A method for detecting related substances in rivaroxaban tablets and its application.

Abstract

This application discloses a method for detecting related substances in rivaroxaban tablets and its application, belonging to the field of pharmaceutical analysis technology. The method employs high-performance liquid chromatography (HPLC) with a column packed with octadecylsilane-bonded silica gel. The column is 150 mm long, 34.0 mm in inner diameter, and has a particle size of 23 μm. Mobile phase A is a 0.095%-0.105% phosphoric acid aqueous solution, and mobile phase B is acetonitrile. Gradient elution is used. The flow rate is 0.95-1.05 ml / min. The column temperature is 3337℃. The detection wavelength is 230-250 nm. The injection volume is 1020 μl. This method is highly specific, sensitive, and accurate, and can simultaneously detect known and unknown impurities, including LFSB1 to LFSB13, in rivaroxaban tablets, making it suitable for the quality control of this formulation.

Description

Technical Field

[0001] This application belongs to the field of pharmaceutical analysis technology, specifically relating to a high-performance liquid chromatography method for the detection of related substances in rivaroxaban tablets. Background Technology

[0002] Rivaroxaban is a highly selective, direct-acting oral anticoagulant that inhibits factor Xa and is widely used for the prevention and treatment of thromboembolic diseases. Various organic impurities may be introduced or generated during its formulation and storage, mainly including unreacted starting materials, synthetic byproducts, intermediates, and degradation products. The presence of these substances may affect the safety and efficacy of the drug; therefore, establishing an analytical method capable of accurately and quantitatively detecting these impurities is crucial for controlling the quality of rivaroxaban tablets.

[0003] Several HPLC methods for the detection of rivaroxaban-related substances have been disclosed in the prior art. For example, Chinese patent application CN110849994A discloses a gradient elution method using phosphoric acid solution and acetonitrile as the mobile phase, but the pH of the mobile phase used is low (1.5-2.0), which may adversely affect the column life. The detection method disclosed in Chinese patent application CN116500173A only targets seven specific impurities, failing to cover all known impurities, especially extremely polar impurities. Methods such as CN105259282A and CN106442831A use ion-pairing reagents, resulting in complex mobile phase systems, cumbersome preparation, less environmentally friendly practices, and higher analytical costs. Methods such as CN108152412A do not adequately study key degradation impurities.

[0004] Therefore, there is an urgent need in this field for a detection method that can simultaneously detect process impurities and degradation impurities in rivaroxaban tablets, has good separation effect and accuracy, and has a simple mobile phase system, is environmentally friendly, cost-controllable, and robust, so as to ensure the safety of clinical medication. Summary of the Invention

[0005] In view of this, the primary objective of this application is to provide a method for detecting related substances in rivaroxaban tablets, which has good specificity, sensitivity, accuracy, and robustness, and can simultaneously detect substances including LFSB. 1 to LFSB Thirteen known impurities, including 13, and other unknown impurities.

[0006] To achieve the above objectives, this application adopts the following technical solution: One aspect of this application discloses a method for detecting related substances in rivaroxaban tablets, which employs high-performance liquid chromatography (HPLC). This method is simple to operate, highly sensitive, specific, and accurate, and can precisely control the content of specific and non-specific impurities in rivaroxaban tablets, thereby effectively achieving quality control of rivaroxaban tablets.

[0007] In this application, the main component of the rivaroxaban tablets tested is rivaroxaban, chemically named 5-chloro-nitro-({(5S)-2-oxo-3-[4-(3-oxo-4-morpholinyl)phenyl]-1,3-azolidin-5-yl}methyl)-2-thiophene-carboxamide, and its structural formula is as follows: .

[0008] The 13 impurities detected and analyzed in this application are shown in Table 1: Table 1. Relevant impurities LFSB-1~LFSB-13

[0009] The chromatographic conditions for the detection method in this application are as follows: Chromatographic column: The stationary phase is octadecylsilane-bonded silica gel (C18).

[0010] In some specific examples, the particle size of the filler is 2-3 μm, for example, it can be any particle size or a range between 2.0 μm, 2.1 μm, 2.2 μm, 2.3 μm, 2.4 μm, 2.5 μm, 2.6 μm, 2.7 μm, 2.8 μm, 2.9 μm, and 3.0 μm.

