Method for detecting isomer impurities in Vonoprazan fumarate starting material and application of method

Abstract

The invention provides a method for detecting isomer impurities in a Vonoprazan fumarate starting material and application of the Vonoprazan fumarate starting material, and the detection method comprises the following steps: (1) mixing a to-be-detected sample with a solvent to obtain a test solution; and (2) carrying out gas chromatography detection on the test solution, and determining the content of the isomer impurities in the sample to be detected according to a detection result. The detection method provided by the invention has a remarkable impurity separation effect, can effectively avoid hydrolysis of the to-be-detected test sample and the isomer impurities, and realizes evaluation and calculation of the content of the isomer impurities in the vonorrhoea fumarate starting material.

Description

Technical Field

[0001] The present invention belongs to the technical field of drug detection, and particularly relates to a method for detecting isomeric impurities in the starting material of vonoprazan fumarate and its application. Background Art

[0002] Vonoprazan is a potassium-competitive acid blocker (PCAB) that can inhibit gastric acid secretion mediated by H + 、K + -ATPase. Potassium-competitive acid blockers are an alternative to proton pump inhibitors in the treatment of acid-related diseases. Different from proton pump inhibitors, potassium-competitive acid blockers are not affected by CYP2C19 gene polymorphism and do not require an acid-resistant formulation. In addition, since vonoprazan can accumulate in the gastric body mucosa, especially in parietal cells, its potency is 350 times stronger than that of the proton pump inhibitor lansoprazole.

[0003] In February 2015, vonoprazan fumarate was first launched in Japan for the treatment of acid-related diseases and as an adjuvant drug for eradicating Helicobacter pylori. In May 2022, the US Food and Drug Administration (FDA) approved vonoprazan fumarate for use in a combination package product containing amoxicillin and clarithromycin to treat Helicobacter pylori infection. Studies have shown that when vonoprazan fumarate, amoxicillin, and clarithromycin are used in combination, the eradication rate of Helicobacter pylori is approximately 90%. The chemical name of vonoprazan fumarate is: 1-[5-(2-fluorophenyl)-1-(pyridine-3-sulfonyl)-1H-pyrrol-3-yl]-N-methylmethanamine monofumarate, and its structural formula is:

[0004]

[0005] In the synthesis route of vonoprazan fumarate, pyridine-3-sulfonyl chloride is used as a key starting material, and its purity plays a decisive role in the impurity level of the final product. During the synthesis of pyridine-3-sulfonyl chloride, there may be two isomers of sulfonation, which will react with thionyl chloride and are likely to introduce isomeric impurities pyridine-2-sulfonyl chloride and pyridine-4-sulfonyl chloride into the product.

[0006]

[0007] Pyridine-2-sulfonyl chloride will participate in the subsequent reactions of the vonoprazan fumarate synthesis process, thereby having a negative impact on the quality of the final product. Given that there are many potential impurities in vonoprazan fumarate itself, the impurities introduced by pyridine-3-sulfonyl chloride undoubtedly further increase the R & D difficulty of vonoprazan fumarate. To effectively simplify the R & D process and improve the quality stability of vonoprazan fumarate, it is particularly important to control the quality of the starting material pyridine-3-sulfonyl chloride from the source.

[0008] CN116699018A discloses a method for simultaneously detecting multiple structurally similar impurity compounds in pyridine-3-sulfonyl chloride, mainly for the determination of impurities pyridine-2-sulfonyl chloride and pyridine-4-sulfonyl chloride in pyridine-3-sulfonyl chloride by high performance liquid chromatography (HPLC). Since acyl chloride compounds are extremely unstable in the presence of water, the compounds detected in this patent are all related degradation products, namely pyridine-3-sulfonic acid, pyridine-2-sulfonic acid, and pyridine-4-sulfonic acid. Due to the differences in the response values of different compounds and the complexity of the reaction process, the conversion rate cannot be proven, and the specific content of impurity pyridine-2-sulfonyl chloride in pyridine-3-sulfonyl chloride cannot be truly reflected.

[0009] CN113390983A discloses a detection method for simultaneously determining pyridine-3-sulfonic acid, methyl pyridine-3-sulfonate, and ethyl pyridine-3-sulfonate in voruprazan fumarate. The detection method used is still HPLC, which is not applicable to the determination of impurity pyridine-2-sulfonyl chloride in pyridine-3-sulfonyl chloride.

[0010] CN118883760A discloses a method for detecting 5-chloropyridine-3-sulfonyl chloride in pyridine-3-sulfonyl chloride, but the isomeric impurity pyridine-2-sulfonyl chloride is not studied and controlled in this patent.

