Method for detecting impurities in Milsartan potassium

Abstract

The invention belongs to the technical field of pharmaceutical analysis, particularly relates to a method for detecting impurities in Milsartan potassium, and provides a convenient, efficient and accurate detection method for solving the problem of detection of the impurities in Milsartan potassium. According to the method, LC-MS is adopted for detection, the content of the impurities in Milsartan potassium can be detected, so that the medication safety is effectively guaranteed, and the detection cost is reduced. The method is convenient, efficient and accurate, completely conforms to the guidance principle verified by methods in Chinese pharmacopoeia in the aspects of system applicability, repeatability, specificity and accuracy, and can be used for quality control of the Milsartan potassium.

Description

Technical Field

[0001] The invention belongs to the technical field of drug analysis, and particularly relates to a method for detecting impurities in asartan potassium. Background Art

[0002] Asartan Potassium is an angiotensin II receptor antagonist (ARB) drug developed by Takeda Pharmaceuticals. It is a prodrug and is indicated for essential hypertension in adults. Lowering blood pressure can reduce the risk of fatal and non-fatal cardiovascular events, mainly reducing the risk of stroke and myocardial infarction. Asartan Potassium can be used alone or in combination with other antihypertensive drugs.

[0003] During the synthesis of Azilsartan Potassium, there may be residual byproducts such as deethyl azilsartan ester and azilsartan ester (hereinafter referred to as "azilsartan impurity G"), so it is necessary to study the residues of deethyl azilsartan ester and azilsartan impurity G in Azilsartan Potassium. Deethyl azilsartan ester, structural formula: , Azilsartan ester (Azilsartan impurity G), structural formula: , which belongs to genotoxic impurities. Genetic (gene) toxic impurities refer to impurities that can cause genotoxicity, including mutagenic impurities and other types of non-mutagenic impurities. Genetic (gene) toxic impurities mainly come from the production process of raw materials or preparations, such as starting materials, reactants, catalysts, reagents, solvents, intermediates, by-products, degradation products, etc. Mutagenic impurities refer to genotoxic impurities that may directly cause DNA damage and DNA mutations at low levels, which may cause cancer. The research and control of genotoxic impurities has always been challenging for quality researchers. In recent years, there have been many cases of CDE supplements on the research of genotoxic impurities. Therefore, its content needs to be strictly controlled during the production process of Asartan Potassium.

[0004] Currently, there are few literature reports on the detection methods of impurities in Asartan Potassium. As a pharmaceutical industry, it is very necessary to strictly control the impurity content in the product. The study of material impurities is helpful for the optimization of drug synthesis process and quality control. The study of the inspection method of impurities in Asartan Potassium provides a good reference for evaluating the quality of Asartan Potassium drugs, and also provides a technical basis for optimizing the synthesis process of Asartan Potassium drugs. It is of great significance to determine the adverse reactions caused by impurities in Asartan Potassium.

[0005] The invention discloses a detection method for determining impurities in asesartan potassium by using LC-MS. By using the LC-MS method for detection, a convenient, efficient and accurate detection method is provided to solve the problem of detecting impurities in asesartan potassium. The method can detect the content of impurities in asesartan potassium, thereby effectively ensuring the safety of medication and facilitating the quality control of asesartan potassium. Summary of the invention

[0006] The invention provides a detection method for determining impurities in ashartan potassium by using LC-MS. In order to solve the problem of detecting impurities in ashartan potassium, a convenient, efficient and accurate detection method is provided. The method can detect the content of impurities in ashartan potassium, thereby effectively ensuring the safety of medication and facilitating the quality control of ashartan potassium. The method is convenient, efficient and accurate, and fully complies with the guiding principles of method verification of the Chinese Pharmacopoeia in terms of system applicability, repeatability, specificity and accuracy, and can be used for the quality control of ashartan potassium.

[0007] To achieve the above object, the present invention provides the following technical solutions: A method for testing impurities in potassium azilsartan, characterized in that the impurities are ethyl azilsartan ester and azilsartan ester, and the ethyl azilsartan ester has the structural formula: , the structural formula of the azilsartan ester is: , the method is to detect by LC-MS, and the detection method comprises the following steps: (1) Prepare solutions including blank solution, sensitivity solution, reference solution and test solution.

