Method for detecting ibuprofen related substances in ibuprofen tablets

Abstract

The invention relates to the field of medicine detection, in particular to a method for detecting ibuprofen related substances in an ibuprofen tablet, the detection method adopts high performance liquid chromatography, and chromatographic conditions comprise that octadecylsilane chemically bonded silica is used as a filler of a chromatographic column; a mobile phase is composed of a mobile phase A and a mobile phase B, and gradient elution is carried out; the mobile phase A is a C6-12 sodium alkane sulfonate solution; the mobile phase B is acetonitrile; the gradient elution procedure comprises a first elution stage, a second elution stage, a third elution stage and a fourth elution stage which are continuously carried out. According to the detection method provided by the invention, the interference of blank auxiliary materials and other related substances can be avoided, the quality detection indexes of the ibuprofen tablets are enriched, and the product quality can be better controlled.

Description

Technical Field

[0001] The present application relates to the field of medical detection, and specifically to a method for detecting ibuprofen-related substances in ibuprofen renin tablets. Background Art

[0002] Ibuprofen tablets are compound preparations, mainly used to temporarily relieve the following symptoms associated with colds or flu, headaches, fever, sinus pressure, nasal congestion, mild body aches, reduce nasal swelling, and temporarily restore free breathing through the nasal cavity. It was first launched in the UK on November 17, 2008, and there is currently no reference preparation imported into my country. There are few methods for determining ibuprofen-related substances in Ibuprofen tablets in the prior art, and most of them are related substance detection methods for some dosage forms of single preparations of each component. Ibuprofen tablets are a compound preparation, and there are many impurities and interacting impurities in the two main components, and there are excipients with strong absorption in the excipients that interfere with impurity detection.

[0003] Therefore, it is urgent to propose an accurate, stable and less interfered method for the detection of ibuprofen-related substances in ibuprofen-containing renin tablets. Summary of the invention

[0004] The purpose of the present invention is to provide a method for detecting ibuprofen-related substances in ibuprofen tablets, so as to better solve the problem of interference detection between blank excipients and impurities in ibuprofen tablets, more effectively control the product quality of ibuprofen tablets, and improve enterprise standards.

[0005] To this end, the embodiment of the present application provides a method for detecting ibuprofen-related substances in ibuprofen renin tablets, wherein the detection method adopts high performance liquid chromatography, and the chromatographic conditions include: the chromatographic column uses octadecylsilane bonded silica gel as a filler; the mobile phase is composed of mobile phase A and mobile phase B and gradient elution is performed; the mobile phase A is C 6-12 Sodium alkanesulfonate solution; the mobile phase B is acetonitrile; and the gradient elution procedure includes a first elution stage, a second elution stage, a third elution stage and a fourth elution stage performed continuously,

[0006] The first elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 75% to 85% to 45% to 55%; the volume ratio of the mobile phase B changes from 15% to 25% to 45% to 55%; and the time of the first elution stage is 15 to 25 minutes,

[0007] wherein the second elution stage is an isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the time of the second elution stage is 25 to 35 minutes,

[0008] The third elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 45% to 55% to 75% to 85%, and the volume ratio of the mobile phase B changes from 45% to 55% to 15% to 25%; and the time of the third elution stage is 1.5 to 2.5 minutes,

[0009] The fourth elution stage is isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the duration of the fourth elution stage is 6 to 10 minutes.

[0010] In some embodiments, the ibuprofen-related substances in the ibuprofen renin tablets include one or more of impurity J, impurity E, impurity N, impurity B, formulation impurity 1, formulation impurity 2, and formulation impurity 3,

[0011] wherein the impurity J is (2RS)-2-[4-(2-methylpropyl)phenyl]propionic acid; the impurity E is 1-[4-(2-methylpropyl)phenyl]acetone; the impurity N is (2RS)-2-[4-(2-ethylpropyl)phenyl]propionic acid; the impurity B is (2RS)-2-[4-(2-butylpropyl)phenyl]propionic acid; the formulation impurity 1 is 3-((R)-1-hydroxy-2-(methylamino)ethyl)phenyl-2-(4-isobutylphenyl)propanoate; the formulation impurity 2 is (R)-1-(3-hydroxyphenyl)-2-(methylamino)ethyl-2-(4-isobutylphenyl)propionic acid ethyl ester; and the formulation impurity 3 is N-((R)-2-hydroxy-2-(3-hydroxyphenyl)ethyl)-2-(4-isobutylphenyl)-N-methylpropionamide.

