in the present invention adopts triethylamine, deionized water and 1-butyl-3-methylimidazole toluenesulfonate in a certain ratio to prepare a cosolvent, the presence of triethylamine in the cosolvent can effectively regulate and increase the overall polarity of the cosolvent, improve the solubility of dihydroergotamine mesylate in the cosolvent, the presence of 1-butyl-3-methylimidazole toluenesulfonate can not only effectively dissolve excess triethylamine, but also contain hydrophilic sulfonic acid groups thereon can provide good The acidic environment promotes the dissolution of ergotamine mesylate. At the same time, the imidazole group with a polar structure on 1-butyl-3-methylimidazole toluenesulfonate can further promote the dissolution of dihydroergotamine mesylate. In addition, the present application creatively replaces the commonly used alkaline regulator triethylamine with an elution enhancer. The elution enhancer not only has a certain alkalinity itself and can play a role in replacing triethylamine, but also has low-polarity alkyl chains and siloxane groups bonded to the imidazole groups thereon, which reduces the overall polarity, making it easier to be eluted, and can further improve the accuracy of high performance liquid chromatography analysis. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for describing the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0018] Figure 1 is a chromatogram of the solution to be tested in Example 1 of the present invention; Figure 2 It is the chromatogram of the solution to be tested in Comparative Example 1 of the present invention. DETAILED DESCRIPTION

[0019] In order to make the implementation methods of the present application easier to understand, the present application will be described in detail below in conjunction with specific examples. These examples are only for illustration and are not limited to the scope of application of the present application.

[0020] The contents of the main raw materials and their components used in the examples are as follows: The dihydroergotamine mesylate raw material is produced by TEVA Czech Industrial Co., Ltd., and the approval number of the product is H20120253. The dihydroergotamine mesylate reference substance is provided by the China Food and Drug Inspection Institute, and the CAS number of the product is 8067-24-1.

[0021] Preparation Examples 1-3 and Comparative Preparation Example 1 provide a method for preparing an elution enhancer.

[0022] Preparation Example 1 The elution enhancer was prepared by the following steps: 1-aminopropyl-3-vinyl imidazole bis trifluoromethanesulfonyl imide salt and platinum catalyst were added to isopropanol, the temperature was raised to 30°C, the speed was controlled to be 500 rpm and stirred for 40 min until uniform, the pH was adjusted to 8 with 0.6M potassium hydroxide aqueous solution, and then the mixed solution a of 1,1,3,3-tetramethyldisiloxane and isopropanol was added dropwise, and the dripping was controlled to be completed within 10 min. After the dripping was completed, the temperature was raised to reflux, the speed was maintained unchanged, and the stirring reaction was continued for 6 h. After the reaction was completed, it was cooled to room temperature , control the rotary evaporation temperature to 63°C to remove isopropanol, wash with anhydrous ethanol and deionized water three times in sequence, and then dry at 50°C to constant weight, wherein the platinum catalyst is an isopropanol solution of chloroplatinic acid with a mass fraction of 1%, the mass ratio of 1-aminopropyl-3-vinylimidazole bistrifluoromethanesulfonyl imide salt, platinum catalyst, isopropanol and mixed solution a is 6:2:30:55, and in the mixed solution a, the mass ratio of 1,1,3,3-tetramethyldisiloxane and isopropanol is 26:1.

