A method for detecting the content of ezetimibe rosuvastatin calcium tablets by reversed phase chromatography system
The main components and impurities in ezetimibebrevisuvastatin calcium tablets were separated by high performance liquid chromatography (HPLC), which solved the problem of poor separation effect in the existing technology and achieved high sensitivity and high accuracy detection effect, which is suitable for the quality control of ezetimibebrevisuvastatin calcium tablets.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SHANXI YUNPENG PHARMA
- Filing Date
- 2026-04-10
- Publication Date
- 2026-06-26
Smart Images
Figure CN122282995A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of pharmaceutical analysis technology, specifically to a method for detecting the content of ezetimibe / suvastatin calcium tablets using a reversed-phase chromatography system. Background Technology
[0002] Ezetimibe, chemically named 1-(4-fluorophenyl)-3(R)-[3-(4-fluorophenyl)-3(S)-hydroxypropyl]-4(S)-(4-hydroxyphenyl)-2-acetidinone; rosuvastatin calcium, chemically named bis-[(E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methanesulfonyl)amino]-pyrimidin-5-yl](3R,5S)-3,5-dihydroxyhept-6-enoic acid] calcium salt (2:1). Ezetimibe-rosuvastatin calcium tablets are a novel lipid-regulating compound preparation, composed of a cholesterol absorption inhibitor and an HMG-CoA reductase inhibitor, with significant clinical value. Developed by Sanofi, this preparation was launched in the EU in 2019, with strengths including 10mg / 10mg, 10mg / 20mg, and 10mg / 40mg.
[0003] During the production and storage of ezetimibe / rosuvastatin calcium tablets, various process impurities and degradation impurities may be generated, including: rosuvastatin impurity B, rosuvastatin impurity C, rosuvastatin impurity D, rosuvastatin impurity K, rosuvastatin impurity M, rosuvastatin impurity N, and ezetimibe impurity A, ezetimibe impurity D, and ezetimibe impurity E. All of these impurities are substances requiring control and must be effectively separated from the main component to ensure drug quality. Among them: Rosuvastatin impurity B is a stereoisomer of rosuvastatin calcium, which is a process by-product and degradation product; Rosuvastatin impurity C is an impurity generated by the degradation of rosuvastatin calcium under light, acid-base or oxidative conditions; Rosuvastatin impurity D is a process impurity and degradation impurity of rosuvastatin calcium; Rosuvastatin impurities K, M, and N are degradation impurities generated during the production or storage of rosuvastatin calcium; Ezetimibe impurities A, D, and E are process impurities and degradation impurities generated during the synthesis, degradation or storage of ezetimibe.
[0004] Currently, this compound preparation is not included in the pharmacopoeias of various countries, and existing detection methods are insufficient to achieve good separation of the main component from multiple impurities, exhibiting deficiencies in specificity, sensitivity, and repeatability. Therefore, developing a content determination method with good separation effect, high accuracy, and wide applicability is of great significance. Summary of the Invention
[0005] Therefore, the purpose of this invention is to provide a reversed-phase chromatography detection method with strong specificity, high sensitivity, wide linear range, good precision and accuracy, and excellent repeatability and robustness, which can simultaneously determine the content of ezetimibe and rosuvastatin calcium in ezetimibe-rosuvastatin calcium tablets and achieve effective separation of the main component and various impurities.