[0011] Since there is currently no effective method for detecting pyridine-2-sulfonyl chloride. Therefore, how to provide a method that can accurately and effectively detect the impurity pyridine-2-sulfonyl chloride in the raw material of pyridine-3-sulfonyl chloride has become an urgent problem to be solved. Summary of the Invention

[0012] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a detection method and its application for isomeric impurities in the starting material of voruprazan fumarate. The detection method provided by the present invention has a significant impurity separation effect, can effectively avoid the hydrolysis of the test sample to be detected and the isomeric impurities, and realizes the evaluation and calculation of the content of isomeric impurities in the starting material of voruprazan fumarate.

[0013] To achieve the purpose of this invention, the following technical solutions are adopted:

[0014] On the one hand, the present invention provides a detection method for isomeric impurities in the starting material of voruprazan fumarate, and the detection method includes the following steps:

[0015] (1) Mix the test sample with a solvent to obtain a test solution;

[0016] (2) Perform gas chromatography detection on the test solution, and determine the content of isomeric impurities in the test sample according to the detection results;

[0017] The starting material of vonoprazan fumarate is pyridine-3-sulfonyl chloride;

[0018] The isomeric impurity is pyridine-2-sulfonyl chloride.

[0019] By adopting a gas-phase method, the above method has a remarkable impurity separation effect, can effectively avoid the hydrolysis of the test sample to be detected and the isomeric impurity, and realizes the evaluation and calculation of the content of the isomeric impurity in the starting material of vonoprazan fumarate.

[0020] Preferably, the solvent in step (1) includes acetonitrile.

[0021] Preferably, the solvent in step (1) is subjected to a drying treatment.

[0022] Preferably, the chromatographic column for the gas chromatography detection in step (2) is an HP-1 capillary chromatographic column.

[0023] Preferably, the carrier gas flow rate for the gas chromatography detection in step (2) is 0.9 - 1.1 mL / min, such as 0.9 mL / min, 0.95 mL / min, 1 mL / min, 1.05 mL / min or 1.1 mL / min, etc., but not limited to the values listed above, and other unlisted values within the above value range are equally applicable.

[0024] Preferably, the inlet temperature for the gas chromatography detection in step (2) is 195 - 205 °C, such as 195 °C, 196 °C, 197 °C, 198 °C, 199 °C, 200 °C, 201 °C, 202 °C, 203 °C, 204 °C or 205 °C, etc., but not limited to the values listed above, and other unlisted values within the above value range are equally applicable.

[0025] Preferably, the detector temperature for the gas chromatography detection in step (2) is 305 - 315 °C, such as 305 °C, 306 °C, 307 °C, 308 °C, 309 °C, 310 °C, 311 °C, 312 °C, 313 °C, 314 °C or 315 °C, etc., but not limited to the values listed above, and other unlisted values within the above value range are equally applicable.

[0026] Preferably, the temperature programming for the gas chromatography detection in step (2) is as follows:

[0027] Taking 35 - 45 °C as the starting temperature, maintaining for 1.5 - 2.5 min, and then heating at a rate of 18 - 22 °C / min to 290 - 310 °C, and maintaining for 4 - 6 min.

[0028] The selection of the above specific reagents and parameters can effectively improve the separation effect of each chromatographic peak and improve the detection accuracy.

[0029] On the other hand, the present invention also provides the application of the detection method as described above in the quality control of voruprazan fumarate.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] The detection method provided by the present invention has a significant separation effect. At the same time, this method quantitatively detects the isomeric impurities involved in the present invention, realizing the evaluation and calculation of the content of isomeric impurities in the starting material of voruprazan fumarate;

[0032] Using the method of the present invention can accurately control the quality of the starting material of voruprazan fumarate, which has the characteristics of strong specificity (resolution greater than 1.5), high sensitivity (quantitation limit can be as low as 2.5 μg / mL, detection limit can be as low as 1.3 μg / mL), high accuracy, good repeatability, and good durability, and can accurately determine the content of isomeric impurities in the starting material of voruprazan fumarate. Description of the Drawings

[0033] Figure 1 It is the gas chromatogram of the blank solvent for the determination of isomers in Example 2;

[0034] Figure 2 It is the gas chromatogram of the sample for the determination of isomers in Example 2;

[0035] Figure 3 It is the gas chromatogram for the localization of isomeric impurities in the determination of isomers in Example 2;

[0036] Figure 4 It is the gas chromatogram of the mixed solution for the determination of isomers in Example 2.

[0037] Figure 5 It is the detection result diagram of the method of the example in Comparative Example 1;

[0038] Figure 6 It is the detection result diagram of the method of Comparative Example 1 in Comparative Example 1;

[0039] Figure 7 It is the detection result diagram of the method of the example in Comparative Example 2;

[0040] Figure 8 It is the detection result diagram of the method of Comparative Example 2 in Comparative Example 2. Detailed Embodiments

[0041] The technical solutions of the present invention will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations of the present invention.

[0042] Example 1 Sample Testing

[0043] This example provides a method for detecting isomeric impurities in the starting material of vorapaxar fumarate, and the specific steps are as follows:

[0044] Preparation of the solvent: Mix acetonitrile and molecular sieve in a ratio of 10% (m / v), let it stand for 24 hours, and filter to obtain.

