Determination method for in-vitro dissolution curve of febuxostat tablet
By using hydrochloric acid solution (pH 1.2) and high-performance liquid chromatography in the dissolution determination of febulista tablets, combined with the dilution step of phosphate buffer solution, the problem that the existing methods cannot distinguish between different sources of febulista tablets was solved, and effective evaluation of the quality of generic drugs and consistency with the original preparation was achieved.
Patent Information
- Application Number
- CN202510167970.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-14
- Publication Date
- 2025-06-20
AI Technical Summary
The existing dissolution method of febulista tablets cannot effectively distinguish febulista tablets from different sources, and cannot accurately evaluate the quality of generic drugs and consistency with the original preparation.
Hydrochloric acid solution (pH 1.2) was used as the dissolution medium, and the dissolution amount was performed by dissolution meter paddle method, and the dissolution amount was determined by high-performance liquid chromatography. Combined with the two-step dilution step of phosphate buffer solution, the in vitro dissolution curve of febusterstat tablets was obtained.
This method can effectively distinguish febulista tablets from different sources, improve the quality of generic drugs and the evaluation ability of consistency with the original preparation, and solve the problem that existing methods cannot distinguish the inherent quality of the products.
Smart Images
Figure CN120177645A_ABST
Abstract
Description
Technical Field
[0001] The present invention discloses a technique in the field of pharmaceutical detection, and relates to a method for determining the in vitro dissolution curve of febuxostat tablets. Background Art
[0002] Hyperuricemia is the cause of various symptoms caused by urate deposition represented by gout. The higher the serum uric acid value, the higher the incidence of gouty arthritis, and it will cause kidney damage. Therefore, various symptoms caused by hyperuricemia should be eliminated early to prevent recurrence and disease progression. In recent years, as one of the lifestyle diseases, there have been more and more reports suggesting its association with hypertension and hyperlipidemia. It is a marker of metabolic syndrome and may also be a risk factor for arteriosclerosis. Therefore, the importance of its treatment is increasing. The treatment of hyperuricemia mainly focuses on reducing uric acid.
[0003] Like allopurinol, febuxostat is also a uric acid-lowering drug that inhibits uric acid production. However, its molecular structure is different from that of allopurinol and purine. It can bind to xanthine oxidase through a non-competitive mechanism and inhibit the activity of xanthine oxidase, thereby achieving the purpose of inhibiting uric acid production. Febuxostat is a specific xanthine oxidase inhibitor that inhibits not only the reduced form but also the oxidized form of xanthine oxidase. Therefore, a smaller dose can exert a greater xanthine oxidase inhibitory effect.
[0004] Febuxostat was developed by Teijin of Japan and was launched in the United States in February 2009 under the trade name ULORIC. The license holder is TAKEDA PHARMACEUTICALS USA INC, and the specifications are 40 mg and 80 mg.
[0005] Dissolution Profile: A dissolution rate-time curve plotted by the dissolution rates obtained at different time points of a preparation. The dissolution curve reflects the in vitro dissolution and dissolution process of a preparation, and the quality of a generic drug can be evaluated by comparing the similarity of the in vitro dissolution curves of the generic preparation and the original preparation.
[0006] Currently, there is little research on the dissolution rate of febuxostat tablets. For the existing dissolution rate method of febuxostat tablets, the dissolution rate of febuxostat tablets is determined by ultraviolet spectrophotometry. This method uses a pH 6.0 buffer solution as the dissolution medium, and the dissolution rate can reach more than 80% at 30 minutes. Referring to the requirements of the "Guidelines for Determination and Comparison of Dissolution Curves of Ordinary Oral Solid Preparations" and the "Q&A on Dissolution Curve Research in the 'Technical Guidelines for Research on Pharmaceutical Changes of Approved Chemical Drugs (Trial)'", this method cannot obtain the dissolution behavior of febuxostat tablets from different sources and distinguish the intrinsic quality of the products. Summary of the Invention
[0007] In view of this, the present application provides a method for determining the in vitro dissolution curve of febuxostat tablets, including the following specific steps: Step 1: Take febuxostat tablets, use hydrochloric acid solution as the dissolution medium, and the pH of the hydrochloric acid solution is 1.2. Perform dissolution according to the paddle method of the dissolution apparatus. At different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with phosphate buffer solution, shake well, and use it as the test sample solution; Step 2: Take febuxostat reference substance, place it in a volumetric flask, dissolve and dilute to the scale with phosphate buffer solution, shake well, accurately measure an appropriate amount, place it in a volumetric flask, add phosphate buffer solution to dilute to the scale, shake well, and use it as the reference substance solution. And the pH of the phosphate buffer solution in this step is 6.8; Step 3: Use high performance liquid chromatography, inject the test sample solution and the reference substance solution into the high performance liquid chromatography respectively, record the chromatogram, and calculate the dissolution amount.