[0011] In some specific examples, the inner diameter of the chromatographic column is 3-4.0 mm, for example, it can be any inner diameter or a range between 3.0 mm, 3.1 mm, 3.2 mm, 3.3 mm, 3.4 mm, 3.5 mm, 3.6 mm, 3.7 mm, 3.8 mm, 3.9 mm, and 4.0 mm.

[0012] As a preferred example, the chromatographic column used has the following specifications: packing material particle size of 3 μm, column length of 150 mm, and inner diameter of 4.0 mm.

[0013] Any chromatographic column in the art that meets the above specifications can be used in this application. Specific examples include the Shim-Pack GIS C18 column or other C18 columns with similar separation performance.

[0014] Mobile phase: Composed of mobile phase A and mobile phase B, using a gradient elution mode. Mobile phase A is an aqueous phosphoric acid solution. In some specific examples, the volume percentage of phosphoric acid in the aqueous phosphoric acid solution is 0.095%-0.105%, for example, it can be any value or a range between 0.095%, 0.096%, 0.097%, 0.098%, 0.099%, 0.100%, 0.101%, 0.102%, 0.103%, 0.104%, and 0.105%. As a preferred example, the volume percentage of phosphoric acid in the aqueous phosphoric acid solution is 0.1%. Mobile phase B is acetonitrile.

[0015] This application avoids damage to the chromatographic column caused by strong acid conditions by using a low-concentration phosphoric acid aqueous solution as the aqueous phase. At the same time, it eliminates the need to adjust the pH value, simplifies the operation steps, and achieves good impurity separation effect and peak shape.

[0016] Gradient elution program: Given the significant polarity differences between rivaroxaban and its impurities, a specific gradient program is employed to achieve effective elution and separation of all components. An exemplary gradient program may include multiple time points and corresponding variations in the ratio of mobile phase A to B to ensure that impurities ranging from highly polar to weakly polar can elute and separate well within a reasonable timeframe. In some specific examples of this application, the gradient elution program is shown in Table 2: Table 2 Gradient elution program

[0017] Flow rate: The flow rate of the method is 0.95-1.05 ml / min, for example, it can be any value or a range between 0.95 ml / min, 0.96 ml / min, 0.97 ml / min, 0.98 ml / min, 0.99 ml / min, 1.00 ml / min, 1.01 ml / min, 1.02 ml / min, 1.03 ml / min, 1.04 ml / min, and 1.05 ml / min. As a preferred example, the flow rate is 1.0 ml / min.

[0018] Column temperature: The column temperature of the method is 33-37℃, for example, it can be any value or a range between 33.0℃, 33.5℃, 34.0℃, 34.5℃, 35.0℃, 35.5℃, 36.0℃, 36.5℃, and 37.0℃. As a preferred example, the column temperature is 35℃.

[0019] Detection wavelength: This application uses an ultraviolet detector for detection, with a detection wavelength of 230-250 nm. For example, it can be any value or a range between 230 nm, 231 nm, 232 nm, 233 nm, 234 nm, 235 nm, 236 nm, 237 nm, 238 nm, 239 nm, 240 nm, 241 nm, 242 nm, 243 nm, 244 nm, 245 nm, 246 nm, 247 nm, 248 nm, 249 nm, and 250 nm. As a preferred example, the detection wavelength is 241 nm.

[0020] Injection volume: The injection volume of the method is 10-20 μl, for example, it can be any value or a range between 10 μl, 11 μl, 12 μl, 13 μl, 14 μl, 15 μl, 16 μl, 17 μl, 18 μl, 19 μl, and 20 μl. As a preferred example, the injection volume is 10 μl.

[0021] It should be understood that the detection method of this application also includes the preparation of related solutions, including solvents, test solutions, control solutions, sensitivity solutions, impurity stock solutions, and system suitability solutions.

[0022] In some specific examples, the solvent used to dissolve and dilute the sample reagent is, in this application, a mixed solvent of 0.01 mol / L phosphoric acid solution and acetonitrile. In some specific examples, the volume ratio is 50:50. The 0.01 mol / L phosphoric acid solution can be prepared by measuring 0.67 ml of phosphoric acid and diluting it with water to 1000 ml.

[0023] In some specific examples, the test solution is prepared as follows: take an appropriate amount of rivaroxaban tablets, grind them into a fine powder, accurately weigh an appropriate amount of the fine powder (approximately equivalent to 25 mg of rivaroxaban), place it in a 50 ml volumetric flask, add solvent to dissolve and dilute to the mark, shake well, filter, and take the filtrate to obtain a test solution containing 0.5 mg of rivaroxaban per 1 ml.

