A method for determining impurities in vardenafil fumarate tablets

Abstract

This invention discloses a method for determining impurities in vonoprazan fumarate tablets, belonging to the field of pharmaceutical testing technology. This invention, through a specific chromatographic column, mobile phase, and gradient elution scheme, can effectively determine the content of impurities A, B, C, and E in vonoprazan fumarate tablets. This method has advantages such as excellent separation effect, good peak shape, strong stability, high sensitivity, and short detection time, enabling accurate and efficient determination of the content of key impurities in vonoprazan fumarate tablets, meeting the requirements of pharmaceutical quality control.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical testing technology, specifically relating to a method for determining impurities in vonoprazan fumarate tablets. Background Technology

[0002] Vonoprazan fumarate tablets are a novel potassium-competitive acid blocker, indicated for the treatment of acid-related disorders such as reflux esophagitis. Impurity detection is crucial for the quality control of this drug, as its presence can affect efficacy and potentially cause safety issues. Impurities A (CAS: 2170020-79-6), B (CAS: 928325-82-0), C (CAS: 1885094-62-1), and E (CAS: 1610043-62-3) are key impurities for quality control, requiring precise qualitative and quantitative analysis.

[0003] High-performance liquid chromatography (HPLC) is a commonly used method for drug impurity detection. Although there have been relevant studies on the detection of impurities in vonoprazan fumarate tablets, current HPLC detection methods are difficult to achieve accurate and efficient detection of the four key impurities mentioned above.

[0004] First, the four impurities exhibit significant differences in polarity and acid-base properties, making it difficult to simultaneously control their retention ranges under conventional high-performance liquid chromatography (HPLC) conditions. This results in low peak resolution for each impurity, failing to meet the pharmacopoeia requirement of ≥1.5. Specifically, impurity E is highly polar, exhibiting almost no retention in the HPLC system and readily eluting at dead time, overlapping with solvent and excipient peaks, thus hindering effective integration. Impurity C is weakly polar, with strong retention capacity, easily exhibiting excessively broad peaks and tailing, and a long elution time exceeding 30 minutes under conventional detection conditions. When using the chromatographic conditions described in Chinese patent application CN 116953129 A, its retention time reaches as long as 43 minutes, and the chromatographic peak is also prone to overlapping with unknown impurities. Impurities A and B are of moderate polarity, with their elution positions close to the main peak (vonoprazan fumarate). However, the content of the main component vonoprazan in the analyte is usually significantly higher than that of the impurities, causing the peaks of impurities A and B to be easily masked by the main peak, making baseline separation impossible and severely impacting detection accuracy. In addition, impurity A is an acidic compound (aspartic acid derivative), while impurities B, C, and E are basic amine compounds. The coexistence of acidic and basic compounds in the same mobile phase system will further lead to tailing and bifurcation of basic impurities, and an increase in peak width and decrease in sensitivity of acidic impurities. At the same time, slight fluctuations in the pH of the mobile phase will cause significant drift in the retention time of the four impurities, further exacerbating the separation difficulty and resulting in unstable separation effect.

[0005] Secondly, conventional mobile phase systems are insufficient to meet the limits of detection (LOD) and quantitation (LOQ) requirements for the four impurities. The control limits for these four impurities in vonoprazan fumarate raw material are relatively low, typically requiring ≤0.1%. However, due to their retention behavior, impurity E exhibits a low response value due to its rapid elution, while impurity C suffers from a reduced peak height due to excessive retention. Conventional mobile phase systems cannot simultaneously accommodate the response sensitivity of all four impurities, making it difficult to meet the specified LOD and LOQ standards and achieve accurate quantification of these four impurities.

