Determination method of chiral isomers in rosuvastatin calcium ezetimibe tablet

Abstract

The invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets. The invention provides a method for determining a chiral isomer in a rosuvastatin calcium ezetimibe tablet, which comprises the following steps: changing an isocratic elution mode into a gradient elution mode, establishing an effective pretreatment method, optimizing a test solution solvent and a high performance liquid chromatography with specific parameters, and determining the chiral isomer in the rosuvastatin calcium ezetimibe tablet. The problems that the solubility difference of rosuvastatin calcium and ezetimibe is large, rosuvastatin calcium is unstable to acid, ezetimibe is unstable to alkali, isocratic elution time is too long and the like are solved. The quality of chiral isomers of rosuvastatin calcium and ezetimibe, which are recorded in European Pharmacopoeia and United States Pharmacopoeia, is controlled; establishing sensitivity and system applicability requirements; the retention time and correction factors of specific impurities are marked, so that the detection result is more accurate.

Description

Technical Field

[0001] The present application belongs to the field of pharmaceutical analysis technology, and specifically relates to a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets. Background Art

[0002] Rosuvastatin calcium is a selective HMG-CoA reductase inhibitor that primarily acts on the liver, increasing the number of receptors on the surface of hepatic LDL cells, promoting LDL absorption and catabolism, and inhibiting VLDL synthesis in the liver, thereby reducing the total number of VLDL and LDL particles. Ezetimibe selectively inhibits intestinal absorption of dietary sterols and bile cholesterol by specifically binding to the Niemann-Pick C1-Like 1 protein on the brush border membrane of intestinal villus epithelial cells.

[0003] As a combination preparation, rosuvastatin calcium ezetimibe tablets not only enhance therapeutic efficacy and achieve synergistic effects, but also simplify treatment plans and improve patient compliance. They also cover multiple disease symptoms, enhancing overall efficacy. Therefore, rosuvastatin calcium ezetimibe tablets have irreplaceable clinical value in the management of hypercholesterolemia and the control of complex symptoms.

[0004] According to the "Technical Guidelines for Quality Control Research of Chiral Drugs" issued by the Drug Review Center of the State Food and Drug Administration, chiral drugs should detect and control the content of chiral isomers throughout the entire R&D cycle. Currently, neither the Chinese Pharmacopoeia nor foreign pharmacopoeias include rosuvastatin calcium ezetimibe tablets, and the import registration standards do not include detection items for chiral isomers. The European Pharmacopoeia's rosuvastatin calcium raw material quality standard includes an analytical method for detecting chiral isomer impurity G, but it is an isocratic elution high-performance liquid chromatography method, and ezetimibe is difficult to elute; the United States Pharmacopoeia's ezetimibe raw material quality standard includes an analytical method for detecting chiral isomers, but this method cannot separate and detect rosuvastatin-related components.

[0005] The existing technology lacks a method capable of simultaneously detecting chiral isomers in rosuvastatin calcium and ezetimibe tablets. Rosuvastatin calcium and ezetimibe have significant solubility differences; rosuvastatin calcium is unstable in acid, while ezetimibe is unstable in base. Furthermore, existing isocratic elution methods require excessively long elution times, making them inadequate for simultaneous detection of both chiral isomers in a compound preparation.

[0006] Therefore, it is of great practical significance to establish a method for simultaneously detecting chiral isomers in rosuvastatin calcium ezetimibe tablets. Summary of the Invention

[0007] Based on this, one embodiment of the present application provides a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets to meet the detection requirements of simultaneously detecting chiral isomers in rosuvastatin calcium ezetimibe tablets.

[0008] On the one hand, the present application provides a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets, comprising the following steps:

[0009] High performance liquid chromatography was used to prepare the test solution, reference solution, sensitivity solution and system suitability solution using phosphate buffer-acetonitrile as solvent.

[0010] The chromatographic conditions include: the filling material of the chromatographic column includes silica gel with cellulose-tris (4-methylbenzoate) coated on the surface; and the detection wavelength is 240nm-244nm.

[0011] Mobile phase A included 0.08%-0.12% trifluoroacetic acid solution, and mobile phase B included acetonitrile, and gradient elution was performed;

[0012] The test solution, the control solution, the system suitability solution and the sensitivity solution were respectively drawn into a high performance liquid chromatograph for analysis and the chromatogram was recorded.

[0013] In one embodiment, the volume ratio of phosphate buffer to acetonitrile in the phosphate buffer-acetonitrile solvent is (58-62):(38-42).

[0014] In one embodiment, the pH value of the phosphate buffer-acetonitrile solvent is 5.0-6.5.

[0015] In one embodiment, the chromatographic column is DAICEL, OJ-3R.

[0016] In one embodiment, the size of the chromatographic column is 4.6 mm×250 mm.

[0017] In one embodiment, the column temperature is 20°C-30°C.

[0018] In one embodiment, the flow rate is 0.4 mL / min-0.6 mL / min.

[0019] In one embodiment, the injection volume is 10 μL-50 μL.

[0020] In one embodiment, the gradient elution procedure comprises:

[0021] 0 min, the volume percentage of the mobile phase A is 27, and the volume percentage of the mobile phase B is 73.

[0022] 50 min, the volume percentage of the mobile phase A is 27, and the volume percentage of the mobile phase B is 73.

[0023] 60 min, the volume percentage of the mobile phase A is 45, and the volume percentage of the mobile phase B is 55.

[0024] 90 min, the volume percentage of the mobile phase A is 45, and the volume percentage of the mobile phase B is 55.

[0025] 90.1 min, the volume percentage of the mobile phase A was 27, and the volume percentage of the mobile phase B was 73.

[0026] 100 min, the volume percentage of the mobile phase A is 27, and the volume percentage of the mobile phase B is 73.