[0012] In some embodiments, the first elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 78% to 82% to 48% to 52%; the volume ratio of the mobile phase B changes from 18% to 22% to 48% to 52%; and the time of the first elution stage is 18 to 22 minutes.

[0013] In some embodiments, the second elution stage is an isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the time of the second elution stage is 28 to 32 minutes.

[0014] In some embodiments, the third elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 48% to 52% to 78% to 82%, and the volume ratio of the mobile phase B changes from 48% to 52% to 18% to 22%; and the time of the third elution stage is 1.8 to 2.2 minutes.

[0015] In some embodiments, the fourth elution stage is an isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the duration of the fourth elution stage is 7 to 9 minutes.

[0016] In some embodiments, the first elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 80% to 50%; the volume ratio of the mobile phase B changes from 20% to 50%; and the time of the first elution stage is 20 minutes.

[0017] In some embodiments, the second elution stage is an isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the time of the second elution stage is 30 minutes.

[0018] In some embodiments, the third elution stage is linear gradient elution, wherein the volume ratio of the mobile phase A changes from 50% to 80%, and the volume ratio of the mobile phase B changes from 50% to 20%; and the time of the third elution stage is 2 minutes.

[0019] In some embodiments, the fourth elution stage is isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the duration of the fourth elution stage is 8 minutes.

[0020] In some embodiments, the mobile phase A is selected from an aqueous solution of sodium octane sulfonate, an aqueous solution of sodium hexane sulfonate, or an aqueous solution of sodium decane sulfonate.

[0021] In some embodiments, the concentration of mobile phase A is 3 to 3.5 g / L.

[0022] In some embodiments, the pH of the mobile phase A is 2.2 to 3.

[0023] In some embodiments, the size of the chromatographic column is 250×4.6 mm, and the filler particle size is 5 μm.

[0024] In some embodiments, the column temperature of the chromatography column is 30 to 40°C.

[0025] In some embodiments, the flow rate of the mobile phase is 0.8 to 1.2 mL / min.

[0026] In some embodiments, the chromatographic conditions further include a detection wavelength of 210 to 220 nm.

[0027] In some embodiments, the concentration of mobile phase A is 3.25 g / L.

[0028] In some embodiments, the pH of mobile phase A is 2.6.

[0029] In some embodiments, the column temperature of the chromatographic column is 35°C.

[0030] In some embodiments, the flow rate of the mobile phase is 1 mL / min.

[0031] In some embodiments, the detection wavelength is 214 nm.

[0032] In some embodiments, the detection method comprises the following steps:

[0033] S1. dissolving the bromelain tablets to be tested in a solvent to prepare a sample solution to be tested;

[0034] S2. The sample solution to be tested was diluted 400 to 600 times to obtain a control solution;

[0035] S3. using the high performance liquid chromatography to detect the sample solution and the control solution; and

[0036] S4. Calculate the weight percentage of each of the ibuprofen-related substances relative to ibuprofen in the ibuprofen tablet to be tested based on the peak area of ​​the ibuprofen-related substances in the sample solution to be tested and the peak area of ​​ibuprofen in the control solution.

[0037] In some embodiments, the solvent comprises acetonitrile and water in a volume ratio of 1:1.

[0038] In some embodiments, the test sample solution comprises ibuprofen at a concentration of 1 to 3 mg / mL.

[0039] The embodiments of the present application achieve the following beneficial effects:

[0040] By optimizing the high performance liquid chromatography, the interference of blank auxiliary material (propyl gallate) and other main component related substances in the detection of ibuprofen related substances in ibuprofen tablets is avoided. The embodiment of the present application provides a method for detecting ibuprofen related substances in ibuprofen tablets with good accuracy, repeatability and sensitivity, which can accurately control the impurities of the preparation, making the quality detection indicators of ibuprofen tablets more comprehensive, which is conducive to better control of product quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0042] Figure 1This is a flow chart of the method for detecting ibuprofen-related substances in ibuprofen renin tablets provided in an embodiment of the present application.

