Method for testing related substances of olmesartan medoxomil amlodipine tablets

Abstract

The invention belongs to the technical field of pharmaceutical analysis, particularly relates to a method for detecting related substances of olmesartan medoxomil amlodipine tablets, provides a convenient, efficient and accurate detection method for solving the problem of detection of the related substances of the olmesartan medoxomil amlodipine tablets, and can detect the content of the related substances of the olmesartan medoxomil amlodipine tablets. According to the method, the quality of the olmesartan medoxomil amlodipine tablet can be effectively controlled, so that the medication safety is effectively guaranteed, the quality control of the olmesartan medoxomil amlodipine tablet is facilitated, the method is convenient, efficient and accurate, the system applicability, repeatability, specificity, precision and accuracy completely meet the guidance principle verified by methods in Chinese pharmacopoeia, and the method can be used for the quality control of the olmesartan medoxomil amlodipine tablet.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for detecting related substances in olmesartan medoxomil and amlodipine tablets. Background Art

[0002] Olmesartan medoxomil and amlodipine tablets are a compound antihypertensive drug composed of an angiotensin II receptor blocker (ARB) olmesartan medoxomil and a calcium channel blocker (CCB) amlodipine. Olmesartan medoxomil is an angiotensin II receptor antagonist (ARB) drug and a prodrug. After entering the gastrointestinal tract, it can be rapidly and completely hydrolyzed into the active metabolite olmesartan, and its absorption is not affected by food. This drug has few interactions and a half-life of up to 13 hours, allowing for once-a-day dosing. Amlodipine is a dihydropyridine calcium antagonist (also known as a calcium ion antagonist or slow channel blocker). It can inhibit the transmembrane entry of calcium ions into vascular smooth muscle and myocardium, and has a stronger effect on vascular smooth muscle cells than on myocardial cells. Clinical studies have confirmed that the combination of olmesartan medoxomil and amlodipine has good antihypertensive effects and is suitable for preparation into a compound preparation. The combined application of the two can not only lower blood pressure but also has the effect of protecting the cardiovascular system.

[0003] Currently, in the current standard for olmesartan medoxomil and amlodipine tablets, high performance liquid chromatography is used to determine the substances in this drug. However, the sensitivity and resolution of this method are relatively poor. There are few literature reports on the current methods for determining the content of related substances, and there are still deficiencies in its determination method. For example, Liu Yanming - HPLC method was used to simultaneously determine the contents of 3 components in compound olmesartan medoxomil tablets, and only 3 of the impurities were detected and verified.

[0004] The purpose of the present invention is to provide a method for detecting related substances in olmesartan medoxomil and amlodipine tablets by high performance liquid chromatography, which can detect 7 related substances in olmesartan medoxomil and amlodipine tablets. Based on the research on the process impurities of olmesartan medoxomil and amlodipine tablets, the inventors of the present invention discovered 7 related substances in olmesartan medoxomil and amlodipine tablets, and their impurities and structural formulas are as follows: Amlodipine: Olmesartan Medoxomil: , Amlodipine Related Substance A: , Olmesartan Medoxomil Impurity 21: , Olmesartan: , Olmesartan Medoxomil Dimer: , Olefin Impurity: .

[0005] Currently, there is no publicly available detection method that can effectively separate the latter 5 of the above 7 impurities, let alone accurately quantitatively determine these potential impurities, which poses a great risk to the drug safety of olmesartan medoxomil and amlodipine tablets.