[0024] In some specific examples, the preparation of the control solution is as follows: accurately measure an appropriate amount of the test solution, place it in a volumetric flask, add solvent to quantitatively dilute it to prepare a solution containing approximately 0.5 μg of rivaroxaban per 1 ml (i.e., 0.2% control solution), shake well, and the solution is obtained.

[0025] In some specific examples, the sensitivity solution is prepared by accurately measuring an appropriate amount of the control solution, placing it in a volumetric flask, and quantitatively diluting it with solvent to prepare a solution of suitable concentration, which is used to investigate the detection limit and quantitation limit of the method.

[0026] In some specific examples, the preparation of impurity control stock solutions is divided into two types to accommodate impurities with different solubilities.

[0027] The impurity reference stock solution 1 was prepared as follows: appropriate amounts of impurity LFSB-1, LFSB-3, LFSB-5, LFSB-6, LFSB-11, and LFSB-13 reference standards were taken, dissolved and diluted with acetonitrile to prepare a solution containing approximately 0.2 mg of each relevant impurity per 1 ml.

[0028] The impurity reference stock solution 2 was prepared as follows: appropriate amounts of impurity LFSB-2, LFSB-4, LFSB-7, LFSB-8, LFSB-9, LFSB-10 and LFSB-12 reference standards were taken, dissolved and diluted with solvent to prepare a solution containing approximately 0.2 mg of each relevant impurity per 1 ml.

[0029] In some specific examples, the system suitability solution is prepared as follows: Take an appropriate amount of rivaroxaban reference standard, place it in a volumetric flask, add an appropriate amount of each impurity reference stock solution, dissolve and dilute with solvent to prepare a solution containing 0.5 mg of rivaroxaban and approximately 1 μg of each impurity per 1 ml, shake well, and the solution is obtained. This solution is used to verify the separation efficiency of the chromatographic system.

[0030] Regarding the system suitability requirements of this application, under the chromatographic conditions described herein, in the system suitability solution chromatogram, the resolution between the rivaroxaban main component peak and adjacent impurity peaks, as well as among all known impurity peaks, should be no less than 1.5. In the sensitivity solution chromatogram, the signal-to-noise ratio (S / N) of the main component peak should be greater than 10. Meeting the above requirements indicates that the chromatographic system is suitable for this detection method.

[0031] For specific quantification, this application uses the principal component comparison method with correction factors to calculate the content of each known impurity. The impurity content is calculated according to the following formula:

[0032] in, A S The peak area of ​​impurities in the test solution. The peak area of ​​the main component in the 0.2% control solution is shown. f n This is the correction factor for each impurity.

[0033] In this application, the correction factors for impurities LFSB-1 to LFSB-13 are known ( f n The values ​​are as follows: 1.2, 1.1, 0.9, 1.9, 0.9, 1.2, 1.3, 1.3, 1.2, 1.0, 1.6, 1.3, 1.5. The correction factor for the remaining unknown impurities is calculated as 1.0.

[0034] Another aspect of this application discloses the application of the detection method described herein in the quality control of rivaroxaban tablets. The detection method of this application can detect a wide variety of impurities, has good classification effects, good specificity and repeatability, and ensures the accuracy and reliability of the detection results, thus providing a solid foundation for the quality control of rivaroxaban tablets.

[0035] This application has at least the following beneficial effects: The rivaroxaban tablets provided in this application can analyze and detect a wide range of related substances, exhibit good method specificity, and show no interference from impurity peaks, the main peak, or blank solvents; they also demonstrate high sensitivity, with impurity detection limits reaching the ng level; and high accuracy, with a recovery rate of 90%. The separation efficiency is between 108% and 108%; it is robust, with minor changes in chromatographic conditions not affecting the separation effect; it is environmentally friendly, the mobile phase does not contain ion-pairing reagents, it is simple to prepare, and it is inexpensive. The related substance detection method provided in this application can provide a good foundation for the quality control of rivaroxaban tablets. Attached Figure Description

[0036] Figure 1 This is a chromatogram of the system suitability solution from Example 1.

[0037] Figure 2 This is the chromatogram of the blank solvent in Example 1.

[0038] Figure 3 This is the chromatogram of the limit of quantitation solution in Example 2.

[0039] Figure 4 The chromatogram of the spiked test solution in Example 5 is shown.