[0006] In summary, the current high-performance liquid chromatography (HPLC) method for detecting impurities A, B, C, and E in vonoprazan fumarate tablets suffers from problems such as low resolution, poor peak shape, long detection time, and the inability to simultaneously meet the limits of detection and quantitation. This makes it difficult to achieve accurate and efficient detection of the four impurities and fails to meet the stringent requirements of drug quality control. Therefore, there is an urgent need for an impurity determination method that can solve the above problems. Summary of the Invention

[0007] In view of the shortcomings of the prior art, the present invention proposes an impurity determination method that can accurately and efficiently determine the content of impurities A, B, C and E in vonoprazan fumarate tablets. This method overcomes the problems of difficult impurity separation, low detection sensitivity, large influence of mobile phase pH and long detection time in the prior art, and meets the strict requirements of drug quality control.

[0008] The technical solution of this invention mainly includes the following: A method for determining impurities in vonoprazan fumarate tablets, wherein the method is high-performance liquid chromatography (HPLC), and the chromatographic conditions for HPLC are as follows: Using a Pursuit 3 C18 or CAPCELL PAK C18 MGH column, add methanesulfonic acid to phosphate buffer, wherein the volume percentage of methanesulfonic acid in the phosphate buffer is not less than 0.1%, and use phosphate buffer-acetonitrile-methanol containing methanesulfonic acid as mobile phase A; and phosphate buffer-acetonitrile-methanol as mobile phase B; perform gradient elution. In the mobile phase A, the volume ratio of phosphate buffer-acetonitrile-methanol is 70:3~5:25~27, and in the mobile phase B, the volume ratio of phosphate buffer-acetonitrile-methanol is 30~35:50:15~20.

[0009] It is necessary to ensure that methanesulfonic acid is added only to mobile phase A and not to mobile phase B in order to accurately control the retention behavior of each impurity peak and achieve effective separation.

[0010] Preferably, the methanesulfonic acid in the phosphate buffer solution has a volume percentage of 0.1% to 0.2%.

[0011] Preferably, the methanesulfonic acid in the phosphate buffer solution is 0.1% by volume.

[0012] Preferably, the concentration of the phosphate buffer solution is 0.0225M to 0.0275M.

[0013] Preferably, the column temperature is 30℃~40℃ during the testing process.

[0014] Preferably, the pH of the mobile phase is 6.6 to 7.0.

[0015] Preferably, the gradient elution procedure is as follows: From 0 min to 10 min, the volume percentage of mobile phase A is 100%, and the volume percentage of mobile phase B is 0%. From 10 min to 50 min, the volume percentage of mobile phase A decreases uniformly from 100% to 0%, while the volume percentage of mobile phase B increases uniformly from 0% to 100%. From 50 min to 55 min, the volume percentage of mobile phase A remains 0%, and the volume percentage of mobile phase B remains 100%. From 55.1 min to 70 min, the volume percentage of mobile phase A is 100%, and the volume percentage of mobile phase B is 0%.

[0016] Preferably, the impurities include impurity A, impurity B, impurity C and impurity E, wherein the CAS number of impurity A is 2170020-79-6, the CAS number of impurity B is 928325-82-0, the CAS number of impurity C is 1885094-62-1 and the CAS number of impurity E is 1610043-62-3.

[0017] Preferably, the chromatographic column has an inner diameter of 4.6 mm, a column length of 100 mm, and a particle size of 3 µm.

[0018] Preferably, in the mobile phase A, the volume ratio of phosphate buffer-acetonitrile-methanol is 70:3:27, and in the mobile phase B, the volume ratio of phosphate buffer-acetonitrile-methanol is 35:50:15.

[0019] Preferably, the chromatographic conditions for the high-performance liquid chromatography are as follows: A Pursuit 3 C18 column was used as the chromatographic column. 0.1% (v / v) methanesulfonic acid was added to phosphate buffer. The mobile phase A was 0.025 mol / L phosphate buffer-acetonitrile-methanol containing 0.1% (v / v) methanesulfonic acid; the mobile phase B was 0.025 mol / L phosphate buffer-acetonitrile-methanol. The pH of the mobile phase was 6.8. Gradient elution was performed at a column temperature of 35 °C. The volume ratio of phosphate buffer-acetonitrile-methanol in mobile phase A is 70:3:27, and the volume ratio of phosphate buffer-acetonitrile-methanol in mobile phase B is 35:50:15.

