Method for detecting related substances in itopride hydrochloride tablet by HPLC (High Performance Liquid Chromatography)

Abstract

The invention relates to a method for detecting related substances in itopride hydrochloride tablets by HPLC (High Performance Liquid Chromatography), and the related substances comprise an impurity A, an impurity B and an impurity C. The method comprises the following steps: step 1, respectively preparing a blank solvent, a blank auxiliary material, a reference solution, a test solution and a system applicability solution; 2, detecting the blank solvent, the blank auxiliary material, the reference substance solution, the test solution and the system applicability solution in the step 1 by adopting HPLC (High Performance Liquid Chromatography), and recording a chromatogram; the quality of the main degraded impurities in the tablet is controlled by the method, and the method has the advantages of high sensitivity, strong specificity, good separation degree, high precision, good reproducibility and good accuracy, and can more conveniently and effectively control the quality of the product.

Description

Technical Field

[0001] The present invention discloses a technology related to pharmaceutical detection, and relates to a detection method for related substances in itopride hydrochloride tablets by HPLC. Background Art

[0002] Itopride hydrochloride is a new type of gastrointestinal prokinetic agent, applicable to various symptoms caused by functional dyspepsia, such as upper abdominal discomfort, postprandial fullness, early satiety, anorexia, nausea, vomiting, etc. It was first successfully developed by Hokuriku Pharmaceutical Co., Ltd. in Japan, and the tablets were approved for marketing in Japan in 1995. Currently, the original research product (specification: 50 mg, trade name: Approval number of domestic drugs: J20160075, manufacturer: ABBOTT LABORATORIES (M) SDN.BHD) has been imported into China, and 9 domestic companies have been approved to market generic drugs of this variety, with the specification of 50 mg for all.

[0003] In order to ensure the quality of itopride hydrochloride tablet raw materials and improve the safety of clinical medication, it is necessary to detect the related substances in itopride hydrochloride tablets. However, in the existing detection methods, some substances cannot be separated in the solvent, so inaccurate detection results will be produced. Summary of the Invention

[0004] In view of this, the present application provides a detection method for related substances in itopride hydrochloride tablets by HPLC, wherein the related substances are impurity A, impurity B, and impurity C, and the method includes the following steps: Step 1: Prepare a blank solvent, blank excipients, reference solution, test solution, and system suitability solution respectively; Step 2: Detect the blank solvent, blank excipients, reference solution, test solution, and system suitability solution in Step 1 by HPLC, and record the chromatogram.

[0005] According to one aspect of the present application, the blank solvent is prepared from potassium dihydrogen phosphate solution and acetonitrile solution.

[0006] According to one aspect of the present application, the preparation method of the blank excipients solution is as follows: Take an appropriate amount of blank excipients, place them in a volumetric flask, add an appropriate amount of blank solvent, sonicate for 30 minutes, dilute to the scale with blank solvent, shake well, filter, and take the subsequent filtrate to obtain the blank excipients solution.

[0007] According to one aspect of the present application, the preparation method of the reference solution is as follows: Take appropriate amounts of itopride hydrochloride reference substance, impurity A reference substance, impurity B reference substance, and impurity C reference substance, place them in a volumetric flask, dissolve and dilute to the scale with blank solvent, shake well to obtain the reference solution.

[0008] According to one aspect of the present application, the system suitability solution is prepared as follows: Take appropriate amounts of itopride hydrochloride reference substance, impurity A reference substance, impurity B reference substance, and impurity C reference substance, place them in a volumetric flask, dissolve with blank solvent and dilute to the scale, shake well to obtain the system suitability solution.

[0009] According to one aspect of the present application, HPLC is used to detect the above-mentioned blank solvent, blank excipients, reference substance solution, test sample solution, and system suitability solution, record the chromatogram, and the detection conditions are as follows: Octadecylsilane chemically bonded silica gel is used as the filler, and a phosphate buffer solution containing sodium heptanesulfonate and acetonitrile are used as the mobile phase.

[0010] According to one aspect of the present application, the phosphate buffer solution containing sodium heptanesulfonate contains 2.5 mmol / L sodium heptanesulfonate and 10 mmol / L potassium dihydrogen phosphate, and the pH value is adjusted to 2.8 - 3.2 with phosphoric acid.

[0011] According to one aspect of the present application, the column temperature of the detection conditions is 25°C - 35°C.

[0012] According to one aspect of the present application, the flow rate of the detection conditions is 0.9 ml / min - 1.1 ml / min, the wavelength of the detection conditions is 221 - 225 nm, and the injection volume is 20 μl.

