A method for detecting the content of related substances in vonoprazan fumarate

Abstract

The present invention provides a kind of detection method of thirteen kinds of related substance contents in vonoprazan fumarate, and high performance liquid chromatography is used for detection, and the chromatographic conditions are: the test solution and the reference solution are injected into the high performance liquid chromatograph, and the chromatographic conditions are: the chromatographic column adopts octadecyl bonded silica gel column; the ultraviolet detector is used, and the detection wavelength is 260nm; the injection volume: 10μl; the column temperature: 30℃‑40℃; the flow rate: 0.8ml / min‑1.0ml / min; the gradient elution method is used, and the mobile phase A: the solution of 0.02mol / L potassium dihydrogen phosphate solution adjusted to pH 8.0 with potassium hydroxide solution and isopropanol mixed with a volume ratio of 95:5; the mobile phase B: the mixed solution of acetonitrile and isopropanol with a volume ratio of 95:5. The method can be used to detect the contents of 13 kinds of related substances in vonoprazan fumarate, and high performance liquid chromatography is used for detection, and the method has high detection sensitivity, and the verification results show that the system applicability and specificity of this method meet the requirements, and the needs of drug production quality monitoring can be met.

Description

Technical Field

[0001] The invention relates to the field of drug analysis, and in particular to a method for detecting the content of related substances in vonoprazan fumarate. Background Art

[0002] The chemical structure of vonoprazan fumarate is 5-(2-fluorophenyl)-N-methyl-1-(3-pyridylsulfonyl)-1H-pyrrole-3-methylamine fumarate. In related studies, it was found that impurities of various structures may be mixed into vonoprazan fumarate APIs, and there are many types. According to the guidelines of ICH (The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use), for impurities present in APIs, it is generally required to limit the impurity content in the APIs to a certain range.

[0003] Regarding the detection method of related substances in vonoprazan fumarate, Chinese patent CN116953129B reports a method for detecting the contents of 12 impurities in vonoprazan fumarate using HPLC, and the chromatographic conditions used are octadecyl bonded silica gel chromatographic column, mobile phase A: phosphate solution-acetonitrile-methanol; mobile phase B: methanol, using gradient elution method. The types of substances detected are 12 related substances such as vonoprazan fumarate impurity A in vonoprazan fumarate. However, the types of related substances that can be detected by this patented method are limited, and all common related substances in vonoprazan fumarate raw materials cannot be detected. Therefore, in order to better control the quality of vonoprazan fumarate raw materials, it is urgent to develop a method that can detect all common related substances in vonoprazan fumarate raw materials. Summary of the invention

[0004] In order to solve the above problems, the present invention provides a method for detecting the content of related substances in vonoprazan fumarate, which can detect the contents of 13 related substances in vonoprazan fumarate. The detection method has high sensitivity, and the system applicability and specificity meet the requirements of the pharmacopoeia, and can be used for the quality control of vonoprazan fumarate raw materials.

[0005] The present invention provides a method for detecting related substances in vonoprazan fumarate, which is analyzed by high performance liquid chromatography. The detection method comprises the following steps: (1) Solution preparation: Take the vonoprazan fumarate reference substance and prepare it into a reference substance solution with a mixed solution of acetonitrile: water = 30:70 (V / V); take the vonoprazan fumarate sample to be tested and prepare it into a test solution with a mixed solution of acetonitrile: water = 30:70 (V / V); (2) The test solution and the reference solution were injected into the high performance liquid chromatograph under the following chromatographic conditions: a chromatographic column filled with octadecyl bonded silica gel; a ghost-buster column (Welch Ghost-Buster Column, 4.6 mm × 50 mm); an ultraviolet detector with a detection wavelength of 260 nm; an injection volume of 10 μl; a column temperature of 30°C-40°C; a flow rate of 0.8 ml / min-1.0 ml / min; a gradient elution method; mobile phase A: a solution of 0.02 mol / L potassium dihydrogen phosphate solution adjusted to pH 8.0 with potassium hydroxide solution and isopropanol in a volume ratio of 95:5; mobile phase B: a solution of acetonitrile and isopropanol in a volume ratio of 95:5.