[0008] According to one aspect of the present application, take febuxostat tablets, use hydrochloric acid solution as the dissolution medium, perform dissolution according to the paddle method of the dissolution apparatus, the amount of hydrochloric acid solution added is 900 ml, and the rotation speed of the paddle method of the dissolution apparatus is 50 revolutions per minute.
[0009] According to one aspect of the present application, at different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with phosphate buffer solution, shake well, and use it as the test sample solution, and the sampling time points are 5 min - 30 min.
[0010] According to one aspect of the present application, take 0.1 ml of the dissolution solution and add 0.9 ml of phosphate buffer solution to use it as the test sample solution.
[0011] According to one aspect of the present application, after taking out the dissolution solution, add the same temperature and the same volume of hydrochloric acid solution.
[0012] According to one aspect of the present application, take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a volumetric flask, dissolve and dilute to the scale with phosphate buffer solution, shake well, accurately measure 5 ml, place it in a volumetric flask, add phosphate buffer solution to dilute to the scale, and shake well.
[0013] According to one aspect of the present application, take febuxostat tablets, use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, perform dissolution according to the paddle method of the dissolution apparatus, the rotation speed of the paddle method is 50 revolutions per minute. At different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with phosphate buffer solution, shake well, and use it as the test sample solution, and the pH of the phosphate buffer solution is 6.0 - 8.0.
[0014] According to one aspect of the present application, the phosphate buffer solution is prepared as follows: Take febuxostat tablets, use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, perform dissolution according to the paddle method of the dissolution apparatus, the rotation speed of the paddle method is 50 revolutions per minute. At different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with the phosphate buffer solution, and shake well to obtain the test solution. Among them, the phosphate buffer solution in the measurement step can also be an acetate buffer solution, and the pH of the acetate buffer solution is 4.5 - 5.5.
[0015] According to one aspect of the present application, the chromatographic conditions of the high-performance liquid chromatography are as follows: The chromatographic column is filled with octadecylsilane-bonded silica gel, the detector is a VWD detector, and the detection wavelength is 317 nm.
[0016] According to one aspect of the present application, the mobile phase is a combination of acetonitrile, water and glacial acetic acid, the column temperature is 40 °C; the flow rate is 2.0 ml / min, the running time is 7 min, and the sample volume is 20 μL.
[0017] The beneficial effects of the present invention: The following specific steps are included: Step 1: Take febuxostat tablets, use hydrochloric acid solution as the dissolution medium, and the pH of the hydrochloric acid solution is 1.2. Perform dissolution according to the paddle method of the dissolution apparatus. At different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with the phosphate buffer solution, and shake well to obtain the test solution; Step 2: Take a febuxostat reference substance, place it in a volumetric flask, dissolve and dilute it to the scale with the phosphate buffer solution, shake well, accurately measure an appropriate amount, place it in a volumetric flask, add the phosphate buffer solution to dilute it to the scale, and shake well to obtain the reference solution. The pH of the phosphate buffer solution in this step is 6.8; Step 3: Use high-performance liquid chromatography, inject the test solution and the reference solution into the high-performance liquid chromatography respectively, record the chromatogram, and calculate the dissolution amount; Through the dissolution results of the raw material drug, combined with the in vitro-in vivo correlation, select the hydrochloric acid solution with a pH of 1.2 as the dissolution medium. The obtained curve has excellent discrimination ability, which is of great significance for guiding the research and development of febuxostat tablet generic preparations and evaluating the quality and efficacy consistency between the preparation and the original preparation, and can obtain the dissolution behavior of febuxostat tablets from different sources and distinguish the internal quality of the products. Description of the Drawings
[0018] Figure 1 Shows the full wavelength scanning diagram of febuxostat;
[0019] Figure 2 Shows the saturated solubility curve diagram of febuxostat in Examples 1 - 7;
[0020] Figure 3 Shows the dissolution curve diagram of the reference preparation 4225B;
[0021] Figure 4 Shows the dissolution curve diagram of the similarity experiment;
[0022] Figure 5 Show the chromatogram of the reference solution in the specificity test. Detailed implementation mode
[0023] The following embodiments are provided to better further understand the present invention, which is not limited to the described optimal implementation mode, and does not constitute a limitation to the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior art features falls within the protection scope of the present invention.
[0024] For those not indicating specific experimental steps or conditions in the embodiments, the operations or conditions of the conventional experimental steps described in the literature in this field can be followed. For the reagents or instruments not indicating the manufacturer, they are all conventional reagent products that can be obtained through commercial purchase.