[0023] Preparation Example 2 The elution enhancer was prepared by the following steps: 1-aminopropyl-3-vinyl imidazole bis trifluoromethanesulfonyl imide salt and platinum catalyst were added to isopropanol, the temperature was raised to 35°C, the speed was controlled to be 550 rpm and stirred for 35 min until uniform, the pH was adjusted to 8.5 with 0.8 M potassium hydroxide aqueous solution, and then the mixed solution a of 1,1,3,3-tetramethyldisiloxane and isopropanol was added dropwise, and the dripping was controlled within 10 min. After the dripping was completed, the temperature was raised to reflux, the speed was maintained unchanged, and the stirring reaction was continued for 7 h. After the reaction was completed, it was cooled to room temperature and the rotation speed was controlled to be constant. The steaming temperature is 65°C to remove isopropanol, and the mixture is washed with anhydrous ethanol and deionized water for 4 times in sequence, and then dried at 55°C to constant weight to obtain an elution enhancer, wherein the platinum catalyst is an isopropanol solution of chloroplatinic acid with a mass fraction of 1%, the mass ratio of 1-aminopropyl-3-vinylimidazole bistrifluoromethanesulfonyl imide salt, the platinum catalyst, isopropanol and the mixed solution a is 8:2.5:60:35, and in the mixed solution a, the mass ratio of 1,1,3,3-tetramethyldisiloxane and isopropanol is 28:1.3.

[0024] Preparation Example 3 The elution enhancer was prepared by the following steps: 1-aminopropyl-3-vinyl imidazole bis trifluoromethanesulfonyl imide salt and platinum catalyst were added to isopropanol, the temperature was raised to 40°C, the speed was controlled to be 600 rpm and stirred for 30 min until uniform, the pH was adjusted to 9.0 with 1.0 M potassium hydroxide aqueous solution, and then the mixed solution a of 1,1,3,3-tetramethyldisiloxane and isopropanol was added dropwise, and the dripping was controlled within 10 min. After the dripping was completed, the temperature was raised to reflux, the speed was maintained unchanged, and the stirring reaction was continued for 8 h. After the reaction was completed, it was cooled to room temperature and the rotation speed was controlled to be constant. The steaming temperature is 67°C to remove isopropanol, and the mixture is washed with anhydrous ethanol and deionized water for 5 times in sequence, and then dried at 60°C to constant weight to obtain an elution enhancer, wherein the platinum catalyst is an isopropanol solution of chloroplatinic acid with a mass fraction of 1%, the mass ratio of 1-aminopropyl-3-vinylimidazole bistrifluoromethanesulfonyl imide salt, the platinum catalyst, isopropanol and the mixed solution a is 10:3:65:40, and in the mixed solution a, the mass ratio of 1,1,3,3-tetramethyldisiloxane and isopropanol is 30:1.6.

[0025] Comparative Preparation Example 1 The elution enhancer was prepared by the following steps: 1-Ethyl-3-methylimidazole bis(trifluoromethanesulfonyl)imide salt and platinum catalyst were added to isopropanol, the temperature was raised to 30°C, the speed was controlled to be 500 rpm and stirred for 40 min until uniform, the pH was adjusted to 8 with 0.6M potassium hydroxide aqueous solution, and then the mixed solution a of 1,1,3,3-tetramethyldisiloxane and isopropanol was added dropwise, and the dripping was controlled to be completed within 10 min. After the dripping was completed, the temperature was raised to reflux, the speed was maintained unchanged, and the stirring reaction was continued for 6 h. After the reaction was completed, it was cooled to room temperature , control the rotary evaporation temperature to 63°C to remove isopropanol, wash with anhydrous ethanol and deionized water three times in sequence, and then dry at 50°C to constant weight, wherein the platinum catalyst is an isopropanol solution of chloroplatinic acid with a mass fraction of 1%, the mass ratio of 1-ethyl-3-methylimidazole bistrifluoromethanesulfonyl imide salt, platinum catalyst, isopropanol and mixed solution a is 6:2:55:30, and in the mixed solution a, the mass ratio of 1,1,3,3-tetramethyldisiloxane and isopropanol is 26:1.

[0026] Examples 1-3 and Comparative Examples 1-4 provide an analytical method for determining the content of dihydroergotamine mesylate raw material drug.