[0045] Preparation of the test solution: Take 0.1 g of pyridine-3-sulfonyl chloride, accurately weigh it, place it in a 20 mL volumetric flask, dissolve it with dried acetonitrile and dilute to the mark, and shake well.

[0046] Preparation of the control solution: Accurately measure 1 mL of the test solution, place it in a 200 mL volumetric flask, dilute it to the mark with dried acetonitrile, and shake well.

[0047] Perform gas chromatography on the above solutions under the following conditions:

[0048] Use a capillary column with 100% dimethyl polysiloxane as the stationary liquid (HP-1, 30 m × 0.320 mm, 0.25 μm); the initial temperature is 40 °C, maintain for 2 minutes, increase the temperature to 300 °C at a rate of 20 °C per minute, and maintain for 5 minutes; the inlet temperature is 200 °C; the detector is a flame ionization detector (FID), the detector temperature is 310 °C, and the split ratio is 20:1; the carrier gas is nitrogen; the carrier gas flow rate is 1.0 mL per minute; the injection volume is 1 μL.

[0049] The results are as follows:

[0050] The relative retention time of pyridine-2-sulfonyl chloride to the main peak is about 0.7, and its content shall not exceed 0.5%.

[0051] After determination of multiple batches of samples, the content of pyridine-2-sulfonyl chloride can meet the requirements.

[0052] Table 1 Determination results of multiple batches of pyridine-3-sulfonyl chloride

[0053] Sample Pyridine-2-sulfonyl chloride (%) Batch 1 ND Batch 2 ND Batch 3 ND

[0054] Note: "ND" means the determination result is less than the detection limit (1.3 μg / mL).

[0055] Example 2 Specificity test

[0056] 2-1. Information of the test substance and impurities

[0057] Table 2 Summary table of information of the test substance and impurities

[0058]

[0059]

[0060] 2 - 2. Preparation of solutions:

[0061] Blank solvent: Acetonitrile (Mix acetonitrile with molecular sieve in a ratio of 10% (m / v), let stand for 24 hours, filter, and obtain.)

[0062] Test solution: Weigh accurately 0.1 g of pyridine - 3 - sulfonyl chloride sample, place it in a 20 - mL volumetric flask, dissolve it with acetonitrile and dilute to the mark, shake well;

[0063] Reference solution: Pipette accurately 1 mL of the test solution into a 200 - mL volumetric flask, dilute to the mark with acetonitrile, and shake well.

[0064] Mixed solution: Weigh accurately an appropriate amount of pyridine - 2 - sulfonyl chloride, dilute it with acetonitrile to a solution containing 500 μg per 1 mL as the impurity stock solution. Weigh accurately about 0.1 g of pyridine - 3 - sulfonyl chloride, place it in a 20 - mL volumetric flask, accurately add 1 mL of the above - mentioned impurity stock solution, dissolve it with acetonitrile and dilute to the mark, shake well, and obtain.

[0065] Pyridine - 2 - sulfonyl chloride impurity localization solution: Pipette accurately 1 mL of the above - mentioned impurity stock solution into a 20 - mL volumetric flask, dilute to the mark with acetonitrile, and shake well. As the localization solution for the impurity pyridine - 2 - sulfonyl chloride.

[0066] Pyridine - 3 - sulfonic acid and pyridine - 4 - sulfonyl chloride impurity localization solutions: Weigh accurately appropriate amounts of pyridine - 3 - sulfonic acid and pyridine - 4 - sulfonyl chloride impurities respectively, dilute them with acetonitrile to solutions containing 500 μg per 1 mL; Pipette accurately 1 mL of each of the above - mentioned impurity stock solutions into different 20 - mL volumetric flasks, dilute to the mark with acetonitrile, and shake well. As the localization solutions for each impurity.

[0067] 2 - 3. Test conditions:

[0068] Instrument: Gas chromatograph;

[0069] Chromatographic column: HP - 1, 30 m × 0.320 mm, 0.25 μm;

[0070] Carrier gas: High - purity nitrogen, flow rate 1.0 mL per minute;

[0071] Injector temperature: 200 °C;

[0072] Detector type: Flame ionization detector (FID);

[0073] Detector temperature: 310 °C;

[0074] Injection volume: 1 μL;

[0075] The split ratio is: 20:1;

[0076] Temperature programming: The initial temperature is 40 °C, maintained for 2 minutes, then heated at a rate of 20 °C per minute to 300 °C and maintained for 5 minutes.

[0077] 2 - 4. Experimental procedures and conclusions:

[0078] Precisely measure 1 μL of each of the above solutions and inject them into the gas chromatograph respectively, and record the chromatograms and results. The relevant chromatograms are shown in Figure 1-4 , and the results are shown in Table 3.

