HPLC Determination of Residual Solvent 3-Pyridine Sulfonic Acid in Vonoprazan Fumarate API

Abstract

This invention provides a high-performance liquid chromatography (HPLC) method for the detection of 3-pyridinesulfonic acid in vonoprazan fumarate raw material. The method mainly includes five steps: preparation of diluent, preparation of test solution, preparation of reference solution, chromatographic conditions, and determination of 3-pyridinesulfonic acid in vonoprazan fumarate. The chromatographic column is packed with octadecylsilane-bonded silica gel, and the mobile phase is a mixture of sodium phosphate solution and acetonitrile in a specific ratio. The detection wavelength is 210 nm, the flow rate is 0.5–1.5 mL / min, the injection volume is 5–50 μL, and the column temperature is 25–40 °C. The detection method provided by this invention for determining 3-pyridinesulfonic acid in vonoprazan fumarate raw material has good accuracy and repeatability, strong specificity, high accuracy, and is simple to operate.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a high performance liquid chromatography method for determining residual solvent 3-pyridine sulfonic acid in a vonoprazan fumarate raw material drug. Background Art

[0002] Vonoprazan is a new type of potassium channel blocker (P-CABs) that inhibits K + For H + -K + -ATPase (proton pump) binding action, prematurely terminates the secretion of gastric acid, and has a strong and lasting inhibitory effect on gastric acid secretion. Vonoprazan fumarate tablets were developed by Takeda Pharmaceutical Company Limited of Japan and were first approved for marketing in Japan in December 2014 for the treatment of non-erosive gastroesophageal reflux disease, duodenal ulcer, gastric ulcer, and erosive esophagitis (cure and maintenance treatment).

[0003] In the synthesis process of the raw material drug, the organic solvent 3-pyridinesulfonic acid is required. This solvent has certain adverse effects on human health and its content needs to be strictly controlled. Summary of the invention

[0004] The object of the present invention is to provide a high performance liquid chromatography detection method for 3-pyridine sulfonic acid in vonoprazan fumarate raw materials, which has good specificity, high sensitivity, accuracy, precision, and strong durability, and can quickly and accurately determine the content of 3-pyridine sulfonic acid in vonoprazan fumarate raw materials, thereby better controlling product quality and improving the safety of clinical medication.

[0005] To achieve the above object, the present invention is characterized by comprising the following steps:

[0006] (1) Preparation of diluent: water.

[0007] (2) Preparation of test solution: Take an appropriate amount of vonoprazan fumarate, place it in a volumetric flask, dissolve it in a 40°C water bath and dilute it to make a solution containing 0.5 mg to 1.5 mg of vonoprazan fumarate per 1 ml, which is used as the test solution.

[0008] (3) Preparation of reference solution: Weigh an appropriate amount of 3-pyridinesulfonic acid and dissolve it in water to make a solution of about 1 μg per 1 ml, which is used as the reference solution.

[0009] (4) Chromatographic conditions:

[0010] According to the high performance liquid chromatography method, a chromatographic column with octadecylsilane bonded silica gel as a filler and a phosphate buffer solution-acetonitrile (the pH value of the phosphate buffer solution is 5.8 to 6.8) as a mobile phase, and the concentration of the phosphate buffer solution is 15 to 40 mmol / L. The following steps are included:

[0011] Detection on high performance liquid chromatography

[0012] Chromatographic column filler: The chromatographic column is filled with octadecylsilane bonded silica gel, the column size is 150 mm in length × 5 mm in inner diameter, and the particle size of the chromatographic column filler is 3 to 5 μm.

[0013] Column temperature: 25℃-40℃

[0014] Flow rate: 0.8mL / min-1.2mL / min

[0015] Detection wavelength: 200~215nm

[0016] Mobile phase: Phase A is phosphate buffer solution-acetonitrile, phase B is acetonitrile

[0017] Diluent: Water

[0018] The present invention establishes a high performance liquid chromatography analysis method for residual solvent 3-pyridinesulfonic acid that may exist in vonoprazan fumarate by testing the preparation methods of diluent, test solution and reference solution and optimizing the chromatographic conditions. The method uses an octadecylsilane bonded silica gel chromatographic column and the diluent is water, which significantly improves the sensitivity and accuracy of the detection method.

[0019] The present invention also verifies the established detection method, including specificity, detection limit and quantification limit, linearity and range, accuracy, precision, solution stability and durability. The verification results are good, indicating that the method can accurately determine the content of 3-pyridine sulfonic acid in the vonoprazan fumarate raw material drug, ensuring that the quality of the product is controllable. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The HPLC chromatogram of the solvent in Example 1 is

[0021] Figure 2 It is the HPLC chromatogram of the reference substance solution in Example 1.