[0006] The present invention provides a convenient, efficient and accurate detection method for solving the problem of detecting related substances in olmesartan medoxomil and amlodipine tablets, which can detect the related substances in olmesartan medoxomil and amlodipine tablets, thus effectively ensuring the safety of drug use and facilitating the quality control of olmesartan medoxomil and amlodipine tablets. Summary of the Invention

[0007] The technical solution of the present invention is as follows: The present invention provides a detection method for determining related substances in olmesartan medoxomil and amlodipine tablets by high performance liquid chromatography. To solve the problem of detecting related substances in olmesartan medoxomil and amlodipine tablets, a convenient, efficient and accurate detection method is provided. This method can detect the content of related substances in olmesartan medoxomil and amlodipine tablets, thus effectively ensuring the safety of drug use and facilitating the quality control of olmesartan medoxomil and amlodipine tablets. This method is convenient, efficient and accurate, and fully complies with the guiding principles of method validation in the Chinese Pharmacopoeia in terms of system suitability, repeatability, specificity, precision and accuracy, and can be used for the quality control of olmesartan medoxomil and amlodipine tablets.

[0008] To achieve the above object, the present invention provides the following technical solution: A method for detecting related substances in olmesartan medoxomil and amlodipine tablets, the detection method comprising the following steps: (1) Preparation of solutions: Prepare blank solution, sensitivity solution, reference solution and test solution respectively.

[0009] (2) Determination method: Use high performance liquid chromatography to determine the related substances in olmesartan medoxomil and amlodipine tablets. After the system is stable, inject blank solution, sensitivity solution, reference solution and test solution respectively, and record the chromatogram; The chromatographic conditions are as follows: Chromatographic column: Octadecylsilyl silica gel as filler, Detection wavelength: 254 nm; Flow rate: 2.0 ± 0.1 ml / min; Column temperature: 60 ± 2 °C; Injection volume: 10 μl, Injector temperature: 5 °C, Mobile phase: Potassium dihydrogen phosphate solution: Acetonitrile system as mobile phase, Gradient elution is used.

[0010] Further, the blank solution (diluent) is 50% acetonitrile; the mobile phase is as follows: Mobile phase A: acetonitrile: buffer solution (15:85); Mobile phase B: acetonitrile: buffer solution (80:20) (Buffer solution: Take a certain amount of potassium dihydrogen phosphate, add water to 1000 ml, adjust the pH to 3.2 with phosphoric acid, and filter through a membrane filter); The reference solution: Take appropriate amounts of olmesartan medoxomil reference substance and amlodipine reference substance and place them in a volumetric flask, add the blank solution to dilute to the scale, and shake well; The sensitivity solution: Take an appropriate amount of the reference solution, place it in a volumetric flask, add the blank solution to dilute to the scale, and shake well; The test solution: Take an appropriate amount of the test sample, weigh it accurately, place it in a volumetric flask, and dilute to the scale with the diluent; The chromatographic column is Waters, SunFire® C8 4.6 mm×100 mm, 3.5 μm or a chromatographic column with equivalent performance.

[0011] Further, the elution gradient is as follows:

[0012] The reference solution is eluted according to the following gradient:

[0013] The method for detecting the related substances of olmesartan medoxomil and amlodipine tablets further includes method validation before detection. The method validation is carried out according to the chromatographic conditions of the formal detection, and the determination results are as follows:

[0014]

[0015] The present invention discloses a method for detecting the related substances of olmesartan medoxomil and amlodipine tablets. To solve the problem of detecting the related substances of olmesartan medoxomil and amlodipine tablets, a convenient, efficient and accurate detection method is provided. This method can detect the content of related substances in olmesartan medoxomil and amlodipine tablets, thus effectively ensuring the safety of drug use. Description of the Drawings

[0016] Figure 1 It is the chromatogram of the blank solution of the related substances of olmesartan medoxomil and amlodipine tablets Figure 2 It is the chromatogram of the test solution of the related substances of olmesartan medoxomil and amlodipine tablets Figure 3 It is the chromatogram of the reference solution of the related substances of olmesartan medoxomil and amlodipine tablets Figure 4 It is the chromatogram of the selectivity solution of the related substances of olmesartan medoxomil and amlodipine tablets Embodiments Example 1

[0017] (1) Prepare the solution: Blank solution (diluent): Acetonitrile: water (100:100) Blank excipient solution: Weigh accurately 943 mg of the mixed excipients in the prescription amount (equivalent to the excipient amount in about 10 tablets), place in a 100 ml volumetric flask, add an appropriate amount of diluent, dissolve by ultrasonic wave, dilute to the scale with diluent, shake well, and centrifuge to obtain the solution.