[0020] The beneficial effects of this invention are: This invention, by selecting Pursuit 3 C18 or CAPCELL PAK C18 MGH columns and optimizing the mobile phase composition and gradient elution program, can effectively control the retention behavior of impurities of different polarities, achieving good separation between impurities E and C, as well as impurities A and B, and the main peak. It solves the separation problem caused by the difference in impurity polarity and their positional relationship with the main peak, resulting in high resolution and significantly improving the accuracy of detection.

[0021] This invention adds a specific proportion of methanesulfonic acid to phosphate buffer solution to precisely control the mobile phase pH at 6.6~7.0, effectively improving the peak shape problem when these four impurities coexist, reducing impurity tailing, bifurcation, and increased impurity peak width, while enhancing the method's tolerance to mobile phase pH fluctuations, stabilizing impurity retention time, ensuring stable separation, and improving the reliability and stability of detection results.

[0022] Using the optimized chromatographic conditions of this invention, while achieving rapid peak elution of impurities and shortening the overall detection time, higher resolution can still be achieved, and the limits of detection (LOD) and quantitation (LOQ) are low, enabling accurate detection of low-content impurities and improving detection efficiency and sensitivity.

[0023] By investigating conditions such as column temperature and buffer salt concentration, the results show that the impurity determination method provided by this invention can maintain good detection performance under certain conditions of variation. It has wide applicability and stability for the determination of impurities in different batches of vonoprazan fumarate tablets, providing a reliable analytical method for drug quality control. Attached Figure Description

[0024] Figure 1 Specificity test (Experiment 1) Chromatogram of diluent.

[0025] Figure 2 Specificity test (Experiment 1) Chromatogram of blank excipient solution.

[0026] Figure 3 Specificity test (Experiment 1): Chromatogram of fumaric acid solution.

[0027] Figure 4 Specificity test (Experiment 1): Chromatogram of impurity A solution.

[0028] Figure 5 Specificity test (Experiment 1): Chromatogram of impurity B solution.

[0029] Figure 6 Specificity test (Experiment 1): Chromatogram of impurity C solution.

[0030] Figure 7 Specificity test (Experiment 1): Chromatogram of impurity E solution.

[0031] Figure 8 Specificity test (Experiment 1) System suitability solution chromatogram.

[0032] Figure 9 Chromatogram of self-control solution in specificity experiment (Experiment 1).

[0033] Figure 10 Specificity test (Experiment 1): Chromatogram of the test solution. Detailed Implementation

[0034] To facilitate a clearer understanding of the technical content of this invention by those skilled in the art, the invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0035] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the experimental materials and reagents used in the following examples are commercially available.

[0036] In the following examples, the 0.0225mol / L, 0.025mol / L, and 0.0275mol / L phosphate buffer solutions were prepared as follows: 3.06g, 3.40g, and 3.74g of potassium dihydrogen phosphate and 8.06g, 8.94g, and 9.85g of disodium hydrogen phosphate dodecahydrate were dissolved in water and diluted to 2000mL.

[0037] The pH of the mobile phase is adjusted using a test solution of phosphoric acid or sodium hydroxide.

[0038] In the following embodiments, the formulation of the vonoprazan fumarate tablets is as follows:

[0039] The vonoprazan fumarate tablets are prepared by the following method: (1) Pretreatment Mannitol and hydroxypropyl cellulose passed through a 40-mesh sieve.

[0040] (2) Ingredients Solution preparation: Weigh 5.26 kg of purified water and place it in a heat-insulated stirring tank. When the material water temperature reaches 50℃, add 66 g of fumaric acid until it is completely dissolved. The fumaric acid aqueous solution is controlled at 55±5℃. Weigh the material after discharge and add the evaporated purified water.

[0041] Weighing: Weigh 29.79 kg of mannitol, 5.979 kg of microcrystalline cellulose (SH-101), 1.314 kg of hydroxypropyl cellulose, and vonoprazan fumarate (the amount of each ingredient is calculated according to the formula).