[0040] Figure 5 Comparison of chromatograms for the durability test in Example 6. Detailed Implementation

[0041] The present application will be further illustrated below with reference to specific embodiments. It should be noted that the specific embodiments below are for illustrative purposes only and do not limit the scope of the present application in any way.

[0042] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for the purpose of describing particular embodiments only and is not intended to be limiting of this application.

[0043] In addition, unless otherwise specified, methods without detailed conditions or steps are conventional methods, and the reagents and materials used are commercially available.

[0044] The specific information on rivaroxaban tablets, rivaroxaban raw materials, blank excipients, and various impurity reference standards used in the examples is shown in Table 3: Table 3 Raw Material and Reagent Information

[0045] Example 1: Specificity Test of Detection Method 1.1 Chromatographic conditions Chromatographic column: Octadecylsilane-bonded silica gel as stationary phase (Shim-Pack GIS C18, 4.0 × 150 mm, 3 μm); 0.1% phosphoric acid solution as mobile phase A; acetonitrile as mobile phase B; flow rate 1.0 mL / min; column temperature 35 °C; detection wavelength 241 nm; injection volume 10 μL. Elution was performed according to the following gradient program:

[0046] 1.2 Solution preparation: (1) Solvent: 0.01 mol / L phosphoric acid solution (measure 0.67 ml of phosphoric acid and dilute with water to 1000 ml) - acetonitrile (50:50, v / v) (2) Impurity reference stock solution 1: Take appropriate amounts of impurity LFSB-1, impurity LFSB-3, impurity LFSB-5, impurity LFSB-6, impurity LFSB-11 and impurity LFSB-12 reference standards, dissolve and dilute with acetonitrile to prepare a solution containing about 0.2 mg of each in 1 ml.

[0047] (3) Impurity reference stock solution 2: Take appropriate amounts of each of the following reference standards: LFSB-2, LFSB-4, LFSB-7, LFSB-8, LFSB-9, LFSB-10, and LFSB-13, dissolve and dilute them with solvent to prepare a solution containing approximately 0.2 mg of each in 1 ml.

[0048] (4) System adaptability solution: Take an appropriate amount of rivaroxaban, place it in a volumetric flask, add an appropriate amount of each impurity control stock solution, dissolve and dilute with solvent to prepare a solution containing 0.5 mg of rivaroxaban and 1 μg of each impurity per 1 ml.

[0049] 1.3 Sample Injection and Detection: Take 10 μl of the system suitability solution and inject it into the high-performance liquid chromatograph (Shimadzu LC-2040C). Perform the injection and analysis according to the chromatographic conditions described in section 1.1, and record the chromatogram. The analytical results are shown in Table 4.

[0050] Table 4 Specificity Test Results

[0051] The analysis results show that this detection method effectively separates rivaroxaban and its 13 impurities, with good separation accuracy and no interference between the main peak and the impurities. Furthermore, Figure 1 The chromatogram of the system-adaptive solution is shown, indicating good separation between impurities, between the main peak and preceding and following peaks, and effective separation of each impurity from adjacent impurities. Figure 2 The blank solvent chromatogram shows that the solvent does not interfere with the detection of various impurities, indicating that the detection method of this application has good specificity.

[0052] Example 2: Limit of Quantitation Test of Detection Method 2.1 Chromatographic conditions: Same as item 1.1 in Example 1.

[0053] 2.2 Solution preparation: The preparation of solvent, impurity control stock solution 1 and impurity control stock solution 2 is the same as in item 1.2 of Example 1.

[0054] Preparation of sensitivity test solution: Take an appropriate amount of the above impurity control stock solution, dilute it stepwise with solvent, and use the signal-to-noise ratio (S / N) of about 10 as the limit of quantitation test solution.

[0055] 2.3 Limit of Quantitation Test: Following the chromatographic conditions set in section 2.1, 10 μl of the limit-of-quantitation (LOQ) test solution was injected into the high-performance liquid chromatograph (HPLC) and the chromatogram was recorded. The LOQ analysis results are shown in Table 5. Table 5 Results of Limit of Quantitation Test

[0056] The results showed that, using the method of Example 1, impurities 1-13 and rivaroxaban could still be separated and detected well at concentrations as low as approximately 1 ng. Furthermore, Figure 3 The chromatograms of the limit of quantitation are provided, showing that each impurity can still be detected at the limit of quantitation level, and the peak shapes of each impurity are good, with good separation between each impurity. This indicates that the detection method of this application has good sensitivity, can effectively detect low-content impurities, eliminate potential drug use risks, and ensure the effectiveness and safety of drug quality.