[0043] Figure 2 This is the test result of the system suitability solution in Example 2 of the present application, wherein the retention times of impurity J, impurity E and ibuprofen are 24.002, 46.124 and 38.096 min, respectively.

[0044] Figure 3 This is the test result of the verification solution in Example 2 of the present application, wherein the chromatographic peaks before the relative retention time of 0.4 min are the peaks of phenylephrine hydrochloride and its related substances, which are ignored; the retention time of impurity J is 23.717 min, the retention time of impurity N is 26.511 min, the retention time of preparation impurity 1 is 32.085 min and 32.637 min, the retention time of preparation impurity 2+3 is 35.223 min, the retention time of impurity B is 39.210 min, the retention time of impurity E is 45.342 min and the retention time of ibuprofen is 37.506 min. DETAILED DESCRIPTION

[0045] The present invention is further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art and are not intended to limit the present invention in any way.

[0046] This application is made based on the following knowledge of the inventor:

[0047] There are few methods disclosed in the prior art for determining ibuprofen-related substances in ibuprofen tablets, and most of them are related substance detection methods for some dosage forms of single preparations of each component. Ibuprofen tablets are compound preparations, and there are many impurities and interacting impurities in the two main components, and there are excipients with strong absorption in the excipients that interfere with impurity detection.

[0048] Wang Peiqi et al. optimized the detection method of related substances of ibuprofen tablets in BP to detect ibuprofen related substances in ibuprofen tablets (R917, A, 1001-249 (2014) 12-1062-05), but failed to effectively control the impurities of the preparation; Li Xiangling et al. used HPLC to simultaneously determine the related substances of each main component in ibuprofen tablets (CN116046948A), but the operation time was long, the separation between impurities and the main peak was not ideal, all impurities had correction factors that were not convenient to calculate, and the quantitative limits of some impurities were high.

[0049] After extensive research, the inventors have optimized the conditions of high performance liquid chromatography to detect ibuprofen-related substances in ibuprofen tablets, avoiding the interference of blank excipients (propyl gallate) and other main ingredient-related substances, and can accurately control the impurities in the preparation. A method for detecting ibuprofen-related substances in ibuprofen tablets with good accuracy, repeatability and sensitivity is provided, which makes the quality inspection indicators of ibuprofen tablets more comprehensive and is conducive to better control of product quality.

[0050] Figure 1 The present invention provides a method for detecting ibuprofen-related substances in ibuprofen-related renin tablets, wherein a liquid chromatography condition capable of distinguishing ibuprofen-related substances from other components in ibuprofen-related renin tablets is proposed. Based on the detection conditions of the high performance liquid chromatography, the present invention uses a self-control method to detect and calculate the weight percentage of various ibuprofen-related substances in ibuprofen-related renin tablets relative to ibuprofen. Figure 1 As shown, the method for detecting ibuprofen-related substances in ibuprofen renin tablets may include the following steps: S1-S4.

[0051] Step S1. dissolving the brexan tablets to be tested in a solvent to prepare a sample solution to be tested.

[0052] In some embodiments, the solvent comprises acetonitrile and water in a 1:1 volume ratio.

[0053] In some embodiments, the test sample solution comprises ibuprofen at a concentration of 1 to 3 mg / mL.

[0054] Step S2: diluting the sample solution to be tested by 400 to 600 times to prepare a control solution.

[0055] Step S3: using the high performance liquid chromatography method to detect the sample solution and the control solution respectively.