[0025] According to one aspect of the present application, a method for determining the in vitro dissolution curve of febuxostat tablets is provided, including the following specific steps: Step 1: Take febuxostat tablets, use hydrochloric acid solution as the dissolution medium, and the pH of the hydrochloric acid solution is 1.2. Perform dissolution according to the paddle method of the dissolution apparatus. At different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with phosphate buffer solution, shake well, and use it as the test solution; Step 2: Take febuxostat reference substance, place it in a volumetric flask, dissolve and dilute it to the scale with phosphate buffer solution, shake well, accurately measure an appropriate amount, place it in a volumetric flask, add phosphate buffer solution to dilute it to the scale, shake well, and use it as the reference solution, and the pH of the phosphate buffer solution in this step is 6.8; Step 3: Use high performance liquid chromatography, inject the test solution and the reference solution into the high performance liquid chromatography respectively, record the chromatogram, and calculate the dissolution amount.
[0026] According to one aspect of Step 1 of the present application, take febuxostat tablets, use hydrochloric acid solution as the dissolution medium, perform dissolution according to the paddle method of the dissolution apparatus, the amount of hydrochloric acid solution added is 900 ml, and the rotation speed of the paddle method of the dissolution apparatus is 50 revolutions per minute.
[0027] According to one aspect of Step 1 of the present application, at different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with phosphate buffer solution, shake well, and use it as the test solution, and the sampling time points are 5 min - 30 min.
[0028] Preferably, the sampling time points include 5 min, 10 min, 15 min, and 30 min.
[0029] According to one aspect of Step 1 of the present application, preferably, take 0.1 ml of the dissolution solution and add 0.9 ml of phosphate buffer solution to use it as the test solution.
[0030] According to one aspect of Step 1 of the present application, after taking out the dissolution solution, add the same volume of hydrochloric acid solution at the same temperature.
[0031] According to one aspect of Step 2 of the present application, preferably, take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a volumetric flask, dissolve it with phosphate buffer solution and dilute to the mark, shake well, accurately measure 5 ml, place it in a volumetric flask, and dilute to the mark with phosphate buffer solution, shake well.
[0032] According to one aspect of Step 1 and Step 2 of the present application, take febuxostat tablets, use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, carry out dissolution according to the paddle method of the dissolution apparatus, the rotation speed of the paddle method is 50 revolutions per minute, at different sampling time points, take an appropriate amount of the dissolution solution, carry out two-step dilution with phosphate buffer solution, shake well, and use it as the test solution, and the pH of the phosphate buffer solution is 6.0 - 8.0.
[0033] Preferably, the pH of the phosphate buffer solution is 6.8.
[0034] According to one aspect of Step 1 and Step 2 of the present application, take febuxostat tablets, use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, carry out dissolution according to the paddle method of the dissolution apparatus, the rotation speed of the paddle method is 50 revolutions per minute, at different sampling time points, take an appropriate amount of the dissolution solution, carry out two-step dilution with phosphate buffer solution, shake well, and use it as the test solution, wherein, the phosphate buffer solution for measuring the steps can also be acetate buffer solution, and the pH of the acetate buffer solution is 4.5 - 5.5.
[0035] Preferably, the phosphate buffer solution (pH 6.8) is prepared according to the description in General Principles 8004, Volume IV of the Chinese Pharmacopoeia 2020 Edition. Take 250 ml of 0.2 mol / L potassium dihydrogen phosphate solution, add 118 ml of 0.2 mol / L sodium hydroxide solution, dilute to 1000 ml with water, shake well, and obtain it.
[0036] According to one aspect of Step 3 of the present application, the chromatographic conditions of high performance liquid chromatography are: the chromatographic column is filled with octadecylsilane-bonded silica gel, the detector is a VWD detector, and the detection wavelength is 317 nm.
[0037] According to one aspect of Step 3 of the present application, the mobile phase is a combination of acetonitrile, water and glacial acetic acid, the column temperature is 40 °C; the flow rate is 2.0 ml / min, the running time is 7 min, and the sample volume is 20 μL.
[0038] The following further describes the present invention in conjunction with embodiments. The described embodiments are only a part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.
[0039] Example 1
[0040] A method for determining the in vitro dissolution curve of febuxostat tablets, comprising the following steps:
[0041] 1. Chromatographic conditions:
[0042] Chromatographic column: TSKgel ODS-80Ts C18 column (4.6 mm x 250 mm, 5 μm); mobile phase: acetonitrile-water-glacial acetic acid (600:400:2); detection wavelength: 317 nm; column temperature: 40 °C; flow rate: 2.0 ml / min; injection volume: 20 μl.
[0043] 2. Solution preparation
[0044] Hydrochloric acid solution with pH 1.2: Take 4 ml of hydrochloric acid solution, dilute it with water to 500 ml, and shake well to obtain.
