Ezetimibe rosuvastatin calcium tablet isomer determination method

Abstract

The invention relates to the technical field of medicine analysis, in particular to a method for determining an ezetimibe rosuvastatin calcium tablet isomer, and rosuvastatin calcium isomer impurities in an ezetimibe rosuvastatin calcium tablet are determined by adopting a high performance liquid chromatography. A chromatographic column is a polysaccharide derivative coating type chiral chromatographic column; a mobile phase A is a 0.08%-0.12% trifluoroacetic acid aqueous solution; the mobile phase B is acetonitrile; carrying out gradient elution; the flow velocity is 0.4-0.6 ml per minute; the column temperature is 23-27 DEG C; the detection wavelength is 240 to 244nm; the sample injection volume is 5-10 microliters. According to the method, separation of rosuvastatin calcium and isomer impurities thereof can be realized, interference of ezetimibe and isomer impurities thereof in a two-component preparation is eliminated, and the isomer impurities of rosuvastatin calcium are accurately detected.

Description

Technical Field

[0001] This application relates to the technical field of pharmaceutical analysis, and particularly relates to a method for determining the isomers of ezetimibe rosuvastatin calcium tablets. Background Art

[0002] Ezetimibe rosuvastatin calcium tablets (English name: Ezetimibe Rosuvastatin Calcium Film-Coated Tablets) are a compound preparation of ezetimibe and rosuvastatin calcium. Ezetimibe is a potent lipid-lowering drug. After oral administration, it is rapidly absorbed and widely conjugated into a pharmacologically active phenolic glucuronide (ezetimibe-glucuronide), and its structure is as follows:

[0003]

[0004] Rosuvastatin calcium is a selective HMG-CoA reductase inhibitor. Ezetimibe rosuvastatin calcium tablets are a lipid-lowering drug with dual effects and can be used to treat chronic diseases caused by excessive blood lipids, such as fat, cholesterol, and triglycerides. Currently, it is on the market in countries such as Japan (licensee: Merck Sharp & Dohme), the Czech Republic (licensee: Sanofi), and Poland, and its structure is as follows:

[0005]

[0006] In order to control the quality of drugs, it is necessary to control the contents of rosuvastatin calcium and ezetimibe isomers. There are many isomeric impurities related to rosuvastatin calcium and ezetimibe. Existing reports only determine the isomers of a single component, and it is impossible to exclude the interference of isomeric impurities of the two components with each other, and the chromatographic conditions are complex. There is no literature report on the detection method of isomers in the ezetimibe rosuvastatin calcium two-component preparation.

[0007] Table 1 Statistical Table of Ezetimibe Isomeric Impurities

[0008]

[0009] Table 2 Statistical Table of Rosuvastatin Calcium Isomeric Impurities

[0010] Summary of the Invention

[0011] The object of the present invention is to overcome the deficiencies of the above-mentioned prior art and establish a method for determining the isomers of ezetimibe rosuvastatin calcium to determine the isomeric impurities of rosuvastatin calcium in ezetimibe rosuvastatin calcium tablets. This method can achieve the separation of rosuvastatin calcium and its isomeric impurities, and exclude the interference of ezetimibe and its isomeric impurities in the two-component preparation, and accurately detect the isomeric impurities of rosuvastatin calcium.

[0012] The present invention provides a method for determining the isomers of ezetimibe rosuvastatin calcium tablets, comprising:

[0013] Determining the rosuvastatin calcium isomer impurity in ezetimibe rosuvastatin calcium tablets by high performance liquid chromatography;

[0014] The chromatographic column is a polysaccharide derivative coated chiral chromatographic column;

[0015] Mobile phase A is an aqueous solution of 0.08% - 0.12% trifluoroacetic acid; mobile phase B is acetonitrile; gradient elution is carried out;

[0016] The flow rate is 0.4 - 0.6 ml per minute; the column temperature is 23 - 27 °C; the detection wavelength is 240 - 244 nm; the injection volume is 5 - 10 μl.

[0017] In some embodiments, the chromatographic column is Daicel CHIRALCEL OJ-3R, 250 mm × 4.6 mm, 3 μm.

[0018] In some embodiments, mobile phase A is an aqueous solution of 0.1% trifluoroacetic acid.

[0019] In some embodiments, the initial ratio of mobile phase A in the gradient elution is 70% - 73%, and mobile phase B is 27% - 30%.