[0027] Example 1 The present embodiment provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug, comprising the following steps: Step S1, taking dihydroergotamine mesylate raw material drug and adding it to anhydrous acetonitrile, controlling the speed to be 350rpm and stirring for 20min until uniform, to obtain a stock solution to be tested with a concentration of 80 μg / mL, accurately measuring 1 mL, placing it in a 20 mL volumetric flask, adding a cosolvent, controlling the temperature to be 6°C, shaking well, and fixing the volume to obtain a 5 μg / mL solution to be tested, wherein the cosolvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.1:100:1.2; Step S2, add the dihydroergoline mesylate reference substance to anhydrous acetonitrile, control the speed to 350 rpm and stir for 20 min until uniform, to obtain a reference stock solution with a concentration of 80 µg / mL, accurately measure 1 mL, put it in a 20 mL volumetric flask, add a co-solvent, control the temperature to 6°C, shake well, and adjust the volume to obtain a 5 µg / mL reference solution, wherein the co-solvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.1:100:1.2; Step S3, centrifuge the test solution and the control solution at a centrifugal force of 15000g for 10 minutes, accurately measure 20μL, inject them into the high performance liquid chromatograph in sequence, and record the chromatogram, wherein the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is the elution enhancer prepared in Preparation Example 1, the volume ratio of phase A, phase B and phase C is 60:40:1.8, the flow rate is 1.5mL / min, the detection wavelength is 280mm, and the column temperature is 30°C.

[0028] Example 2 The present embodiment provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug, comprising the following steps: Step S1, taking dihydroergotamine mesylate raw material drug and adding it to anhydrous acetonitrile, stirring for 15 min at a speed of 400 rpm until uniform, to obtain a stock solution with a concentration of 90 µg / mL to be tested, accurately measuring 1 mL, placing it in a 20 mL volumetric flask, adding a cosolvent, controlling the temperature to 7°C, shaking well, and fixing the volume to obtain a 6 µg / mL test solution, wherein the cosolvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.3:100:1.4; Step S2, add the dihydroergotamine mesylate bulk drug reference substance to anhydrous acetonitrile, control the speed to 400 rpm and stir for 15 min until uniform, to obtain a reference stock solution with a concentration of 90 μg / mL, accurately measure 1 mL, put it in a 20 mL volumetric flask, add a cosolvent, control the temperature to 7°C, shake well, and adjust the volume to obtain a 6 μg / mL reference solution, wherein the cosolvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.3:100:1.4; Step S3, centrifuge the test solution and the photo solution at a centrifugal force of 17500g for 8 minutes, accurately measure 20μL, inject them into the high performance liquid chromatograph in sequence, and record the chromatogram, wherein the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is the elution enhancer prepared in Preparation Example 2, the volume ratio of phase A, phase B and phase C is 70:42:2.0, the flow rate is 1.5mL / min, the detection wavelength is 280mm, and the column temperature is 30℃.

[0029] Example 3 The present embodiment provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug, comprising the following steps: Step S1, taking dihydroergotamine mesylate raw material and adding it to anhydrous acetonitrile, stirring for 10 min at a speed of 450 rpm until uniform, to obtain a stock solution to be tested with a concentration of 100 µg / mL, accurately measuring 1 mL, placing it in a 20 mL volumetric flask, adding a cosolvent, controlling the temperature to 8°C, shaking well, and fixing the volume to obtain a 7 µg / mL solution to be tested, wherein the cosolvent is a mixture of triethylamine, deionized water, and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.5:100:1.6; Step S2, add the dihydroergoline mesylate reference substance to anhydrous acetonitrile, control the speed to 450 rpm and stir for 10 min until uniform, to obtain a reference stock solution with a concentration of 100 µg / mL, accurately measure 1 mL, put it in a 20 mL volumetric flask, add a cosolvent, control the temperature to 8°C, shake well, and adjust the volume to obtain a 7 µg / mL reference solution, wherein the cosolvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.5:100:1.6; Step S3, centrifuge the test solution and the control solution at a centrifugal force of 20000g for 5 minutes, accurately measure 20μL, inject them into the high performance liquid chromatograph in sequence, and record the chromatogram, wherein the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is the elution enhancer prepared in Preparation Example 3, the volume ratio of phase A, phase B and phase C is 80:45:2.2, the flow rate is 1.5mL / min, the detection wavelength is 280mm, and the column temperature is 30°C.

