Detection method for determining N-nitroso-Vonoprasan in Vonoprasan fumarate

Abstract

The invention discloses a detection method for determining N-nitroso-Vonoprasan in Vonoprasan fumarate, and belongs to the technical field of medicine detection.The detection method comprises the steps that a solution is prepared, and detection is conducted; the detection comprises the following steps: determining a test solution and a reference solution by adopting a liquid chromatography-mass spectrometry method, recording a chromatogram, and calculating the content of N-nitroso-Vonorazan according to the chromatogram and an external standard method; according to the method, a triple quadrupole rod liquid phase mass spectrometer is adopted; a chromatographic column adopts octadecyl silane bonded silica gel as a filling agent, the specification of the chromatographic column is Poroshell 120EC-C18, 1.9 mu m, 50 mm * 3.0 mm; a mobile phase is composed of a mobile phase A and a mobile phase B, and gradient elution is adopted; the ultra-high pressure chromatographic column is adopted, the operation time is shorter, the signal-to-noise ratio is higher, the detection sensitivity is higher, the detection cost is lower, and the detection requirement can be met on a low-end triple quadrupole mass spectrum.

Description

Technical Field

[0001] The present invention relates to the technical field of drug detection, and more specifically to a detection method for determining N-nitroso-vonoprazan in vonoprazan fumarate. Background Art

[0002] Vonoprazan fumarate tablets are mainly used to treat reflux esophagitis and eradicate Helicobacter pylori. They are potassium ion competitive acid blockers (P-CAB for short) and a new type of acid-suppressing drug.

[0003] N-nitroso-vonoprazan (NVP) is a nitrosamine impurity detected in vonoprazan fumarate API and its tablets, with the structural formula:

[0004] The formation of N-nitroso-vonoprazan impurities is caused by the nitrosation reaction between the secondary amine functional group in the vonoprazan fumarate raw material and the nitrite in the excipients, which leads to the formation of N-nitroso-vonoprazan impurities.

[0005] N-nitroso-vonoprazan contains the warning structure of sulfonate esters, which poses a carcinogenic risk. According to the ICH M7 guidelines, N-nitroso-vonoprazan impurities need to be strictly controlled. Since the recommended daily intake of N-nitroso-vonoprazan is 96 ng / day and the maximum daily dose of vonoprazan fumarate tablets is 40 mg / day, the limit of N-nitroso-vonoprazan in vonoprazan fumarate is 2.4 ppm. However, the detection limit of conventional liquid phase ultraviolet detection method is higher than 10 ppm, so the conventional liquid phase ultraviolet detection method cannot meet the detection sensitivity requirements.

[0006] When the liquid chromatography-mass spectrometry (LC-MS / MS) method was used to detect and quantify N-nitroso-vonoprazan impurities, it was found that the detection limit of the liquid chromatography-mass spectrometry (LC-MS / MS) method on a low-resolution mass spectrometer was 0.036 ppm, which can fully meet the control requirements of N-nitroso-vonoprazan in fumarate vonoprazan tablets. However, the existing liquid chromatography-mass spectrometry (LC-MS / MS) method has the following problems when detecting N-nitroso-vonoprazan in fumarate vonoprazan tablets: first, the liquid chromatography-mass spectrometry (LC-MS / MS) method has a low response on a low-end triple quadrupole mass spectrometer; second, the detection results of the liquid chromatography-mass spectrometry (LC-MS / MS) method are greatly affected by the mass spectrometry resolution; third, the liquid chromatography-mass spectrometry (LC-MS / MS) method has a long running time and high detection cost.

[0007] Chinese patent CN118566356A discloses a method for detecting the genotoxic impurity N-nitroso-vonoprazan in vonoprazan fumarate, which can determine the N-nitroso-vonoprazan impurity in vonoprazan fumarate, and has the advantages of high selectivity, good sensitivity, and a quantitative limit as low as 0.18 ppm. In addition, the test and separation time is short and the detection efficiency is high. However, the detection method has a low response on a low-end triple quadrupole mass spectrometer, the detection result is greatly affected by the mass spectrometry resolution, and the running time is long, and the detection cost is high. Summary of the invention

[0008] In view of the shortcomings of the prior art, the present invention provides a detection method for determining N-nitroso-vonoprazan in vonoprazan fumarate, which uses an ultra-high pressure chromatographic column with shorter running time, higher signal-to-noise ratio, higher detection sensitivity, and lower detection cost, and can also meet detection requirements on a low-end triple quadrupole mass spectrometer.