[0018] Sensitivity solution: Accurately pipette 5.0 ml of the reference solution into a 20 ml volumetric flask, dilute to the scale with diluent, and shake well to obtain the solution. (The concentration of olmesartan medoxomil is about 1.0 μg / ml; the concentration of amlodipine is about 0.25 μg / ml) Prepare olmesartan medoxomil stock solution, amlodipine stock solution, amlodipine related substance A stock solution ①, olmesartan impurity 21 stock solution ①, olmesartan dimer stock solution ①, olmesartan stock solution ①, olefin impurity stock solution ①: Respectively take olmesartan medoxomil, amlodipine, amlodipine related substance A, olmesartan impurity 21, olmesartan dimer, olmesartan, olefin impurity reference substances and place them in volumetric flasks. After dissolving by ultrasonic wave with diluent, dilute to the scale and shake well to prepare olmesartan medoxomil stock solution: 400 μg / ml, amlodipine stock solution: 100 μg / ml, amlodipine related substance A stock solution ①: 3100 μg / ml, olmesartan impurity 21 stock solution ①: 1000 μg / ml, olmesartan dimer stock solution ①: 1000 μg / ml, olmesartan stock solution ①: 1000 μg / ml, olefin impurity stock solution ①: 1000 μg / ml.

[0019] Prepare olmesartan medoxomil identification solution, amlodipine identification solution, amlodipine related substance A identification solution, olmesartan impurity 21 identification solution, olmesartan dimer identification solution, olmesartan identification solution, olefin impurity identification solution: Respectively take 1.0 ml of olmesartan medoxomil stock solution, amlodipine stock solution, amlodipine related substance A stock solution ①, olmesartan impurity 21 stock solution ①, olmesartan dimer stock solution ①, olmesartan stock solution ①, olefin impurity stock solution ① and place them in different volumetric flasks. Dissolve with diluent and dilute to the scale, shake well to prepare olmesartan medoxomil identification solution: 4 μg / ml, amlodipine identification solution: 1 μg / ml, amlodipine related substance A identification solution: 2 μg / ml, olmesartan impurity 21 identification solution: 4 μg / ml, olmesartan dimer identification solution: 4 μg / ml, olmesartan identification solution: 30 μg / ml, olefin impurity identification solution: 12 μg / ml Mixed Impurity Reference Stock Solution: Weigh accurately 100 mg of olmesartan reference substance and 20 mg of olefin impurity reference substance, and place them in a 100-ml volumetric flask. Accurately pipette 9.5 ml of amlodipine related substance A stock solution ①, 6.6 ml of olmesartan impurity 21 stock solution ①, and 6.6 ml of olmesartan dimer stock solution ①. Add an appropriate amount of diluent, shake and sonicate to dissolve, then dilute to the scale with diluent and mix well to obtain the solution. (Olmesartan: 1000 μg / ml, Olefin: 400 μg / ml, Amlodipine related substance A: 67 μg / ml, Olmesartan impurity 21: 132 μg / ml, Olmesartan dimer: 132 μg / ml) Test Solution (Raw Material): Weigh accurately 200 mg of olmesartan medoxomil raw material and 69 mg of amlodipine besylate raw material, place them in a 100-ml volumetric flask, add diluent to 70% of the volumetric flask, shake and sonicate if necessary to dissolve, then dilute to the scale with diluent and mix well to obtain the solution.