[0006] Specifically, the gradient elution method is performed according to the following table:

[0007] Specifically, the chromatographic column is Agilent Eclipse XDB-C18, 4.6 mm×250 mm, 5.0 μm.

[0008] Specifically, the related substances are the following 13 structures:

[0009] Specifically, the results of the detection method are calculated according to the following formula: ; Wherein, Ru: peak area of ​​each impurity in the test solution; ms: the amount of vonoprazan fumarate in the reference solution (mg); Rs: vonoprazan peak area in reference solution; C: the content of vonoprazan fumarate in the reference solution; m: amount of test sample (mg); 100: The ratio of the dilution factor of the reference solution to the dilution factor of the test solution; F: Correction factor. The validation of the method is summarized as follows:

[0010]

[0011]

[0012] Beneficial effects: The present invention provides a method for detecting related substances in vonoprazan fumarate, which can be used to detect the contents of 13 related substances in vonoprazan fumarate. The detection is performed by high performance liquid chromatography. The method has high detection sensitivity. The verification results show that the system applicability and specificity of this method meet the requirements and can meet the needs of drug production quality monitoring. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 is the sensitivity solution chromatogram; Figure 2 is the chromatogram of the reference solution; Figure 3 is a selective solution chromatogram; Figure 4 For selective solution chromatogram continued; Figure 5 is the precision and repeatability solution chromatogram; Figure 6 For precision and repeatability, the solution chromatogram is continued in 1; Figure 7 For precision and repeatability solution chromatogram continued 2; Figure 8 For precision and repeatability, the solution chromatogram is continued in 3; Fig. 9 The chromatogram is a comparative example. DETAILED DESCRIPTION Example 1

[0014] System suitability Solution preparation: Diluent: acetonitrile: water = 30:70 (V / V); Blank solution: diluent; Reference solution: Prepare vonoprazan fumarate reference solution (vonoprazan fumarate concentration: 14.9046 μg / ml).

[0015] Sensitivity solution: Prepare with vonoprazan fumarate reference substance (vonoprazan fumarate concentration: 0.7452 μg / ml) The above solution was injected, and the chromatographic conditions were as follows: chromatographic column: Agilent Eclipse XDB-C18 with octadecyl bonded silica gel as filler, 4.6mm×250mm, 5.0μm; UV detector, detection wavelength: 260nm; injection volume: 10μl; column temperature: 30℃; flow rate: 0.8ml / min; mobile phase A: 0.02mol / L potassium dihydrogen phosphate solution, pH adjusted to 8.0 with potassium hydroxide solution: isopropanol = 95:5; mobile phase B: acetonitrile: isopropanol = 95:5 (V / V), gradient elution. The gradient elution was performed according to the following gradient:

[0016] The sensitivity solution chromatogram is as follows Figure 1 As shown, the chromatogram of the reference solution is Figure 2 shown.

[0017] The system suitability test results are shown in Table 1 below.

[0018]

[0019] In the chromatogram of the sensitive solution, the signal-to-noise ratio of the chromatographic peak height of Vonoprazan was 18.6; in the chromatogram of the reference solution, the theoretical plate number calculated based on the Vonoprazan peak was 97995, which met the standard requirements. (The standard stipulates that: in the chromatogram of the sensitive solution, the signal-to-noise ratio of the chromatographic peak height of Vonoprazan should be no less than 10; in the chromatogram of the reference solution, the theoretical plate number calculated based on the Vonoprazan peak should be no less than 5000.) The system stability test results of the obtained reference solution are shown in Table 2 below.

[0020]

[0021] From the above experiments, it can be seen that the recovery rate of the main peak of vonoprazan meets the standard requirements.

[0022] Example 2 Specificity experiment Solution preparation: Diluent: acetonitrile: water = 30:70 (V / V); Blank solution: diluent.

[0023] Preparation of test solution: Prepare the test solution of vonoprazan fumarate (vonoprazan fumarate concentration: 1.5064 μg / ml)

[0024] The above solution was injected, and the chromatographic conditions were the same as those in Example 1; the chromatogram of the reference solution was shown in FIG2 , the chromatograms of the selective solution were shown in FIG3 and FIG4 , and the obtained specificity detection results were shown in the following table.