[0009] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows: A method for determining N-nitroso-vonoprazan in vonoprazan fumarate comprises: preparing a solution, detecting; The solution preparation comprises using the fine powder of vonoprazan fumarate tablets as a raw material to prepare the test solution, and using the N-nitroso-vonoprazan reference substance as a raw material to prepare the reference substance solution; The detection adopts liquid chromatography-mass spectrometry to measure the test solution and the reference solution, records the chromatogram, and calculates the content of N-nitroso-vonoprazan according to the chromatogram by external standard method; In the detection, the liquid chromatography-mass spectrometry method uses a triple quadrupole liquid mass spectrometer; The injection volume of the test solution and the reference solution was 5 μL respectively; The chromatographic conditions of the liquid chromatography-mass spectrometry method are as follows: the chromatographic column uses octadecylsilane bonded silica gel as a filler; The mobile phase consists of mobile phase A and mobile phase B, mobile phase A is 0.09-0.11% formic acid solution, and mobile phase B is acetonitrile; Flow rate: 0.35-0.45 mL / min; Injection volume: 5 μL; Column temperature: 25-35°C; Gradient elution was used, and the degree of gradient elution was as follows: the total volume of the mobile phase was 100%, at 0-3 min, the volume of mobile phase A was 59-61%, and the volume of mobile phase B was 39-41%; at 3-5 min, the volume of mobile phase A decreased from 59-61% to 4-6%, and the volume of mobile phase B increased from 39-41% to 94-96%; at 5-7 min, the volume of mobile phase A was 4-6%, and the volume of mobile phase B was 94-96%; at 7-12 min, the volume of mobile phase A was 59-61%, and the volume of mobile phase B was 39-41%; Further, in the preparation solution, the preparation method of the test solution is: accurately weighing the fine powder of vonola fumarate raw tablets, adding it into a centrifuge tube, adding a solvent to dissolve, vortexing, shaking, centrifuging, filtering the supernatant, and obtaining the filtrate; In the preparation of the test solution, the solvent is acetonitrile-water, wherein acetonitrile: water = 5:5; The vortex time was 1 min, the shaking time was 10 min, and the filter membrane used for filtration was a 0.22 μm organic nylon 66 filter membrane; The concentration of vonoprazan fumarate in the test solution is 1.8-2.2 mg / mL; The preparation method of the reference substance solution is as follows: accurately weighing the N-nitroso-vonoprazan reference substance, dissolving it in a solvent, and quantitatively diluting it to prepare the reference substance solution; In the preparation of the reference solution, the solvent is acetonitrile-water, wherein acetonitrile:water=5:5; The concentration of the N-nitroso-vonoprazan reference substance in the reference substance solution is 6.48-7.92 ng / mL; Further, in the detection, the chromatographic conditions of the liquid chromatography-mass spectrometry are as follows: the specifications of the chromatographic column are Poroshell 120EC-C18, 1.9 μm, 50 mm×3.0 mm; Further, in the detection, the mass spectrometry conditions of the liquid chromatography-mass spectrometry method are: ion source type: HeatedESI+; Positive ion spray voltage: 3500V; Ion transfer tube temperature: 325°C; Evaporation temperature: 350℃.

[0010] Sheath gas flow: 50Arb; Auxiliary gas flow: 10Arb; Quantitative ion pair (M / Z): 375→315, collision voltage: 10 V, tube voltage: 84 V; Qualifying ion pair (M / Z): 375→173, collision voltage: 27 V, tube voltage: 84 V; Sheath gas and auxiliary gas are high-purity nitrogen; The collision gas was high-purity argon; Mass spectrometer switching valve: 0-1.8min, the switching valve opens, the effluent from the chromatographic column enters the waste liquid, 1.8-5min enters the mass spectrometer collection, and after 5min, the switching valve opens, and the effluent from the chromatographic column enters the waste liquid.