[0042] Preparation: Add half of the mannitol, vonoprazan fumarate, microcrystalline cellulose (SH-101), hydroxypropyl cellulose, and the remaining half of the mannitol to a high-efficiency mixing granulator in the following order for premixing (settings: paddle speed 100 rpm, blade speed 1200 rpm). After 600 seconds, a mixed powder is obtained. Then, prepare the soft material by keeping the blade speed constant (blade speed 1200 rpm) and setting the paddle speed to 90 rpm. Add the prepared fumarate aqueous solution to the high-efficiency mixing granulator within 40-60 seconds. After 180 seconds (including the 40-60 seconds of liquid addition), discharge the material. Visually inspect the soft material; it should be free of foreign matter, and it should clump together when squeezed but crumble easily when poked.

[0043] (3) Granulation Add the prepared soft material to the hopper of a vibrating pellet mill and granulate it using a 2.5mm round-hole stainless steel sieve to obtain wet pellets. Check the appearance of the wet pellets; they should be uniform in size, appropriately compact, uniform in color, and free of foreign matter. Then, dry them.

[0044] (4) Drying Wet granules are fed into a fluidized bed granulator. The inlet air temperature is set to 60~75℃ and the fan frequency to 20~50Hz. Drying is paused when the material temperature is ≥45℃. A sample is taken to test the moisture content. The material is discharged after the moisture content is ≤1.0% (using a rapid moisture analyzer), resulting in white or off-white dried granules.

[0045] (5) Granulation and total blending Granulation: Add the dried granules to a 24-mesh manual sieve. The granules below 24 mesh are qualified granules. The coarse granules above 24 mesh are granulated using a pulverizer and granulator (1.5mm sieve). The granulated granules and the granules below 24 mesh are dry granules.

[0046] Final Mixing: Weigh out the prescribed amount of croscarmellose sodium based on the yield of the dry granules (if the yield of some granules is not less than 98.0%, weigh out the prescribed amount of croscarmellose sodium directly). First, add half of the dry granules to the hopper mixer, then add the croscarmellose sodium, and finally add the remaining dry granules to the hopper mixer. Adjust the speed of the hopper mixer to 8 rpm and mix for 20 minutes. Next, weigh out the prescribed amount of magnesium stearate based on the yield of the dry granules and add it to the mixer (if the yield of some granules is not less than 98.0%, weigh out the prescribed amount of magnesium stearate directly). Adjust the speed of the mixer to 8 rpm and mix for 5 minutes. Then stop the machine and discharge the material to obtain the final mixed granules (i.e., intermediate product), which are white or off-white in appearance.

[0047] It is obtained after tableting, coating, and aluminum-plastic packaging.

[0048] The vonoprazan fumarate described in the formulation of this invention is the active pharmaceutical ingredient (API) for vonoprazan fumarate tablets. This API can be a commercially available product or synthesized using conventional processes.

[0049] In a specific embodiment of the present invention, the raw materials synthesized using conventional processes (Yu, Q.-Y.; Zeng, H.; Yao, K.; Li, J.-Q.; Liu, Y. Novel and Practical Synthesis of Vonoprazan Fumarate. Synth. Commun. 2017, 47, 1169–1174.) are selected, and the synthesis process is as follows: After alkaline hydrolysis, the carboxylic acid group of fluoroarylpyrrole carboxylic acid ester is activated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), and then coupled with methylamine to generate an amide. After removing the NH proton from the pyrrole ring with n-butyllithium, it reacts with 3-pyridinesulfonyl chloride to generate a sulfonamide. Subsequently, the amide is reduced to an amine, fumaric acid is added, and it is recrystallized from methanol to obtain vonoprazan fumarate.

[0050] In the following examples, impurity A, CAS number: 2170020-79-6; molecular formula: C 21 H 20 FN3O6S; Molecular weight: 461.46; English name: 2-[[5-(2-fluorophenyl)-1-pyridin-3-ylsulfonylpyrrol-3-yl]methyl-methylamino]butanedioic acid.