[0027] In one embodiment, the method for preparing the system suitability solution comprises: taking a rosuvastatin calcium reference substance, a rosuvastatin impurity G reference substance, an ezetimibe reference substance, and an S,S,R-ezetimibe reference substance, mixing them, and then dissolving them.

[0028] In one embodiment, the chiral isomer comprises at least one of the following isomers:

[0029] Rosuvastatin impurity G: (3S,5R,6E)-7-[4-(4-fluorophenyl)-2-(N-methylmethanesulfonamido)-6-(isopropyl)pyrimidin-5-yl]-3,5-dihydroxyheptenoic acid.

[0030] R,R,S-Ezetimibe: (3R,4S)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0031] R,R,R-Ezetimibe: (3R,4R)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0032] S,S,S-Ezetimibe: (3S,4S)-1-(4-fluorophenyl)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0033] S,S,R-Ezetimibe: (3S,4R)-1-(4-fluorophenyl)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0034] R,S,R-Ezetimibe: (3S,4R)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0035] In one embodiment, the resolution of the rosuvastatin peak and the rosuvastatin impurity G peak is not less than 1.5.

[0036] The resolution of the ezetimibe peak and the S,S,R-ezetimibe peak must be no less than 1.5.

[0037] In one embodiment, the retention time of rosuvastatin impurity G is 40.7 min; the retention time of R,R,S-ezetimibe is 78.0 min; the retention time of R,R,R-ezetimibe is 72.7 min; the retention time of S,S,S-ezetimibe is 70.8 min; the retention time of S,S,R-ezetimibe is 79.1 min; and the retention time of R,S,R-ezetimibe is 79.1 min.

[0038] The present application provides a method for determining chiral isomers in rosuvastatin calcium and ezetimibe tablets, comprising changing the isocratic elution mode to gradient elution, establishing an effective pretreatment method, optimizing the test solution solvent, and a high-performance liquid chromatography method with specific parameters. The method solves the problems of large solubility differences between rosuvastatin calcium and ezetimibe, instability of rosuvastatin calcium to acid, instability of ezetimibe to base, and excessively long isocratic elution time. The method also performs quality control on the chiral isomers of rosuvastatin calcium and ezetimibe listed in the European Pharmacopoeia and the United States Pharmacopoeia, establishes sensitivity and system suitability requirements, and annotates the retention times and correction factors of specific impurities to make the detection results more accurate. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more fully understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present application. Those skilled in the art can also derive other drawings based on these drawings without inventive effort.

[0040] Figure 1 Chromatograms showing the detection of rosuvastatin impurity G, rosuvastatin, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, R,S,R-ezetimibe, and ezetimibe using the method of Example 1 of the present application;

[0041] Figure 2 This is the concentration and peak area trend graph of rosuvastatin;

[0042] Figure 3 This is a trend diagram of the concentration and peak area of ​​rosuvastatin impurity G;

[0043] Figure 4 This is a trend graph of the concentration and peak area of ​​ezetimibe;

[0044] Figure 5 This is a trend graph of the concentration and peak area of ​​S,S,S-ezetimibe;

[0045] Figure 6 This is a trend graph of the concentration and peak area of ​​R,R,R-ezetimibe;

[0046] Figure 7 This is the concentration and peak area trend graph of R,R,S-ezetimibe;

[0047] Figure 8 This is the trend graph of the concentration and peak area of ​​S,S,R-ezetimibe;

[0048] Figure 9 This is the concentration and peak area trend graph of R,S,R-ezetimibe;

[0049] Figure 10 Chromatograms showing the detection of rosuvastatin impurity G, rosuvastatin, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, R,S,R-ezetimibe, and ezetimibe using the method of Example 3 of the present application;

[0050] Figure 11 Chromatograms showing the detection of rosuvastatin impurity G, rosuvastatin, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, R,S,R-ezetimibe, and ezetimibe using the method of Example 4 of the present application;

[0051] Figure 12 Chromatograms of rosuvastatin impurity G, rosuvastatin, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, R,S,R-ezetimibe, and ezetimibe detected by the method of Comparative Example 2 of the present application;

[0052] Figure 13 The chromatograms of rosuvastatin impurity G, rosuvastatin, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, R,S,R-ezetimibe, and ezetimibe were detected using the method of Comparative Example 3 of the present application. DETAILED DESCRIPTION

[0053] Below in conjunction with embodiment and example, the application is described in further detail.Should be understood that these embodiment and example are only used to illustrate the application and are not used to limit the scope of the application, and the purpose of providing these embodiment and example is to make the understanding of the disclosure of the application more thorough and comprehensive.It should also be understood that the application can be implemented in many different forms, is not limited to the embodiment and example described herein, and those skilled in the art can make various changes or modifications without violating the connotation of the application, and the equivalent form obtained also falls within the protection scope of the application.In addition, in the description hereinafter, a large amount of specific details are given in order to provide a more complete understanding of the application, and it should be understood that the application can be implemented without one or more of these details.

[0054] 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.

[0055] the term

[0056] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0057] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the arbitrary and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical and" and also undoubtedly includes technical solutions connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution of all being connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the four-item combination of A, B, C, and D (that is, the technical solution of all being connected by "logical AND").

[0058] In this application, "plurality", "multiple", "multiple times", "multiples", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0059] As used herein, "combination thereof", "any combination thereof", "any combination thereof" and the like include all suitable combinations of any two or more of the listed items.

[0060] In this application, the terms "optionally," "optional," and "optional" mean optional or dispensable, i.e., they refer to either option being selected from two parallel options: "with" or "without." If a technical solution contains multiple "optional" clauses, each "optional" clause is independent unless otherwise specified and there are no contradictions or constraints.