[0020] Reference Solution: Accurately pipette 1.0 ml of the reference stock solution into a 100-ml volumetric flask, dilute to the scale with diluent and mix well to obtain the solution. (Concentration of olmesartan medoxomil is about 4 μg / ml; Concentration of amlodipine is about 1 μg / ml) Test Solution (Preparation): Take 10 olmesartan medoxomil and amlodipine tablets, place them in a 100-ml volumetric flask, add 20 ml of water, sonicate for 5 min to dissolve the coating material, immediately add 20 ml of acetonitrile, then add 30 ml of diluent, sonicate for about 5 min to dissolve, dilute to the scale with diluent and mix well, centrifuge (at a speed of 8000 for 5 minutes), and take the supernatant. (Concentration of olmesartan medoxomil is about 2 mg / ml; Concentration of amlodipine is about 0.5 mg / ml) Selectivity Solution (Raw Material): Weigh accurately 200 mg of olmesartan medoxomil raw material and 69 mg of amlodipine besylate raw material, place them in a 100-ml volumetric flask, shake and sonicate if necessary to dissolve, pipette 3.0 ml of the mixed impurity reference stock solution, then dilute to the scale with diluent and mix well to obtain the solution.

[0021] Selectivity Solution (Raw Material): Weigh accurately 200 mg of olmesartan medoxomil raw material and 69 mg of amlodipine besylate raw material, place them in a 100-ml volumetric flask, shake and sonicate if necessary to dissolve, pipette 3.0 ml of the mixed impurity reference stock solution, then dilute to the scale with diluent and mix well to obtain the solution.

[0022] Selective solution (preparation): Take 10 olmesartan medoxomil and amlodipine tablets, place them in a 100 ml volumetric flask, add 20 ml of water, ultrasonicate for 5 minutes to dissolve the coating material, immediately add 20 ml of acetonitrile, then add 30 ml of diluent, ultrasonicate for about 5 minutes to dissolve, transfer 3.0 ml of the mixed impurity reference stock solution, dilute to the mark with diluent, shake well, centrifuge (at a speed of 8000 for 5 minutes), and take the supernatant.

[0023] Acid-degraded test solution (preparation): Take 10 olmesartan medoxomil and amlodipine tablets, place them in a 100 ml volumetric flask, add 5 ml of 0.1 mol / L hydrochloric acid solution, shake well to wet thoroughly, let stand at room temperature for 10 minutes, add 5 ml of 0.1 mol / L sodium hydroxide solution to neutralize, add 10 ml of acetonitrile, then add diluent to dissolve and dilute to the mark, shake well, centrifuge (at a speed of 8000 for 5 minutes), and take the supernatant; prepare the acid blank solution, acid-degraded blank excipient solution, acid-degraded test solution (olmesartan medoxomil), acid-degraded test solution (amlodipine), acid-degraded test solution (amlodipine + blank excipient), and acid-degraded test solution (olmesartan medoxomil + blank excipient) in the same way.

[0024] Alkali-degraded test solution (preparation): Take 10 olmesartan medoxomil and amlodipine tablets, place them in a 100 ml volumetric flask, add 5 ml of 0.01 mol / L sodium hydroxide, shake well to wet thoroughly, let stand at room temperature for 10 minutes, add 5 ml of 0.01 mol / L hydrochloric acid solution to neutralize, add 10 ml of acetonitrile, then add diluent to dissolve and dilute to the mark, shake well, centrifuge (at a speed of 8000 for 5 minutes), and take the supernatant; prepare the alkali blank solution, alkali-degraded blank excipient solution, alkali-degraded test solution (olmesartan medoxomil), alkali-degraded test solution (amlodipine), alkali-degraded test solution (amlodipine + blank excipient), and alkali-degraded test solution (olmesartan medoxomil + blank excipient) in the same way.

[0025] Oxidation-degraded test solution (preparation): Take 10 olmesartan medoxomil and amlodipine tablets, place them in a 100 ml volumetric flask, add 5 ml of 3% hydrogen peroxide solution, shake well to wet thoroughly, let stand at room temperature for 2 hours, add 5 ml of acetonitrile, add diluent to dissolve and dilute to the mark, shake well, centrifuge (at a speed of 8000 for 5 minutes), and take the supernatant; prepare the oxidation blank solution, oxidation-degraded blank excipient solution, oxidation-degraded test solution (olmesartan medoxomil), oxidation-degraded test solution (amlodipine), oxidation-degraded test solution (amlodipine + blank excipient), and oxidation-degraded test solution (olmesartan medoxomil + blank excipient) in the same way.