[0056] In some embodiments, the ibuprofen-related substances in the ibuprofen renin tablets to be tested include one or more of impurities J, impurity E, impurity N, impurity B, formulation impurity 1, formulation impurity 2, and formulation impurity 3, impurity J is: (2RS)-2-[4-(2-methylpropyl)phenyl]propionic acid; impurity E is 1-[4-(2-methylpropyl)phenyl]acetone; impurity N is (2RS)-2-[4-(2-ethylpropyl)phenyl]propionic acid; impurity B is (2RS)-2-[ Preparation impurity 1 is 3-((R)-1-hydroxy-2-(methylamino)ethyl)phenyl-2-(4-isobutylphenyl)propionate; Preparation impurity 2 is (R)-1-(3-hydroxyphenyl)-2-(methylamino)ethyl-2-(4-isobutylphenyl)propionic acid ethyl ester; Preparation impurity 3 is N-((R)-2-hydroxy-2-(3-hydroxyphenyl)ethyl)-2-(4-isobutylphenyl)-N-methylpropionamide. Detailed information on the aforementioned related substances is shown in Table 1 below.

[0057] Table 1 Information on related substances of Buluorenin Tablets

[0058]

[0059]

[0060] In some embodiments, the detection method adopts high performance liquid chromatography, and the chromatographic conditions include: the chromatographic column uses octadecylsilane bonded silica gel as a filler; the mobile phase consists of mobile phase A and mobile phase B and performs gradient elution; the mobile phase A is C 6-12 The invention relates to a sodium alkanesulfonate solution; the mobile phase B is acetonitrile; and the gradient elution procedure includes a first elution stage, a second elution stage, a third elution stage and a fourth elution stage which are performed continuously, wherein the first elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 75% to 85% to 45% to 55%; the volume ratio of the mobile phase B changes from 15% to 25% to 45% to 55%; and the time of the first elution stage is 15 to 25 minutes, wherein the second elution stage is an isocratic elution, wherein the mobile phase A and the mobile phase B are The volume ratio of mobile phase A and mobile phase B remains unchanged; and the time of the second elution stage is 25 to 35 minutes, wherein the third elution stage is linear gradient elution, wherein the volume ratio of mobile phase A changes from 45% to 55% to 75% to 85%, and the volume ratio of mobile phase B changes from 45% to 55% to 15% to 25%; and the time of the third elution stage is 1.5 to 2.5 minutes, wherein the fourth elution stage is isocratic elution, wherein the volume ratio of mobile phase A and mobile phase B remains unchanged; and the time of the fourth elution stage is 6 to 10 minutes.

[0061] In some embodiments, the first elution stage is a linear gradient elution, wherein the volume ratio of mobile phase A changes from 78% to 82% to 48% to 52%; the volume ratio of mobile phase B changes from 18% to 22% to 48% to 52%; and the time of the first elution stage is 18 to 22 minutes.

[0062] In some embodiments, the second elution stage is an isocratic elution, in which the volume ratio of mobile phase A and mobile phase B remains unchanged; and the time of the second elution stage is 28 to 32 minutes.

[0063] In some embodiments, the third elution stage is a linear gradient elution, wherein the volume ratio of mobile phase A changes from 48% to 52% to 78% to 82%, and the volume ratio of mobile phase B changes from 48% to 52% to 18% to 22%; and the time of the third elution stage is 1.8 to 2.2 minutes.

[0064] In some embodiments, the fourth elution stage is an isocratic elution, in which the volume ratio of mobile phase A and mobile phase B remains unchanged; and the duration of the fourth elution stage is 7 to 9 minutes.

[0065] In some embodiments, the first elution stage is a linear gradient elution, wherein the volume ratio of mobile phase A changes from 80% to 50%; the volume ratio of mobile phase B changes from 20% to 50%; and the time of the first elution stage is 20 minutes.

[0066] In some embodiments, the second elution stage is an isocratic elution, in which the volume ratio of mobile phase A and mobile phase B remains unchanged; and the time of the second elution stage is 30 minutes.

[0067] In some embodiments, the third elution stage is linear gradient elution, wherein the volume ratio of mobile phase A changes from 50% to 80%, and the volume ratio of mobile phase B changes from 50% to 20%; and the time of the third elution stage is 2 minutes.

[0068] In some embodiments, the fourth elution stage is an isocratic elution, in which the volume ratio of mobile phase A and mobile phase B remains unchanged; and the time of the fourth elution stage is 8 minutes.