[0011] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention provides a method for detecting N-nitroso-vonoprazan in vonoprazan fumarate according to the product characteristics and the properties of the compound, using a triple quadrupole liquid mass spectrometer for determination and gradient elution, and the blank solvent does not interfere with the detection of N-nitroso-vonoprazan. By optimizing the conditions, the sensitivity and accuracy of N-nitroso-vonoprazan are further improved and meet the limit requirements, while the detection cost is reduced. (2) The detection method of the present invention for determining N-nitroso-vonoprazan in vonoprazan fumarate uses an ultra-high pressure chromatographic column Poroshell 120EC-C18, 1.9 μm, 50 mm×3.0 mm, which has a shorter running time, a sharper peak shape, and a higher signal-to-noise ratio when detecting N-nitroso-vonoprazan; (3) The detection method of the present invention for determining N-nitroso-vonoprazan in vonoprazan fumarate can be applied to a low-resolution triple quadrupole mass spectrometer, with faster analysis speed, better specificity, lower detection cost and higher accuracy, so as to facilitate the detection and control of N-nitroso-vonoprazan impurities, which is conducive to the popularization of the detection method of N-nitroso-vonoprazan. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 is the liquid chromatogram of the blank solution in Example 2; Figure 2 It is the liquid chromatogram of the reference substance in Example 2. DETAILED DESCRIPTION

[0013] In order to more clearly understand the above-mentioned purpose, features and advantages of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be noted that the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

[0014] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from the description. Therefore, the protection scope of the present invention is not limited to the embodiments disclosed below.

[0015] Example 1 This embodiment provides a method for detecting N-nitroso-vonoprazan in vonoprazan fumarate, specifically: 1. Instrument: A triple quadrupole liquid mass spectrometer was used for detection. The chromatographic column was filled with octadecylsilane bonded silica gel, with specifications of Poroshell 120EC-C18, 1.9 μm, 50 mm × 3.0 mm; Reagents: Methanol is of mass spectrometry grade, formic acid is of mass spectrometry grade, and water is ultrapure water; The manufacturer of N-nitroso-vonoprazan reference substance is SINCO, the batch number is 23-07-2612, and the purity is 98.75%.

[0016] 2. Preparation of solution: Preparation of test solution: Take an appropriate amount of vonoprazan fumarate tablet powder (equivalent to 20 mg of vonoprazan fumarate), accurately weigh it, put it in a 50 mL centrifuge tube, add 10 mL of solvent to dissolve it, vortex for 1 min, shake it for 10 min, centrifuge it, take the supernatant and filter it with a 0.22 μm organic nylon 66 filter membrane, and take the filtrate to obtain the solution; The solvent is acetonitrile-water (5:5); Preparation of reference solution: accurately weigh an appropriate amount of N-nitroso-vonoprazan reference substance, dissolve it in solvent and quantitatively dilute it to make a reference solution containing 7.2 ng of N-nitroso-vonoprazan reference substance per 1 mL of solvent; The solvent is acetonitrile-water (5:5).

[0017] 3. Liquid chromatography and mass spectrometry conditions: Flow rate: 0.4 mL / min; Injection volume: 5 μL; Column temperature: 30°C; Mobile phase A: 0.1% formic acid; Mobile phase B: acetonitrile; Gradient elution was used, and the total volume of the mobile phase was 100%. 0-3min, the volume percentage of the mobile phase A is 60%, and the volume percentage of the mobile phase B is 40%; 3-5min, the volume percentage of the mobile phase A decreases from 60% to 5%, and the volume percentage of the mobile phase B increases from 40% to 95%; 5-7min, the volume percentage of the mobile phase A is 5%, and the volume percentage of the mobile phase B is 95%; 7-12 min, the volume percentage of the mobile phase A is 60%, and the volume percentage of the mobile phase B is 40%; Ion source type: Heated ESI+; Positive ion spray voltage: 3500V; Ion transfer tube temperature: 325°C; Evaporation temperature: 350℃.

[0018] Sheath gas flow: 50Arb; Auxiliary gas flow: 10Arb; Quantitative ion pair (M / Z): 375→315, collision voltage: 10 V, tube voltage: 84 V; Qualifying ion pair (M / Z): 375→173, collision voltage: 27 V, tube voltage: 84 V; Sheath gas and auxiliary gas are high-purity nitrogen; The collision gas was high-purity argon; Mass spectrometer switching valve: 0-1.8min, the switching valve opens, the effluent from the chromatographic column enters the waste liquid, 1.8-5min enters the mass spectrometer collection, and after 5min, the switching valve opens, and the effluent from the chromatographic column enters the waste liquid.

[0019] 4. Detection: Accurately pipette 5 μL of the test solution and the reference solution respectively, inject into a triple quadrupole liquid mass spectrometer, record the chromatogram, and calculate the content of N-nitroso-vonoprazan according to the chromatogram using the external standard method.