[0030] Comparative Example 1 This comparative example provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug, comprising the following steps: Step S1, taking dihydroergotamine mesylate raw material drug and adding it to anhydrous acetonitrile, stirring for 20 min at a speed of 350 rpm until uniform, to obtain a stock solution to be tested with a concentration of 80 µg / mL, accurately measuring 1 mL, placing it in a 20 mL volumetric flask, adding a cosolvent, controlling the temperature to 6°C, shaking well, and fixing the volume to obtain a 5 µg / mL solution to be tested, wherein the cosolvent is a mixture of triethylamine, deionized water and methanesulfonic acid in a mass ratio of 1.1:100:1.2; Step S2, add the dihydroergoline mesylate reference substance to anhydrous acetonitrile, control the speed to 350 rpm and stir for 20 min until uniform, to obtain a reference stock solution with a concentration of 80 µg / mL, accurately measure 1 mL, place it in a 20 mL volumetric flask, add a cosolvent, control the temperature to 6°C, shake well, and dilute to obtain a 5 µg / mL reference solution; Step S3, centrifuge the test solution and the control solution at a centrifugal force of 15000g for 10 minutes, accurately measure 20μL, inject them into the high performance liquid chromatograph in sequence, and record the chromatogram, wherein the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is the elution enhancer prepared in Preparation Example 1, the volume ratio of phase A, phase B and phase C is 60:40:1.8, the flow rate is 1.5mL / min, the detection wavelength is 280mm, and the column temperature is 30°C.

[0031] Comparative Example 2 This comparative example provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug, comprising the following steps: Step S1, taking dihydroergotamine mesylate raw material drug and adding it to anhydrous acetonitrile, controlling the speed to be 350rpm and stirring for 20min until uniform, to obtain a stock solution to be tested with a concentration of 80 μg / mL, accurately measuring 1 mL, placing it in a 20 mL volumetric flask, adding a cosolvent, controlling the temperature to be 6°C, shaking well, and fixing the volume to obtain a 5 μg / mL test solution, wherein the cosolvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole acetate in a mass ratio of 1.1:100:1.2; Step S2, add the dihydroergotamine mesylate reference substance to anhydrous acetonitrile, control the speed to 350 rpm and stir for 20 min until uniform, to obtain a reference stock solution with a concentration of 80 µg / mL, accurately measure 1 mL, put it in a 20 mL volumetric flask, add a co-solvent, control the temperature to 6°C, shake well, and adjust the volume to obtain a 5 µg / mL reference solution, wherein the co-solvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazolium acetate in a mass ratio of 1.1:100:1.2; Step S3, centrifuge the test solution and the control solution at a centrifugal force of 15000g for 10 minutes, accurately measure 20μL, inject them into the high performance liquid chromatograph in sequence, and record the chromatogram, wherein the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is the elution enhancer prepared in Preparation Example 1, the volume ratio of phase A, phase B and phase C is 60:40:1.8, the flow rate is 1.5mL / min, the detection wavelength is 280mm, and the column temperature is 30°C.