[0025]

[0026]

[0027] From the above experiments, it can be seen that the blank solution has no interference with the detection of each component, and the minimum separation between each component and the adjacent peak in the selective solution is 1.5, which meets the standard requirements. (The standard stipulates that the blank solution should have no interference with the detection of each component, and the separation between each component and the adjacent peak in the selective solution should be ≥1.2.) Example 3

[0028] Precision experiment Solution preparation: diluent is the same as in Example 1; Test solution: 1.5170mg / ml Reference solution: 15.0090 mg / ml Preparation of precision and repeatability solution: Take impurity AM samples, respectively prepare them into solutions with a concentration of 300 mg / ml with diluent, take 5 ml of the obtained solution, mix and dilute to 100 ml to obtain a mixed stock solution; weigh 75 mg of vonoprazan fumarate sample, mix with 5 ml of the mixed stock solution and dilute to 100 ml to obtain precision solution Precision, prepare 6 portions in parallel in the same way to obtain Precision 1-6;

[0029] The above solution was injected, and the chromatographic conditions were the same as those in Example 1; the chromatograms of the precision repeatability experiment are shown in Figures 5 and 6. Figure 6 , as shown in Figure 7 and Figure 8; the experimental results are shown in the following table:

[0030] From the experimental results, it can be seen that the RSD of each impurity meets the standard requirements. (The standard stipulates: in the determination results of 6 test solution (spiked): if the impurity content is less than LOQ, the RSD is not calculated; if the impurity content is ≥ LOQ, the RSD should be ≤10.0%.) Comparative Example 1

[0031] The precision solution of Comparative Example 1 was tested under the following conditions: The chromatographic column uses Agilent Eclipse XDB-C18 with octadecyl bonded silica as filler, 4.6mm×250mm, 5.0μm; uses ultraviolet detector, detection wavelength: 260nm; injection volume: 10μl; column temperature: 30℃; flow rate: 0.8ml / min; mobile phase A: 0.02mol / L potassium dihydrogen phosphate solution, pH adjusted to 6.5 with potassium hydroxide solution: isopropanol = 95:5; mobile phase B: acetonitrile: isopropanol = 95:5 (V / V), gradient elution. The gradient elution is performed according to the following gradient:

[0032] The test results show that the separation between impurities E, A and G is poor, and the separation that does not meet the test requirements is not less than 1.5; Fig. 9 shown.

Claims

1. A method for detecting the content of related substances in vonoprazan fumarate, using high performance liquid chromatography for analysis, characterized in that: The detection method can detect 13 related substances contained in vonoprazan fumarate, and the 13 related substances are as follows: The detection method comprises the following steps: (1) Solution preparation: Take the vonoprazan fumarate reference substance and prepare it into a reference substance solution with a mixed solution of acetonitrile and water in a volume ratio of 30:70; take the vonoprazan fumarate sample to be tested and prepare it into a test solution with a mixed solution of acetonitrile and water in a volume ratio of 30:70; (2) Inject the test solution and the reference solution into a high performance liquid chromatograph under the following chromatographic conditions: the chromatographic column is an octadecyl bonded silica gel column; An ultraviolet detector was used with a detection wavelength of 260 nm; injection volume: 10 μl; column temperature: 30°C-40°C; flow rate: 0.8 ml / min-1.0 ml / min; a gradient elution method was adopted, mobile phase A: a solution of 0.02 mol / L potassium dihydrogen phosphate solution whose pH value was adjusted to 8.0 with potassium hydroxide solution and isopropanol in a volume ratio of 95:5; mobile phase B: a mixed solution of acetonitrile and isopropanol in a volume ratio of 95:

5.

2. The detection method according to claim 1, characterized in that The gradient elution was performed according to the following gradient: 。 3. The detection method according to claim 1, characterized in that The chromatographic conditions also include a ghost peak trapping column Welch Ghost-Buster Column, 4.6 mm×50 mm.

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