[0061] In this application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0062] In this application, when referring to a numerical interval (i.e., a numerical range), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, as well as each numerical value between the two numerical endpoints. Unless otherwise specified, when a numerical interval refers only to an integer within the numerical interval, it includes the two endpoint integers of the numerical range, as well as each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as t is an integer selected from 1-10, indicating that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0063] Rosuvastatin calcium (Rosuvastatin Calciμm), chemically known as (3R,5S,6E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-(N-methylmethanesulfonamido)-5-pyrimidinyl]-3,5-dihydroxy-6-heptenoic acid calcium salt (2:1), is a selective HMG-CoA reductase inhibitor. HMG-CoA reductase inhibitors are the rate-limiting enzyme that converts 3-hydroxy-3-methylglutaryl-CoA to mevalonate, a precursor of cholesterol. Rosuvastatin's primary site of action is the liver, the target organ for cholesterol lowering. Rosuvastatin increases the number of surface receptors on hepatic LDL cells, promoting LDL absorption and catabolism, and inhibiting hepatic synthesis of very low density lipoprotein (VLDL), thereby reducing the total number of VLDL and LDL particles.

[0064] Ezetimibe, whose chemical name is 1-(4-fluorophenyl)-3(R)-[3-(4-fluorophenyl)-3(S)-hydroxypropyl]-4(S)-(4-hydroxyphenyl)-2-azetidinone, selectively inhibits the intestinal absorption of sterols (cholesterol and plant sterols) in food and cholesterol in bile by specifically binding to the Niemann-PickC1-Like1 protein (NPC1L1, which plays a key role in the intestinal absorption of sterols) on the brush border membrane of the villi epithelial cells of the small intestine, especially the jejunum.

[0065] On the one hand, the present application provides a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets, comprising the following steps:

[0066] High performance liquid chromatography was used to prepare the test solution, reference solution, sensitivity solution and system suitability solution in phosphate buffer-acetonitrile as solvent.

[0067] The chromatographic conditions include: the filling material of the chromatographic column includes silica gel with cellulose-tris (4-methylbenzoate) coated on the surface; and the detection wavelength is 240nm-244nm.

[0068] Mobile phase A included 0.08%-0.12% trifluoroacetic acid solution, and mobile phase B included acetonitrile, and gradient elution was performed.

[0069] Separately draw the test solution, control solution, system suitability solution and sensitivity solution, inject them into the high performance liquid chromatograph, analyze them, and record the chromatogram.

[0070] In one embodiment, the volume ratio of phosphate buffer to acetonitrile in the phosphate buffer-acetonitrile solvent is (58-62):(38-42).

[0071] In one embodiment, the pH value of the phosphate buffer-acetonitrile solvent is 5.0-6.5.

[0072] In one embodiment, the chromatographic column is DAICEL, OJ-3R.

[0073] In one embodiment, the size of the chromatographic column is 4.6 mm×250 mm.

[0074] In one embodiment, the column temperature is 20°C-30°C.

[0075] In one embodiment, the flow rate is 0.4 mL / min-0.6 mL / min.

[0076] In one embodiment, the injection volume is 10 μL-50 μL.

[0077] Specifically, the following steps are included:

[0078] (1) Prepare the test solution, reference solution, sensitivity solution, and system suitability solution using phosphate buffer-acetonitrile (60:40) as solvent;

[0079] (2) Chromatographic conditions: The chromatographic column was filled with silica gel coated with cellulose tris(4-methylbenzoate), with a size of 4.6 mm × 250 mm and a filler particle size of 3 μm; the column temperature was 20°C–30°C; the detection wavelength was 242 nm; the mobile phase A was 0.08%–0.12% trifluoroacetic acid solution, and the mobile phase B was acetonitrile, with a flow rate of 0.4 mL / min–0.6 mL / min for gradient elution;

[0080] (3) Accurately pipette 10-50 μl of the test solution, control solution, system suitability solution, and sensitivity solution respectively, inject them into the high performance liquid chromatograph, analyze, and record the chromatogram.

[0081] In the above step (1), the solvent is prepared by adding 600 mL of 10 m mol / L NaHPO4 solution to 400 mL of acetonitrile, mixing well, and adjusting the pH to 5.0-6.5 with 10% phosphoric acid solution or 1 mol / L sodium hydroxide.

[0082] The test solution is prepared as follows: take an appropriate amount of fine powder of the product, accurately weigh it, add a solvent to dissolve it by ultrasonication and dilute it into a solution containing approximately 0.2 mg to 2 mg of rosuvastatin and ezetimibe per 1 mL.

[0083] The control solution was prepared by accurately measuring an appropriate amount of the test solution and diluting it with solvent to form a solution containing approximately 0.4 μg each of rosuvastatin and ezetimibe per 1 mL.

[0084] The sensitivity solution was prepared as follows: an appropriate amount of the control solution was accurately taken and diluted with solvent to form a mixed solution containing approximately 0.1 μg of rosuvastatin and 0.1 μg of ezetimibe per 1 mL.

[0085] The system suitability solution is prepared as follows: appropriate amounts of rosuvastatin calcium reference substance, rosuvastatin impurity G reference substance, ezetimibe reference substance, and S,S,R-ezetimibe reference substance are dissolved in solvent and diluted to prepare a mixed solution containing approximately 0.2 mg of rosuvastatin, 0.4 μg of rosuvastatin impurity G, 0.2 mg of ezetimibe, and 0.4 μg of S,S,R-ezetimibe per 1 mL.

[0086] In one embodiment, the solvent has a pH of 5.7.

[0087] The specific preparation process is as follows: Solvent: Weigh 0.936g of sodium dihydrogen phosphate, add 600mL of water to dissolve, then add 400mL of acetonitrile, mix well, and adjust the pH to 5.7 with 10% phosphoric acid solution or 1mol / L sodium hydroxide.

[0088] Test solution: Take about 300 mg of the fine powder of this product, accurately weigh it, put it into a 50 mL volumetric flask, add 25 mL of solvent, sonicate for 10 minutes to dissolve it, let it cool, add solvent to dilute to the scale, shake well, centrifuge at 10,000 rpm for 5 minutes, and take the supernatant.

[0089] Control solution: Accurately measure 100 μl of the test solution into a 50 mL volumetric flask, then add solvent to dilute to the mark and shake well.