[0045] Acetate buffer solution with pH 4.5: Weigh about 1.5 g of crystalline sodium acetate, dilute it with water to 500 ml, adjust the pH value to 4.5 with glacial acetic acid, and shake well to obtain.
[0046] Aqueous solution: Take purified water and degas it to obtain.
[0047] Reference solution: Take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a 100 ml volumetric flask, dissolve it with phosphate buffer solution (pH 6.8) and dilute it to the scale, shake well, accurately measure 5 ml, place it in a 100 ml volumetric flask, and dilute it to the scale with phosphate buffer solution (pH 6.8), shake well.
[0048] Preparation of test solution: Take febuxostat tablets, place them in a volumetric flask, use 900 ml of hydrochloric acid solution with pH 1.2 as the dissolution medium, perform dissolution according to the paddle method of the dissolution apparatus, the rotation speed of the paddle method is 50 revolutions per minute, at different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with acetate buffer solution with pH 4.5, place it in a constant temperature shaking water bath at room temperature for 24 h, filter, and take the subsequent filtrate as the test solution.
[0049] Perform determination according to the chromatographic conditions, and calculate the solubility of febuxostat by the external standard method.
[0050] Example 2
[0051] A method for determining the in vitro dissolution curve of febuxostat tablets, comprising the following steps:
[0052] 1. Chromatographic conditions:
[0053] Chromatographic column: TSKgel ODS-80Ts C18 column (4.6 mm x 250 mm, 5 μm); Mobile phase: acetonitrile - water - glacial acetic acid (600:400:2); Detection wavelength: 317 nm; Column temperature: 40 °C; Flow rate: 2.0 ml / min; Injection volume: 20 μl.
[0054] 2. Solution preparation
[0055] Hydrochloric acid solution with pH 1.2: Take 4 ml of hydrochloric acid solution, dilute it with water to 500 ml, shake well, and it is ready.
[0056] Acetate buffer solution with pH 5.5: Weigh about 3 g of crystalline sodium acetate, dilute it with water to 500 ml, adjust the pH value to 5.5 with glacial acetic acid, shake well, and it is ready.
[0057] Aqueous solution: Take purified water and degas it, and it is ready.
[0058] Reference solution: Take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a 100 ml volumetric flask, dissolve and dilute it to the mark with phosphate buffer solution (pH 6.8), shake well, accurately measure 5 ml, place it in a 100 ml volumetric flask, and dilute it to the mark with phosphate buffer solution (pH 6.8), shake well.
[0059] Preparation of test solution: Take febuxostat tablets, place them in a volumetric flask, use 900 ml of hydrochloric acid solution with pH 1.2 as the dissolution medium, carry out dissolution according to the paddle method of the dissolution apparatus, the rotation speed of the paddle method is 50 revolutions per minute, at different sampling time points, take an appropriate amount of the dissolution solution, carry out two-step dilution with acetate buffer solution with pH 5.5, place it in a constant temperature shaking water bath at room temperature for 24 h, filter, and take the subsequent filtrate as the test solution.
[0060] Determination is carried out according to the chromatographic conditions, and the solubility of febuxostat is calculated by the external standard method.
[0061] Example 3
[0062] A method for determining the in vitro dissolution curve of febuxostat tablets, comprising the following steps:
[0063] 1. Chromatographic conditions:
[0064] Chromatographic column: TSKgel ODS-80Ts C18 column (4.6 mm x 250 mm, 5 μm); Mobile phase: acetonitrile - water - glacial acetic acid (600:400:2); Detection wavelength: 317 nm; Column temperature: 40 °C; Flow rate: 2.0 ml / min; Injection volume: 20 μl.
[0065] 2. Solution preparation
[0066] Hydrochloric acid solution of pH 1.2: Take 4 ml of hydrochloric acid solution, dilute it with water to 500 ml, shake well, and you will get it.
[0067] Phosphate buffer solution of pH 6.0: Weigh about 3.4 g of potassium dihydrogen phosphate and about 0.1 g of disodium hydrogen phosphate, dilute with water to 500 ml, adjust the pH value to 6.0 with sodium hydroxide solution, shake well, and you will get it.
[0068] Aqueous solution: Take purified water and degas it to get it.
[0069] Reference solution: Take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a 100-ml volumetric flask, dissolve and dilute to the mark with phosphate buffer solution (pH 6.8), shake well, accurately measure 5 ml, place it in a 100-ml volumetric flask, and dilute to the mark with phosphate buffer solution (pH 6.8), shake well.