[0069] In some embodiments, mobile phase A is selected from an aqueous solution of sodium octane sulfonate, an aqueous solution of sodium hexane sulfonate, or an aqueous solution of sodium decane sulfonate.

[0070] In some embodiments, the concentration of mobile phase A is 3 to 3.5 g / L.

[0071] In some embodiments, the concentration of mobile phase A is 3.25 g / L.

[0072] In some embodiments, the pH of mobile phase A is 2.2-3.

[0073] In some embodiments, the pH of mobile phase A is 2.6.

[0074] In some embodiments, the size of the chromatographic column is 250×4.6 mm, and the filler particle size is 5 μm.

[0075] In some embodiments, the column temperature of the chromatography column is 30 to 40°C.

[0076] In some embodiments, the column temperature of the chromatography column is 35°C.

[0077] In some embodiments, the flow rate of the mobile phase is 0.8 to 1.2 mL / min.

[0078] In some embodiments, the flow rate of the mobile phase is 1 mL / min.

[0079] In some embodiments, the chromatographic conditions further include a detection wavelength of 210 to 220 nm.

[0080] In some embodiments, the detection wavelength is 214 nm.

[0081] Step S4. Calculate the weight percentage of each of the ibuprofen-related substances relative to ibuprofen in the ibuprofen tablet to be tested according to the peak area of ​​the ibuprofen-related substances in the sample solution to be tested and the peak area of ​​ibuprofen in the control solution.

[0082] In the present application embodiment, ibuprofen tablets are ground and dissolved in a solvent to obtain a sample solution containing about 1 to 3 mg / mL of ibuprofen at a concentration. Due to the large difference in the content of ibuprofen and ibuprofen related substances (i.e., impurity J, impurity E, impurity N, impurity B, formulation impurity 1, formulation impurity 2, and formulation impurity 3, etc.) in the sample solution to be tested, the weight percentage of various ibuprofen related substances relative to ibuprofen is calculated directly based on the peak area of ​​ibuprofen related substances in the solution to be tested, and there is a large error. Therefore, the present application embodiment is further quantitatively diluting the sample solution to be tested by 400 to 600 times to obtain a diluted control solution, and using the peak area of ​​ibuprofen in the control solution multiplied by the dilution multiple as the ibuprofen peak area in the solution to be tested, to calculate the weight percentage (detection amount) of various ibuprofen related substances in the solution to be tested relative to ibuprofen, to reduce the error caused by the large difference in the content of ibuprofen and its related substances.

[0083] The specific calculation method is:

[0084] According to the chromatographic analysis results, the chromatographic peak areas of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, and preparation impurity 2+3 in the sample solution to be tested are recorded as A J , A E , A N , A B , A制剂1 , A 制剂2+3 The chromatographic peak area of ​​ibuprofen in the control solution is multiplied by the dilution factor (in some embodiments, the dilution factor is 400 to 600 times, such as 425 times, 450 times, 475 times, 500 times, 525 times, 550 times, 575 times, etc.) and is recorded as A. 1 .

[0085] Detection amount of impurity J = A J / A 1 ×100%;

[0086] Detection amount of impurity E = A E / A 1 )×100%;

[0087] Detection amount of impurity N = A N / A 1 ×100%;

[0088] Detection amount of impurity B = A B / A 1 ×100%;

[0089] Detection amount of impurity 1 in the preparation = A 制剂1 / A 1 ×100%;

[0090] Detection amount of preparation impurities 2+3 = A 制剂2+3 / A 1 ×100%.

[0091] The following examples are used to further illustrate the advantages and features of the present method, but are not intended to limit the present invention. The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are performed according to the techniques or conditions described in the literature in the art or according to the product instructions.

[0092] The materials and reagents used in the following examples, unless otherwise specified, can be obtained from commercial sources. Unless otherwise specified, the quantitative analysis experiments in the following examples were repeated three times, and the results were averaged.