[0090] Sensitivity solution: Accurately measure 5 mL of the control solution into a 20 mL volumetric flask, add solvent to dilute to the scale, and shake well.

[0091] System suitability solution: Weigh approximately 2 mg each of rosuvastatin impurity G reference substance and S,S,R-ezetimibe reference substance into the same 100 mL volumetric flask, dissolve with solvent and dilute to the scale, shake well; measure 1 mL from each solution into a 50 mL volumetric flask, then weigh approximately 10 mg each of rosuvastatin calcium reference substance and ezetimibe reference substance into the 50 mL volumetric flask, dissolve with solvent and dilute to the scale, shake well.

[0092] In one embodiment, the gradient elution procedure comprises:

[0093] At 0 min, the volume percentage of mobile phase A was 27, and the volume percentage of mobile phase B was 73; at 50 min, the volume percentage of mobile phase A was 27, and the volume percentage of mobile phase B was 73; at 60 min, the volume percentage of mobile phase A was 45, and the volume percentage of mobile phase B was 55; at 90 min, the volume percentage of mobile phase A was 45, and the volume percentage of mobile phase B was 55; at 90.1 min, the volume percentage of mobile phase A was 27, and the volume percentage of mobile phase B was 73; at 100 min, the volume percentage of mobile phase A was 27, and the volume percentage of mobile phase B was 73.

[0094] In one embodiment, the method for preparing the system suitability solution comprises:

[0095] Take rosuvastatin calcium reference substance, rosuvastatin impurity G reference substance, ezetimibe reference substance and S,S,R-ezetimibe reference substance, mix and dissolve.

[0096] In one embodiment, the resolution of the rosuvastatin peak and the rosuvastatin impurity G peak is not less than 1.5.

[0097] The resolution of the ezetimibe peak and the S,S,R-ezetimibe peak must be no less than 1.5.

[0098] In one embodiment, the retention time of rosuvastatin impurity G is 40.7 min; the retention time of R,R,S-ezetimibe is 78.0 min; the retention time of R,R,R-ezetimibe is 72.7 min; the retention time of S,S,S-ezetimibe is 70.8 min; the retention time of S,S,R-ezetimibe is 79.1 min; and the retention time of R,S,R-ezetimibe is 79.1 min.

[0099] The sensitivity requirement is: in the chromatogram of the sensitivity solution, the signal-to-noise ratio of the rosuvastatin peak and the ezetimibe peak must not be less than 10.

[0100] The calculation method is: self-control method with correction factor added.

[0101] In one embodiment, the chiral isomer comprises at least one of the following isomers:

[0102] Rosuvastatin impurity G: (3S,5R,6E)-7-[4-(4-fluorophenyl)-2-(N-methylmethanesulfonamido)-6-(isopropyl)pyrimidin-5-yl]-3,5-dihydroxyheptenoic acid.

[0103] R,R,S-Ezetimibe: (3R,4S)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0104] R,R,R-Ezetimibe: (3R,4R)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0105] S,S,S-Ezetimibe: (3S,4S)-1-(4-fluorophenyl)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0106] S,S,R-Ezetimibe: (3S,4R)-1-(4-fluorophenyl)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0107] R,S,R-Ezetimibe: (3S,4R)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

[0108] The relevant limit requirements for the above substances are as follows, as shown in Table A:

[0109] Table A

[0110] Impurity name Retention time approx. Correction Factor limit Rosuvastatin Impurity G 40.7 minutes 1.2 0.2％ R,R,S-ezetimibe 78.0min 1.0 0.4％ R,R,R-ezetimibe 72.7 minutes 1.0 0.2％ S,S,S-ezetimibe 70.8 minutes 1.0 0.2％ S,S,R-ezetimibe+R,S,R-ezetimibe 79.1min 1.0 0.2％

[0111] The embodiments of the present application will be described in detail below with reference to the examples. It should be understood that these examples are intended to illustrate the present application only and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manuals or conventional conditions in this area, or according to the conditions recommended by the manufacturer, or with reference to experimental methods known in the art.

[0112] In the following specific examples, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument testing accuracy or operational accuracy are allowed.

[0113] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the above-mentioned processes does not mean the order of execution. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0114] Example 1

[0115] This embodiment provides a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets, as follows:

[0116] Chromatographic column: DAICEL, OJ-3R, 3μm, 4.6*250mm;

[0117] Mobile phase A: 0.1% trifluoroacetic acid solution;

[0118] Mobile phase B: acetonitrile;

[0119] Column temperature: 25°C;

[0120] Flow rate: 0.5 mL / min;

[0121] Detection wavelength: 242nm;

[0122] Injection volume: 50 μL

[0123] The elution gradient is shown in Table 1 below:

[0124] Table 1

[0125] Time min Mobile phase A% Mobile phase B% 0 27 73 50 27 73 60 45 55 90 45 55 90.1 27 73 100 27 73

[0126] This example is a representative embodiment of the present invention. The chromatographic conditions in this example were validated with reference to the 2020 edition of the Chinese Pharmacopoeia 9101 Analytical Method Validation Guidelines. The content includes:

[0127] 1. Specificity test

[0128] Solution preparation:

[0129] Rosuvastatin standard solution: weigh 10.09 mg of rosuvastatin calcium reference substance into a 50 mL volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and obtain.

[0130] Ezetimibe localizing solution: Weigh 10.22 mg of ezetimibe reference substance into a 50 mL volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and obtain.

[0131] Stock solutions of each impurity: Weigh 1.27 mg, 1.54 mg, 1.32 mg, 1.14 mg, 1.26 mg, and 1.35 mg of rosuvastatin impurity G reference substance, R,R,S-ezetimibe reference substance, R,R,R-ezetimibe reference substance, S,S,S-ezetimibe reference substance, and S,S,R-ezetimibe reference substance respectively into different 20 mL volumetric flasks, add acetonitrile to dissolve and dilute to the scale, shake well, and obtain.