[0022] Figure 3 The HPLC chromatogram of the vonoprazan fumarate test solution in Example 1 is shown in FIG. DETAILED DESCRIPTION

[0023] The technical solution of the present invention is described below in conjunction with specific examples, but does not constitute any limitation to the present invention.

[0024] Example 1

[0025] Specificity test

[0026] Adopt the method in the present invention.

[0027] Chromatographic column: GL Science Inertsil ODS-3C18 (4.6mm×150mm, 5μm)

[0028] Mobile Phase:

[0029] Mobile phase A: 0.025 mol / L sodium phosphate solution (pH 6.8)-acetonitrile (98:2)

[0030] Mobile phase B: acetonitrile

[0031] Linear Gradient:

[0032]

[0033] Flow rate: 1.0ml / min

[0034] Column temperature: 30°C

[0035] Detection wavelength: 210nm

[0036] Injection volume: 20 μl

[0037] Experimental steps:

[0038] Blank solvent: water

[0039] Preparation of system suitability solution: accurately weigh valsartan reference substance and enantiomer reference substance, dilute them with diluent to make each ml contain about 1 mg of valsartan and 1 μg of enantiomer, as system suitability solution;

[0040] Preparation of sample solution: Take an appropriate amount of vonoprazan fumarate, place it in a volumetric flask, dissolve it in a 40°C water bath and dilute it to make a solution containing 0.5 mg to 1.5 mg of vonoprazan fumarate per 1 ml, which is used as the test solution.

[0041] Preparation of reference solution: Take an appropriate amount of 3-pyridinesulfonic acid reference substance, dissolve it in water and quantitatively dilute it to make a solution containing about 1 μg / ml. Take each of the above solutions and inject them into the high performance liquid chromatograph respectively, and record the chromatogram.

[0042] The results of blank solvent are shown in the attached Figure 1 .

[0043] The results of the system suitability solution are attached. Figure 2 .

[0044] The results of the sample solution are shown in the attached Figure 3 .

[0045] Results: The blank solvent did not interfere with the detection of vonoprazan fumarate and its 3-pyridine sulfonic acid, and the method had good specificity.

[0046] Example 2

[0047] Limit of detection and limit of quantitation tests.

[0048] The chromatographic method in Example 1 was used.

[0049] Experimental steps:

[0050] Blank solvent: water

[0051] Detection limit and quantification limit solution: Take an appropriate amount of 3-pyridinesulfonic acid reference substance, add water to prepare a series of concentration solutions, and measure according to the law. The quantification limit and detection limit of the substance are determined by the signal-to-noise ratio. The results are shown in the table below:

[0052] Limit of Quantitation Results

[0053] name Limit of quantification concentration (μg / mL) S / N Equivalent to sample concentration (1mg / ml) (%) 3-Pyridinesulfonic acid 0.0062 22.4 0.006

[0054] Detection limit results

[0055] name Limit of quantification concentration (μg / mL) S / N Equivalent to sample concentration (1mg / ml) (%) 3-Pyridinesulfonic acid 0.0205 5.4 0.02

[0056] Example 3

[0057] Linear Relationship and Range

[0058] The chromatographic method in Example 1 was used.

[0059] Experimental steps:

[0060] Blank solvent: water

[0061] Linear solution: Take an appropriate amount of 3-pyridinesulfonic acid reference substance, dissolve it in water and prepare a series of concentration solutions, and measure according to the method.

[0062] Results: Within the concentration range of 0.0205 μg / ml to 2.0533 μg / ml, the peak area and concentration of TAK8-A had a good linear relationship, the correlation coefficient r was greater than 0.995, and the Y-axis intercept deviation was less than 25%; the linear results were good.

[0063] Example 3

[0064] Recovery test

[0065] The chromatographic method in Example 1 was used.

[0066] Experimental steps:

[0067] Accurately weigh an appropriate amount of vonoprazan fumarate, place it in a volumetric flask, accurately add an appropriate amount of 3-pyridinesulfonic acid stock solution, dissolve it with water and dilute it to the scale, prepare solutions with relative concentrations of 50%, 100%, and 150%, shake well, and use them as test solutions. Prepare 3 copies of each concentration in parallel. Accurately measure an appropriate amount of 3-pyridinesulfonic acid stock solution, quantitatively dilute it with water to prepare a solution of appropriate concentration, and use it as a reference solution. Accurately measure 20μl of each of the above solutions, inject them into the liquid chromatograph, and record the chromatogram and peak area. Calculate the recovery rate.