[0070] Preparation of test solution: Take an excessive amount of febuxostat raw material, place it in a 50-ml volumetric flask, add hydrochloric acid solution of pH 1.2, acetate buffer solution of pH 6.0 and aqueous solution respectively to make up the volume, place it in a constant-temperature shaking water bath at room temperature for 24 h, filter, take the subsequent filtrate, and determine it according to the chromatographic conditions. The solubility of febuxostat is calculated by the external standard method.
[0071] Example 4
[0072] A method for determining the in vitro dissolution curve of febuxostat tablets, comprising the following steps:
[0073] 1. Chromatographic conditions:
[0074] Chromatographic column: TSKgel ODS-80Ts C18 column (4.6 mm x 250 mm, 5 μm); The mobile phase is acetonitrile-water-glacial acetic acid (600:400:2); The detection wavelength is 317 nm; The column temperature is 40 °C; The flow rate is 2.0 ml / min; The injection volume is 20 μl.
[0075] 2. Solution preparation
[0076] Hydrochloric acid solution of pH 1.2: Take 4 ml of hydrochloric acid solution, dilute it with water to 500 ml, shake well, and you will get it.
[0077] Phosphate buffer solution of pH 6.8: Weigh about 3.4 g of potassium dihydrogen phosphate and about 0.5 g of disodium hydrogen phosphate, dilute with water to 500 ml, adjust the pH value to 6.8 with sodium hydroxide solution, shake well, and you will get it.
[0078] Aqueous solution: Take purified water and degas it to get it.
[0079] Reference solution: Take 11 mg of febuxostat reference substance dried at 105°C for 4 hours, place it in a 100-ml volumetric flask, dissolve it with phosphate buffer (pH 6.8) and dilute to the mark, shake well. Accurately measure 5 ml and place it in a 100-ml volumetric flask, add phosphate buffer (pH 6.8) to dilute to the mark, and shake well.
[0080] Preparation of test solution: Take febuxostat tablets, place them in a volumetric flask, use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, carry out dissolution according to the paddle method of the dissolution apparatus, the rotation speed of the paddle method is 50 revolutions per minute. At different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with phosphate buffer (pH 6.8), place it in a constant-temperature shaking water bath at room temperature for 24 h, filter, and take the subsequent filtrate as the test solution.
[0081] Determination is carried out according to the chromatographic conditions, and the solubility of febuxostat is calculated by the external standard method.
[0082] Example 5
[0083] A method for determining the in vitro dissolution curve of febuxostat tablets, comprising the following steps:
[0084] 1. Chromatographic conditions:
[0085] Chromatographic column: TSKgel ODS-80Ts C18 column (4.6 mm x 250 mm, 5 μm); The mobile phase is acetonitrile - water - glacial acetic acid (600:400:2); The detection wavelength is 317 nm; The column temperature is 40°C; The flow rate is 2.0 ml / min; The injection volume is 20 μl.
[0086] 2. Solution preparation
[0087] Hydrochloric acid solution with pH 1.2: Take 4 ml of hydrochloric acid solution, dilute it with water to 500 ml, shake well, and obtain.
[0088] Phosphate buffer with pH 8.0: Weigh about 3.4 g of potassium dihydrogen phosphate and about 0.9 g of disodium hydrogen phosphate, dilute with water to 500 ml, adjust the pH value to 8.0 with sodium hydroxide solution, shake well, and obtain.
[0089] Aqueous solution: Take purified water and degas it to obtain.
[0090] Reference solution: Take 11 mg of febuxostat reference substance dried at 105°C for 4 hours, place it in a 100-ml volumetric flask, dissolve it with phosphate buffer (pH 6.8) and dilute to the mark, shake well. Accurately measure 5 ml and place it in a 100-ml volumetric flask, add phosphate buffer (pH 6.8) to dilute to the mark, and shake well.
[0091] Preparation of the test solution: Take febuxostat tablets and place them in a volumetric flask. Use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, and perform dissolution according to the paddle method of the dissolution apparatus. The rotation speed of the paddle method is 50 revolutions per minute. At different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with phosphate buffer solution with a pH of 8.0, place it in a constant temperature shaking water bath at room temperature for 24 h, filter, and take the subsequent filtrate as the test solution.
[0092] Perform the determination according to the chromatographic conditions, and calculate the solubility of febuxostat by the external standard method.
[0093] Example 6
[0094] A method for determining the in vitro dissolution curve of febuxostat tablets, comprising the following steps:
[0095] 1. Chromatographic conditions:
[0096] Chromatographic column: TSKgel ODS-80Ts C18 column (4.6 mm x 250 mm, 5 μm); Use acetonitrile-water-glacial acetic acid (600:400:2) as the mobile phase; Detection wavelength is 317 nm; Column temperature is 40 °C; Flow rate is 2.0 ml / min; Injection volume is 20 μl.