[0093] Instruments and reagents used in the examples of this application:

[0094] Instrument: Shimadzu LC-20AT high performance liquid chromatograph

[0095] Chromatographic column: C18 (Kromasil 4.6×250mm, 5μm)

[0096] Reagents: Ibuprofen (China Food and Drug Inspection Institute 100179-201707); Ibuprofen impurity J (China Food and Drug Inspection Institute 510108-202102); Ibuprofen impurity E (China Food and Drug Inspection Institute 100722-202105); Ibuprofen impurity N (Pharmaceu tical chemistry laboratory Co. Ltd 22-IBU-21-95); Ibuprofen impurity B (Toron Research Chemicals 5-SUM-76-1); Preparation impurity 1 (Baonuo Synthesis 20161025-15); Preparation impurity 2 (Baonuo Synthesis 20161025-16); Preparation impurity 3 (GP BNF-NN-161026); Acetonitrile (chromatographic grade FISHER); Sodium octane sulfonate (chromatographic grade FISHER); Phosphoric acid (analytical grade).

[0097] Samples to be tested: Ibuprofen tablets produced by Beijing New Leading Pharmaceutical Technology Development Co., Ltd., with the specification of each tablet containing 200 mg of ibuprofen and 10 mg of phenylephrine hydrochloride.

[0098] Example 1

[0099] 1. Preparation of system suitability solution

[0100] Standard samples of impurity J, impurity E and ibuprofen were taken respectively, accurately weighed, and solvent was added to prepare system suitability solutions containing 4 μg, 4 μg and 2 mg of impurity J, impurity E and ibuprofen per 1 ml of solution, respectively.

[0101] 2. Preparation of sample solution

[0102] The sample to be tested was dissolved in a solvent (acetonitrile-water solution with a volume ratio of 1:1) to prepare a sample solution containing approximately 2 mg of ibuprofen per 1 ml of the solution.

[0103] Specific steps: Take 10 ibuprofen tablets as the sample to be tested, grind them into powder, mix them evenly, take about 217 mg of ibuprofen tablet fine powder, accurately weigh it, put it in a 50 ml volumetric bottle, add an appropriate amount of solvent, ultrasonicate for 5 minutes, cool it, dilute it to the scale with solvent, shake it evenly, filter it, and take the filtrate to obtain the sample solution to be tested containing about 2 mg / ml of ibuprofen.

[0104] 3. Preparation of control solution

[0105] The sample solution to be tested was diluted 500 times with a solvent (acetonitrile-water solution with a volume ratio of 1:1) to prepare a control solution.

[0106] Specific steps: accurately measure 2 ml of the sample solution to be tested, place it in a 100 ml volumetric flask, dilute it to the scale with solvent, shake well, accurately measure 5 ml, place it in a 50 ml volumetric flask, dilute it to the scale with solvent, shake well, and you will get the control sample solution.

[0107] 4. Preparation of verification solution

[0108] Impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2, preparation impurity 3 and ibuprofen standard were taken respectively, accurately weighed, and solvent (acetonitrile-water solution with a volume ratio of 1:1) was added to prepare verification solutions containing 4μg, 5μg, 3μg, 4μg, 3μg, 3μg, 3μg, 2mg of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2, preparation impurity 3 and ibuprofen per 1ml of solution.

[0109] Example 2

[0110] The system suitability solution and verification solution prepared in Example 1 were used to verify the suitability of the chromatographic conditions. The test results were as follows: Figure 2 and 3 shown.

[0111] The chromatographic conditions were as follows:

[0112] Chromatographic column: C18 (Kromasil 4.6×250 mm, 5 μm).

[0113] Column temperature: 35°C.

[0114] Detection wavelength: 214nm.

[0115] Mobile phase flow rate: 1 mL / min.

[0116] Mobile phase A: sodium octane sulfonate aqueous solution with a pH of 2.6 and a concentration of 3.25 g / L; mobile phase B: acetonitrile.

[0117] The elution program is shown in Table 2 below, wherein 0 to 20 min is the first elution stage; 20 to 50 min is the second elution stage; 50 to 52 min is the third elution stage, and 52 to 60 min is the fourth elution stage.