[0006] To achieve the aforementioned objectives, the technical solution adopted is as follows: A method for detecting the content of ezetimibe / suvastatin calcium tablets using reversed-phase chromatography, employing high-performance liquid chromatography (HPLC) with 0.1% trifluoroacetic acid solution as mobile phase A, acetonitrile as mobile phase B, and methanol as mobile phase C; gradient elution is performed using a chromatographic column packed with octadecylsilane-bonded silica gel; the detection wavelength is 242 nm, and the injection volume is 10 μL. The gradient elution procedure is as follows: 0.0 min to 5.0 min, mobile phase A is 55%, mobile phase B is 35%, and mobile phase C is 10%; 5.01 min, mobile phase A was adjusted to 65%, mobile phase B to 35%, and mobile phase C to 0%; From 5.01 min to 10.0 min, mobile phase A was adjusted to 50%, mobile phase B to 50%, and mobile phase C to 0%. From 10.0 min to 12.0 min, maintain mobile phase A at 50%, mobile phase B at 50%, and mobile phase C at 0%. 12.01 min, the recovery rates of mobile phase A were 55%, mobile phase B was 35%, and mobile phase C was 10%. From 12.01 min to 15.0 min, maintain mobile phase A at 55%, mobile phase B at 35%, and mobile phase C at 10%. This method can separate and detect the following impurities: rosuvastatin impurity B, rosuvastatin impurity C, rosuvastatin impurity D, rosuvastatin impurity K, rosuvastatin impurity M, rosuvastatin impurity N, ezetimibe impurity A, ezetimibe impurity D, and ezetimibe impurity E. The structures of each impurity are as follows: RSF impurity B, RSF impurity, CRSF impurity D The structural formula of RSF impurity B is: ; The structural formula of RSF impurity C is: ; The structural formula of RSF impurity D is: ; The structural formula of RSF impurity K is: ; The structural formula of RSF impurity M is: ; The structural formula of RSF impurity N is: ; The structural formula of impurity A in YZ is: ; The structural formula of impurity D in YZ is: ; The structural formula of YZ impurity E is: .
[0007] As a further improvement of the present invention, the high performance liquid chromatography flow rate is 1.1 mL / min to 1.3 mL / min.
[0008] As a further improvement of the present invention, the high performance liquid chromatography flow rate is 1.2 mL / min.
[0009] As a further improvement of the present invention, the high performance liquid chromatography column temperature is 25℃~35℃.
[0010] As a further improvement of the present invention, the high performance liquid chromatography column temperature is 30°C.
[0011] As a further improvement of the present invention, the chromatographic column is an ACEA UltraCore2.5 SuperC18 with dimensions of 4.6 mm × 150 mm and a diameter of 2.5 μm, or a chromatographic column with equivalent performance.
[0012] As a further improvement of the present invention, the content of ezetimibe is calculated using the external standard method according to the following formula: Ezetimibe content % Where: m 依折对 The sample weight of ezetimibe reference standard; P 依折对 The content of ezetimibe reference standard; A 依折对 V represents the peak area of the ezetimibe peak in the reference solution. 对 This refers to the dilution factor of the reference solution; A 样 V represents the peak area of ezetimibe in the test solution; 样 C is the dilution factor of the test solution; 标 This represents the labeled content of ezetimibe in the test solution.
[0013] As a further improvement of the present invention, the calcium content of rosuvastatin is calculated using the external standard method according to the following formula: ; Where: m瑞舒伐对 This refers to the sample weight of rosuvastatin calcium reference standard; P 瑞舒伐对 This refers to the content of rosuvastatin calcium reference standard; A 瑞舒伐对 V represents the peak area of rosuvastatin in the reference solution. 对 The dilution factor of the reference solution is denoted by f; f is the response factor; A 样 V represents the peak area of rosuvastatin in the test solution. 样 C is the dilution factor of the test solution; 标 This represents the labeled content of rosuvastatin calcium in the test solution.