[0026] High-temperature degradation test solution (preparation): Take 10 olmesartan medoxomil and amlodipine tablets that have been placed in an oven at 60°C for more than 15 days, put them in a 100-ml volumetric flask, add 20 ml of water, ultrasonicate for 5 minutes to dissolve the coating material, immediately add 20 ml of acetonitrile, then add 30 ml of diluent, ultrasonicate for about 5 minutes to dissolve, add diluent to dissolve and dilute to the mark, shake well, centrifuge (rotation speed 8000, 5 minutes), and take the supernatant; prepare the high-temperature degradation blank excipient solution, high-temperature degradation test solution (olmesartan medoxomil), high-temperature degradation test solution (amlodipine), high-temperature degradation test solution (amlodipine + blank excipient), and high-temperature degradation test solution (olmesartan medoxomil + blank excipient) in the same way.

[0027] Light degradation test solution (preparation): Take 10 olmesartan medoxomil and amlodipine tablets that have been placed in a strong light irradiation test chamber (4500 ± 500 Lux, 90 μW / cm2) for more than 15 days, put them in a 100-ml volumetric flask, add 20 ml of water, ultrasonicate for 5 minutes to dissolve the coating material, immediately add 20 ml of acetonitrile, then add 30 ml of diluent, ultrasonicate for about 5 minutes to dissolve, add diluent to dissolve and dilute to the mark, shake well, centrifuge (rotation speed 8000, 5 minutes), and take the supernatant; prepare the light degradation blank excipient solution, light degradation test solution (olmesartan medoxomil), light degradation test solution (amlodipine), light degradation test solution (amlodipine + blank excipient), and light degradation test solution (olmesartan medoxomil + blank excipient) in the same way.

[0028] The chromatographic conditions are as follows: Chromatographic column: Octadecylsilyl silica gel as the filler (Waters, SunFire® C8 4.6 mm × 100 mm, 3.5 μm or a chromatographic column with equivalent efficiency) Wavelength: 254 nm Flow rate: 2.0 ml / min Injection tray temperature: 5°C Column temperature: 60°C Injection volume: 10 μl Buffer solution: Take 2.04 g of potassium dihydrogen phosphate, add water to 1000 ml, adjust the pH to 3.2 with phosphoric acid, and filter through a 0.45-μm filter membrane.

[0029] Mobile phase A: Acetonitrile: Buffer solution (15:85) Mobile phase B: Acetonitrile: Buffer solution (80:20) Perform elution according to the following gradient:

[0030] The reference solution is run using the following gradient:

[0031] Determination method: After the system is stable, inject 1 - 3 needles of blank solution, 1 needle of sensitivity solution, 6 needles of reference solution, and 1 needle of test solution. Inject 1 needle of reference solution at the 8 - needle injection interval and at the end of the sequence.

[0032] (1) Calculate the content of amlodipine - related impurities.

[0033] Content (%) =

[0034] AR—Average peak area of amlodipine in the reference solution AS—Peak area of amlodipine - related substance A or other impurities attributed to amlodipine in the test solution mR—Sampling amount of amlodipine besylate reference substance in the reference solution (mg) N—Sampling amount of the sample (tablets) Q—Specification of amlodipine RRF—Relative response factor CR—Content of amlodipine besylate reference substance DR—Dilution factor of the reference solution Ds—Dilution factor of the test solution M1—Molecular weight of amlodipine, 408.88 M2—Molecular weight of amlodipine besylate, 567.05 (2) Calculate the content of olmesartan medoxomil - related impurities.