[0020] Example 2 Specificity Test Blank solution: acetonitrile-water (5:5).

[0021] Preparation of reference solution: Take an appropriate amount of N-nitroso-vonoprazan reference and dilute with solvent to obtain a 7.2 ng / mL solution; The solvent is acetonitrile-water (5:5).

[0022] Take 5 μL of blank solution and reference solution respectively, and record the liquid chromatogram according to the liquid chromatography analysis conditions, mass spectrometry conditions and detection method in Example 1; the liquid chromatogram of the blank solution obtained is shown in Figure 1 The liquid chromatogram of the obtained reference solution is shown in Figure 2 .

[0023] Verification results: The solvent blank had no interference with the determination of N-nitroso-vonoprazan.

[0024] Example 3 System adaptability experiment Preparation of reference solution: Take an appropriate amount of N-nitroso-vonoprazan reference and dilute with solvent to obtain a 7.2 ng / ml solution; The solvent is acetonitrile-water (5:5).

[0025] The reference solution was sampled six times continuously, and the liquid chromatogram was recorded according to the liquid chromatography analysis conditions, mass spectrometry conditions and detection method in Example 1.

[0026] Verification results: The RSD of the peak area of ​​the reference solution after six consecutive injections was 7.5%.

[0027] Example 4 Repeatability Experiment Preparation of reference solution: Take an appropriate amount of N-nitroso-vonoprazan reference and dilute with solvent to obtain a 7.2 ng / mL solution; The solvent is acetonitrile-water (5:5).

[0028] Preparation of reproducible solution: Take an appropriate amount of fine powder of vonoprazan fumarate tablets (equivalent to 20 mg of vonoprazan fumarate), accurately weigh it, put it in a 50 mL centrifuge tube, add 10 mL of reference solution to dissolve it, vortex for 1 min, shake it for 10 min, centrifuge it, filter the supernatant, and take the filtrate to obtain the solution; Prepare 6 replicate solutions as described above.

[0029] Determination: Take a repetitive solution injection, record the liquid chromatogram according to the liquid chromatography analysis conditions, mass spectrometry conditions and detection method in Example 1, refer to the liquid chromatogram in Example 3, and calculate the content of N-nitroso-vonoprazan according to the external standard method.

[0030] Verification results: The RSD of the content of six samples was 9.19%.

[0031] Example 5 Intermediate Precision Experiment On another day of the repetitive experiment of Example 4, another laboratory technician prepared the repetitive solution six times in succession according to the preparation method of the repetitive experiment of Example 4.

[0032] Determination: Take a repetitive solution injection, record the chromatogram according to the liquid chromatography analysis conditions, mass spectrometry conditions and detection method in Example 1, refer to the liquid chromatogram in Example 3, and calculate the content of N-nitroso-vonoprazan according to the external standard method.

[0033] Verification results: The RSD of the six samples was 12.52%; the RSD of the repeatability and intermediate precision for 12 times was 11.04% Example 6 Linearity Experiment Weigh appropriate amounts of N-nitroso-vonoprazan reference substance respectively, and prepare linear solutions 1-5 with concentrations of 0.72 ng / mL, 2.88 ng / mL, 5.76 ng / mL, 7.2 ng / mL, and 14.4 ng / mL with solvent; The solvent is acetonitrile-water (5:5).

[0034] Determination: Linear solutions No. 1-5 were sampled separately, and the chromatograms were recorded according to the liquid chromatography analysis conditions, mass spectrometry conditions and detection method in Example 1.

[0035] Verification results: In the concentration range of 0.72ng / mL-14.4ng / mL, the linear equation Y=157.2320X-50.2747 was obtained, the correlation coefficient R was 0.9993, the RSD of the response factor was 5.59%, and the ratio of the Y-axis intercept to the 100% response value was 4.69%.

[0036] Example 7 Accuracy Experiment Preparation of reference solution: Take an appropriate amount of N-nitroso-vonoprazan (NVP) reference substance and dilute it with solvent to a solution of 7.2 ng / ml; The solvent is acetonitrile-water (5:5).

[0037] Preparation of sample blank solution: Take an appropriate amount of vonoprazan fumarate tablet powder (equivalent to 20 mg of vonoprazan fumarate), accurately weigh it, put it in a 50 mL centrifuge tube, add 10 mL of solvent to dissolve it, vortex for 1 min, shake it for 10 min, centrifuge it, filter the supernatant, and take the filtrate to obtain the solution; The solvent is acetonitrile-water (5:5).