[0068] Results: The recovery rates of 3-pyridinesulfonic acid at each concentration were in line with the requirements of the pharmacopoeia. The accuracy of this method was good. The specific data are shown in the table below:

[0069] Valsartan enantiomer recovery results

[0070]

[0071] The recoveries at the limit values ​​of 50%, 100% and 150% were as follows: the recovery of 3-pyridinesulfonic acid was 102.1% to 103.7%, respectively. The results showed that the method had good accuracy.

[0072] Example 4 Durability test of the method of the present invention

[0073] The tolerance of the test method was investigated by changing parameters such as flow rate, column temperature, and different brands of chromatographic columns. Only one parameter was changed in each test, and the results were compared with the results when the parameters were not changed. No 3-pyridinesulfonic acid was detected. The specific results are shown in the table below:

[0074]

[0075] Example 5 Sample detection test

[0076] 1. Instruments and conditions:

[0077] High performance liquid chromatograph: Thermofisher Dixonex U3000, UV detector

[0078] Chromatographic column: GL Science Inertsil ODS-3C18 (4.6mm×150mm, 5μm)

[0079] Mobile Phase:

[0080] Mobile phase A: 0.025 mol / L sodium phosphate solution (pH 6.8)-acetonitrile (98:2)

[0081] Mobile phase B: acetonitrile

[0082] Linear Gradient:

[0083]

[0084] Flow rate: 1.0ml / min

[0085] Column temperature: 30°C

[0086] Injection volume: 20 μl

[0087] Detection wavelength: 210nm

[0088] 2. Experimental methods and results

[0089] (1) Preparation of diluent: water.

[0090] (2) Preparation of test solution: Take an appropriate amount of vonoprazan fumarate, place it in a volumetric flask, dissolve it in a 40°C water bath and dilute it to make a solution containing 0.5 mg to 1.5 mg of vonoprazan fumarate per 1 ml, which is used as the test solution.

[0091] (3) Preparation of reference solution: Weigh an appropriate amount of 3-pyridinesulfonic acid and dissolve it in water to make a solution of about 1 μg per 1 ml, which is used as the reference solution.

[0092] Take 20 μl of each of the above solutions, inject them into the liquid chromatograph in sequence, and record the chromatogram.

[0093] Table 4 Vonoprazan fumarate 3-pyridine sulfonic acid detection experimental results

[0094] batch number 3-Pyridinesulfonic acid 12828-105-200001 Not detected 12828-105-200002 Not detected 12828-105-200003 Not detected

Claims

1. A high performance liquid chromatography method for detecting 3-pyridine sulfonic acid in vonoprazan fumarate raw material, characterized in that The following steps are involved: (1) Preparation of diluent: water. (2) Preparation of test solution: Take an appropriate amount of vonoprazan fumarate, place it in a volumetric flask, dissolve it in a 40°C water bath and dilute it to make a solution containing 0.5 mg to 1.5 mg of vonoprazan fumarate per 1 ml, which is used as the test solution. (3) Preparation of reference solution: Weigh an appropriate amount of 3-pyridinesulfonic acid and dissolve it in water to make a solution of about 1 μg per 1 ml, which is used as the reference solution. (4) The chromatographic column of the high performance liquid chromatography method is: an octadecylsilane bonded silica gel filled chromatographic column; the mobile phase is: a phosphate buffer solution, and the pH value of the mobile phase is 5.8 to 6.

8. The 3-pyridinesulfonic acid in the vonoprazan fumarate raw material in the test solution was detected by high performance liquid chromatography, and then the content of 3-pyridinesulfonic acid in the vonoprazan fumarate raw material was calculated by external standard method based on the concentration-peak area of ​​the 3-pyridinesulfonic acid reference substance.

2. The detection method according to claim 1, characterized in that: The reverse phase chromatography column in step (1) is a GL Science Inertsil chromatography column.

3. The detection method according to claim 1, characterized in that: The size of the chromatographic column of the high performance liquid chromatography in step (1) is 150 mm in length and 5 mm in inner diameter, and the particle size of the chromatographic column filler is 3 to 5 μm.

4. The detection method according to claim 1, characterized in that: The phosphate buffer solution in step (1) is prepared by mixing sodium phosphate with water, and then adjusting the pH with aqueous ammonia to obtain a phosphate buffer solution; the concentration of the phosphate buffer solution is 15 to 40 mmol / L.

5. The detection method according to claim 1, characterized in that: The concentration of the phosphate buffer solution is 25 mmol / L.

6. The detection method according to claim 1, characterized in that: The column temperature of the high performance liquid chromatography in the step is 25-40°C.

7. The detection method according to claim 1, characterized in that: The flow rate of the high performance liquid chromatography in the step is 0.8 to 1.2 mL / min.

8. The detection method according to claim 1, characterized in that: The detection wavelength of the high performance liquid chromatography in the step is 200-215 nm.

Images