[0020] In some embodiments, the gradient elution program is as follows:

[0021] Time (min) Mobile Phase A (%) Mobile Phase B (%) 0 72 28 35 72 28 38 35 65 50 35 65 50.1 72 28 60 72 28 .

[0022] In some embodiments, the rosuvastatin calcium isomer impurity is impurity F, and the structure of impurity F is as follows:

[0023]

[0024] In some embodiments, the flow rate is 0.5 ml per minute.

[0025] In some embodiments, the column temperature is 25 °C.

[0026] In some embodiments, the detection wavelength is 242 nm.

[0027] In some embodiments, the injection volume is 5 μl.

[0028] Beneficial effects:

[0029] The present invention establishes a method for determining the ezetimibe rosuvastatin calcium isomers to determine the rosuvastatin calcium isomer impurities in ezetimibe rosuvastatin calcium tablets. This method can achieve the separation of rosuvastatin calcium and its isomer impurities, and exclude the interference of ezetimibe and its isomer impurities in the two-component preparation. It has high accuracy, good stability, strong durability, can realize the detection of rosuvastatin calcium isomer impurities in the two-component preparation, the resolution between impurity F and rosuvastatin can reach 5.36, has strong specificity, high detection efficiency and low cost. Description of the Drawings

[0030] In order to more clearly illustrate the technical solutions of the present application, the accompanying drawings required for use in the embodiments will be briefly introduced below. Obviously, for those of ordinary skill in the art, other accompanying drawings can also be obtained based on these drawings without creative efforts.

[0031] Figure 1 It is the detection spectrum of rosuvastatin calcium isomer impurities in the system suitability solution;

[0032] Figure 2 It is the detection spectrum of rosuvastatin calcium isomer impurities at a flow rate of 0.4 ml per minute;

[0033] Figure 3 It is the detection spectrum of rosuvastatin calcium isomer impurities at a flow rate of 0.6 ml per minute;

[0034] Figure 4 It is the detection spectrum of rosuvastatin calcium isomer impurities at a column temperature of 23 °C;

[0035] Figure 5 It is the detection spectrum of rosuvastatin calcium isomer impurities at a column temperature of 27 °C;

[0036] Figure 6 It is the detection spectrum of rosuvastatin calcium isomer impurities at a detection wavelength of 240 nm;

[0037] Figure 7 It is the detection spectrum of rosuvastatin calcium isomer impurities at a detection wavelength of 244 nm;

[0038] Figure 8 It is the detection spectrum of rosuvastatin calcium isomer impurities with mobile phase A of 0.08% trifluoroacetic acid aqueous solution;

[0039] Figure 9 It is the detection spectrum of rosuvastatin calcium isomer impurities with mobile phase A of 0.12% trifluoroacetic acid aqueous solution;

[0040] Figure 10 It is the detection spectrum of rosuvastatin calcium isomer impurities with an injection volume of 10 μl;

[0041] Figure 11 It is the chromatogram for detecting impurities of rosuvastatin calcium isomers with the proportion of mobile phase A in gradient elution being 73%;

[0042] Figure 12 It is the chromatogram for detecting impurities of rosuvastatin calcium isomers with the proportion of mobile phase A in gradient elution being 70%. Specific embodiments

[0043] To enable those skilled in the art to understand the features and effects of the present invention, the following is a general description and definition of the terms and phrases mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used herein shall have the ordinary meanings understood by those skilled in the art for the present invention. In case of conflicts, the definitions in this specification shall prevail.

[0044] The theories or mechanisms described and disclosed herein, whether right or wrong, shall not limit the scope of the present invention in any way, that is, the content of the present invention can be implemented without being limited by any specific theory or mechanism.

[0045] In this article, all features defined in the form of numerical ranges or percentage ranges, such as numerical values, quantities, contents, and concentrations, are only for the sake of brevity and convenience. Accordingly, the description of numerical ranges or percentage ranges should be regarded as having covered and specifically disclosed all possible sub-ranges and individual numerical values within the ranges (including integers and fractions).

[0046] In this article, for the sake of concise description, all possible combinations of all technical features in each embodiment or example are not described. Therefore, as long as there are no contradictions in the combinations of these technical features, the technical features in each embodiment or example can be combined arbitrarily, and all possible combinations should be considered as the scope described in this specification.