[0035] Content (%) =

[0036] AR—Average peak area of olmesartan medoxomil in the reference solution; AS—Peak area of olmesartan impurity 21, olmesartan, olefin impurity, olmesartan dimer or other impurities attributed to olmesartan medoxomil in the test solution mR—Sampling amount of olmesartan medoxomil reference substance in the reference solution (mg) N—Sampling amount of the sample (tablets) Q—Specification of olmesartan medoxomil RRF—Relative response factor CR—Content of olmesartan medoxomil reference substance DR—Dilution factor of the reference solution DS—Dilution factor of the test solution (3) When calculating unknown impurities, the relative response factor is 1.00 for all.

[0037] (4) Total impurities = Content of each known impurity (impurities other than olmesartan) + Content of each other impurity.

[0038] Example 2 Linear Six concentrations are taken within the concentration range of LOQ to 200% for each component to prepare six test solutions. A linear regression is performed with concentration and peak area, and it is required to be linear.

[0039] Solution preparation: Inject the sample according to Example 1. The test results are shown in Table 2 below.

[0040]

[0041] Precision of Example 3 Repeatability is achieved by measuring six test sample solutions, and it is required that the differences in the six measurement results meet the acceptable criteria.

[0042] Solution preparation: Inject the sample according to Example 1. The test results are shown in Table 3 below.

[0043]

[0044] Intermediate precision is achieved by another inspector measuring six spiked test sample solutions at different times and with different instruments. The measurement results of the six spiked test sample solutions meet the acceptable criteria. It is required that the total of 12 measurement results, including the six measurement results of repeatability, meet the acceptable criteria.

[0045] Durability of Example 4 For the solution stability study, the control solution and the test sample solution are placed at room temperature and 5°C respectively during the test period to detect the law of the change of the test results over time, providing a basis for the placement conditions of the test sample solution during detection.

[0046] Solution preparation: Inject the sample according to Example 1. The test results are shown in Table 4 below.

[0047]

[0048] Matters not covered by this invention are well-known technologies.

[0049] The above embodiments are only for illustrating the technical concept and features of the present invention, and the purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly. It should not be used to limit the protection scope of the present invention. Any equivalent changes or modifications made according to the spirit and essence of the present invention should be covered within the protection scope of the present invention.

Claims

1. A method for detecting related substances in olmesartan medoxomil and amlodipine tablets, characterized in that, The detection method includes the following steps: (1) Preparation of solutions: Prepare a blank solution, a sensitivity solution, a reference solution, and a test solution respectively; (2) Determination method: Use high performance liquid chromatography to determine the related substances of olmesartan medoxomil and amlodipine tablets. After the system is stable, inject the blank solution, the sensitivity solution, the reference solution, and the test solution respectively, and record the chromatogram. The chromatographic conditions are as follows: Chromatographic column: Octadecylsilyl silica gel bonded silica gel as the filler, detection wavelength: 254 nm; Flow rate: 2.0 ± 0.1 ml / min; Column temperature: 60 ± 2 °C; Injection volume: 10 μl, injector temperature: 5 °C, mobile phase: potassium dihydrogen phosphate solution: acetonitrile system as the mobile phase, gradient elution is used.

2. The detection method according to claim 1, characterized in that: The blank solution (diluent) is 50% acetonitrile; The mobile phase is: Mobile phase A: acetonitrile: buffer solution (15:85), Mobile phase B: acetonitrile: buffer solution (80:20) (Buffer solution: Take a certain amount of potassium dihydrogen phosphate, add water to 1000 ml, adjust the pH to 3.2 with phosphoric acid, and filter through a membrane filter); The reference solution: Take appropriate amounts of olmesartan medoxomil reference substance and amlodipine reference substance and place them in a volumetric flask, add the blank solution to dilute to the scale and shake well; The sensitivity solution: Take an appropriate amount of the reference solution and place it in a volumetric flask, add the blank solution to dilute to the scale and shake well; The test solution: Take an appropriate amount of the test sample, accurately weigh it, place it in a volumetric flask, and dilute to the scale with the diluent; The chromatographic column is Waters, SunFire® C8 4.6 mm × 100 mm, 3.5 μm or a chromatographic column with equivalent performance; The elution gradient is: 。

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