[0014] The beneficial effects of this invention are as follows: Compared with the prior art, this invention uses a 0.05% trifluoroacetic acid-acetonitrile-methanol gradient elution system, which not only effectively separates rosuvastatin and ezetimibe, as well as other known impurities, but also exhibits good peak symmetry. It can also rapidly detect ezetimibe and rosuvastatin content below 3 / 100 of the sample concentration. Furthermore, it demonstrates significant advantages in specificity, limit of quantitation, limit of detection, linear range, and repeatability. It offers advantages such as a more environmentally friendly detection method, wider applicability, high precision and accuracy, good repeatability, and good system suitability, providing an important methodological reference for the quality control of ezetimibe and rosuvastatin calcium drugs. Attached Figure Description
[0015] The accompanying drawings, which form part of this application, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings: Figure 1 For the wiring diagram of Yizhemai; Figure 2 Linear graph of rosuvastatin calcium; Figure 3 It involves checking the blank solvent liquid chromatogram; Figure 4 It involves checking the liquid chromatogram of blank excipients; Figure 5 It involves examining the liquid chromatogram of an ezetimibe solution; Figure 6 It involves examining the liquid chromatogram of rosuvastatin solution; Figure 7 It checks for impurities interfering with the liquid chromatogram; Figure 8 It involves checking the liquid chromatogram of the sample solution. Detailed Implementation
[0016] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present invention will now be described in detail with reference to the accompanying drawings and embodiments.
[0017] To enable those skilled in the art to better understand the present application, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present application, and not all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort should fall within the scope of protection of the present application.
[0018] According to the detection method described in this invention, the content of ezetimibe / suvastatin calcium tablets is mainly detected, and the external standard method is used for calculation.
[0019] The detection method described in this invention uses a Thermo Fisher Scientific / Waters high-performance liquid chromatograph and an ACEUltraCore2.5 SuperC18 column (4.6×150mm, 2.5μm) in its implementation.
[0020] The present invention will be further illustrated below with reference to the embodiments.
[0021] Example 1: Chromatographic conditions and chromatographic system of the detection method of the present invention Instrument: Thermo Fisher Scientific / Waters High Performance Liquid Chromatography (HPLC) Chromatographic column: ACE UltraCore 2.5 Super C18 column (4.6×150mm, 2.5μm) Mobile phase A: 0.1% trifluoroacetic acid Mobile phase B: Acetonitrile Mobile phase C: Methanol The gradient procedure is as follows: Based on volume ratio, gradient elution was performed using a C18 column. The elution program was as follows: elution was initiated with mobile phase A at 55%, mobile phase B at 35%, and mobile phase C at 10%, and continued for 5.0 minutes; the proportion of mobile phase A was increased while the proportion of mobile phase C was decreased, and at 5.01 minutes, the proportion of mobile phase A was adjusted to 65% and the proportion of mobile phase C to 0%; the proportion of mobile phase A was decreased while the proportion of mobile phase B was increased, and at 10.0 minutes, the proportion of mobile phase A was adjusted to 50% and the proportion of mobile phase B to 50%; elution was continued for 12.0 minutes; at 12.01 minutes, the proportion of mobile phase A was adjusted to 55%, mobile phase B to 35%, and mobile phase C to 10%; elution was continued at this mobile phase ratio for 15.0 minutes.
[0022] Flow rate: 1.2 mL / min; Column temperature: 30℃; Detection wavelength: 242nm; Injection volume: 10 μL; Test solution: Take 5 tablets of this product, place them in a 250ml volumetric flask, add an appropriate amount of solvent, sonicate and shake for 5 minutes to dissolve ezetimibe and rosuvastatin, dilute to the mark with solvent, and shake well; centrifuge at 10,000 rpm for 10 minutes, filter the supernatant, and collect the filtrate.
[0023] Reference solution: Accurately weigh ezetimibe and rosuvastatin calcium reference standards, dissolve them in solvent and dilute quantitatively to prepare a solution containing approximately 200 μg per ml.
[0024] Calculation method: Ezetimibe content %, Where: m 依折对 The sample weight of ezetimibe reference standard; P 依折对 The content of ezetimibe reference standard; A 依折对 V represents the peak area of the ezetimibe peak in the reference solution. 对 This refers to the dilution factor of the reference solution; A 样 V represents the peak area of ezetimibe in the test solution; 样 C is the dilution factor of the test solution; 标 This represents the labeled content of ezetimibe in the test solution.