[0097] 2. Solution preparation
[0098] Hydrochloric acid solution with a pH of 1.2: Take 4 ml of hydrochloric acid solution, dilute it with water to 500 ml, and shake well to obtain.
[0099] Aqueous solution: Take purified water and degas it to obtain.
[0100] Reference solution: Take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a 100 ml volumetric flask, dissolve and dilute it to the mark with phosphate buffer solution (pH 6.8), shake well, accurately measure 5 ml, place it in a 100 ml volumetric flask, and dilute it to the mark with phosphate buffer solution (pH 6.8), and shake well.
[0101] Preparation of the test solution: Take febuxostat tablets and place them in a volumetric flask. Use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, and perform dissolution according to the paddle method of the dissolution apparatus. The rotation speed of the paddle method is 50 revolutions per minute. At different sampling time points, take an appropriate amount of the dissolution solution, filter, and take the subsequent filtrate as the test solution.
[0102] Perform the determination according to the chromatographic conditions, and calculate the solubility of febuxostat by the external standard method.
[0103] Example 7
[0104] A method for determining the in vitro dissolution curve of febuxostat tablets, comprising the following steps:
[0105] 1. Chromatographic conditions:
[0106] The chromatographic column is a TSKgel ODS-80Ts C18 column (4.6 mm x 250 mm, 5 μm); the mobile phase is acetonitrile-water-glacial acetic acid (600:400:2); the detection wavelength is 317 nm; the column temperature is 40 °C; the flow rate is 2.0 ml / min; the injection volume is 20 μl.
[0107] 2. Solution preparation
[0108] Aqueous solution: Take purified water and degas it to obtain the solution.
[0109] Reference solution: Take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a 100 ml volumetric flask, dissolve and dilute to the mark with phosphate buffer solution (pH 6.8), shake well, accurately measure 5 ml, place it in a 100 ml volumetric flask, and dilute to the mark with phosphate buffer solution (pH 6.8), shake well.
[0110] Preparation of test solution: Take febuxostat tablets, place them in a volumetric flask, use the aqueous solution as the dissolution medium, perform dissolution according to the paddle method of the dissolution apparatus, the rotation speed of the paddle method is 50 revolutions per minute, at different sampling time points, take an appropriate amount of the dissolution solution, filter, and take the subsequent filtrate as the test solution.
[0111] Determination is carried out according to the chromatographic conditions, and the solubility of febuxostat is calculated by the external standard method.
[0112] The results show that the saturated solubilities of febuxostat in pH 1.2 hydrochloric acid solution, pH 4.5 acetate buffer solution, pH 5.5 acetate buffer solution, pH 6.0 phosphate buffer solution, pH 6.8 phosphate buffer solution, pH 8.0 phosphate buffer solution, and aqueous solution are 0.00001581 mg / ml, 0.00243 mg / ml, 0.04 mg / ml, 0.08 mg / ml, 1.6 mg / ml, 8.1 mg / ml, and 0.0057 mg / ml respectively.
[0113] The results indicate that febuxostat has pH dependence. In the pH range of 1.2 - 8.0 medium, the solubility is positively correlated with the pH value. The higher the pH value, the higher the saturated solubility, and the solubility is relatively small in acidic media. Regarding the establishment of a dissolution test method with discrimination
[0114] 1. Selection of dissolution medium
[0115] According to the requirements of the "General Principles for the Determination and Comparison of Dissolution Curves of Oral Solid Dosage Forms" and the "Q&A on Dissolution Curve Research in the 'Technical Guidelines for the Research on Pharmaceutical Changes of Listed Chemical Drugs (Trial)'", and combined with the results of the saturated solubility of febuxostat, select the pH 1.2 hydrochloric acid solution under "2 Solution preparation" as the most discriminatory dissolution curve.
[0116] 2. Determination of the Curve
[0117] Take the reference preparation (batch number: 4225B, febuxostat tablets of TEIJIN PHARMA LIMITED, trade name: Febuxostat Tablets / Feburic, launched in China in September 2018, specification: 40 mg), 12 tablets. According to the dissolution and release determination method (the first method in General Principles 0931, Volume IV of Chinese Pharmacopoeia 2020 Edition), conduct a dissolution experiment in hydrochloric acid medium at pH 1.2 (temperature: 37 °C), with the medium volume of 900 ml and a rotation speed of 50 revolutions per minute. After 5 min, 10 min, 15 min, and 30 min, respectively take 10 ml of the sample, and simultaneously replenish the same temperature and volume of the dissolution medium. Then, accurately transfer 0.1 ml of the sample solution and add 0.9 ml of phosphate buffer solution (pH 6.8) to prepare the test solution. Take an appropriate amount of the reference substance and prepare the reference solution according to the "2. Solution Preparation" section. Take the above two solutions and conduct the determination according to the "1. Chromatographic Conditions". Calculate the cumulative dissolution at each time point by the external standard method and draw the dissolution curve.