[0032] Comparative Example 3 This comparative example provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug, comprising the following steps: Step S1, taking dihydroergotamine mesylate raw material drug and adding it to anhydrous acetonitrile, controlling the speed to be 350rpm and stirring for 20min until uniform, to obtain a stock solution to be tested with a concentration of 80 μg / mL, accurately measuring 1 mL, placing it in a 20 mL volumetric flask, adding a cosolvent, controlling the temperature to be 6°C, shaking well, and fixing the volume to obtain a 5 μg / mL solution to be tested, wherein the cosolvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.1:100:1.2; Step S2, add the dihydroergoline mesylate reference substance to anhydrous acetonitrile, control the speed to 350 rpm and stir for 20 min until uniform, to obtain a reference stock solution with a concentration of 80 µg / mL, accurately measure 1 mL, put it in a 20 mL volumetric flask, add a co-solvent, control the temperature to 6°C, shake well, and adjust the volume to obtain a 5 µg / mL reference solution, wherein the co-solvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.1:100:1.2; Step S3, centrifuge the test solution and the control solution at a centrifugal force of 15000g for 10 minutes, accurately measure 20µL, inject them into the high performance liquid chromatograph in sequence, and record the chromatogram, wherein the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is triethylamine, the volume ratio of phase A, phase B and phase C is 60:40:1.8, the flow rate is 1.5mL / min, the detection wavelength is 280mm, and the column temperature is 30°C.

[0033] Comparative Example 4 This comparative example provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug, comprising the following steps: Step S1, taking dihydroergotamine mesylate raw material drug and adding it to anhydrous acetonitrile, controlling the speed to be 350rpm and stirring for 20min until uniform, to obtain a stock solution to be tested with a concentration of 80 μg / mL, accurately measuring 1 mL, placing it in a 20 mL volumetric flask, adding a cosolvent, controlling the temperature to be 6°C, shaking well, and fixing the volume to obtain a 5 μg / mL solution to be tested, wherein the cosolvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.1:100:1.2; Step S2, add the dihydroergoline mesylate reference substance to anhydrous acetonitrile, control the speed to 350 rpm and stir for 20 min until uniform, to obtain a reference stock solution with a concentration of 80 µg / mL, accurately measure 1 mL, put it in a 20 mL volumetric flask, add a co-solvent, control the temperature to 6°C, shake well, and adjust the volume to obtain a 5 µg / mL reference solution, wherein the co-solvent is a mixture of triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.1:100:1.2; Step S3, centrifuge the test solution and the control solution at a centrifugal force of 15000g for 10 minutes, accurately measure 20μL, inject them into the high performance liquid chromatograph in sequence, and record the chromatogram, wherein the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is the elution enhancer prepared in comparative preparation example 1, the volume ratio of phase A, phase B and phase C is 60:40:1.8, the flow rate is 1.5mL / min, the detection wavelength is 280mm, and the column temperature is 30°C.

[0034] Comparative Example 5 This comparative example provides an analytical method for determining the content of dihydroergotamine mesylate raw material drug, comprising the following steps: Step S1, take dihydroergotamine mesylate raw material and add it to anhydrous acetonitrile, control the speed to 350rpm and stir for 20min until uniform, to obtain a stock solution with a concentration of 80 μg / mL to be tested, accurately measure 1 mL, put it in a 20 mL volumetric flask, add a hydrochloric acid solution with a volume fraction of 0.6%, control the temperature to 6°C, shake well, and dilute to obtain a 5 μg / mL test solution; Step S2, add the dihydroergotamine mesylate reference substance to anhydrous acetonitrile, control the speed to 350 rpm and stir for 20 min until uniform, to obtain a reference stock solution with a concentration of 80 μg / mL, accurately measure 1 mL, put it in a 20 mL volumetric flask, add a 0.6% hydrochloric acid solution by volume, control the temperature to 6°C, shake well, and dilute to obtain a 5 μg / mL reference solution; Step S3, centrifuge the test solution and the control solution at a centrifugal force of 15000g for 10 minutes, accurately measure 20μL, inject them into the high performance liquid chromatograph in sequence, and record the chromatogram, wherein the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is the elution enhancer prepared in Preparation Example 1, the volume ratio of phase A, phase B and phase C is 60:40:1.8, the flow rate is 1.5mL / min, the detection wavelength is 280mm, and the column temperature is 30°C.