[0057] Example 3: Linearity and Correction Factor Test of Detection Method 3.1 Chromatographic conditions: Same as item 1.1 in Example 1. 3.2 Solution preparation: The solvent is as described in section 1.2 of Example 1.

[0058] Take appropriate amounts of rivaroxaban reference standard and impurities LFSB-1 to LFSB-13, dissolve and dilute to the mark with solvent to prepare solutions of different linear concentrations, as shown in Table 6: Table 6 Linear solutions (unit: μg / ml)

[0059] 3.3 Linearity and Correction Factor Test Following the chromatographic conditions set in section 3.1, 10 μl of solutions with different linear concentrations were accurately measured and injected into the liquid chromatograph, and the chromatograms were recorded. The results are shown in Table 7. Table 7. Results of Linearity and Correction Factor Tests

[0060] The results showed that, using the method of Example 1, impurities LFSB-1 to LFSB-13 and rivaroxaban exhibited good linearity within the limit of quantitation to approximately 10 μg / ml, with a wide linear range. This indicates that the testing method of this application has good linearity, can quantitatively detect impurity content, and has high accuracy. The correction factors were all between 0.2 and 5.0; therefore, the principal component comparison method with correction factors can be used to calculate the content of each impurity for related substances testing.

[0061] Specifically, the formula for calculating impurity content is:

[0062] in, A S The peak area of ​​impurities in the test solution. The peak area of ​​the main component in the 0.2% control solution is shown. f n This is the correction factor for each impurity.

[0063] Example 4: Precision Test of Detection Method 4.1 Chromatographic conditions: Same as item 1.1 in Example 1. 4.2 Solution preparation: (1) Solvent: 0.01 mol / L phosphoric acid solution (measure 0.67 ml of phosphoric acid and dilute with water to 1000 ml) - acetonitrile (50:50, v / v).

[0064] (2) Blank excipient solution: Weigh 825 mg of blank excipient accurately, place it in a 50 ml volumetric flask, add solvent to dissolve and quantitatively dilute to the mark, shake well, and the solution is obtained.

[0065] (3) System suitability solution: Weigh 50 mg of rivaroxaban accurately, place it in a 100 ml volumetric flask, add 0.5 ml of each impurity reference stock solution, dissolve and quantitatively dilute to the mark with solvent, shake well, and the solution is ready.

[0066] (4) Sensitivity solution: Accurately measure 5 ml of the control solution, place it in a 20 ml volumetric flask, dilute with solvent to the mark, and shake well to obtain the solution.

[0067] (5) Test solution: Take 20 rivaroxaban tablets, grind them finely in an agate mortar, weigh about 850 mg (approximately equivalent to 25 mg of rivaroxaban), place them in a 50 ml volumetric flask, add solvent to dissolve and quantitatively dilute to the mark, shake well, and collect the filtrate. (Prepare 6 parallel portions) (6) Control solution: Accurately measure 1 ml of the test solution and place it in a 100 ml volumetric flask. Dilute to the mark with solvent and shake well. Accurately transfer 2 ml of the above solution to a 10 ml volumetric flask, dilute to the mark with solvent, and shake well. (Prepare 6 parallel portions) 4.3 Precision Test Following the chromatographic conditions set in section 4.1, accurately measure 10 μl each of the solvent, system suitability solution, sensitivity solution, control solution, and test solution, and inject them into the liquid chromatograph, recording the chromatograms. The results are shown in Table 8. Table 8 Precision Test Results

[0068] The results showed that, using the method of Example 1, the average contents of impurity LFSB-5, other largest unknown single impurities, and total impurities in the six test samples determined by the first person were 0.015%, 0.020%, and 0.035%, respectively, which were less than 0.05%, so no RSD was calculated. The average contents of impurity LFSB-5, other largest unknown single impurities, and total impurities in the six test samples determined by the second person were 0.013%, 0.019%, and 0.045%, respectively, which were less than 0.05%, so no RSD was calculated. The average contents of impurity LFSB-5, other largest unknown single impurities, and total impurities in the 12 samples were 0.014%, 0.020%, and 0.040%, respectively, which were less than 0.05%, so no RSD was calculated. Because the impurity content was less than 0.05%, the absolute difference between the contents measured by the two individuals was not compared. These results indicate that the detection method provided in this application has good precision.