[0118] The results are shown in Figure 1 , and the specific data are shown in Table 1:
[0119] Table 1 Summary Table of Dissolution Curve Results of Reference Preparation 4225B
[0120]
[0121]
[0122] The average dissolution of the reference preparation 4225B in hydrochloric acid medium at pH 1.2 is 32.1% at 5 min, 40.3% at 10 min, and the average dissolutions at subsequent 15 min and 30 min are 42.0% and 43.0% respectively. The dissolution differences at the latter three points are less than 5%, and the dissolution platform has been reached at 30 min. Moreover, there are no inflection points and sudden release in the curve in this medium. Therefore, it is considered that the hydrochloric acid medium at pH 1.2 is suitable for evaluating the consistency between the reference and the originator.
[0123] Similarity Experiment
[0124] The test articles provided in this example are the self-made preparation and the reference preparation. Other chromatographic conditions, preparation of the test solution, preparation of the reference solution, etc. are the same as those in the determination of the curve.
[0125] The test articles selected this time are the self-made preparation - batch 24051001 and the reference preparation - batch 4225B. The cumulative dissolution of this product at different time points is shown in Table 2, and the dissolution curve atlas is shown in Figure 4 :
[0126] Summary Table of Cumulative Dissolution Data
[0127]
[0128] Specificity Test
[0129] Blank Solvent: Take 0.1 ml of the dissolution medium, add 0.9 ml of phosphate buffer solution (pH 6.8), mix well, and you will get it.
[0130] Blank Excipient Solution: Take about 23 mg of blank excipients, place them in a 100 ml volumetric flask, dilute to the mark with the dissolution medium, shake well, then accurately measure 0.1 ml, add 0.9 ml of phosphate buffer solution (pH 6.8), mix well, and you will get it.
[0131] Test Solution: Take the sample solution of the reference preparation in Example 2 at 30 minutes of dissolution as the test solution.
[0132] Reference Solution: Take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a 100 ml volumetric flask, dissolve and dilute to the mark with phosphate buffer solution (pH 6.8), shake well, accurately measure 5 ml, place it in a 100 ml volumetric flask, dilute to the mark with phosphate buffer solution (pH 6.8), and shake well.
[0133] Accurately measure 20 μL of each of the above solutions respectively, inject them into the liquid chromatography system, and record the chromatogram. The results are shown in Table 3:
[0134] Table 3 Dissolution Methodology Validation - Specificity Results
[0135]
[0136] The results show that neither the blank solvent nor the blank excipients interfere with the detection of dissolution.
[0137] Solution Stability
[0138] Test Solution: Take the sample solution of the reference preparation in Example 2 at 30 minutes of dissolution as the test solution.
[0139] Reference Solution: Take 11 mg of febuxostat reference substance dried at 105 °C for 4 hours, place it in a 100 ml volumetric flask, dissolve and dilute to the mark with phosphate buffer solution (pH 6.8), shake well, accurately measure 5 ml, place it in a 100 ml volumetric flask, dilute to the mark with phosphate buffer solution (pH 6.8), and shake well.
[0140] Accurately measure 20 μL of each of the above solutions respectively, and inject them into the liquid chromatography system at 5.5 h, 9.5 h, 17.5 h, 25.5 h, and 30 h respectively, and record the chromatogram. The results are shown in Table 4:
[0141] Table 4 Dissolution Methodology Validation - Results of Solution Temperature Stability
[0142]
[0143] The results showed that when the reference solution and the test solution were placed at room temperature (10 - 30°C) for 30 h, the maximum deviations were 0.38% and 1.18% respectively, both less than 5%. Therefore, the reference solution and the test solution were stable when placed at room temperature for 30 h.
[0144] In this application, through the following steps: Step 1: Take febuxostat tablets, use hydrochloric acid solution as the dissolution medium, and the pH of the hydrochloric acid solution is 1.2. Perform dissolution according to the paddle method of the dissolution apparatus. At different sampling time points, take an appropriate amount of the dissolution solution, perform two-step dilution with phosphate buffer solution, and shake well to obtain the test solution; Step 2: Take febuxostat reference substance, place it in a volumetric flask, dissolve and dilute to the mark with phosphate buffer solution, shake well, accurately measure an appropriate amount, place it in a volumetric flask, add phosphate buffer solution to dilute to the mark, and shake well to obtain the reference solution, and the pH of the phosphate buffer solution in this step is 6.8; Step 3: Use high performance liquid chromatography. Inject the test solution and the reference solution into the high performance liquid chromatography respectively, record the chromatogram, and calculate the dissolution amount; Through the dissolution results of the active pharmaceutical ingredient, combined with the in vitro-in vivo correlation, select pH 1.2 hydrochloric acid solution as the dissolution medium. The obtained curve has excellent discrimination ability, which is of great significance for guiding the research and development of generic febuxostat tablets and evaluating the quality and efficacy consistency between the generic preparation and the original preparation, and solves the technical problem of being unable to obtain the dissolution behavior of febuxostat tablets from different sources and distinguishing the intrinsic quality of products.