[0035] Performance Testing Using the above-mentioned test method, the dihydroergoline sulfonate raw materials in Examples 5-7 and Comparative Examples 1-5 were tested in turn by high performance liquid chromatography, and the specific test data are shown in Table 1 below: Table 1 Chromatographic analysis parameters of each component of dihydroergotamine mesylate raw material drug in Examples 1-3 and Comparative Examples 1-5

[0036]

[0037] Note: Component 1 is dihydroergocriptine mesylate, component 2 is α-dihydroergocryptine mesylate, component 3 is dihydroergocriptine mesylate, and component 4 is β-dihydroergocryptine mesylate.

[0038] As can be seen from Table 1, the minimum separation degree between the components in the table is greater than 1.5, the tailing factor is less than 1.09, and the number of theoretical plates is greater than 10000. Therefore, the chromatographic system can effectively separate the four components in the dihydroergoline mesylate bulk drug, meeting the relevant requirements of component ratio inspection and separation degree. At the same time, compared with Comparative Examples 1-5, the number of theoretical plates between the components in the chromatographic analysis method of Example 1-3 is larger, the tailing factor is smaller, and the separation degree is greater, which shows that the separation effect of dihydroergocorin mesylate, dihydroergoline mesylate and dihydroergocryptine mesylate (α type and β type) obtained by the determination method of the content of dihydroergoline mesylate bulk drug in Example 1-3 is better than that of Comparative Examples 1-5, the separation accuracy is higher, and the method reliability is higher.

[0039] by Figure 1 and Figure 2 For example, from the figure, it can be seen that compared with Figure 2 , Figure 1 The peak distances between the components are wider and the peak shapes are sharper, which indicates that Example 1 has a better separation effect than Comparative Example 1.

[0040] The results of the above test can be explained as follows: compared with Comparative Examples 1-2, the synergy of the imidazole group and the sulfonic acid group contained in the co-solvent in the embodiment improves the accuracy of component determination. At the same time, compared with Comparative Examples 3 and 4, the elution enhancer in the embodiment can not only replace triethylamine well, but also the alkyl chain and siloxane group thereon reduce the overall polarity of the system, making the results of component determination more accurate.

[0041] Repeatability test: Take dihydroergotamine mesylate reference substance and add it to anhydrous acetonitrile, control the speed to 350rpm and stir for 20min until uniform, to obtain 5 μg / mL reference substance solution, refer to the chromatographic conditions of Example 1, accurately measure 50μL of reference substance solution and inject it into the liquid chromatograph, inject continuously 6 times, the results are shown in Table 2; Table 2 Repeatability test results

[0042] Note: Component 1 is dihydroergocriptine mesylate, component 2 is α-dihydroergocryptine mesylate, component 3 is dihydroergocriptine mesylate, and component 4 is β-dihydroergocryptine mesylate.

[0043] It can be seen from Table 2 that the peak area of ​​each component in the table has good repeatability, which proves that the detection method in the embodiment of the present invention has good reliability and repeatability.

[0044] The cosolvents in Examples 1-3 and Comparative Examples 1-2 were used as dissolution media, respectively. The dihydroergotamine mesylate bulk drug was stirred at a speed of 100 rpm for 120 min using a paddle method. The dissolution of the dihydroergotamine mesylate bulk drug was determined, and the specific test results are shown in Table 3 below. Table 3 Performance test of solubility of co-solvent in Examples 1-3 and Comparative Examples 1-2

[0045] As shown in Table 3 above, compared with Comparative Examples 1-2, the solubility of the cosolvents in Examples 1-3 of the present application to the dihydroergoline mesylate bulk drug is more significant. By comparing Comparative Examples 1-2 with Example 1, it can be seen that the synergy of the imidazole group and the sulfonic acid group in 1-butyl-3-methylimidazole toluenesulfonate in the cosolvent in the embodiment can promote the dissolution of the dihydroergoline mesylate bulk drug.