[0069] Example 5: Accuracy Test of Detection Method 5.1 Chromatographic conditions: Same as item 1.1 in Example 1.

[0070] 5.2 Solution preparation: The solvent, impurity stock solution 1, and impurity stock solution 2 were prepared according to item 1.2 of Example 1.

[0071] Rivaroxaban stock solution: Accurately weigh 4 mg of rivaroxaban reference standard, place it in a 20 ml volumetric flask, dissolve it in acetonitrile and dilute quantitatively to the mark, then shake well.

[0072] Mixed reference solution: Accurately transfer 0.5 ml of each impurity stock solution and rivaroxaban stock solution into a 100 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0073] Mixed impurity stock solution: Accurately transfer 5 ml of each impurity reference stock solution into the same 100 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0074] Test solution: Take 20 rivaroxaban tablets, grind them finely in an agate mortar, weigh about 850mg (about equivalent to 25mg of rivaroxaban), put them in a 50ml volumetric flask, add solvent to dissolve and dilute to the mark, shake well, and take the filtrate.

[0075] Sample preparation: Take 20 rivaroxaban tablets, grind them finely in an agate mortar, weigh out about 850mg (about equivalent to 25mg of rivaroxaban), make 12 portions, place them in different 50ml volumetric flasks, add solvent to dissolve, make 3 portions into one group, make 4 groups in total, add 1.2ml, 2.5ml, 5ml and 12.5ml of mixed impurity stock solution to each group respectively, dilute to the mark with solvent, shake well, and collect the filtrate.

[0076] 5.3 Accuracy Test Following the chromatographic conditions set in section 5.1, accurately measure 10 μl of each of the above solutions and inject them into the liquid chromatograph, recording the chromatograms. The results are shown in Table 9: Table 9 Accuracy Test Results

[0077] The results showed that the average recoveries of impurities LFSB-1 to LFSB-13 were within the limit concentration range of 90% to 108% at concentrations of 0.048% to 0.5%, and the RSDs of the recoveries for both samples were less than 10%, indicating that the detection method of this application has good recovery and high accuracy. Furthermore... Figure 4 The chromatogram at a 0.5% spike concentration is shown. It can be seen that the separation between each impurity and between the main peak and the preceding and following peaks are all effective, indicating that the detection method of this application can effectively control the quality of rivaroxaban tablets.

[0078] Example 6: Robustness Test of the Detection Method 6.1 Chromatographic conditions: Based on the conditions in section 1.1 of Example 1, the column temperature (±2℃), flow rate (±0.05ml / min), mobile phase A concentration (±0.005%), initial mobile phase ratio (±1%), and different batches of the same brand of chromatographic columns were changed. It should be noted that the changed conditions are all single variables, and other conditions not specified are the same as those in section 1.1 of Example 1.

[0079] 6.2 Solution preparation: Solvent: 0.01 mol / L phosphoric acid solution (measure 0.67 ml of phosphoric acid and dilute with water to 1000 ml) - acetonitrile (50:50, v / v) Blank excipient solution: Weigh 825 mg of blank excipient accurately, place it in a 50 ml volumetric flask, add solvent to dissolve and dilute quantitatively to the mark, shake well to obtain the solution.

[0080] System suitability solution: Weigh 50 mg of rivaroxaban accurately, place it in a 100 ml volumetric flask, add 0.5 ml of each impurity stock solution, dissolve and quantitatively dilute to the mark with solvent, shake well, and the solution is ready.

[0081] 6.3 Durability Test Following the chromatographic conditions set in section 6.1, accurately measure 10 μl of each of the above solutions and inject them separately into the liquid chromatograph, recording the chromatograms. The chromatograms are shown below. Figure 5 As shown in Table 10: Table 10 Durability Test Results

[0082] The results showed that by slightly changing the column temperature (±2℃), flow rate (±0.05ml / min), mobile phase A phosphoric acid concentration (±0.005%), initial mobile phase ratio (±1%), and using different batches of the same brand of chromatographic column, compared with the conditions in Example 1, the separation of impurities and between the main peak and adjacent peaks in the system adaptability solution chromatogram met the requirements. This method has good robustness.