[0025] , Where: m 瑞舒伐对 This refers to the sample weight of rosuvastatin calcium reference standard; P 瑞舒伐对 This refers to the content of rosuvastatin calcium reference standard; A 瑞舒伐对 V represents the peak area of rosuvastatin in the reference solution. 对 The dilution factor of the reference solution is denoted by f; f is the response factor; A 样 V represents the peak area of rosuvastatin in the test solution. 样 C is the dilution factor of the test solution; 标 This represents the labeled content of rosuvastatin calcium in the test solution.
[0026] Example 2: Specificity test of the detection method described in this invention Blank solvent: Acetonitrile-water (50:50); Reference solution: Accurately weigh ezetimibe and rosuvastatin calcium reference standards, dissolve them in solvent and dilute quantitatively to prepare a solution containing approximately 200 μg per ml, in duplicate.
[0027] Test solution: Take 5 tablets of this product, place them in a 250ml volumetric flask, add an appropriate amount of solvent, sonicate and shake for 5 minutes to dissolve ezetimibe and rosuvastatin, dilute to the mark with solvent, and shake well; centrifuge at 10,000 rpm for 10 minutes, filter the supernatant, and collect the filtrate.
[0028] Impurity stock solution: Dissolve and dilute an appropriate amount of RSF-impurity B in acetonitrile to prepare a solution containing approximately 0.2 mg per ml, which serves as the RSF-impurity B stock solution. Prepare stock solutions of RSF-impurity C, RSF-impurity D, RSF-impurity M, RSF-impurity N, YZ-impurity A, YZ-impurity D, YZ-impurity E, and YZ-impurity F in the same manner. Take an appropriate amount of RSF-impurity K, dissolve and dilute it in acetonitrile-water (50:50) to prepare a solution containing approximately 0.2 mg per ml, which serves as the RSF-impurity K stock solution.
[0029] Impurity location solution: Take 1.0 ml of each impurity stock solution and place it in a 100 ml volumetric flask. Add acetonitrile-water (50:50) to dissolve and dilute to prepare a solution containing approximately 2 μg per ml. Impurity Mixture Solution: Weigh 10 mg each of ezetimibe and rosuvastatin calcium reference standards accurately. Take 0.5 ml of each impurity stock solution and place them in the same 50 ml volumetric flask. Dissolve and dilute them separately with acetonitrile-water (50:50) to prepare a solution containing approximately 2 μg of each impurity per ml. Under the above chromatographic conditions, accurately measure 10 μl each of the solvent, test solution, reference solution, RSF-impurity B, RSF-impurity C, RSF-impurity D, RSF-impurity M, RSF-impurity N, YZ-impurity A, YZ-impurity D, YZ-impurity E, YZ-impurity F, RSF-impurity K solution, and mixed impurity solution, and inject them into the liquid chromatograph, recording the chromatograms. The results are shown in Table 1 below.
[0030] Table 1 Results of specificity test Conclusion: Blank solvent and blank excipient do not interfere with impurity detection. The localization solutions for each impurity can be effectively separated from rosuvastatin and ezetimibe. The theoretical plate number based on rosuvastatin is greater than 5000, and the tailing factor is less than 2.0. The theoretical plate number based on ezetimibe is greater than 5000, and the tailing factor is less than 2.0. The specificity is good.
[0031] Example 3: Test of the limit of quantitation and limit of detection of the detection method described in this invention The reference solution was serially diluted with solvent to prepare a limit of quantitation solution with a signal-to-noise ratio of approximately 10:1. 10 μl of the solution was accurately measured and injected sequentially into the liquid chromatograph, and the chromatograms were recorded. The experimental results are shown in Table 2-3 below.
[0032] Table 2 Results of Limit of Quantitation Tests for Ezetimibe and Rosuvastatin Calcium Table 3. Precision results of the limit of quantitation test for ezetimibe and rosuvastatin calcium. The results showed that the rosuvastatin solution concentration with a quantitation limit was 0.04998 μg / ml, the signal-to-noise ratio was 13.5–17.8, and the RSD of the 6-shot quantitation limit was 7.8%, less than 10.0%; the ezetimibe solution concentration with a quantitation limit was 0.005063 μg / ml, the signal-to-noise ratio was 10.9–14.4, and the RSD of the 6-shot quantitation limit was 4.7%, less than 10.0%; demonstrating the good sensitivity of the present invention.