[0008] (2) Determination method: LC-MS was used to determine the content of impurities in mesartan potassium. After the system was stable, blank solution, sensitivity solution, reference solution and test solution were added respectively, and the chromatogram was recorded. The chromatographic conditions are as follows: Chromatographic column: octadecylsilane bonded silica gel as filler, injection volume: 0.2-1.0µl, column temperature: 30±5℃, flow rate: 0.4±0.1ml / min, ammonium acetate solution as mobile phase A, acetonitrile as mobile phase B, gradient elution; The mass spectrometry conditions were as follows: ion source type: ESI, drying gas temperature: 300 °C, drying gas flow rate: 8 L / min, nebulizer gas pressure: 45 psi, sheath gas temperature: 350 °C, sheath gas flow rate: 11 L / min, capillary voltage: 3500 V, polarity: positive, acquisition mode: SIM, switching time setting of the switch valve: 0 min: to Waste; 3.0 min: to MS; 9.0 min: to Waste; .

[0009] Further, the elution gradient is: .

[0010] Further, the mobile phase A: 20 mM ammonium acetate solution; The blank solution: diluent; The volume ratio of the diluent: acetonitrile: water is 60:40; The reference substance solution: take appropriate amounts of deethyl azilsartan ester stock solution and azilsartan ester stock solution respectively, place them in a volumetric flask, add diluent to dilute to the scale, and shake well; Desethyl Azilsartan Ester Stock Solution: Take an appropriate amount of desethyl Azilsartan Ester Reference Standard, place it in a volumetric flask, add diluent to dissolve and dilute to the scale, and shake well; The azilsartan ester stock solution: take an appropriate amount of azilsartan ester reference substance, place it in a volumetric flask, add diluent to dissolve and dilute to the scale, and shake well; The sensitivity solution: take appropriate amounts of deethyl azilsartan ester stock solution and azilsartan ester stock solution respectively, place them in a volumetric flask, add diluent to dilute to the scale, and shake well; Test solution: Take an appropriate amount of the test sample and place it in a volumetric flask, add an appropriate amount of diluent to dissolve it, then dilute it to the scale with the diluent and shake well; the chromatographic column is Agilent Eclipse plus C18 RRHD 3.0×150mm, 1.8 μm.

[0011] The invention discloses a method for detecting impurities in lasartan potassium, and provides a convenient, efficient and accurate detection method to solve the problem of detecting impurities in lasartan potassium. The method can detect the content of impurities in lasartan potassium, thereby effectively ensuring the safety of medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is the blank solution spectrum of impurities in Asartan Potassium; Figure 2 This is the impurity sensitivity solution spectrum of Asartan Potassium; Figure 3 This is the selective solution spectrum of impurities in Asartan Potassium; Figure 4 This is the spectrum of the impurity reference solution in Asartan Potassium; Figure 5 This is the spectrum of the impurity test solution in Asartan Potassium. DETAILED DESCRIPTION

[0013] The present invention is further illustrated by the following examples, but are not intended to limit the present invention.

[0014] Example 1: Determination of impurities in Asartan potassium (1) Preparation of solution: Ammonium acetate: AR and above; Water: ultrapure water; Acetonitrile: HPLC and above; Diluent: acetonitrile: water = 60:40 (V / V); Blank solution: diluent; Deethyl Azilsartan Ester Stock Solution: Take about 25 mg of deethyl Azilsartan Ester Reference Material, weigh accurately, place in a 50 ml volumetric flask, add diluent to dissolve and dilute to scale, shake well; accurately measure 750 µl of the above solution, place in a 10 ml volumetric flask, add diluent to dilute to scale, shake well. (Concentration: 37.5 µg / ml) Azilsartan Impurity G Stock Solution: Take about 25 mg of Azilsartan Impurity G Reference Substance, weigh accurately, place in a 50 ml volumetric flask, add diluent to dissolve and dilute to scale, shake well; accurately measure 750 µl of the above solution, place in a 10 ml volumetric flask, add diluent to dilute to scale, shake well. (Concentration: 37.5 µg / ml) Reference substance stock solution: Accurately measure 500µl of deethyl azilsartan ester stock solution and azilsartan impurity G stock solution, place in a 20ml volumetric flask, add diluent to dilute to the mark, and shake well. (Concentration: deethyl azilsartan ester 937.5ng / ml, azilsartan impurity G 937.5ng / ml) Reference solution: Accurately measure 500µl of reference stock solution and place it in a 50ml volumetric flask, add diluent to dilute to the mark, and shake well. (Concentration: deethyl azilsartan ester 9.375ng / ml, azilsartan impurity G 9.375ng / ml) Sensitivity stock solution: Accurately measure 100µl of deethyl azilsartan ester stock solution and 50µl of azilsartan impurity G stock solution, place in a 20ml volumetric flask, add diluent to dilute to the mark, and shake well. (Concentration: deethyl azilsartan ester 187.5ng / ml, azilsartan impurity G 93.75ng / ml) Sensitivity solution: Accurately measure 500µl of sensitivity stock solution and place it in a 50ml volumetric flask, add diluent to dilute to the mark, and shake well. (Concentration: deethyl azilsartan ester 1.875ng / ml, azilsartan impurity G 0.9375ng / ml) Test solution: Take about 25 mg of the test sample, weigh accurately, place in a 50 ml volumetric flask, add diluent to dissolve and dilute to the scale, shake well. (Concentration 0.5 mg / ml) (2) Chromatographic conditions: Instrument: Agilent InfinityII 1290 UHPLC+Agilent 6470 ESI-QQQ-MS, electronic analytical balance.