[0079] Table 3 Localization results of isomeric impurities in the starting material of voruprazan fumarate

[0080]

[0081]

[0082] Conclusion:

[0083] The above samples were separated using a gas chromatograph. From the test results, it can be seen that:

[0084] The blank solvent was detected, and it was found that the baseline of the gas chromatogram was stable. Except for the acetonitrile peak, there were no other characteristic peaks, indicating that the blank solvent did not interfere with the determination of impurities in the test solution.

[0085] From the chromatogram of the localization solution, the retention time of the characteristic peak of pyridine - 3 - sulfonyl chloride in the starting material of voruprazan fumarate was 8.713 min.

[0086] The retention time of pyridine - 2 - sulfonyl chloride was: 6.090 min; when the mixed solution was used for determination, the resolution between pyridine - 3 - sulfonyl chloride and pyridine - 2 - sulfonyl chloride was far greater than 1.5, and the resolution was good.

[0087] Under the conditions of this invention, neither pyridine - 3 - sulfonic acid nor pyridine - 4 - sulfonyl chloride showed peaks and did not interfere with the determination of the isomeric impurity pyridine - 2 - sulfonyl chloride.

[0088] From the above experimental detection results, it can be seen that the determination method provided by this invention shows a significant separation effect and good specificity.

[0089] Example 3 Selection of chromatographic column

[0090] Solution preparation: Prepare the mixed solution using the method of Example 2.

[0091] On the basis of Example 2, change the type of chromatographic column, keep other chromatographic conditions unchanged, and detect the isomeric impurity pyridine - 2 - sulfonyl chloride in the starting material of voruprazan fumarate.

[0092] Among them, the capillary column in Example 2 is: HP-1, 30 m × 0.320 mm, 0.25 μm;

[0093] Change the column type of Example 2:

[0094] The capillary column of Method 1 is: HP-1, 30 m × 0.320 mm, 0.25 μm;

[0095] The capillary column of Method 2 is: polyethylene glycol (PEG-20M), 30 m × 0.32 mm, 0.33 μm;

[0096] The capillary column of Method 3 is: HP-1, 30 m × 0.320 mm, 0.25 μm (from a different batch of the same manufacturer as the capillary column of Method 1)

[0097] Other parameters and steps are the same as those in Example 2. Accurately measure 1 μL of the mixed solution and inject it into the gas chromatograph respectively, record the results, and the results are shown in Table 4.

[0098] Table 4 Results of the investigation of isomeric impurities in the starting materials of voruprazan fumarate

[0099]

[0100] Conclusion:

[0101] Using the HP-1, 30 m × 0.320 mm, 0.25 μm capillary column in Method 1, the resolution between the starting material and the isomeric peak is greater than 1.5, showing good resolution.

[0102] Using the polyethylene glycol (PEG-20M), 30 m × 0.32 mm, 0.33 μm capillary column in Method 2, the starting material and its isomeric peak completely overlap.

[0103] Using the HP-1, 30 m × 0.320 mm, 0.25 μm (the same type but different batch as in Example 2) capillary column in Method 3, the resolution between the starting material and the isomeric peak is greater than 1.5, showing good resolution.

[0104] Using the method of the present invention, a capillary column with 100% dimethyl polysiloxane (or similar polarity) as the stationary liquid as the chromatographic column, such as the HP-1, 30 m × 0.320 mm, 0.25 μm capillary column, can simultaneously separate the starting materials of voruprazan fumarate and its isomeric impurities. However, after changing to the polyethylene glycol (PEG-20M), 30 m × 0.32 mm, 0.33 μm capillary column, the chromatographic peaks of related substances cannot be completely separated.

[0105] Example 4 High-temperature destructive test

[0106] The high-temperature forced degradation test is a rigorous analytical method designed to simulate relatively extreme conditions, such as high-temperature conditions, to accelerate the destruction of samples. The core purpose of this test is to deeply explore the degradation products of samples under severe conditions, and particularly focus on the separation of the main peak of pyridine-3-sulfonyl chloride from the isomeric peak of pyridine-2-sulfonyl chloride. By carefully analyzing the characteristics of these degradation products and the separation effect, we can comprehensively evaluate the effectiveness and applicability of the adopted analytical method, ensuring that it can accurately and reliably reflect the true quality status of samples in practical applications.

[0107] 4-1. Solution preparation:

[0108] Diluent: Acetonitrile.

[0109] Undegraded solution: Take 0.1 g of pyridine-3-sulfonyl chloride sample, accurately weigh it, place it in a 20 mL volumetric flask, dissolve it with acetonitrile and dilute to the mark, and shake well.

[0110] High-temperature degradation solution: Take about 0.1 g of this product, accurately weigh it, place it in a 20 mL volumetric flask, destroy it at 60 °C for 1 hour, take it out, let it cool, dissolve it with the solvent and dilute to the mark, and shake well to obtain.