[0038] Preparation of accuracy test solution No. 1 (quantitative limit concentration): Weigh an appropriate amount of fine powder of vonoprazan fumarate tablets (equivalent to 20 mg of vonoprazan fumarate), accurately weigh, and dissolve in a 50 mL centrifuge tube, add 10 mL of the 0.72 ng / mL solution in the linear experiment item of Example 6, vortex for 1 min, shake for 10 min, centrifuge, filter the supernatant, and obtain the filtrate; Prepare 3 solutions according to the above method.

[0039] Preparation of accuracy test solution No. 2: Take an appropriate amount of fine powder of vonoprazan fumarate tablets (equivalent to 20 mg of vonoprazan fumarate), accurately weigh it, put it into a 50 ml centrifuge tube, add 10 ml of the 7.2 ng / ml solution in the linear experiment of Example 6 to dissolve it, vortex for 1 min, shake it for 10 min, centrifuge it, filter the supernatant, and take the filtrate to obtain the solution; Prepare 3 solutions according to the above method.

[0040] Preparation of accuracy test solution No. 3: Weigh an appropriate amount of fine powder of vonoprazan fumarate tablets (equivalent to 20 mg of vonoprazan fumarate), accurately weigh, and dissolve in a 50 mL centrifuge tube, add 10 mL of the 14.4 ng / mL solution in the linear experiment of Example 6, vortex for 1 min, shake for 10 min, centrifuge, filter the supernatant, and obtain the filtrate; Prepare 3 solutions according to the above method.

[0041] Determination: Take sample blank solution, accuracy test solution No. 1, accuracy test solution No. 2, and accuracy test solution No. 3 and inject them respectively, and record the chromatograms.

[0042] Verification results: average recovery rate 91.5%.

[0043] Example 8 Detection Limit and Quantification Limit Experiment Weigh the N-nitroso-vonoprazan reference substance, dilute it with solvent to different concentrations and inject it. The limit of quantitation is 0.36ppm. The RSD of the peak area of ​​the limit of quantitation solution injected six times is 2.41%, and the detection limit is 0.036ppm. The solvent is acetonitrile-water (5:5).

[0044] Example 9 Durability Test Preparation of reference solution: Take an appropriate amount of N-nitroso-vonoprazan (NVP) reference substance and dilute it with solvent to a solution of 7.2 ng / ml; The solvent is acetonitrile-water (5:5).

[0045] Preparation of durability solution: Weigh an appropriate amount of vonoprazan fumarate tablet powder (equivalent to 20 mg of vonoprazan fumarate), accurately weigh it, put it into a 50 mL centrifuge tube, add 10 mL of 7.2 ng / ml reference solution to dissolve it, vortex for 1 min, shake it for 10 min, centrifuge it, filter the supernatant, and take the filtrate to obtain the final product.

[0046] Determination: Detect the solution once according to the original chromatographic conditions, changing the column temperature from 30°C to 28°C and 32°C respectively, record the chromatogram, and calculate the content of N-nitroso-vonoprazan according to the external standard method using the reference solution under the system suitability item.

[0047] Verification results: The ratio of the durability solution content measured after changing the parameters to the content measured under normal conditions is between 0.8-1.2.

[0048] Example 10 Solution stability test Preparation of reference solution: Take an appropriate amount of N-nitroso-vonoprazan reference and dilute it with solvent to a solution of 7.2 ng / mL; The solvent is acetonitrile-water (5:5).

[0049] Preparation of sample solution: Take an appropriate amount of vonoprazan fumarate tablet powder (equivalent to 20 mg of vonoprazan fumarate), accurately weigh it, put it in a 50 mL centrifuge tube, add 10 mL of 7.2 ng / mL reference solution to dissolve it, vortex for 1 min, shake for 10 min, centrifuge, filter the supernatant, and take the filtrate.

[0050] Determination: Take reference solution and sample solution and inject them at different time periods, and record the chromatograms.

[0051] Verification results: The RSD of the peak area of ​​the reference solution within 10 hours was 5.63%; the RSD of the peak area of ​​the sample solution within 10 hours was 8.26%.