[0013] According to one aspect of the present application, the structural formula of itopride hydrochloride:

[0014]

[0015] The structural formula of impurity A is:

[0016]

[0017] The structural formula of impurity B is:

[0018]

[0019] The structural formula of impurity C is:

[0020]

[0021] Advantages of the present invention: Step 1: Prepare blank solvent, blank excipients, reference substance solution, test sample solution, and system suitability solution respectively; Step 2: Use HPLC to detect the blank solvent, blank excipients, reference substance solution, test sample solution, and system suitability solution in Step 1, and record the chromatogram; Through the above steps, the main degradation impurities in the tablet are quality controlled. This method has high sensitivity, strong specificity, good resolution, high precision, good reproducibility, and good accuracy, and can more conveniently and effectively control the quality of this product. Description of the Drawings

[0022] Figure 1 Show the structural formula of itopride hydrochloride;

[0023] Figure 2 Show the structural formula of impurity A;

[0024] Figure 3 Show the structural formula of impurity B;

[0025] Figure 4 Show the structural formula of impurity C;

[0026] Figure 5 Show the blank solvent chromatogram in Example 1 of the present application;

[0027] Figure 6 Show the blank excipient chromatogram in Example 1 of the present application;

[0028] Figure 7 Show the system suitability chromatogram in Example 1 of the present application;

[0029] Figure 8 Show the reference solution chromatogram in Example 1 of the present application;

[0030] Figure 9 Show the test solution chromatogram in Example 1 of the present application;

[0031] Figure 10 Show the test solution chromatogram in Example 2 of the present application;

[0032] Figure 11 Show the reference solution chromatogram in Example 2 of the present application;

[0033] Figure 12 Show the test solution chromatogram in Example 3 of the present application;

[0034] Figure 13 Show the reference solution chromatogram in Example 3 of the present application. Detailed implementation manners

[0035] The following examples are provided to better further understand the present invention, which are not limited to the best implementation manners, and do not constitute a limitation to the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior art features falls within the protection scope of the present invention.

[0036] For those examples where specific experimental steps or conditions are not indicated, the operations or conditions of the conventional experimental steps described in the literature in the art can be followed. For reagents or instruments whose manufacturers are not indicated, they are all conventional reagent products that can be obtained through commercial purchase.

[0037] According to one aspect of the present application, a detection method for related substances in itopride hydrochloride tablets by HPLC is provided. Among them, the related substances are impurity A, impurity B, and impurity C, including the following steps: Step 1: Prepare a blank solvent, blank excipients, reference solution, test solution, and system suitability solution respectively; Step 2: Detect the blank solvent, blank excipients, reference solution, test solution, and system suitability solution in Step 1 by HPLC, and record the chromatogram.

[0038] Further, in Step 1, the blank solvent is prepared from potassium dihydrogen phosphate solution and acetonitrile solution.

[0039] Further, in Step 1, the preparation of the blank excipients solution is as follows: Take an appropriate amount of blank excipients, place them in a volumetric flask, add an appropriate amount of blank solvent, sonicate for 30 minutes, dilute to the scale with blank solvent, shake well, filter, and take the subsequent filtrate to obtain the blank excipients solution.

[0040] Further, in Step 1, the preparation of the reference solution is as follows: Take appropriate amounts of itopride hydrochloride reference substance, impurity A reference substance, impurity B reference substance, and impurity C reference substance, place them in a volumetric flask, dissolve and dilute to the scale with blank solvent, and shake well to obtain the reference solution.

[0041] Further, in Step 1, the preparation of the system suitability solution is as follows: Take appropriate amounts of itopride hydrochloride reference substance, impurity A reference substance, impurity B reference substance, and impurity C reference substance, place them in a volumetric flask, dissolve and dilute to the scale with blank solvent, and shake well to obtain the system suitability solution.

[0042] According to one aspect of the present application, the above blank solvent, blank excipients, reference solution, test solution, and system suitability solution are detected by HPLC, and the chromatogram is recorded. The detection conditions are that octadecylsilane chemically bonded silica gel is used as the filler, and a phosphate buffer solution containing sodium heptanesulfonate and acetonitrile are used as the mobile phase.

[0043] Preferably, in Step 2, the phosphate buffer solution containing sodium heptanesulfonate contains 2.5 mmol / L sodium heptanesulfonate and 10 mmol / L potassium dihydrogen phosphate, and the pH value is adjusted to 2.8 - 3.2 with phosphoric acid.