[0111] 4-2. Experimental procedures and conclusions:

[0112] Precisely measure 1 μL of each of the above solutions and inject them into the gas chromatograph respectively. The chromatographic conditions are the same as those in Example 2, record the chromatogram, and the results are shown in Table 5 below.

[0113] Table 5 Results of the degradation experiment of the starting material of voruprazan fumarate

[0114]

[0115]

[0116] Conclusion:

[0117] From the above results, it can be seen that under high-temperature conditions, the resolution between the main peak and the adjacent peak in each degraded test sample is not less than 1.5, the resolution between each impurity peak is not less than 1.2, the tailing factor of the main peak USP tailing is 0.5 - 2.0, the theoretical plate number of the main peak is not less than 5000; the material balance meets the range of 90.0% - 110.0%. This indicates that the method of this invention patent has good specificity.

[0118] Example 5 Linear and correction factor test

[0119] Instrument 1:

[0120] 5-1. Solution preparation and linear results:

[0121] Diluent: Acetonitrile.

[0122] Stock solution: Accurately weigh 5 mg each of the impurity pyridine-2-sulfonyl chloride and pyridine-3-sulfonyl chloride reference substances, place them in the same 20 mL volumetric flask, dissolve with a diluent and dilute to the mark, shake well, and use as the reference stock solution. (250 μg / mL).

[0123] Preparation of linear solutions: Prepare linear solutions with corresponding concentrations using the linear stock solution according to the following table, mix well and label.

[0124] Table 6 Preparation of linear solutions

[0125]

[0126] The linear results are shown in Table 7-8.

[0127] Table 7 Linear test results of pyridine-3-sulfonyl chloride

[0128]

[0129] Table 8 Linear test results of pyridine-2-sulfonyl chloride

[0130]

[0131] The results show that:

[0132] For pyridine-3-sulfonyl chloride in the concentration range of 5.2740 μg / mL to 52.7396 μg / mL, which is equivalent to 20% to 200% of the limit concentration, the linear equation is y = 83.7521x - 190.2683, the correlation coefficient r is 0.997, greater than 0.990, the absolute value of the y-axis intercept is equivalent to 9.5% of the peak area of the 100% limit concentration solution, less than 25%, all meet the requirements, and the linear relationship is good. The sum of squared residuals is 48794.

[0133] For pyridine-2-sulfonyl chloride in the concentration range of 2.5405 μg / mL to 56.4545 μg / mL, which is equivalent to 9% to 200% of the limit concentration, the linear equation is y = 86.1898x - 11.0080, the correlation coefficient r is 0.9996, greater than 0.990, the absolute value of the y-axis intercept is equivalent to 0.45% of the peak area of the 100% limit concentration solution, less than 25%, all meet the requirements, and the linear relationship is good. The sum of squared residuals is 10410.

[0134] Instrument 2:

[0135] By different experimenters, on different dates, using different instruments, refer to "Instrument 1" under "Example 5" for the linear experiment, and the results are shown in Table 9-10.

[0136] Table 9 Linear test results of pyridine-3-sulfonyl chloride 2

[0137]

[0138] Table 10 Pyridine-2-sulfonyl chloride linearity 2 test results

[0139]

[0140]

[0141] 5-2. Calibration factor results and conclusions:

[0142] Based on the determination results of Instrument 1 and Instrument 2, the calibration factor of pyridine-2-sulfonyl chloride was calculated, and the results are shown in Table 11.

[0143] Table 11 Calibration factor results

[0144] Impurity name First time Second time Average value RD / % Pyridine-2-sulfonyl chloride 0.97 0.87 0.92 5.5

[0145] Conclusion:

[0146] Through the analysis of the above linear solutions, it was found that:

[0147] The relative calibration factors of the impurity pyridine-2-sulfonyl chloride were 0.97 and 0.87 respectively, the average value was 0.92, and the RD value was 5.5%, not greater than 10%, meeting the requirements. The calibration factor of the impurity pyridine-2-sulfonyl chloride was within the range of 0.9 - 1.1, so no calibration factor was required.

[0148] Example 6 Quantitative limit and detection limit tests

[0149] 6-1. Solution preparation:

[0150] Diluent: Acetonitrile.

[0151] Quantitative limit solution: Take 0.9 mL of the stock solution in Example 5, place it in the same 100 mL volumetric flask, dilute it to the mark with the diluent, and shake well. Prepare 6 parallel portions.

[0152] Detection limit solution: Take 0.9 mL of the stock solution in Example 5, place it in the same 200 mL volumetric flask, dilute it to the mark with the diluent, and shake well. Prepare 3 parallel portions.

[0153] 6-2. Experimental procedures and conclusions:

[0154] Precisely measure 1 μL of each of the above solutions and inject them into the gas chromatograph respectively. The chromatographic conditions are the same as those in Example 3. Record the chromatograms and results, and the results are shown in Tables 12 - 13.