[0033] Example 4: Linearity and range detection of the detection method described in this invention Solvent: Acetonitrile-water (50:50) 200% linear stock solution: Weigh 20 mg of rosuvastatin calcium and ezetimibe respectively, accurately, place them in a 50 ml volumetric flask, dilute to the mark with solvent, and shake well.
[0034] The 200% stock solution was serially diluted to prepare linear test solutions (concentrations from 10% to 200%). 10 μl of each linear test solution was injected into the liquid chromatograph, and the chromatograms were recorded. Linear regression was performed on the peak area against the concentration of the test solution, and the results are as follows: 1. Linear 1 Linear solution 6: i.e., 200% linear mother liquor; Take 67.5 ml of the linear solution, place it in a 10 ml volumetric flask, dilute with solvent and bring to volume, shake well, and this is linear solution 5 (150%). Take 65.0 ml of the linear solution, place it in a 10 ml volumetric flask, dilute with solvent and bring to volume, shake well, and this is linear solution 4 (100%). Take 65.0 ml of the linear solution, place it in a 20 ml volumetric flask, dilute with solvent and bring to volume, shake well, and this is linear solution 3 (50%). Take 65.0 ml of the linear solution, place it in a 50 ml volumetric flask, dilute with solvent and bring to volume, shake well, and this is linear solution 2 (20%). Take 65.0 ml of the linear solution, place it in a 100 ml volumetric flask, dilute with solvent and bring to volume, shake well, and this is linear solution 1 (10%). Accurately measure 10 μl of each linear solution and inject it into the liquid chromatograph, then record the chromatogram. The results are shown in Table 4 below.
[0035] Table 4 Results of Izzy wiring and range tests Remark: .
[0036] Table 5 Results of Rosuvastatin Calcium Linearity and Range Tests Note: .
[0037] Conclusions: For ezetimibe, within the concentration range of 10%–200% relative to the test sample solution, the linear equation is y = 17,799.3988x + 7,731.7013, with a correlation coefficient r of 0.9999 and a y-intercept of 0.2% of the 100% response value, indicating a good linear relationship. For rosuvastatin calcium, within the concentration range of 10%–200% relative to the test sample solution, the linear equation is y = 20,746.9207x + 3,421.7966, with a correlation coefficient r of 0.9999 and a y-intercept of 0.1% of the 100% response value, also indicating a good linear relationship.
[0038] Example 5: Detection of the injection precision of the detection method described in this invention The reference solution was injected six times repeatedly to examine the injection precision. The results are shown in the table below.
[0039] Table 6 Results of injection precision test (n=6); The results showed that the peak area RSD of rosuvastatin was 0.18% and that of ezetimibe was 0.10%, both less than 2.0%, indicating that the method had good injection precision.
[0040] Example 6: Detection of the repeatability of the detection method described in this invention Blank solvent: acetonitrile-water (50:50).
[0041] Test solution: Take 5 tablets of this product, place them in a 250ml volumetric flask, add an appropriate amount of solvent, sonicate and shake for 5 minutes to dissolve ezetimibe and rosuvastatin, dilute to the mark with solvent, and shake well; centrifuge at 10,000 rpm for 10 minutes, filter the supernatant, and collect the filtrate. Perform 6 replicates.
[0042] Reference solution: Accurately weigh ezetimibe and rosuvastatin calcium reference standards, dissolve them in solvent and dilute quantitatively to prepare a solution containing approximately 200 μg per ml, in duplicate.
[0043] Accurately transfer 10 μl of each of the above sample solutions into the high-performance liquid chromatograph and record the chromatograms. The results are shown in the table below.