[0051] Impurity B, CAS No.: 928325-82-0; Molecular Formula: C 17 H 17 N3O2S·C4H4O4; Molecular weight: 443.47; English name: N-methyl-1-(5-phenyl-1-pyridin-3-ylsulfonylpyrrol-3-yl)methanaminemonofumarate.

[0052] Impurity C, CAS No.: 1885094-62-1; Molecular Formula: C 18 H 18 FN3O2S; Molecular weight: 359.42; English name: 1-[5-(2-fluorophenyl)-1-pyridin-3-ylsulfonylpyrrol-3-yl]-N,N-dimethylmethanamine.

[0053] Impurity E, CAS No.: 1610043-62-3; Molecular Formula: C 12 H 13FN2; Molecular weight: 204.24; English name: 1-[5-(2-fluorophenyl)-1H-pyrrol-3-yl]-N-methylmethanamine.

[0054] Example 1 Diluent: Water-acetonitrile (volume ratio 3:1).

[0055] Test solution: Take vonoprazan fumarate tablets, place them in a volumetric flask, add an appropriate amount of diluent, shake for about 30 minutes to allow the tablets to completely disintegrate, then dilute to the mark with diluent to prepare a solution containing about 0.3 mg of vonoprazan per 1 mL. Shake well, filter, and take the filtrate as the test solution.

[0056] Self-reference solution: Accurately measure 1 mL of the test solution, place it in a 100 mL volumetric flask, dilute to the mark with diluent, and shake well.

[0057] System suitability solution: Take impurities A, B, C, and E, and dilute with diluent to prepare a mixed solution containing impurities A, C, and E at a concentration of 750 µg / mL and impurity B at a concentration of 1000 µg / mL. This is used as the impurity stock solution. Separately, take approximately 8 mg of vonoprazan fumarate reference standard, place it in a 25 mL volumetric flask, add an appropriate amount of diluent to dissolve it, then add 1 mL of the impurity stock solution, and dilute to the mark with diluent. Shake well.

[0058] Chromatographic conditions: A Pursuit 3 C18 column (4.6 mm × 100 mm, 3 µm) was used; 0.1% (v / v) methanesulfonic acid was added to phosphate buffer; mobile phase A consisted of 0.025 mol / L phosphate buffer-acetonitrile-methanol (v / v ratio 70:3:27) containing 0.1% (v / v) methanesulfonic acid; mobile phase B consisted of 0.025 mol / L phosphate buffer-acetonitrile-methanol (v / v ratio 35:50:15); mobile phase pH was 6.8. Gradient elution was performed according to the table below. The flow rate was 1.0 mL / min; the injection volume was 20 µL; the column temperature was 35 °C; and the detection wavelength was 230 nm.

[0059] Table 1 Elution gradient

[0060] Examples 2 to 9 The chromatographic conditions for Examples 2 to 9 are shown in Table 2. Other conditions are the same as in Example 1.

[0061] Table 2

[0062] Example 10 In this example, the mobile phase ratio is adjusted, while other conditions are the same as in Example 1.

[0063] The mobile phase A was 0.025 mol / L phosphate buffer containing 0.1% (v / v) methanesulfonic acid, acetonitrile, and methanol (volume ratio 70:5:25); the mobile phase B was 0.025 mol / L phosphate buffer, acetonitrile, and methanol (volume ratio 30:50:20).

[0064] Example 11 (Comparative Example) In this example, the mobile phase ratio is adjusted, while other conditions are the same as in Example 1.

[0065] The mobile phase A was 0.025 mol / L phosphate buffer containing 0.1% (v / v) methanesulfonic acid, acetonitrile, and methanol (volume ratio 80:3:17); the mobile phase B was 0.025 mol / L phosphate buffer, acetonitrile, and methanol (volume ratio 25:50:25).