[0132] Impurity localization solution: Measure 1 mL of the stock solution of each component into different 50 mL volumetric flasks, add solvent to dilute to the scale, and shake well.

[0133] Mixed solution: Weigh 10.15 mg of rosuvastatin calcium reference substance and 10.11 mg of ezetimibe reference substance into a 50 mL bottle, add 1 mL of each impurity stock solution, dissolve with solvent and dilute to the scale, shake well, and obtain.

[0134] Measurement and results:

[0135] Accurately measure 50 μL of each of the above-mentioned positioning solutions and inject them into the high performance liquid chromatograph. The retention times of rosuvastatin impurity G, rosuvastatin, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, R,S,R-ezetimibe and ezetimibe are 40.707min, 44.611min, 70.821min, 72.728min, 77.970min, 79.052min, 79.058min and 80.382min, respectively. Take 50 μL of the mixed solution and inject it into the high performance liquid chromatograph, record the chromatogram, and the results are shown in FIG. Figure 1 The specific separation conditions are shown in Table 2 below:

[0136] Table 2

[0137] In mixed solution Retention time min Separation from the front peak Rosuvastatin Impurity G 40.697 No front peak Rosuvastatin 44.616 2.0 S,S,S-ezetimibe 70.823 20.9 R,R,R-ezetimibe 72.738 5.1 R,R,S-ezetimibe 77.976 10.9 S,S,R-ezetimibe+R,S,R-ezetimibe 79.052 1.9 Ezetimibe 80.384 2.1

[0138] It can be seen that the chromatographic conditions under this example can effectively distinguish the chromatographic peaks of rosuvastatin impurity G, rosuvastatin, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, R,S,R-ezetimibe, and ezetimibe, and the separation degree between each component is not less than 1.5, and the peak shape is good, which is conducive to identification and positioning, indicating that this method has good specificity.

[0139] 2. Linearity Test and Correction Factor

[0140] Solution preparation:

[0141] Linearity stock solution A: Accurately weigh 5.452 mg, 8.166 mg, 6.906 mg, 4.170 mg, 3.992 mg, 7.968 mg, and 4.974 mg of rosuvastatin, ezetimibe, rosuvastatin impurity G, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, and S,S,R-ezetimibe reference respectively, into a 200 mL volumetric flask, dissolve in solvent, dilute to the mark, and shake well.

[0142] 10%, 50%, 100%, 150%, 200% linear solution A: Accurately measure 0.2, 1, 2, 3, and 4 mL of linear stock solution A into a 100 mL volumetric flask, add solvent to dilute to the scale, and shake well.

[0143] Linear stock solution B: Accurately weigh 4.396 mg of R,S,R-ezetimibe reference substance into a 200 mL volumetric flask, dissolve in solvent and dilute to the mark, then shake well.

[0144] 10%, 50%, 100%, 150%, 200% linear solution B: Accurately measure 0.2, 1, 2, 3, and 4 mL of linear stock solution B into a 100 mL volumetric flask, add solvent to dilute to the scale, and shake well.

[0145] Measurement and results:

[0146] Accurately measure 50 μL of each of the above linear solutions and inject them into the high performance liquid chromatograph to record the chromatogram. The results are shown in Tables 3 to 10 below. Figure 2-Figure 9 As shown:

[0147] Table 3

[0148]

[0149] Table 4

[0150]

[0151] Table 5

[0152]

[0153] Table 6

[0154]

[0155]

[0156] Table 7

[0157]

[0158] Table 8

[0159]

[0160] Table 9

[0161]

[0162] Table 10

[0163]

[0164] It can be seen that rosuvastatin, rosuvastatin impurity G, ezetimibe, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe and R,S,R-ezetimibe have good linearity in the ranges of 0.052μg / mL-1.030μg / mL, 0.070μg / mL-1.381μg / mL, 0.082μg / mL-1.630μg / mL, 0.042μg / mL-0.820μg / mL, 0.040μg / mL-0.791μg / mL, 0.078μg / mL-1.565μg / mL, 0.050μg / mL-0.994μg / mL and 0.044μg / mL-0.876μg / mL, respectively.

[0165] Based on the slopes of the linear regression equations of rosuvastatin, rosuvastatin impurity G, ezetimibe, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, and R,S,R-ezetimibe, the correction factors of rosuvastatin impurity G, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, and R,S,R-ezetimibe were calculated, which are 1.2, 0.97, 0.96, 0.99, 1.0, and 1.0, respectively, all within the range of 0.8-1.2. Impurity peak areas do not need to be corrected during sample testing, that is, the peak area correction factor can be calculated as 1.0.

[0166] 3. Sensitivity test

[0167] Solution preparation: Take 10% linear solutions A and B in the linear test and dilute them step by step to obtain mixed reference solutions of different concentrations.

[0168] Determination and Results: Accurately measure 50 μL of the mixed reference solution of different concentrations and inject it into the high performance liquid chromatograph. Record the chromatogram and calculate the signal-to-noise ratio of each component peak. The limit of quantification is set as a signal-to-noise ratio of not less than 10, and the limit of detection is set as not less than 3. The results are shown in Table 11 below:

[0169] Table 11

[0170]

[0171] From the above results, it can be seen that this method has high sensitivity.

[0172] 4. Repeatability test

[0173] Solution preparation: Test solution: Take about 300 mg of the product fine powder, accurately weigh it, put it into a 50 mL volumetric flask, add 25 mL of solvent, sonicate for 10 minutes to dissolve, let it cool, add solvent to dilute to the scale, shake well, centrifuge at 10,000 rpm for 5 minutes, and take the supernatant; prepare 6 parallel portions.

[0174] Control solution: Accurately measure 100 μL of each test solution into a 50 mL volumetric flask, then add solvent to dilute to the scale and shake well.