[0083] It should be noted that this application is not limited to the above-described embodiments. The above embodiments are merely examples, and any embodiments with the same structure and effect as the technical concept within the scope of this application are included in the technical scope of this application. Furthermore, various modifications that can be conceived by those skilled in the art to the embodiments, and other ways of constructing by combining some of the constituent elements of the embodiments, without departing from the spirit of this application, are also included in the scope of this application.

Claims

1. A method for detecting related substances in rivaroxaban tablets, characterized in that, High-performance liquid chromatography (HPLC) was used for detection, and the chromatographic conditions were as follows: Column: The packing material is octadecylsilane-bonded silica gel; Mobile phase: Mobile phase A is an aqueous solution of phosphoric acid, and mobile phase B is acetonitrile; gradient elution; Flow rate: 0.95 1.05 ml / min; Column temperature: 33 37℃; Detection wavelength: 230 250nm; Injection volume: 10 20μl.

2. The detection method as described in claim 1, characterized in that, The specifications of the chromatographic column are: particle size 2-3μm, column length 150mm, inner diameter 3.0-4.0mm; Preferably, the chromatographic column has the following specifications: particle size 3μm, column length 150mm, and inner diameter 4.0mm.

3. The detection method as described in claim 1, characterized in that, The volume percentage of the phosphoric acid aqueous solution in the mobile phase A is 0.095%. 0.105%; preferably 0.1%.

4. The detection method as described in claim 1, characterized in that, The flow rate is 1.0 ml / min; the column temperature is 35°C; and the detection wavelength is 241 nm.

5. The detection method as described in claim 1, characterized in that, The gradient elution program is as follows: 0-2 min, 96% A; 2-10 min, 96% A → 77% A; 10-20 min, 77% A; 20-40 min, 77% A → 60% A; 40-55 min, 60% A; 55-55.1 min, 60% A → 96% A; 55.1-75 min, 96% A.

6. As claimed in claim 1 5. The detection method according to any one of the claims is characterized by further comprising the following steps: Preparation of test solution: Take an appropriate amount of rivaroxaban tablets, grind them into a fine powder, accurately weigh an appropriate amount of the fine powder, add solvent to dissolve and dilute to 50 ml, shake well, filter, take the filtrate, and prepare a test solution containing 0.5 mg of rivaroxaban per 1 ml. Preparation of the control solution: Accurately measure the test solution, dilute it with solvent to prepare a 0.2% control solution, and shake well; Preparation of sensitivity solution: Accurately measure the control solution, dissolve and dilute to the mark with solvent, and shake well; Preparation of impurity reference stock solution 1: Take appropriate amounts of impurities LFSB-1, LFSB-3, LFSB-5, LFSB-6, LFSB-11, and LFSB-13, dissolve and dilute them with acetonitrile to prepare a solution containing 0.2 mg of each relevant impurity per 1 ml; Preparation of impurity reference stock solution 2: Take appropriate amounts of impurities LFSB-2, LFSB-4, LFSB-7, LFSB-8, LFSB-9, LFSB-10, and LFSB-13, dissolve and dilute them with solvent to prepare a solution containing 0.2 mg of each relevant impurity per 1 ml; Preparation of system suitability solution: Take rivaroxaban reference standard, add stock solutions of each impurity reference standard, dissolve and dilute with solvent to prepare a solution containing 0.5 mg rivaroxaban and 1 μg of each impurity per 1 ml, and shake well; The solvent is a mixed solvent prepared by mixing 0.01 mol / L phosphoric acid solution and acetonitrile at a volume ratio of 50:

50.

7. The detection method as described in claim 1, characterized in that, The formula for calculating the content of the impurities is as follows: in, A S The peak area of ​​impurities in the test solution. The peak area of ​​the main component in the 0.2% control solution is shown. f n This is the correction factor for each impurity.

8. The detection method as described in claim 1, characterized in that, The impurities include LFSB. 1 to LFSB 13, with correction factors of 1.2, 1.1, 0.9, 1.9, 0.9, 1.2, 1.3, 1.3, 1.2, 1.0, 1.6, 1.3, and 1.5 respectively; the correction factor for unknown impurities is 1.

0.

9. The detection method as described in claim 1, characterized in that, In the results of the detection method, the separation degree between each known impurity peak and between the main component peak and the preceding and following peaks should not be less than 1.5, and the signal-to-noise ratio of the main component peak in the sensitivity solution should be greater than 10.

10. As claimed in claim 1 Application of the detection method described in any one of the nine items in the quality control of rivaroxaban tablets.

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