[0044] Preferably, in Step 2, the column temperature of the detection conditions is 25°C - 35°C.

[0045] Preferably, in Step 2, the flow rate of the detection conditions is 0.9 ml / min - 1.1 ml / min.

[0046] Preferably, in Step 2, the wavelength of the detection conditions is 221 - 225 nm, and the injection volume is 20 μl.

[0047] The present invention will be further described below in conjunction with embodiments. The described embodiments are only a part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts fall within the protection scope of the present invention.

[0048] Example 1

[0049] A detection method for related substances in itopride hydrochloride tablets by HPLC includes the following steps:

[0050] Step 1: Prepare a blank excipient solution: Take an appropriate amount of the blank excipient of this product (equivalent to about 25 mg of itopride hydrochloride), place it in a 50-ml volumetric flask, add 30 ml of solvent, ultrasonically treat for 30 minutes, cool to room temperature, dilute to the mark with the solvent, shake well, filter, and take the subsequent filtrate to obtain the solution.

[0051] Step 2: Prepare a system suitability solution: Take appropriate amounts of the reference substances of itopride hydrochloride, impurity A, impurity B, and impurity C, accurately weigh them, dissolve and dilute with the solvent to prepare a solution containing about 1 μg per 1 ml respectively, shake well to obtain the solution. Use the system suitability solution to detect the resolution, and the resolution between each chromatographic peak should meet the requirements.

[0052] Step 3: Prepare a test solution: Take an appropriate amount of the finely ground powder of this product (equivalent to about 25 mg of itopride hydrochloride), place it in a 50-ml volumetric flask, add 30 ml of solvent, ultrasonically treat for 30 minutes, cool to room temperature, dilute to the mark with the solvent, shake well, filter, and take the subsequent filtrate to obtain the solution.

[0053] Step 4: Prepare a reference solution: Take appropriate amounts of the reference substances of itopride hydrochloride, impurity A, impurity B, and impurity C, accurately weigh them, dissolve and dilute with the solvent to prepare a solution containing about 1 μg per 1 ml respectively, shake well to obtain the solution.

[0054] Step 5: Obtain the chromatograms of the test solution and the reference solution under qualified chromatographic separation conditions, and determine the content of each impurity by the external standard method.

[0055] In some embodiments of the present invention, the solvent described in the above Step 1, Step 2, Step 3, and Step 4 is potassium dihydrogen phosphate solution (take 1.36 g of potassium dihydrogen phosphate, add water to 1000 ml to dissolve, adjust the pH value to 3.0 with phosphoric acid, shake well) - acetonitrile (4:1).

[0056] Chromatographic conditions:

[0057] Chromatographic column: Packed with octadecylsilyl silica gel, using Thermo gold AQ C18, 4.6 mm × 250 mm, 5 μm; with phosphate buffer solution (take 1.36 g of potassium dihydrogen phosphate and 0.505 g of 1-heptanesulfonic acid sodium, add 1000 ml of water to dissolve, adjust the pH value to 3.0 with phosphoric acid, shake well, then obtain)-acetonitrile (4:1) as the mobile phase; the flow rate is 1.0 ml per minute; the column temperature is 30 °C; the detection wavelength is 223 nm; the injection volume is 20 μl.

[0058] Example 2

[0059] A detection method for related substances in itopride hydrochloride tablets by HPLC, comprising the following steps:

[0060] Step 1: Prepare the system suitability solution: Take appropriate amounts of itopride hydrochloride, impurity A, impurity B and impurity C reference substances, weigh accurately, dissolve and dilute with a solvent to make a solution containing about 1 μg per 1 ml respectively, shake well, then obtain. Use the system suitability solution to detect the resolution, and the resolution between each chromatographic peak should meet the requirements.

[0061] Step 2: Prepare the test solution: Take an appropriate amount of the finely ground powder of this product (equivalent to about 25 mg of itopride hydrochloride), place it in a 50 ml volumetric flask, add 30 ml of the solvent, ultrasonically treat for 30 minutes, cool to room temperature, dilute to the mark with the solvent, shake well, filter, and take the subsequent filtrate, then obtain.

[0062] Step 3: Obtain the chromatograms of the test solution and the reference solution under qualified chromatographic separation conditions, and determine the content of each impurity by the external standard method.