[0175] Determination and results: Accurately measure 50 μL of the above test solution and control solution and inject them into the high performance liquid chromatograph, record the chromatogram, and calculate the impurity content. The results are shown in Table 12 below:

[0176] Table 12

[0177]

[0178] It can be seen that the RSDs of the individual impurity contents detected in the six test solutions were all lower than 10.0%, indicating that this method has good repeatability.

[0179] 5. Intermediate precision test

[0180] Two repeat tests were conducted by different testers at different times. The results are shown in Table 13 below:

[0181] Table 13

[0182]

[0183] It can be seen that in the two repeatability tests, in each of the 6 test solutions, the RSD of the detected single impurity content was lower than 10.0%, and the relative deviation of the detected single impurity content mean was lower than 10.0%; the RSD of the detected single impurity content in a total of 12 test solutions in the two repeatability tests was lower than 10.0%, and the intermediate precision of this method was good.

[0184] 6. Accuracy test

[0185] Solution preparation: Linear stock solution A: Accurately weigh 5.452 mg, 8.166 mg, 6.906 mg, 4.170 mg, 3.992 mg, 7.968 mg, and 4.974 mg of rosuvastatin, ezetimibe, rosuvastatin impurity G, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, and S,S,R-ezetimibe reference respectively, place them in the same 200 mL volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0186] 50% / 100% / 150% Recovery Solution A: Accurately weigh 300 mg of fine powder into a 50 mL volumetric flask. Accurately add 0.1, 0.5, 1, and 1.5 mL of linear stock solution A. Add 25 mL of solvent and sonicate for 10 minutes to dissolve. Cool, dilute to volume with solvent, shake well, centrifuge at 10,000 rpm for 5 minutes, and collect the supernatant. Prepare three replicates.

[0187] Linear stock solution B: Accurately weigh 4.396 mg of R,S,R-ezetimibe reference substance into a 200 mL volumetric flask, dissolve in solvent and dilute to the mark, then shake well.

[0188] 50% / 100% / 150% Recovery Solution B: Accurately weigh 300 mg of fine powder into a 50 mL volumetric flask. Accurately add 0.1 / 0.5 / 1 / 1.5 mL of linear stock solution B. Add 25 mL of solvent and sonicate for 10 minutes to dissolve. Cool, dilute to volume with solvent, shake well, centrifuge at 10,000 rpm for 5 minutes, and collect the supernatant. Prepare three replicates.

[0189] Control solution: Accurately measure 100 μL of each recovery solution into a 50 mL volumetric flask, then add solvent to dilute to the scale and shake well.

[0190] Determination and results: Accurately measure 50 μL of the above recovery solution and control solution and inject them into the high performance liquid chromatograph, record the chromatogram, and calculate the impurity content. The results are shown in Table 14 below:

[0191] Table 14

[0192]

[0193]

[0194]

[0195] It can be seen that when the added amounts of rosuvastatin impurities G, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, and R,S,R-ezetimibe were 10%, 50%, 100%, and 150%, respectively, the recoveries were all within the range of 90%-110%, and the RSDs of the recoveries at each concentration were all below 10%, indicating that the method has good accuracy.

[0196] 7. Solution stability test

[0197] Solution preparation: Test solution: Take about 300 mg of the fine powder of this product, accurately weigh it, put it into a 50 mL volumetric flask, add 25 mL of solvent, sonicate for 10 minutes to dissolve it, let it cool, add solvent to dilute to the scale, shake well, centrifuge at 10,000 rpm for 5 minutes, and take the supernatant.

[0198] Determination and results: After being placed at room temperature for different periods of time, 10 μL of the test solution was accurately measured and injected into a high performance liquid chromatograph. The chromatogram was recorded and the impurity content was calculated by normalization. The results are shown in Table 15 below:

[0199] Table 15

[0200]

[0201] It can be seen that when the test solution is placed at room temperature for 1, 2, 3, 5, and 7 days, the maximum relative deviation of the single impurity content compared with the time 0 is less than 10%, and the test solution is stable after being placed at room temperature for at least 7 days.

[0202] 8. Durability test

[0203] The chromatographic conditions provided in this example were used as standard conditions. On this basis, other parameters were kept unchanged, and the column temperature ±5°C, flow rate ±0.1 mL / min, trifluoroacetic acid concentration ±10%, initial acetonitrile ratio ±2%, and detection wavelength ±2 nm were used as durability conditions.

[0204] The same batch of rosuvastatin ezetimibe tablets was tested under standard conditions and various durability conditions. The test solution and control solution were prepared according to "IV. Repeatability Test". The results are shown in Table 16 below:

[0205] Table 16

[0206]

[0207] It can be seen that the maximum relative deviation of the detected single impurity content compared with the standard conditions for the same batch of rosuvastatin ezetimibe tablets under various durability conditions was less than 10%, and this method has good durability.

[0208] During the verification process of this embodiment:

[0209] The source of the rosuvastatin ezetimibe tablets is Yaoda Pharmaceutical Co., Ltd., specifications: rosuvastatin 10 mg & ezetimibe 10 mg, batch number: 250529.

[0210] The source of the rosuvastatin calcium reference substance: China Food and Drug Inspection Institute; content: 98.4% (calculated as rosuvastatin calcium); batch number: 101028-202405.

[0211] The source of the rosuvastatin impurity G reference substance: China Food and Drug Inspection Institute; content: not marked, calculated as 100%; batch number: 101454-202402.

[0212] The source of the ezetimibe reference substance: USP reference substance; content: 99.8%; batch number: R075K0.

[0213] The S,S,S-ezetimibe reference substance was obtained from Guangzhou Jiatu Technology Co., Ltd.; the content was 98.3%; and the batch number was 0331-RG-0044.

[0214] The R,R,R-ezetimibe reference substance is from Guangzhou Jiatu Technology Co., Ltd.; content: 99.1%; batch number: 0122-RG-0012.