[0047] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

[0048] Conventional instrument and equipment in the art are used in the following examples. For the experimental methods without specific conditions noted in the following examples, they are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. Various raw materials are used in the following examples. Unless otherwise stated, commercially available products are used, and their specifications are conventional specifications in the art. In the specification of the present invention and the following examples, unless otherwise specified, "%" represents weight percentage, "parts" represents weight parts, and the ratios represent weights.

[0049] The following further illustrates a method for determining ezetimibe rosuvastatin calcium isomers provided by this application in combination with specific embodiments.

[0050] Example 1:

[0051] Method for determining ezetimibe rosuvastatin calcium isomers

[0052] 1. Chromatographic conditions

[0053] Chromatographic column: Daicel CHIRALCEL OJ-3R, 250 mm × 4.6 mm, 3 μm; Detector: DAD

[0054] Mobile phase: 0.1% trifluoroacetic acid aqueous solution as mobile phase A, and acetonitrile as mobile phase B;

[0055] Flow rate: 0.5 ml per minute;

[0056] Column temperature: 25 °C; Detection wavelength is 242 nm; Injection volume is 5 μl;

[0057] Elution mode: Gradient elution, and perform gradient elution according to the following table:

[0058] Table 3 Program of gradient elution

[0059] Time (min) Mobile Phase A (%) Mobile Phase B (%) 0 72 28 35 72 28 38 35 65 50 35 65 50.1 72 28 60 72 28

[0060] 2. Solution preparation

[0061] Solvent: Take 800 ml of acetonitrile and 200 ml of water, mix well, and degas by ultrasonic wave.

[0062] Impurity stock solution:

[0063] Take about 16 mg of impurity F reference substance, weigh accurately, place it in a 100 ml volumetric flask, add solvent to dissolve it ultrasonically and dilute to the scale, and shake well; accurately measure 5 ml of this solution, place it in a 50 ml volumetric flask, and dilute to the scale with solvent, and shake well to obtain.

[0064] Take about 10 mg each of impurity A, impurity B, impurity C, impurity D, and impurity E reference substances, weigh accurately, place them in a 50 ml volumetric flask, add solvent to dissolve them ultrasonically and dilute to the scale, and shake well; accurately measure 1 ml of this solution, place it in a 50 ml volumetric flask, and dilute to the scale with solvent, and shake well to obtain.

[0065] System suitability solution: Take about 25 mg of rosuvastatin calcium reference substance and about 10 mg of ezetimibe reference substance, weigh accurately, place them in a 25 ml volumetric flask, add an appropriate amount of solvent to dissolve them ultrasonically, and then accurately measure 2.5 ml of each of the above impurity stock solutions, dilute to the scale with solvent, and shake well as the system suitability solution.

[0066] Reference substance stock solution:

[0067] Take about 13 mg of rosuvastatin calcium reference substance, accurately weigh it, place it in a 25-ml volumetric flask, add the solvent, dissolve it by ultrasonic wave and dilute it to the scale, shake well, and you will get it.

[0068] Reference substance solution: Accurately measure 1 ml of the reference substance stock solution, place it in a 50-ml volumetric flask, dilute it to the scale with the solvent, shake well, and you will get it (about 10 μg / ml of rosuvastatin), and prepare 2 portions in parallel (starting from weighing).

[0069] Test sample solution:

[0070] Take 5 tablets of ezetimibe rosuvastatin calcium tablets, place them in a 50-ml volumetric flask, add an appropriate amount of solvent (about 70% of the volumetric flask volume), sonicate for 30 minutes until dispersed, cool to room temperature, dilute it to the scale with the solvent, shake well; filter through a 0.22-μm PTFE membrane filter, discard the first 3 ml of the initial filtrate, and take the subsequent filtrate as the test sample solution, and prepare 2 portions in parallel (starting from taking this product).

[0071] 3. Results

[0072] The system suitability is as Figure 1 shown. The resolution between impurity F and rosuvastatin is 5.36, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomer impurities do not interfere with the peaks of rosuvastatin and its isomers.

[0073] Example 2:

[0074] Determination method for ezetimibe rosuvastatin calcium isomers

[0075] 1. The flow rate is 0.4 ml per minute, and the other conditions are the same as in Example 1.

[0076] 2. Results

[0077] The system suitability is as Figure 2 shown. The resolution between impurity F and rosuvastatin is 4.54, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomer impurities do not interfere with the peaks of rosuvastatin and its isomers.