[0044] Table 7 Repeatability Test Results The results showed that the average detection rate of rosuvastatin calcium in the six samples was 100.9%, with an RSD of 0.48% and an RSD of less than 2.0%; the average detection rate of ezetimibe was 105.7%, with an RSD of 0.16% and an RSD of less than 2.0%; therefore, the method had good repeatability.
[0045] Example 7: Accuracy testing of the detection method described in this invention Solvent: Acetonitrile-water (40:60).
[0046] Reference solutions: Weigh 20 mg of ezetimibe and rosuvastatin calcium reference standards respectively, accurately, dissolve in solvent and dilute quantitatively to prepare a solution containing approximately 200 μg per ml, in duplicate.
[0047] 50% Horizontal Spiking Solution: Accurately weigh 10 mg each of ezetimibe and rosuvastatin calcium reference standards, and approximately 280 mg of blank excipient. Place them in a 100 ml volumetric flask, add solvent, sonicate to dissolve and dilute to the mark. Centrifuge at 10,000 rpm for 10 minutes, collect the supernatant, filter, and collect the filtrate. Prepare three parallel aliquots.
[0048] 100% Horizontal Spiking Test Solution: Accurately weigh 10 mg each of ezetimibe and rosuvastatin calcium reference standards, and approximately 280 mg of blank excipient. Place them in a 50 ml volumetric flask, add solvent, sonicate to dissolve and dilute to the mark, centrifuge at 10,000 rpm for 10 minutes, collect the supernatant, filter, and collect the filtrate. Prepare 3 parallel aliquots.
[0049] 150% Horizontal Spiking Solution: Accurately weigh 15 mg each of ezetimibe and rosuvastatin calcium reference standards, and approximately 421 mg of blank excipient. Place them in a 50 ml volumetric flask, add solvent, sonicate to dissolve and dilute to the mark. Centrifuge at 10,000 rpm for 10 minutes, collect the supernatant, filter, and collect the filtrate. Prepare three parallel aliquots.
[0050] Accurately inject 10 μL of each of the above sample solutions into the high-performance liquid chromatograph and record the chromatograms. Calculate the recovery rate using the self-comparison method.
[0051] Recovery rate calculation formula: Recovery rate (%) = ×100% Table 8 Results of the rosuvastatin calcium accuracy test Conclusion: The average recovery rates of rosuvastatin calcium at various concentrations ranged from 98% to 101%, with an RSD of 0.62% < 2.0%, indicating good method accuracy.
[0052] Table 9. Results of the Accuracy Test of Ezekiel Conclusion: The average recovery rate of ezetimibe at various concentrations was 98%–101%, with an RSD of 0.56% < 2.0%, indicating good method accuracy.
[0053] Example 8: Detection of solution stability using the detection method described in this invention Blank solvent: acetonitrile-water (50:50).
[0054] The stability of the test solution and reference solution was assessed using the test solution, self-reference solution, and system suitability solution under the "Specificity" section. These solutions were injected at different time points after preparation under selected chromatographic conditions to examine changes in peak area. The specific results are as follows: Table 10 Stability results of the test solution Table 11 Stability results of ezetimibe reference solution Table 12. Stability results of rosuvastatin calcium reference solution The results showed that the percentage change of peak area within 33 hours for the test solutions was 0.81% for ezetimibe and 1.27% for rosuvastatin. The maximum percentage change of peak area for ezetimibe reference standard was 0.95%, and the maximum percentage change of peak area for rosuvastatin calcium reference standard was 0.49%. All results were less than 2.0%, indicating good solution stability.
[0055] Example 10: Dependency test of the detection method described in this invention Blank solution, test solution, and control solution were prepared according to standard methods and subjected to different chromatographic conditions to investigate the robustness of the method. Specific conditions such as column temperature and flow rate were investigated, as detailed below.
[0056] Table 13 Durability Test Conditions The test results under different conditions are shown in the table below.
[0057] Table 14 Durability Test Results Conclusion: Under all conditions, the theoretical plate number and tailing factor of the reference solution met the requirements, and there were no significant changes in any of the index parameters. The determination of rosuvastatin calcium and ezetimibe content in the test solution was relatively consistent, with RSDs of 0.23% and 0.16%, respectively. The method has good robustness.