[0066] Example 12 (Comparative Example) In this example, trifluoroacetic acid was added to the mobile phase, and other conditions were the same as in Example 1.

[0067] Example 13 (Comparative Example) In this example, formic acid was added to the mobile phase, and other conditions were the same as in Example 1.

[0068] Example 14 (Comparative Example) In this example, no methanesulfonic acid was added to the mobile phase, and other conditions were the same as in Example 1.

[0069] Example 15 (Comparative Example) The elution gradient in this example is shown in Table 3, and other conditions are the same as in Example 1.

[0070] Table 3 Elution gradient

[0071] Test results for each embodiment: Table 4

[0072] Table 4 shows that the determination of impurities in vonoprazan fumarate tablets under different conditions exhibits certain differences. Under the chromatographic conditions of Examples 1-8 and Example 10, the theoretical plate number and peak resolution fluctuated to some extent, but the results were significantly better than those of Examples 9 and Examples 11-15, still meeting the detection requirements and suitable for the detection of impurities in vonoprazan fumarate tablets.

[0073] Different chromatographic columns have different effects on the determination results. The theoretical plate number, tailing factor and resolution of Example 1 (Pursuit 3 C18 column) and Example 8 (CAPCELL PAK C18 MGH column) are better than those of Example 9 (Ultra Aq - C18 column), indicating that different chromatographic columns have different stationary phase properties and different separation capabilities and selectivity for impurities in vonoprazan fumarate tablets.

[0074] Furthermore, the mobile phase additives, mobile phase ratio, and elution gradient have a significant impact on the determination results. The determination results of Example 1 (mesylic acid) are superior to those of Examples 12 (trifluoroacetic acid), 13 (formic acid), and 14 (without mesylic acid) in terms of theoretical plate number and peak resolution, indicating that mesylic acid, as a mobile phase additive, can better improve peak shape and separation in the determination of impurities in vonoprazan fumarate tablets. The determination results of Example 1 under the elution gradient are significantly better than those of Example 15, and the determination results of Examples 1 and 10 under the mobile phase are also significantly better than those of Example 11, demonstrating that a suitable elution gradient and mobile phase ratio can more effectively separate impurities and improve the accuracy and sensitivity of detection.

[0075] In summary, the chromatographic conditions used in this invention, especially those in Examples 1-8 and 10, demonstrate good performance in the determination of impurities in vonoprazan fumarate tablets.

[0076] Example 17 Specificity Experiment Test Method 1: Prepare solutions as shown in Table 5. Take the diluent, blank excipient solution, fumaric acid positioning solution, system suitability solution, reference solution, stock solutions of each impurity solution, and test solution, and inject them for determination using the chromatographic conditions of Example 1. Investigate the retention time of the main peak for each solution.

[0077] Table 5

[0078] Table 6

[0079] The test results showed that no chromatographic peaks were detected in the diluent, and it did not interfere with other solutions. A fumaric acid peak was detected in the blank excipient solution, but it did not interfere with the main components of any of the impurity solutions. The retention time shifts of the corresponding chromatographic peaks in the impurity localization solutions and system suitability solutions were all less than 3% of the retention times of the corresponding components. Fumaric acid and vonoprazan peaks were detected in the test solution, and the retention time shifts were all less than 4% of the retention times of the corresponding components, which were basically consistent.

[0080] Test Method 2: In accordance with the General Chapter 9101 of Part IV of the Chinese Pharmacopoeia 2020, the blank excipients and test solutions were subjected to appropriate forced degradation tests, and test samples under each forced degradation condition were taken and detected using the chromatographic conditions of Example 1.

[0081] Take vonoprazan fumarate tablets into a mortar and grind them into a fine powder for later use.

[0082] ① Undegraded solution: Weigh 150 mg of vonoprazan fumarate tablets into a 50 mL volumetric flask, dissolve and dilute to the mark with diluent, and shake well. Weigh the same weight of blank excipient and perform the same procedure to obtain the undegraded blank solution.