[0046] This specific embodiment is only an explanation of the present invention, and it is not a limitation of the present invention. After reading this specification, those skilled in the art can make non-creative modifications to this embodiment as needed, but as long as it is within the scope of the claims of the present invention, it is protected by the patent law.

Claims

1. An analytical method for measuring the content of dihydroergoline mesylate bulk drug, characterized in that, The following steps are involved: Step S1, taking dihydroergotamine mesylate raw material and adding it to anhydrous acetonitrile to obtain a stock solution to be tested, accurately measuring 1 mL, adding a co-solvent, shaking well, and fixing the volume to obtain a solution to be tested; Step S2, taking dihydroergotamine mesylate reference substance and adding it to anhydrous acetonitrile to obtain a reference stock solution, accurately measuring 1 mL, adding a co-solvent, shaking well, and fixing the volume to obtain a reference solution; Step S3, using high performance liquid chromatography to determine the content of dihydroergotamine mesylate bulk drug; The auxiliary solvent is prepared by mixing triethylamine, deionized water and 1-butyl-3-methylimidazole p-toluenesulfonate in a mass ratio of 1.1-1.5:100:1.2-1.

6.

2. The analytical method for measuring the content of dihydroergoline mesylate bulk drug according to claim 1, characterized in that, In step S1, the concentration of the stock solution to be tested is 80-100 µg / mL, and the concentration of the solution to be tested is 5-7 µg / mL.

3. A method for analyzing the content of dihydroergoline mesylate bulk drug according to claim 1, characterized in that: In step S2, the concentration of the control stock solution is 80-100 µg / mL, and the concentration of the control solution is 5-7 µg / mL.

4. The analytical method for measuring the content of dihydroergoline mesylate bulk drug according to claim 1, characterized in that, In step S3, the specific process of the high performance liquid chromatography is: after the test solution and the control solution are centrifuged, 20 μL are accurately measured and injected into the high performance liquid chromatograph in sequence, and the chromatogram is recorded.

5. The analytical method for measuring the content of dihydroergoline mesylate bulk drug according to claim 4, characterized in that, The centrifugal treatment conditions are: relative centrifugal force of 15000-20000g, and centrifugal time of 5-10min.

6. The analytical method for measuring the content of dihydroergoline mesylate bulk drug according to claim 1, characterized in that, The mobile phase includes phase A, phase B and phase C, phase A is water, phase B is anhydrous acetonitrile, phase C is an elution enhancer, and the volume ratio of phase A, phase B and phase C is 60-80:40-45:1.8-2.

2.

7. The analytical method for measuring the content of dihydroergoline mesylate bulk drug according to claim 6, characterized in that, The elution enhancer was prepared by the following steps: Add 1-aminopropyl-3-vinylimidazole bistrifluoromethanesulfonyl imide salt and platinum catalyst to isopropanol, heat to 30-40°C, stir evenly, adjust pH to 8-9, then add a mixed solution a of 1,1,3,3-tetramethyldisiloxane and isopropanol dropwise, and control the dripping to be completed within 10 minutes. After the dripping is completed, heat to reflux, continue to stir and react for 6-8 hours. After the reaction is completed, cool to room temperature, rotary evaporate, wash, and dry to obtain an elution enhancer.

8. The analytical method for measuring the content of dihydroergoline mesylate bulk drug according to claim 7, characterized in that, The mass ratio of the 1-aminopropyl-3-vinyl imidazole bistrifluoromethanesulfonyl imide salt, the platinum catalyst, isopropanol and the mixed solution a is 6-10:2-3:55-65:30-40. In the mixed solution a, the mass ratio of 1,1,3,3-tetramethyldisiloxane and isopropanol is 26-30:1-1.

6.

9. The analytical method for measuring the content of dihydroergoline mesylate bulk drug according to claim 1, characterized in that, In the high performance liquid chromatography, the flow rate of the mobile phase is 1.5 mL / min, the detection wavelength is 280 mm, and the column temperature is 30°C.

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