[0052] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may also have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for determining N-nitroso-vonoprazan in vonoprazan fumarate, characterized in that: include: Prepare solutions and test; The detection adopts liquid chromatography-mass spectrometry to measure the test solution and the reference solution, records the chromatogram, and calculates the content of N-nitroso-vonoprazan by peak area according to the external standard method based on the chromatogram; In the detection, the liquid chromatography-mass spectrometry method uses a triple quadrupole liquid mass spectrometer; The chromatographic conditions of the liquid chromatography-mass spectrometry method are as follows: the chromatographic column uses octadecylsilane bonded silica gel as a filler; The mobile phase consists of mobile phase A and mobile phase B, mobile phase A is 0.09-0.11% formic acid solution, and mobile phase B is acetonitrile; Flow rate: 0.35-0.45 mL / min; Injection volume: 5 μL; Column temperature: 25-35°C; Gradient elution was adopted, and the degree of gradient elution was as follows: the total volume of mobile phase was 100%, at 0-3 min, the volume of mobile phase A was 59-61%, and the volume of mobile phase B was 39-41%; at 3-5 min, the volume of mobile phase A decreased from 59-61% to 4-6%, and the volume of mobile phase B increased from 39-41% to 94-96%; at 5-7 min, the volume of mobile phase A was 4-6%, and the volume of mobile phase B was 94-96%; at 7-12 min, the volume of mobile phase A was 59-61%, and the volume of mobile phase B was 39-41%.

2. The method for detecting N-nitroso-vonoprazan in vonoprazan fumarate according to claim 1, characterized in that: The solution preparation method uses the fine powder of vonoprazan fumarate tablets as a raw material to prepare the test solution, and uses the N-nitroso-vonoprazan reference substance as a raw material to prepare the reference substance solution.

3. The method for detecting N-nitroso-vonoprazan in vonoprazan fumarate according to claim 1, characterized in that: In the test, the injection volume of the test solution and the reference solution was 5 μL respectively.

4. The method for detecting N-nitroso-vonoprazan in vonoprazan fumarate according to claim 2, characterized in that: In the preparation of the solution, the preparation method of the test solution is: accurately weigh the fine powder of vonola raw fumarate tablets, add it into a centrifuge tube, add a solvent to dissolve, vortex, shake, centrifuge, filter the supernatant, and obtain the filtrate; In the preparation of the test solution, the solvent is acetonitrile-water, wherein acetonitrile: water = 5:5; The vortex time was 1 min, the shaking time was 10 min, and the filter membrane used for filtration was a 0.22 μm organic nylon 66 filter membrane; The concentration of vonoprazan fumarate in the test solution is 1.8-2.2 mg / mL.

5. The method for detecting N-nitroso-vonoprazan in vonoprazan fumarate according to claim 2, characterized in that: In the prepared solution, the preparation method of the reference solution is: accurately weighing the N-nitroso-vonoprazan reference substance, adding a solvent to dissolve it and quantitatively diluting it to prepare the reference solution; In the preparation of the reference solution, the solvent is acetonitrile-water, wherein acetonitrile:water=5:5; The concentration of the N-nitroso-vonoprazan reference substance in the reference substance solution is 6.48-7.92 ng / mL.

6. The method for detecting N-nitroso-vonoprazan in vonoprazan fumarate according to claim 1, characterized in that: In the detection, the chromatographic conditions of the liquid chromatography-mass spectrometry are as follows: the specifications of the chromatographic column are Poroshell120EC-C18, 1.9 μm, 50 mm×3.0 mm.

7. The method for detecting N-nitroso-vonoprazan in vonoprazan fumarate according to claim 1, characterized in that: In the detection, the mass spectrometry conditions of the liquid chromatography-mass spectrometry method are as follows: ion source type: Heated ESI+; Positive ion spray voltage: 3500V; Ion transfer tube temperature: 325°C; Evaporation temperature: 350℃; Sheath gas flow: 50Arb; Auxiliary gas flow: 10Arb; Quantitative ion pair (M / Z): 375→315, collision voltage: 10 V, tube voltage: 84 V; Qualifying ion pair (M / Z): 375→173, collision voltage: 27 V, tube voltage: 84 V; Sheath gas and auxiliary gas are high-purity nitrogen; The collision gas was high-purity argon; Mass spectrometer switching valve: 0-1.8min, the switching valve opens, the effluent from the chromatographic column enters the waste liquid, 1.8-5min enters the mass spectrometer collection, and after 5min, the switching valve opens, and the effluent from the chromatographic column enters the waste liquid.

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