[0058] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, component splitting or combination, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A method for detecting the content of ezetimibe / bupresuastatin calcium tablets using a reversed-phase chromatography system, characterized in that: High-performance liquid chromatography (HPLC) was used with 0.1% trifluoroacetic acid solution as mobile phase A, acetonitrile as mobile phase B, and methanol as mobile phase C. Gradient elution was performed using a column packed with octadecylsilane-bonded silica gel. The detection wavelength was 242 nm, and the injection volume was 10 μL. The gradient elution procedure is as follows: 0.0 min to 5.0 min, mobile phase A is 55%, mobile phase B is 35%, and mobile phase C is 10%; 5.01 min, mobile phase A was adjusted to 65%, mobile phase B to 35%, and mobile phase C to 0%; From 5.01 min to 10.0 min, mobile phase A was adjusted to 50%, mobile phase B to 50%, and mobile phase C to 0%. From 10.0 min to 12.0 min, maintain mobile phase A at 50%, mobile phase B at 50%, and mobile phase C at 0%. 12.01 min, the recovery rates of mobile phase A were 55%, mobile phase B was 35%, and mobile phase C was 10%. From 12.01 min to 15.0 min, maintain mobile phase A at 55%, mobile phase B at 35%, and mobile phase C at 10%.
2. The method for detecting the content of ezetimibe / bresuvastatin calcium tablets using a reversed-phase chromatography system according to claim 1, characterized in that: The high-performance liquid chromatography flow rate is 1.1 mL / min to 1.3 mL / min.
3. The method for detecting the content of ezetimibe / bresuvastatin calcium tablets using a reversed-phase chromatography system according to claim 2, characterized in that: The high-performance liquid chromatography flow rate was 1.2 mL / min.
4. The method for detecting the content of ezetimibe / bresuvastatin calcium tablets using a reversed-phase chromatography system according to claim 1, characterized in that: The high-performance liquid chromatography column temperature is 25℃~35℃.
5. The method for detecting the content of ezetimibe / bresuvastatin calcium tablets using a reversed-phase chromatography system according to claim 4, characterized in that: The high-performance liquid chromatography column temperature is 30℃.
6. The method for detecting the content of ezetimibe / bresuvastatin calcium tablets using a reversed-phase chromatography system according to claim 1, characterized in that: The chromatographic column was an ACEA UltraCore 2.5 Super C18, with dimensions of 4.6 mm × 150 mm and a diameter of 2.5 μm.
7. The method for detecting the content of ezetimibe / brevisuvastatin calcium tablets using a reversed-phase chromatography system according to claim 1, characterized in that, The content of ezetimibe was calculated using the external standard method according to the following formula: Ezetimibe content % Where: m 依折对 The sample weight of ezetimibe reference standard; P 依折对 The content of ezetimibe reference standard; A 依折对 V represents the peak area of the ezetimibe peak in the reference solution. 对 This refers to the dilution factor of the reference solution; A 样 V represents the peak area of ezetimibe in the test solution; 样 C is the dilution factor of the test solution; 标 This represents the labeled content of ezetimibe in the test solution.
8. The method for detecting the content of ezetimibe / brevisuvastatin calcium tablets using a reversed-phase chromatography system according to claim 1, characterized in that, The calcium content of rosuvastatin was calculated using the external standard method according to the following formula: ； Where: m 瑞舒伐对 This refers to the sample weight of rosuvastatin calcium reference standard; P 瑞舒伐对 This refers to the content of rosuvastatin calcium reference standard; A 瑞舒伐对 V represents the peak area of rosuvastatin in the reference solution. 对 The dilution factor of the reference solution is denoted by f; f is the response factor; A 样 V represents the peak area of rosuvastatin in the test solution. 样 C is the dilution factor of the test solution; 标 This represents the labeled content of rosuvastatin calcium in the test solution.