[0145] As described above, it is only the preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the scope disclosed by the present invention, according to the technical solution and its concept of the present invention, makes equivalent substitutions or changes, and all should be covered within the protection scope of the present invention.
Claims
1. A method for determining the in vitro dissolution curve of febuxostat tablets, characterized in that: The specific steps include: Step 1: Take Febuxostat tablets, use hydrochloric acid solution as the dissolution medium, and the pH of the hydrochloric acid solution is 1.2, dissolve according to the dissolution apparatus paddle method, take out an appropriate amount of the dissolution solution at different sampling time points, dilute it in two steps with phosphate buffer, shake well, and use it as the test solution; Step 2: Take the Febuxostat reference substance, place it in a volumetric bottle, dissolve it with phosphate buffer and dilute it to the scale, shake well, accurately measure it, place it in a volumetric bottle, add phosphate buffer to dilute it to the scale, shake well, as the reference substance solution, and the pH of the phosphate buffer in this step is 6.8; Step 3: Using HPLC, inject the test solution and reference solution into HPLC respectively, record the chromatogram, and calculate the dissolution amount.
2. The method for determining the in vitro dissolution curve of febuxostat tablets according to claim 1, characterized in that: Take Febuxostat tablets, use hydrochloric acid solution as the dissolution medium, and dissolve them according to the dissolution apparatus paddle method. The amount of hydrochloric acid solution added is 900 ml, and the speed of the dissolution apparatus paddle method is 50 rpm.
3. The method for determining the in vitro dissolution curve of febuxostat tablets according to claim 2, characterized in that: At different sampling time points, take out an appropriate amount of the dissolution solution, dilute it in two steps with phosphate buffer, shake it well, and use it as the test solution. The sampling time point is 5min-30min.
4. The method for determining the in vitro dissolution curve of febuxostat tablets according to claim 3, characterized in that: Take out 0.1 ml of the dissolution solution and add 0.9 ml of phosphate buffer as the test solution.
5. The method for determining the in vitro dissolution curve of febuxostat tablets according to claim 3, characterized in that: After taking out the dissolution solution, add the same volume of hydrochloric acid solution at the same temperature.
6. The method for determining the in vitro dissolution curve of febuxostat tablets according to claim 4, characterized in that: Take 11 mg of Febuxostat reference substance dried at 105°C for 4 hours, place it in a volumetric flask, dissolve and dilute to the scale with phosphate buffer, shake well, accurately measure 5 ml, place it in a volumetric flask, add phosphate buffer to dilute to the scale, and shake well.
7. The method for determining the in vitro dissolution curve of febuxostat tablets according to any one of claims 1 to 6, characterized in that: Take Febuxostat tablets, use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, and dissolve according to the paddle method of the dissolution apparatus. The paddle speed is 50 rpm. At different sampling time points, take out an appropriate amount of the dissolution solution, dilute it in two steps with phosphate buffer, shake it well, and use it as the test solution. The pH of the phosphate buffer is 6.0-8.
0.
8. The method for determining the in vitro dissolution curve of febuxostat tablets according to claim 7, characterized in that: Take Febuxostat tablets, use 900 ml of hydrochloric acid solution with a pH of 1.2 as the dissolution medium, and dissolve according to the paddle method of the dissolution instrument. The paddle speed is 50 rpm. At different sampling time points, take out an appropriate amount of the dissolution solution, dilute it in two steps with phosphate buffer, shake it well, and use it as the test solution. The phosphate buffer in the test step can also be an acetate buffer, and the pH of the acetate buffer is 4.5-5.
5.
9. The method for determining the in vitro dissolution curve of febuxostat tablets according to claim 8, characterized in that: The chromatographic conditions of the high performance liquid chromatography method are as follows: the chromatographic column is filled with octadecylsilane bonded silica gel, the detector is a VWD detector, and the detection wavelength is 317 nm.
10. The method for determining the in vitro dissolution curve of febuxostat tablets according to claim 9, characterized in that: The mobile phase was a combination of acetonitrile, water and glacial acetic acid, the column temperature was 40°C; the flow rate was 2.0 ml / min, the running time was 7 min, and the sample volume was 20 μL.