[0083] ② Strong acid degradation solution: Weigh 150 mg of vonoprazan fumarate tablet powder into a 50 mL volumetric flask, add 20 mL of diluent and 5 mL of 1 mol / L hydrochloric acid solution, and shake well. After standing for 3 days, add 5 mL of 1 mol / L sodium hydroxide solution to neutralize to neutral (pH 5-7 on pH paper), add diluent to the mark, and shake well. Weigh the same weight of blank excipient and perform the same procedure to prepare a blank solution for strong acid degradation.

[0084] ③ Strong alkali degradation solution: Weigh 150 mg of vonoprazan fumarate tablet powder into a 50 mL volumetric flask, add 20 mL of diluent and 5 mL of 1 mol / L sodium hydroxide solution, and shake well. After standing for 1 hour, add 5 mL of 1 mol / L hydrochloric acid solution to neutralize to neutral (pH 5-7 on pH paper), add diluent to the mark, and shake well. Weigh the same weight of blank excipient and perform the same procedure to prepare a blank solution for strong alkali degradation.

[0085] ④ Strong oxidizing degradation solution: Weigh 150 mg of vonoprazan fumarate tablets into a 50 mL volumetric flask, add 20 mL of diluent and approximately 2 mL of 30% H2O2, and shake well. After standing for 3 days, dry the solution, then add diluent to dissolve it to the mark and shake well. Weigh the same weight of blank excipient and perform the same procedure to prepare a strong oxidizing degradation blank solution.

[0086] ⑤ High-temperature destruction solution: Weigh out 150 mg of vonoprazan fumarate tablet powder and place it at 90℃ for 3 days. Transfer the powder to a 50 mL volumetric flask, dissolve and dilute to the mark with diluent, and shake well. Then, take a blank excipient and repeat the same procedure to prepare the high-temperature destruction blank solution.

[0087] ⑥ Photodegradation solution: Take fine powder of vonoprazan fumarate tablets and expose it to white light irradiation ≥10000 lx and ultraviolet irradiation ≥90 μW / cm². 2After standing for 5 days under the specified conditions, weigh 150 mg into a 50 ml volumetric flask, dissolve and dilute to the mark with diluent, and shake well. Then take the blank excipient and perform the same operation to prepare the light-induced destruction blank solution. Filter each of the above solutions through a 0.45 μm filter membrane, discard 1 mL of the initial filtrate, and inject the subsequent filtrate into the sample.

[0088] Record the purity and resolution of the chromatographic peaks for each sample. Calculate the mass balance of vonoprazan and impurities in each degradation solution. If any chromatographic peak is present in each solution (except for the fumaric acid peak), calculate the peak area ratio using the following formula: Peak area ratio = Σ(A1…An) / M. Where: A1…An: the sum of the areas of all chromatographic peaks in the test solution; M: the sample weight of the test solution. Based on the above peak area ratio results, calculate the mass balance results under each condition. Mass balance = Peak area ratio result under each degradation condition / Peak area ratio result under the non-degradation condition × 100%. The results are shown in Tables 7 and 8.

[0089] Table 7

[0090] Table 8

[0091] Conclusion: No chromatographic peaks were detected in the undegraded blank solution and the degraded blank solutions. The resolution of all known impurities in the undegraded test solution and the degraded test solutions was not less than 5.11, and the peak purity was not less than 99.7%. The mass balance of vonoprazan and impurities in each degradation solution was calculated, and the mass balance results under all degradation conditions were between 97% and 102%.

[0092] Example 18: Limit of Quantitation and Limit of Detection Take the vonoprazan fumarate reference solution and the stock solutions of each impurity, dilute them stepwise, inject and test them. Confirm that the signal-to-noise ratio (S / N) of each bee is ≥10, which is the limit of quantitation (LOQ) concentration solution of that component. Prepare 6 LQ solutions according to the same method at the same concentration and inject them for testing.

[0093] Dilute the solution according to the above method for preparing the limit of quantitation solution to the limit of detection concentration, and confirm that its S / N ≥ 3, which is the limit of detection concentration solution.