[0063] In some embodiments of the present invention, the solvents described in Step 1 and Step 2 above are potassium dihydrogen phosphate solution (take 1.36 g of potassium dihydrogen phosphate, add 1000 ml of water to dissolve, adjust the pH value to 3.0 with phosphoric acid, shake well)-acetonitrile (4:1).

[0064] Chromatographic conditions:

[0065] Chromatographic column: Packed with octadecylsilyl silica gel, using Thermo gold AQ C18, 4.6 mm × 250 mm, 5 μm; with phosphate buffer solution (take 1.36 g of potassium dihydrogen phosphate and 0.505 g of 1-heptanesulfonic acid sodium, add 1000 ml of water to dissolve, adjust the pH value to 3.0 with phosphoric acid, shake well, then obtain)-acetonitrile (4:1) as the mobile phase; the flow rate is 1.0 ml per minute; the column temperature is 25 °C; the detection wavelength is 223 nm; the injection volume is 20 μl.

[0066] Example 3

[0067] An HPLC method for detecting related substances in itopride hydrochloride tablets, comprising the following steps:

[0068] Step 1: Prepare the system suitability solution: Take appropriate amounts of itopride hydrochloride, impurity A, impurity B, and impurity C reference substances, accurately weigh them, dissolve and dilute with a solvent to prepare a solution containing about 1 μg per 1 ml respectively, shake well to obtain. Use the system suitability solution to detect the resolution, and the resolution between each chromatographic peak should meet the requirements.

[0069] Step 2: Prepare the test solution: Take an appropriate amount of the finely ground powder of this product (equivalent to about 25 mg of itopride hydrochloride), place it in a 50 ml volumetric flask, add 30 ml of the solvent, ultrasonically treat for 30 minutes, cool to room temperature, dilute to the mark with the solvent, shake well, filter, and take the subsequent filtrate to obtain.

[0070] Step 3: Obtain the chromatograms of the test solution and the reference solution under qualified chromatographic separation conditions, and determine the content of each impurity by the external standard method.

[0071] In some embodiments of the present invention, the solvents described in Step 1 and Step 2 above are potassium dihydrogen phosphate solution (take 1.36 g of potassium dihydrogen phosphate, add 1000 ml of water to dissolve, adjust the pH value to 2.8 with phosphoric acid, shake well) - acetonitrile (4:1).

[0072] Chromatographic conditions:

[0073] Chromatographic column: Octadecylsilane chemically bonded silica gel is used as the filler, using Thermo gold AQ C18, 4.6 mm × 250 mm, 5 μm; phosphate buffer solution (take 1.36 g of potassium dihydrogen phosphate and 0.505 g of 1-heptanesulfonic acid sodium, add 1000 ml of water to dissolve, adjust the pH value to 3.0 with phosphoric acid, shake well to obtain) - acetonitrile (4:1) is used as the mobile phase; the flow rate is 1.0 ml per minute; the column temperature is 30 °C; the detection wavelength is 223 nm; the injection volume is 20 μl.

[0074] It can be seen that the itopride hydrochloride tablets (batch number: batch 24071101; self-made) were detected according to the method of Example 1, and the detection results were that impurity A was 0.01%, impurity C was 0.01%, impurity B and single impurities were not detected, and the total amount of impurities was 0.02%, meeting the regulations of the pharmacopoeia regarding the related substances of this variety.

[0075] The reference preparation of itopride hydrochloride tablets (batch number: 35836YQ1 batch; trade name: Risù) was tested according to the method of Example 1. The test results showed that the impurity C was 0.02%, the single impurity was 0.01%, neither impurity A nor impurity B was detected, and the total impurity was 0.03%, meeting the requirements of the pharmacopoeia for the related substances of this variety. The itopride hydrochloride tablets (batch number: 24071101 batch; self-made) were tested according to the method of Example 2. The test results showed that the impurity A was 0.01%, the impurity C was 0.01%, neither impurity B nor the single impurity was detected, and the total impurity was 0.02%, meeting the requirements of the pharmacopoeia for the related substances of this variety.

[0076] The reference preparation of itopride hydrochloride tablets (batch number: 35836YQ1 batch; trade name: Risù) was tested according to the method of Example 2. The test results showed that the impurity C was 0.02%, the single impurity was 0.01%, neither impurity A nor impurity B was detected, and the total impurity was 0.03%, meeting the requirements of the pharmacopoeia for the related substances of this variety. The itopride hydrochloride tablets (batch number: 24071101 batch; self-made) were tested according to the method of Example 3. The test results showed that the impurity A was 0.01%, the impurity C was 0.01%, neither impurity B nor the single impurity was detected, and the total impurity was 0.02%, meeting the requirements of the pharmacopoeia for the related substances of this variety.