[0155] Table 12 Investigation results of the quantitative limit and detection limit of the starting material of vonoprazan fumarate and its impurities

[0156]

[0157]

[0158] Conclusion:

[0159] Using the above separation conditions, the limits of quantitation (LOQ) and detection limits (LOD) of the starting material of vonoprazan fumarate and its impurities were detected, and the results are as follows:

[0160] The LOQ of the starting material pyridine-3-sulfonyl chloride was 5.2740 μg / mL, equivalent to 20% of the limit concentration. The signal-to-noise ratios of 6 injections of the LOQ solution were between 15 and 19, all within the range of 9 - 20. The RSD of the peak areas of 6 injections of the LOQ solution was 14.5%, less than 15.0%. The LOD was 2.6370 μg / mL, equivalent to 10% of the limit concentration. The signal-to-noise ratios of 3 injections of the LOD solution were between 8 and 9, all greater than 3, meeting the requirements.

[0161] The LOQ of the starting material impurity pyridine-2-sulfonyl chloride was 2.5405 μg / mL, equivalent to 9% of the limit concentration. The signal-to-noise ratios of 6 injections of the LOQ solution were all 10, all within the range of 9 - 20. The RSD of the peak areas of 6 injections of the LOQ solution was 3.7%, less than 15.0%. The LOD was 1.2702 μg / mL, equivalent to 4.5% of the limit concentration. The signal-to-noise ratios of 3 injections of the LOD solution were between 5 and 6, all greater than 3, meeting the requirements.

[0162] The test results show that the experimental method can meet the requirements for the simultaneous separation and quality control of pyridine-2-sulfonyl chloride in the starting material of vonoprazan fumarate, with good sensitivity.

[0163] Example 7 Recovery Test

[0164] 7-1. Solution Preparation:

[0165] Diluent: Acetonitrile.

[0166] Impurity Stock Solution: Take 10 mg of the impurity pyridine-2-sulfonyl chloride reference substance, accurately weigh it, place it in a 20 mL volumetric flask, dissolve it with the diluent and dilute to the mark, shake well, and use it as the reference stock solution. Prepare 2 parallel portions. (500 μg / mL).

[0167] Accuracy Reference Solution: Accurately measure 1 mL of the impurity stock solution, place it in a 20 mL volumetric flask, dilute it to the mark with acetonitrile, and shake well. Prepare 2 parallel portions (25 μg / mL).

[0168] Accuracy Test Solution (Background Solution): Take 100 mg of this product, accurately weigh it, place it in a 20 mL volumetric flask, dissolve it with the diluent and dilute to the mark, and shake well.

[0169] Accuracy solution: R1 - 50% solution: Take 100 mg of this product, weigh accurately, place it in a 20 mL volumetric flask, accurately add 0.5 mL of the impurity reference stock solution, dissolve with the diluent and dilute to the mark, and shake well. (Prepare 3 parallel portions)

[0170] R2 - 100% solution: Take 100 mg of this product, weigh accurately, place it in a 20 mL volumetric flask, accurately add 1.0 mL of the impurity reference stock solution, dissolve with the diluent and dilute to the mark, and shake well. (Prepare 3 parallel portions)

[0171] R3 - 150% solution: Take 100 mg of this product, weigh accurately, place it in a 20 mL volumetric flask, accurately add 1.5 mL of the impurity reference stock solution, dissolve with the diluent and dilute to the mark, and shake well. (Prepare 3 parallel portions)

[0172] Control solution: Accurately measure 1 mL each of the above test solution and accuracy solution respectively, place them in a 200 mL volumetric flask, dilute to the mark with the diluent, and shake well.

[0173] 7 - 2. Experimental procedures and conclusions:

[0174] Accurately measure 1 μL of each of the above solutions and inject them into the gas chromatograph respectively. The chromatographic conditions are the same as those in Example 3. Record the chromatograms and results. The results are shown in the following table. The relative standard deviation calculation formula is as follows:

[0175]

[0176] Table 14 Determination of impurities in the starting material of voruprazan fumarate - Recovery results

[0177]

[0178] Conclusion:

[0179] By measuring the 50%, 100%, and 150% accuracy solutions (equivalent to the limit concentration), it was found that when calculated by the external standard method and the self - control method respectively, the recoveries of the 9 samples of impurities were between 90.4% - 95.6% and 101.0% - 105.7% respectively, all within the range of 90% - 108%. The RSD values of the recoveries of the 9 samples were 1.9% and 1.8% respectively, both less than 7.5%, meeting the requirements and showing good accuracy.

[0180] Example 8 Repeatability test

[0181] Examine the change in the impurity content by repeatedly measuring the same batch of samples to verify and obtain the method repeatability.

[0182] 8 - 1. Solution preparation:

[0183] Diluent: Acetonitrile;

[0184] Stock solution of impurity reference substance: Weigh accurately 10 mg of pyridine-2-sulfonyl chloride reference substance for impurities, place it in a 20 mL volumetric flask, dissolve and dilute to the mark with diluent, and shake well to obtain the stock solution of impurity reference substance (500 μg / mL).