[0215] The R,R,S-ezetimibe reference substance was sourced from Guangzhou Jiatu Technology Co., Ltd.; content: 98.2%; batch number: 1119-RF-0012.

[0216] The S,S,R-ezetimibe reference substance is from Guangzhou Jiatu Technology Co., Ltd.; content: 99.9%; batch number: 0328-RG-0049.

[0217] The R,S,R-ezetimibe reference substance is from Guangzhou Jiatu Technology Co., Ltd.; content: 99.6%; batch number: 1202-RF-0071.

[0218] Example 2

[0219] This example provides a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets, which is as follows: injection volume: 10 μL / 25 μL / 50 μL, and other chromatographic conditions are the same as in Example 1.

[0220] The preparation of high / medium / low concentration test sample solution is as follows: take about 1.5g / 600mg / 300mg of the fine powder of the product, accurately weigh, put it into a 50mL volumetric flask, add 25ml of solvent, sonicate for 10min to dissolve, let cool, add solvent to dilute to the scale, shake well, centrifuge at 10000rpm for 5min, and take the supernatant.

[0221] The preparation of the control solution was the same as that in Example 1.

[0222] Determination: 10 μL, 25 μL, and 50 μL of high, medium, and low concentration test solutions and corresponding control solutions were injected into the high performance liquid chromatograph, and the chromatograms were recorded. The impurity content in the test solution was calculated. The results are shown in Table 17 below:

[0223] Table 17

[0224]

[0225] It can be seen that the test results are basically consistent when different concentrations of test solution are injected in different volumes.

[0226] Example 3

[0227] This embodiment provides a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets, which is as follows: the elution gradient is shown in Table 18 below:

[0228] Table 18

[0229] Time min Mobile phase A% Mobile phase B% 0 20 73 50 20 73 60 45 55 90 45 55 90.1 20 73 100 20 73

[0230] The remaining chromatographic conditions were the same as those in Example 1, and the preparation of the localization solutions for each component and the preparation of the mixed solution were the same as those in Example 1.

[0231] Determination: Take 50 μL of each positioning solution and inject it into the high performance liquid chromatograph. The retention times of rosuvastatin, ezetimibe, rosuvastatin impurity G, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe and R,S,R-ezetimibe are 55.885min, 79.439min, 50.807min, 71.273min, 73.031min, 77.354min, 78.232min and 78.298min, respectively. Take 10 μL of the mixed solution and inject it into the high performance liquid chromatograph. Record the chromatogram results. Figure 10 .

[0232] Example 4

[0233] This example provides a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets, which is as follows. The elution gradient is shown in Table 19 below:

[0234] Table 19

[0235] Time min Mobile phase A% Mobile phase B% 0 35 73 50 35 73 60 45 55 90 45 55 90.1 35 73 100 35 73

[0236] The remaining chromatographic conditions were the same as those in Example 1, and the preparation of the localization solutions for each component and the preparation of the mixed solution were the same as those in Example 1.

[0237] Determination: Take 50 μL of each positioning solution and inject it into the high performance liquid chromatograph. The retention times of rosuvastatin, ezetimibe, rosuvastatin impurity G, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe and R,S,R-ezetimibe are 28.522 min, 76.486 min, 26.422 min, 67.873 min, 69.531 min, 74.154 min, 75.199 min and 75.232 min, respectively. Take 10 μL of the mixed solution and inject it into the high performance liquid chromatograph. Record the chromatogram. The results are shown in Figure 11 .

[0238] Under the chromatographic conditions of Example 1, Example 3, and Example 4, the chromatographic peaks of rosuvastatin, ezetimibe, rosuvastatin impurity G, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, and R,S,R-ezetimibe can be effectively distinguished. The chromatographic peaks of each component and the adjacent chromatographic peaks can be well separated, and the peak shape is good, which is conducive to identification and positioning, indicating that the chromatographic conditions of Example 1, Example 3, and Example 5 are all highly specific.

[0239] Comparative Example 1

[0240] This comparative example provides the European Pharmacopoeia rosuvastatin isomer method, which is as follows:

[0241] Mobile phase: acetonitrile-0.1% trifluoroacetic acid solution (75:25);

[0242] Isocratic elution mode;

[0243] The remaining chromatographic conditions were the same as in Example 1.

[0244] The preparation of the positioning solutions of each component and the preparation of the mixed solution are the same as those in Example 1.

[0245] Determination: 50 μL of each positioning solution was injected into the high performance liquid chromatograph, and it was found that ezetimibe and its isomers could not be eluted under this condition.

[0246] It can be seen that under the chromatographic conditions of this comparative example, due to the large difference in the polarity of rosuvastatin and ezetimibe molecules, isocratic elution is difficult to complete the elution of all components, and gradient elution is necessary.

[0247] Comparative Example 2

[0248] This comparative example provides the USP ezetimibe isomer method, as follows:

[0249] Chromatographic column: DAICEL, OH-RH, 3μm, 4.6*150mm; 2 in series;

[0250] Mobile phase: acetonitrile-water (45:55); isocratic elution;

[0251] Column temperature: 5°C;

[0252] Flow rate: 0.35 mL / min;

[0253] Detection wavelength: 248nm;

[0254] The remaining chromatographic conditions were the same as in Example 1;

[0255] The preparation of the positioning solutions of each component and the preparation of the mixed solution are the same as those in Example 1;

[0256] Determination: 50 μL of each positioning solution was injected into the HPLC instrument, and it was found that the rosuvastatin peak and the rosuvastatin impurity G peak were completely inseparable. Figure 12 .

[0257] It can be seen that under the chromatographic conditions of this comparative example, it is impossible to simultaneously detect the isomers of rosuvastatin calcium.

[0258] Comparative Example 3

[0259] This comparative example provides a method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets, wherein the solvent is changed to acetonitrile. The chromatographic conditions are the same as those in Example 1.