[0077] The reference preparation of itopride hydrochloride tablets (batch number: 35836YQ1 batch; trade name: Risù) was tested according to the method of Example 3. The test results showed that the impurity C was 0.02%, the single impurity was 0.01%, neither impurity A nor impurity B was detected, and the total impurity was 0.03%, meeting the requirements of the pharmacopoeia for the related substances of this variety.

[0078] In this application, through Step 1: preparing blank solvent, blank excipients, reference solution, test solution and system suitability solution respectively; Step 2: using HPLC to test the blank solvent, blank excipients, reference solution, test solution and system suitability solution in Step 1 and recording the chromatogram; through the above steps, the main degradation impurities in this tablet are quality controlled. This method has high sensitivity, strong specificity, good resolution, high precision, good reproducibility and good accuracy, and can more conveniently and effectively control the quality of this product, solving the technical problem that in the existing detection methods, some substances cannot be separated in the solvent, resulting in inaccurate detection results.

[0079] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the scope disclosed by the present invention, according to the technical solution and its concept of the present invention, making equivalent replacements or changes, shall be covered by the protection scope of the present invention.

Claims

1. A method for detecting related substances in itopride hydrochloride tablets by HPLC, wherein: The relevant substances are impurities A, B and C, and the method is characterized in that it includes the following steps: Step 1: Prepare blank solvent, blank excipient, reference solution, test solution and system suitability solution respectively; Step 2: Use HPLC to test the blank solvent, blank excipient, reference solution, test solution and system suitability solution in step 1, and record the chromatogram.

2. The method for determining related substances in itopride hydrochloride tablets by HPLC according to claim 1, characterized in that: The blank solvent was composed of potassium dihydrogen phosphate solution and acetonitrile solution.

3. The method for detecting related substances in itopride hydrochloride tablets by HPLC according to claim 2, characterized in that: The blank auxiliary material solution was prepared as follows: appropriate amount of blank auxiliary material was taken, placed in a volumetric flask, appropriate amount of blank solvent was added, ultrasonicated for 30 minutes, diluted to the scale with blank solvent, shaken well, filtered, and the filtrate was taken to obtain the blank auxiliary material solution.

4. The method for detecting related substances in itopride hydrochloride tablets by HPLC according to claim 3, characterized in that: The reference solution is prepared as follows: take appropriate amounts of itopride hydrochloride reference substance, impurity A reference substance, impurity B reference substance and impurity C reference substance, place them in a volumetric flask, add blank solvent to dissolve and dilute to the scale, shake well, and obtain the reference solution.

5. The method for detecting related substances in itopride hydrochloride tablets by HPLC according to claim 4, characterized in that: To prepare the system suitability solution, take appropriate amounts of itopride hydrochloride reference substance, impurity A reference substance, impurity B reference substance and impurity C reference substance, place them in a volumetric flask, add blank solvent to dissolve and dilute to the scale, shake well, and obtain the system suitability solution.

6. The method for determining related substances in itopride hydrochloride tablets by HPLC according to any one of claims 1 to 5, characterized in that: The blank solvent, blank excipient, reference solution, test solution and system suitability solution were detected by HPLC, and the chromatograms were recorded. The detection conditions were as follows: octadecylsilane bonded silica gel was used as a filler, and phosphate buffer containing sodium heptane sulfonate and acetonitrile were used as mobile phases.

7. The method for detecting related substances in itopride hydrochloride tablets by HPLC according to claim 6, characterized in that: The phosphate buffer containing sodium heptane sulfonate contains 2.5 mmol / L sodium heptane sulfonate and 10 mmol / L potassium dihydrogen phosphate, and the pH value is adjusted to 2.8-3.2 with phosphoric acid.

8. The method for detecting related substances in itopride hydrochloride tablets by HPLC according to claim 7, characterized in that: The column temperature of the detection condition is 25℃~35℃.

9. The method for detecting related substances in itopride hydrochloride tablets by HPLC according to claim 8, characterized in that: The flow rate of the detection condition is 0.9 ml / min to 1.1 ml / min, the wavelength of the detection condition is 221 to 225 nm, and the injection volume is 20 ul.

10. The method for detecting related substances in itopride hydrochloride tablets by HPLC according to claim 9, characterized in that: The structural formula of itopride hydrochloride: The structural formula of impurity A is: The structural formula of impurity B is: The structural formula of impurity C is:

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