[0094] Detection results: The detection limit concentrations of vonoprazan with impurities A, B, C and E are approximately 0.05 μg / mL, and the quantification limit concentrations are approximately 0.15 μg / mL.

[0095] The above description is only a part of the embodiments of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention shall fall within the protection scope of the present invention.

Claims

1. A method for determining impurities in vonoprazan fumarate tablets, wherein the impurities include impurity A, impurity B, impurity C, and impurity E, wherein the CAS number of impurity A is 2170020-79-6, the CAS number of impurity B is 928325-82-0, the CAS number of impurity C is 1885094-62-1, and the CAS number of impurity E is 1610043-62-3, and the determination method is high-performance liquid chromatography, characterized in that... The chromatographic conditions for the high-performance liquid chromatography are as follows: Using a Pursuit 3 C18 or CAPCELL PAK C18 MGH column, add methanesulfonic acid to phosphate buffer, wherein the volume percentage of methanesulfonic acid in the phosphate buffer is not less than 0.1%, and use phosphate buffer-acetonitrile-methanol containing methanesulfonic acid as mobile phase A; and phosphate buffer-acetonitrile-methanol as mobile phase B; perform gradient elution. In the mobile phase A, the volume ratio of phosphate buffer-acetonitrile-methanol is 70:3~5:25~27, and in the mobile phase B, the volume ratio of phosphate buffer-acetonitrile-methanol is 30~35:50:15~20.

2. The impurity determination method according to claim 1, characterized in that, The methanesulfonic acid in the phosphate buffer solution is 0.1% to 0.2% by volume.

3. The impurity determination method according to claim 1, characterized in that, The methanesulfonic acid in the phosphate buffer solution is 0.1% by volume.

4. The impurity determination method according to claim 1, characterized in that, The concentration of the phosphate buffer solution is 0.0225M~0.0275M.

5. The impurity determination method according to claim 1, characterized in that, During the testing process, the column temperature was 30℃~40℃.

6. The impurity determination method according to claim 1, characterized in that, The pH of the mobile phase is 6.6 to 7.

0.

7. The impurity determination method according to claim 1, characterized in that, The gradient elution procedure is as follows: From 0 min to 10 min, the volume percentage of mobile phase A is 100%, and the volume percentage of mobile phase B is 0%. From 10 min to 50 min, the volume percentage of mobile phase A decreases uniformly from 100% to 0%, while the volume percentage of mobile phase B increases uniformly from 0% to 100%. From 50 min to 55 min, the volume percentage of mobile phase A remains 0%, and the volume percentage of mobile phase B remains 100%. From 55.1 min to 70 min, the volume percentage of mobile phase A is 100%, and the volume percentage of mobile phase B is 0%.

8. The impurity determination method according to claim 1, characterized in that, The chromatographic column has an inner diameter of 4.6 mm, a column length of 100 mm, and a particle size of 3 µm.

9. The impurity determination method according to claim 1, characterized in that, In mobile phase A, the volume ratio of phosphate buffer-acetonitrile-methanol is 70:3:27, and in mobile phase B, the volume ratio of phosphate buffer-acetonitrile-methanol is 35:50:

15.

10. The impurity determination method according to claim 1, characterized in that, The chromatographic conditions for the high-performance liquid chromatography are as follows: A Pursuit 3 C18 column was used as the chromatographic column. 0.1% (v / v) methanesulfonic acid was added to phosphate buffer. The mobile phase A was 0.025 mol / L phosphate buffer-acetonitrile-methanol containing 0.1% (v / v) methanesulfonic acid; the mobile phase B was 0.025 mol / L phosphate buffer-acetonitrile-methanol. The pH of the mobile phase was 6.

8. Gradient elution was performed at a column temperature of 35 °C. The volume ratio of phosphate buffer-acetonitrile-methanol in mobile phase A is 70:3:27, and the volume ratio of phosphate buffer-acetonitrile-methanol in mobile phase B is 35:50:15.

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