[0260] The solvent was changed to acetonitrile;

[0261] The preparation of the positioning solutions of each component and the preparation of the mixed solution are the same as those in Example 1;

[0262] Determination: 50 μL of each positioning solution was injected into the HPLC instrument. It was found that there was a solvent effect. The peaks of rosuvastatin and ezetimibe were deformed, which seriously interfered with the normal detection. Figure 13 .

[0263] It can be seen that the solvent in this comparative example is not applicable.

[0264] In summary:

[0265] 1. Existing methods cannot meet the requirements for detecting the chiral isomer content in rosuvastatin ezetimibe tablets:

[0266] (1) The current pharmaceutical preparation standards do not include the isomer method of rosuvastatin ezetimibe tablets, and no foreign pharmacopoeias include rosuvastatin ezetimibe tablets.

[0267] (2) The European Pharmacopoeia and the United States Pharmacopoeia have included isomer methods for rosuvastatin calcium and ezetimibe, respectively. However, these methods are only applicable to single-ingredient APIs and are not applicable to this compound preparation.

[0268] 2. The analytical method of the present application can simultaneously detect rosuvastatin impurities G, S,S,S-ezetimibe, R,R,R-ezetimibe, R,R,S-ezetimibe, S,S,R-ezetimibe, and R,S,R-ezetimibe, thereby effectively separating the relevant impurities from rosuvastatin and ezetimibe, and can further realize qualitative and quantitative detection, which is beneficial to the quality control of rosuvastatin ezetimibe tablets.

[0269] The embodiments described above only express several implementation methods of the present application, which are convenient for understanding the technical solutions of the present application in a specific and detailed manner, but they cannot be understood as limiting the scope of protection of the patent application. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present application, several variations and improvements can be made, which all fall within the scope of protection of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the scope of protection of the present application. It should also be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided in the present application are all within the scope of protection of the claims attached to the present application. Therefore, the scope of protection of the patent application of this application shall be based on the content of the attached claims, and the description can be used to interpret the content of the claims.

Claims

1. A method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets, characterized in that: The following steps are involved: High performance liquid chromatography was used to prepare the test solution, reference solution, sensitivity solution and system suitability solution using phosphate buffer-acetonitrile as solvent. The chromatographic conditions include: the filler of the chromatographic column includes silica gel coated with cellulose-tris(4-methylbenzoate); the detection wavelength is 240nm-244nm; Mobile phase A included 0.08%-0.12% trifluoroacetic acid solution, and mobile phase B included acetonitrile, and gradient elution was performed; The test solution, the control solution, the system suitability solution and the sensitivity solution were respectively drawn into a high performance liquid chromatograph for analysis and the chromatogram was recorded.

2. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to claim 1, characterized in that: The volume ratio of phosphate buffer to acetonitrile in the phosphate buffer-acetonitrile solvent is (58-62):(38-42).

3. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to claim 1, characterized in that: The pH value of the phosphate buffer-acetonitrile solvent is 5.0-6.

5.

4. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to claim 1, characterized in that: The chromatographic column meets one or more of the following conditions: (1) The chromatographic column is DAICEL, OJ-3R; (2) The specifications of the chromatographic column are 4.6 mm × 250 mm; (3) Column temperature is 20°C-30°C; (4) The flow rate is 0.4 mL / min-0.6 mL / min.

5. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to claim 1, characterized in that: The injection volume is 10μL-50μL.

6. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to claim 1, characterized in that: The gradient elution procedure includes: 0 min, the volume percentage of the mobile phase A is 27, and the volume percentage of the mobile phase B is 73; 50 min, the volume percentage of the mobile phase A is 27, and the volume percentage of the mobile phase B is 73; 60 min, the volume percentage of mobile phase A is 45, and the volume percentage of mobile phase B is 55; 90 min, the volume percentage of the mobile phase A is 45, and the volume percentage of the mobile phase B is 55; 90.1 min, the volume percentage of the mobile phase A is 27, and the volume percentage of the mobile phase B is 73; 100 min, the volume percentage of the mobile phase A is 27, and the volume percentage of the mobile phase B is 73.

7. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to any one of claims 1 to 6, characterized in that: The method for preparing the system suitability solution includes: Take rosuvastatin calcium reference substance, rosuvastatin impurity G reference substance, ezetimibe reference substance and S,S,R-ezetimibe reference substance, mix and dissolve.

8. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to any one of claims 1 to 6, characterized in that: The chiral isomers include at least one of the following isomers: Rosuvastatin impurity G: (3S,5R,6E)-7-[4-(4-fluorophenyl)-2-(N-methylmethanesulfonamido)-6-(isopropyl)pyrimidin-5-yl]-3,5-dihydroxyheptenoic acid; R,R,S-Ezetimibe: (3R,4S)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone; R,R,R-Ezetimibe: (3R,4R)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone; S,S,S-Ezetimibe: (3S,4S)-1-(4-fluorophenyl)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone; S,S,R-ezetimibe: (3S,4R)-1-(4-fluorophenyl)-3-[(S)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone; R,S,R-Ezetimibe: (3S,4R)-1-(4-fluorophenyl)-3-[(R)-3-(4-fluorophenyl)-3-hydroxypropyl]-4-(4-hydroxyphenyl)-2-azetidinone.

9. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to claim 8, characterized in that: The resolution of the rosuvastatin peak and the rosuvastatin impurity G peak must be no less than 1.5; and / or, The resolution of the ezetimibe peak and the S,S,R-ezetimibe peak must be no less than 1.

5.

10. The method for determining chiral isomers in rosuvastatin calcium ezetimibe tablets according to claim 8, characterized in that: The retention time of rosuvastatin impurity G is 40.7 min; the retention time of R,R,S-ezetimibe is 78.0 min; the retention time of R,R,R-ezetimibe is 72.7 min; the retention time of S,S,S-ezetimibe is 70.8 min; the retention time of S,S,R-ezetimibe is 79.1 min; the retention time of R,S,R-ezetimibe is 79.1 min.

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