[0078] Example 3:

[0079] Determination method for ezetimibe rosuvastatin calcium isomers

[0080] 1. The flow rate is 0.6 ml per minute, and the other conditions are the same as in Example 1.

[0081] 2. Results

[0082] The system suitability is as Figure 3As shown, the resolution between impurity F and rosuvastatin is 4.23, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities do not interfere with the peak emergence of rosuvastatin and its isomers.

[0083] Example 4:

[0084] Determination method for ezetimibe rosuvastatin calcium isomers

[0085] 1. The column temperature is 23 °C, and the other conditions are the same as in Example 1.

[0086] 2. Results

[0087] System suitability is as Figure 4 shown, the resolution between impurity F and rosuvastatin is 4.27, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities do not interfere with the peak emergence of rosuvastatin and its isomers.

[0088] Example 5:

[0089] Determination method for ezetimibe rosuvastatin calcium isomers

[0090] 1. The column temperature is 27 °C, and the other conditions are the same as in Example 1.

[0091] 2. Results

[0092] System suitability is as Figure 5 shown, the resolution between impurity F and rosuvastatin is 4.43, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities do not interfere with the peak emergence of rosuvastatin and its isomers.

[0093] Example 6:

[0094] Determination method for ezetimibe rosuvastatin calcium isomers

[0095] 1. The detection wavelength is 240 nm, and the other conditions are the same as in Example 1.

[0096] 2. Results

[0097] System suitability is as Figure 6 shown, the resolution between impurity F and rosuvastatin is 4.46, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities do not interfere with the peak emergence of rosuvastatin and its isomers.

[0098] Example 7:

[0099] Determination method for ezetimibe rosuvastatin calcium isomers

[0100] 1. The detection wavelength was 244 nm, and the other conditions were the same as in Example 1.

[0101] 2. Results

[0102] The system suitability was as Figure 7 shown. The resolution between impurity F and rosuvastatin was 4.41, that is, rosuvastatin and its isomers could be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities did not interfere with the peak emergence of rosuvastatin and its isomers.

[0103] Example 8:

[0104] Determination method for ezetimibe rosuvastatin calcium isomers

[0105] 1. Mobile phase: 0.08% trifluoroacetic acid aqueous solution was used as mobile phase A, and acetonitrile was used as mobile phase B. The other conditions were the same as in Example 1.

[0106] 2. Results

[0107] The system suitability was as Figure 8 shown. The resolution between impurity F and rosuvastatin was 4.46, that is, rosuvastatin and its isomers could be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities did not interfere with the peak emergence of rosuvastatin and its isomers.

[0108] Example 9:

[0109] Determination method for ezetimibe rosuvastatin calcium isomers

[0110] 1. Mobile phase: 0.12% trifluoroacetic acid aqueous solution was used as mobile phase A, and acetonitrile was used as mobile phase B. The other conditions were the same as in Example 1.

[0111] 2. Results

[0112] The system suitability was as Figure 9 shown. The resolution between impurity F and rosuvastatin was 4.42, that is, rosuvastatin and its isomers could be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities did not interfere with the peak emergence of rosuvastatin and its isomers.

[0113] Example 10:

[0114] Determination method for ezetimibe rosuvastatin calcium isomers

[0115] 1. The injection volume was 10 μl, and the other conditions were the same as in Example 1.

[0116] 2. Results

[0117] The system suitability was asFigure 10 As shown, the resolution between impurity F and rosuvastatin is 4.42, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities do not interfere with the peak emergence of rosuvastatin and its isomers.

[0118] Example 11:

[0119] Determination method for ezetimibe rosuvastatin calcium isomers

[0120] 1. Elution mode: gradient elution, and gradient elution is carried out according to the following table:

[0121] Table 4 Program of gradient elution

[0122] Time (min) Mobile Phase A (%) Mobile Phase B (%) 0 73 27 40 73 27 45 25 75 50 25 75 50.1 73 27 55 73 27

[0123] Other conditions are the same as those in Example 1.

[0124] 2. Results

[0125] System suitability is as Figure 11 shown, the resolution between impurity F and rosuvastatin is 4.67, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities do not interfere with the peak emergence of rosuvastatin and its isomers.