[0185] Test solution (spiked): Weigh accurately 100 mg of this product, place it in a 20 mL volumetric flask, accurately add 1 mL of the stock solution of impurity reference substance, dissolve and dilute to the mark with diluent, and shake well to obtain (prepared in 6 parallel portions).

[0186] Control solution: Accurately measure 1 mL of the test solution (spiked), place it in a 200 mL volumetric flask, dilute to the mark with diluent, and shake well.

[0187] 8-2. Experimental procedures and conclusions:

[0188] Accurately measure 1 μL each of the above-mentioned test solutions of Sample 1 to Sample 6 and the control solution, and inject them into the gas chromatograph respectively. The chromatographic conditions are the same as in Example 3. Record the chromatograms and results; Replace different experimenters, conduct the experiment repeatedly on different dates using different instruments to investigate the intermediate precision. The results are shown in Table 15.

[0189] Table 15 Precision test results of pyridine-2-sulfonyl chloride in the starting material of vonoprazan fumarate

[0190]

[0191]

[0192] Conclusion:

[0193] The RSD value of the content of pyridine-2-sulfonyl chloride in 6 replicates of the samples for repeatability is 3.5%, and the RSD value of the content of 6 samples for intermediate precision is 2.9%, both less than 5.0%; The RSD value of the content of 12 samples for precision is 6.6%, all less than 7.5%, all meeting the requirements, and the precision is good.

[0194] Example 9 Robustness test

[0195] By slightly changing the method parameters, investigate the change in system suitability and the tolerance of the measurement results not being affected. The robustness of this product was investigated by changing the column flow rate (±0.1 mL / min), changing the inlet temperature (±5 °C), changing the detector temperature (±5 °C), and replacing the chromatographic columns of the same model but different batches on the basis of the detection conditions in Example 3. The influence of different change situations on the determination results of pyridine-2-sulfonyl chloride. The solution preparation and other chromatographic conditions are the same as in Example 3. The changes in robustness parameters and results are shown in Tables 16-18.

[0196] Table 16 Change table of parameters for the verification of related substances - robustness

[0197]

[0198] Table 17 Related Substances Method Validation - Robustness - System Suitability Results

[0199]

[0200]

[0201] Table 18 Related Substances Method Validation - Robustness - Test Solution Determination Results

[0202]

[0203] Conclusion:

[0204] The results of the robustness test showed that, on the basis of Example 2, when adjusting the parameters and under normal conditions, for the reference solution, the maximum RSD value of the main peak peak area was 5.0%, all less than 10.0%, and the maximum RSD value of the retention time was 0.15%, all less than 1.0%, all meeting the requirements.

[0205] When adjusting the parameters and under normal conditions, the maximum RSD / RD value of the content of impurity pyridine - 2 - sulfonyl chloride was 6.5%, not greater than 7.5%, and the robustness was good.

[0206] Comparative Example 1

[0207] Referring to the detection conditions and methods of CN116699018A, the specificity and other aspects of the examples of the present invention were investigated.

[0208] The difference between the method of Comparative Example 1 and Example 1 was that Example 1 used gas chromatography, and the reagents used were dried to avoid the influence of the presence of water on the stability of pyridine - 3 - sulfonyl chloride and its impurity pyridine - 2 - sulfonyl chloride, that is, the chromatographic peaks shown were all the original peaks of the two; the method of the comparative example used high - performance liquid chromatography, and both the mobile phase and the solvent contained water.

[0209] The comparison of the chromatographic conditions of the present invention and the comparative document is shown in Table 19 below.

[0210] Table 19 Comparison of the Method of the Present Invention and the Comparative Document Method

[0211]

[0212] Referring to the detection conditions and methods of the comparative document, the localization and specificity of the impurity reference solution and the mixed solution of the examples of the present invention were investigated. The specific preparation is as follows:

[0213] Impurity (pyridine-2-sulfonyl chloride) stock solution: Weigh an appropriate amount of pyridine-2-sulfonyl chloride accurately, and dilute it with 5% acetonitrile solution to a solution containing 500 μg per 1 mL as the impurity stock solution.

[0214] Mixed solution: Weigh approximately 0.1 g of pyridine-3-sulfonyl chloride accurately, place it in a 20 mL volumetric flask, accurately add 1 mL of the above-mentioned impurity stock solution, dissolve it with 5% acetonitrile and dilute to the scale, shake well to obtain.

[0215] The results are as Figure 5-6 shown. In the examples, the resolution between pyridine-3-sulfonyl chloride and pyridine-2-sulfonyl chloride is good, and the stability of the compound can be maintained; in Comparative Example 1, the above compound is converted into pyridine-3-sulfonic acid and pyridine-2-sulfonic acid, and the resolution fails to reach 1.5.