[0015] Chromatographic column: Agilent Eclipse plus C18 RRHD 3.0×150mm, 1.8 μm or equivalent Injection volume: 0.5 μl Column temperature: 30°C Flow rate: 0.4ml / min Mobile phase A: 20 mM ammonium acetate solution Mobile phase B: acetonitrile

[0016]

[0017] Switching time setting of on / off valve: 0min: to Waste; 3.0min: to MS; 9.0min: to Waste.

[0018]

[0019] Note: The switching time of the mass spectrometer switch valve can be appropriately adjusted according to the actual situation of the instrument. The purpose is to avoid collecting the signal of mesartan potassium and ensure the response of the signals of each impurity component.

[0020] (3) Steps After the system is stable, inject 1 injection of blank solution (2 to 3 injections if necessary), 1 injection of sensitivity solution, 6 injections of reference solution, 1 injection of blank solution, 1 injection of test solution, and record the spectrum.

[0021] Require The S / N value of each component in the sensitivity solution should not be less than 10; the RSD of the peak area of ​​each component in the reference solution should not be greater than 10.0%.

[0022] calculate

[0023] Where: RU: peak area of ​​each component in the spectrum of the test solution; RS: 6 for the average peak area of ​​each component in the spectrum of the reference solution; CS: concentration of each component in the reference solution (ng / ml); n: dilution factor of the test sample, 50; M: Sample volume of the test product, mg limit

[0024] The method for detecting impurities in Asartan Potassium also includes method validation before detection. The method validation is performed according to the chromatographic conditions of the formal test, and the results are as follows:

[0025]

[0026] Example 2: System Applicability System suitability is achieved by evaluating the S / N value of each component in the sensitivity solution and the RSD of the peak area of ​​each component in the reference solution. It is required that the S / N value of each component in the sensitivity solution should be no less than 10; the RSD of the peak area of ​​each component in the reference solution should be no more than 10.0%.

[0027]

[0028]

[0029] Example 3: Specificity The specificity of the method is achieved by determining whether the blank solution interferes with the detection of each component and the separation degree between each component and adjacent impurity peaks in the selective solution. It is required that the blank solution should have no interference with the detection of each component and the separation degree between each component and adjacent impurity peaks in the selective solution should be no less than 1.5.

[0030]

[0031] Example 4: Precision Repeatability is achieved by measuring the RSD of the results of each component in 6 test solutions (spiked), and it is required that the RSD of the results of each component in 6 test solutions (spiked) should not be greater than 10.0%.

[0032]

[0033]

[0034] Example 5: Limit of Detection and Limit of Quantitation The detection limit is obtained by testing the ratio of the response signal to the noise to be no less than 3:1; the quantification limit is obtained by testing the ratio of the response signal to the noise to be no less than 10:1. At the LOQ concentration level, 6 quantification limit test solutions were repeatedly examined. 去乙基阿齐沙坦酯化物 Should not be greater than 5.625ppm, S / N≥10; LOQ 阿齐沙坦杂质G It should be no greater than 5.625ppm, S / N≥10; the RSD of the unit concentration peak area of ​​each component in the 6 LOQ solutions should be no greater than 20.0%; the S / N of each component in the LOD solution should be no less than 3, and LOD<LOQ.

[0035]

[0036]

[0037] Example 6: Linearity and Range Six points were evenly selected within the range of LOQ concentration to 150% of the limit concentration, and the concentration was used as the horizontal axis and the peak area as the vertical axis to draw a curve. It is required that each component should be linear within the range of LOQ concentration to 150% of the limit concentration, and the square of the linear correlation coefficient R (R 2 ) should be no less than 0.99, and the absolute value of the y-intercept should be within 25% of the 100% concentration peak area.