[0126] Example 12:

[0127] Determination method for ezetimibe rosuvastatin calcium isomers

[0128] 1. Elution mode: gradient elution, and gradient elution is carried out according to the following table:

[0129] Table 5 Program of gradient elution

[0130] Time (min) Mobile Phase A (%) Mobile Phase B (%) 0 70 30 30 70 30 35 48 52 50 48 52 50.1 70 30 60 70 30

[0131] Other conditions are the same as those in Example 1.

[0132] 2. Results

[0133] System suitability is as Figure 12 shown, the resolution between impurity F and rosuvastatin is 4.25, that is, rosuvastatin and its isomers can be well separated by the determination method of the present invention, and ezetimibe and its isomeric impurities do not interfere with the peak emergence of rosuvastatin and its isomers.

[0134] Experimental example 1:

[0135] System suitability test

[0136] Take appropriate amounts of rosuvastatin calcium reference substance, impurity F, ezetimibe reference substance, impurity A, impurity B, impurity C, impurity D, and impurity E reference substances, dissolve and dilute with a solvent to prepare a solution containing approximately 1 mg of rosuvastatin, 1.5 μg of impurity F, 0.4 mg of ezetimibe, and 0.4 μg each of impurity A, impurity B, impurity C, impurity D, and impurity E per 1 ml. The elution order is successively impurity F, rosuvastatin, and ezetimibe and its isomers do not interfere with the elution of rosuvastatin and its isomers.

[0137] Inject for determination, and the results of the system suitability test are shown in Table 6.

[0138] Table 6 Results of System Suitability Test

[0139] Peak Area % Resolution Tailing Factor Impurity F 0.16 2.28 0.92 Rosuvastatin 99.84 N.A 1.03

[0140] Experimental Example 2:

[0141] Robustness Test

[0142] Use the 100% test solution as the test solution to investigate the influence of minor changes in chromatographic conditions (column temperature, flow rate, acid amount of mobile phase A, mobile phase ratio, detection wavelength, injection volume) on the detection results, so as to evaluate the robustness of the method. The results are shown in Tables 7 and 8.

[0143] Table 7 Results of Solution Stability in Robustness Test

[0144]

[0145] As shown in Table 7, rosuvastatin and impurity F are stable within 24 hours at room temperature.

[0146] Table 8 Influence of Chromatographic Conditions in Robustness Test on the Resolution of Rosuvastatin and Impurity F

[0147]

[0148] The data in Tables 7 and 8 show that minor changes in the chromatographic conditions of the present invention have no obvious influence on the separation of rosuvastatin and impurity F, and the resolution under each condition is >1.5, meeting the resolution requirements for the determination of rosuvastatin isomers.

[0149] Although the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Any person familiar with this technology can make various modifications and decorations without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention should be defined by the claims.

Claims

1. A method for determining the isomers of ezetimibe and rosuvastatin calcium tablets, characterized in that, Comprising: Determining the rosuvastatin calcium isomer impurity in ezetimibe and rosuvastatin calcium tablets by high performance liquid chromatography; The chromatographic column is a polysaccharide derivative-coated chiral chromatographic column; Mobile phase A is an aqueous solution of 0.08% - 0.12% trifluoroacetic acid; mobile phase B is acetonitrile; gradient elution is carried out; The flow rate is 0.4 - 0.6 ml per minute; the column temperature is 23 - 27 °C; the detection wavelength is 240 - 244 nm; the injection volume is 5 - 10 μl.

2. The method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, wherein The chromatographic column is Daicel CHIRALCEL OJ-3R, 250 mm × 4.6 mm, 3 μm.

3. The method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, wherein The mobile phase A is an aqueous solution of 0.1% trifluoroacetic acid.

4. The method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, wherein The initial ratio of the gradient elution for mobile phase A is 70% - 73%, and for mobile phase B is 27% - 30%.

5. A method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, characterized in that, The program of the gradient elution is as follows: 。 6. The method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, wherein The rosuvastatin calcium isomer impurity is impurity F, and the structure of impurity F is as follows:

7. The method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, characterized in that, The flow rate is 0.5 ml per minute.

8. A method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, characterized in that, The column temperature is 25 °C.

9. The method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, wherein The detection wavelength is 242 nm.

10. The method for determining the isomers of ezetimibe and rosuvastatin calcium tablets according to claim 1, wherein, The injection volume is 5 μl.

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