[0216] Conclusion:

[0217] From the comparative analysis results, it can be seen that under the detection conditions of Comparative Example 1, pyridine-3-sulfonyl chloride and the impurity pyridine-2-sulfonyl chloride are converted into pyridine-3-sulfonic acid and pyridine-2-sulfonic acid, and there is a partial overlap between the two chromatographic peaks, and the resolution fails to reach 1.5. The examples can not only maintain the stability of the sulfonyl chloride sample but also achieve good separation of pyridine-3-sulfonyl chloride and pyridine-2-sulfonyl chloride.

[0218] Comparative Example 2

[0219] Refer to the detection conditions and methods of CN118883760A to conduct a specificity investigation on the examples of the present invention.

[0220] The difference between the method of Comparative Example 2 and Example 1 is that there are differences in the chromatographic conditions, and the polarities of the capillary chromatographic columns used are different; in addition, the solvent used in Comparative Example 2 is not dehydrated, which may not affect the detection of impurities in the comparative example, but will have a significant impact on the isomeric impurities involved in the present invention.

[0221] The comparison of the chromatographic conditions between the present invention and the comparative document is shown in Table 20 below.

[0222] Table 20 Comparison of the method of the present invention and the comparative document method

[0223]

[0224]

[0225] Refer to the detection conditions and methods of Comparative Example 2 to conduct localization and specificity investigations on the impurity reference solution and the mixed solution of the examples of the present invention. The specific preparations are as follows:

[0226] Impurity (pyridine-2-sulfonyl chloride) stock solution: Weigh an appropriate amount of pyridine-2-sulfonyl chloride accurately, and dilute it with 5% acetonitrile solution to make a solution containing 500 μg per 1 mL as the impurity stock solution.

[0227] Mixed solution: Weigh 0.1 g of pyridine-3-sulfonyl chloride accurately, place it in a 20 mL volumetric flask, accurately add 1 mL of the above-mentioned impurity stock solution, dissolve it with 5% acetonitrile and dilute to the scale, and shake well to obtain it.

[0228] The results are as Figure 7-8 shown. In the examples, the resolution between pyridine-3-sulfonyl chloride and pyridine-2-sulfonyl chloride is good, and the stability of the compound can be maintained; in Comparative Example 2, a capillary column with completely different polarity from that in the examples is used, resulting in almost complete overlap of pyridine-3-sulfonyl chloride and pyridine-2-sulfonyl chloride (around 17 min), and due to the poor separation effect, the reproducibility of the measurement results is poor.

[0229] Conclusion:

[0230] It can be seen from the comparative analysis of the results that under the detection conditions of Comparative Example 2, the chromatographic peaks of pyridine-3-sulfonyl chloride and the impurity pyridine-2-sulfonyl chloride almost completely overlap, and good separation cannot be achieved. The examples can achieve good separation of pyridine-3-sulfonyl chloride and pyridine-2-sulfonyl chloride.

[0231] The applicant declares that the present invention uses the above examples to illustrate the detection method and its application of isomeric impurities in the starting materials of voruprazan fumarate of the present invention, but the present invention is not limited to the above examples, that is, it does not mean that the present invention must rely on the above examples to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent substitution of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and the disclosure scope of the present invention.

[0232] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0233] In addition, it should be noted that in the various specific technical features described in the above specific embodiments, they can be combined in any suitable way without conflict. In order to avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A method for detecting isomeric impurities in a vonoprazan fumarate starting material, characterized in that: The detection method comprises the following steps: (1) Mixing the sample to be tested with a solvent to obtain a test solution; (2) subjecting the test sample solution to gas chromatography detection, and determining the isomeric impurity content in the test sample based on the detection results; The starting material of vonoprazan fumarate is pyridine-3-sulfonyl chloride; The isomeric impurity is pyridine-2-sulfonyl chloride.

2. The detection method according to claim 1, characterized in that: The solvent in step (1) comprises acetonitrile.

3. The detection method according to claim 1 or 2, characterized in that: The solvent in step (1) is dried.

4. The detection method according to any one of claims 1 to 3, characterized in that: The chromatographic column for gas chromatography detection in step (2) is an HP-1 capillary chromatographic column.

5. The detection method according to any one of claims 1 to 4, characterized in that: The carrier gas flow rate of the gas chromatography detection in step (2) is 0.9-1.1 mL / min.

6. The detection method according to any one of claims 1 to 5, characterized in that: The injection port temperature of the gas chromatography detection in step (2) is 195-205°C.

7. The detection method according to any one of claims 1 to 6, characterized in that: The detector temperature of the gas chromatography detection in step (2) is 305-315°C.

8. The detection method according to any one of claims 1 to 7, characterized in that: The temperature rise program of the gas chromatography detection in step (2) is as follows: The starting temperature was 35-45°C and maintained for 1.5-2.5 min, then the temperature was increased to 290-310°C at a rate of 18-22°C / min and maintained for 4-6 min.

9. Use of the detection method according to any one of claims 1 to 8 in the quality control of vonoprazan fumarate.

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