[0038]

[0039]

[0040] Example 7: Accuracy Accuracy is achieved through the recovery rate between the measured concentration and the theoretical concentration of the measured component and the total RSD of the recovery rate (n=9). It is required that the recovery rate of each component in the accuracy solution with LOQ concentration, 100% limit concentration, and 150% limit concentration should be between 70.0% and 130.0%, and the total RSD of the recovery rate (n=9) should not be greater than 20.0%.

[0041]

[0042]

[0043] Example 8: Durability The reference solution, test solution and selective solution were placed at room temperature for a period of time before being injected, and the change pattern of the test results over time was investigated to provide a reference for the placement time of the reference solution and test solution during the test.

[0044] Requirements: Compared with the reference solution at 0 hr, the recovery rate of each component of the reference solution during the room temperature inspection is between 80.0% and 120.0%, and there is no obvious trend of change, then the reference solution is stable during the room temperature inspection; If the test solution has the component to be tested detected at 0hr, and the test solution is placed at room temperature for a period of time, the change value of the test result is within 20% of the limit, and there is no obvious change trend, then the test solution is stable during the room temperature investigation; if the test solution has not detected the component to be tested at 0hr, and the test solution has been placed at room temperature for a period of time, and still no components are detected, then the test solution is stable during the room temperature investigation; When the selective solution is placed at room temperature for a period of time, the recovery rate of each component is between 80.0% and 120.0%, and there is no obvious change trend, which means that the selective solution is stable during the room temperature investigation.

[0045]

[0046]

[0047]

Claims

1. A method for testing impurities in Asartan Potassium, characterized in that: The impurities are ethyl azilsartan ester and azilsartan ester, and the structural formula of the ethyl azilsartan ester is: , the structural formula of the azilsartan ester is: , the method is to detect by LC-MS, and the detection method comprises the following steps: (1) Prepare solutions, including blank solution, sensitivity solution, reference solution and test solution; (2) Determination method: LC-MS was used to determine the content of impurities in mesartan potassium. After the system was stable, blank solution, sensitivity solution, reference solution and test solution were added respectively, and the chromatogram was recorded. The chromatographic conditions are as follows: Chromatographic column: octadecylsilane bonded silica gel as filler, injection volume: 0.2-1.0µl, column temperature: 30±5℃, flow rate: 0.4±0.1ml / min, ammonium acetate solution as mobile phase A, acetonitrile as mobile phase B, gradient elution; The mass spectrometry conditions were as follows: ion source type: ESI, drying gas temperature: 300 °C, drying gas flow rate: 8 L / min, nebulizer gas pressure: 45 psi, sheath gas temperature: 350 °C, sheath gas flow rate: 11 L / min, capillary voltage: 3500 V, polarity: positive, acquisition mode: SIM, switching time setting of the switch valve: 0 min: to Waste; 3.0 min: to MS; 9.0 min: to Waste; 。 2. The inspection method according to claim 1, characterized in that: The elution gradient is: 。 3. The inspection method according to claim 2, characterized in that: The mobile phase A: 20 mM ammonium acetate solution; The blank solution: diluent; The diluent: The volume ratio of acetonitrile: water is 60:40; The reference substance solution: take appropriate amounts of deethyl azilsartan ester stock solution and azilsartan ester stock solution respectively, place them in a volumetric flask, add diluent to dilute to the scale, and shake well; Desethyl Azilsartan Ester Stock Solution: Take an appropriate amount of desethyl Azilsartan Ester Reference Standard, place it in a volumetric flask, add diluent to dissolve and dilute to the scale, and shake well; The azilsartan ester stock solution: take an appropriate amount of azilsartan ester reference substance, place it in a volumetric flask, add diluent to dissolve and dilute to the scale, and shake well; The sensitivity solution: take appropriate amounts of deethyl azilsartan ester stock solution and azilsartan ester stock solution respectively, place them in a volumetric flask, add diluent to dilute to the scale, and shake well; Test solution: Take an appropriate amount of the test sample and place it in a volumetric flask, add an appropriate amount of diluent to dissolve it, then dilute it to the scale with the diluent and shake well; the chromatographic column is Agilent Eclipse plus C18 RRHD 3.0×150mm, 1.8 μm.

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