[0118] Table 2

[0119]

[0120] Test results:

[0121] In the test results of system suitability solution ( Figure 2) can be seen that the chromatographic peaks of impurity J, impurity E and ibuprofen are completely separated, and the retention times of impurity J, impurity E and ibuprofen are 24.002, 46.124 and 38.096 min, respectively.

[0122] In the test results of the verification solution ( Figure 3 ) It can be seen that the chromatographic peak before the relative retention time of 0.4min is the peak of phenylephrine hydrochloride related substances, which can be ignored; the retention time of impurity J is 23.717min, the retention time of impurity N is 26.511min, the retention time of preparation impurity 1 is 32.085min and 32.637min, the retention time of preparation impurity 2+3 is 35.223min, the retention time of impurity B is 39.210min, the retention time of impurity E is 45.342min and the retention time of ibuprofen is 37.506min. The ibuprofen impurity is completely separated from the phenylephrine hydrochloride impurity; the preparation impurity 2 overlaps with the preparation impurity 3; the preparation impurity 1 is a double peak. The theoretical plate number is high, the peak shape is good, and the baseline is stable.

[0123] Example 3 Repeatability

[0124] Since the types of impurities contained in the ibuprofen tablets, the sample to be tested used in the embodiment of the present application, are relatively small, in order to demonstrate that the detection method provided in the embodiment of the present application can simultaneously separate and detect 7 impurities related to ibuprofen in the ibuprofen tablets, this embodiment prepares a mixed test solution containing impurities J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2 and preparation impurity 3 at concentrations of approximately 4 μg / ml, 5 μg / ml, 3 μg / ml, 4 μg / ml, 3 μg / ml, 3 μg / ml, 3 μg / ml, 2 mg / ml, and ibuprofen, and the mixed test solution is diluted 500 times to prepare a self-control solution of the mixed test solution. Six portions of each solution are prepared in parallel and detected according to the chromatographic conditions of Example 2.

[0125] The specific calculation method is:

[0126] According to the chromatographic analysis results, the chromatographic peak areas of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, and preparation impurity 2+3 in the mixed test solution are recorded as A J , A E , A N , A B , A 制剂1 , A 制剂2+3 The chromatographic peak area of ​​ibuprofen in the self-control solution of the mixed test solution multiplied by the dilution factor of 500 is recorded as A 1 .

[0127] Detection amount of impurity J = A J / A 1 ×100%;

[0128] Detection amount of impurity E = A E / A 1 )×100%;

[0129] Detection amount of impurity N = A N / A 1 ×100%;

[0130] Detection amount of impurity B = A B / A 1 ×100%;

[0131] Detection amount of impurity 1 in the preparation = A 制剂1 / A 1 ×100%;

[0132] Detection amount of preparation impurities 2+3 = A 制剂2+3 / A 1 ×100%.

[0133] Test results: The detection amount of impurity J was 0.13% (RSD was 1.73%), the detection amount of impurity E was 0.36% (RSD was 1.62%), the detection amount of impurity N was 0.07% (RSD was 1.67%), the detection amount of impurity B was 0.07% (RSD was 0.88%), the detection amount of preparation impurity 1 was 0.05% and 0.02% (RSD was 2.09% and 8.07%), and the detection amount of preparation impurity 2+3 was 0.32% (RSD was 1.74%).

[0134] Example 4 Intermediate Precision

[0135] According to the method of Example 3, 12 mixed test sample solutions were prepared in parallel, and the detection was carried out according to the chromatographic conditions of Example 2.

[0136] Test results: The detection amount of impurity J was 0.13% (RSD was 8.45%), the detection amount of impurity E was 0.36% (RSD was 4.17%), the detection amount of impurity N was 0.07% (RSD was 5.01%), the detection amount of impurity B was 0.07% (RSD was 11.46%), the detection amount of preparation impurity 1 was 0.05% and 0.02% (RSD was 5.89% and 4.91%), and the detection amount of preparation impurity 2+3 was 0.32% (RSD was 4.46%).

[0137] Example 5 Recovery

[0138] Reagent preparation:

[0139] Take the standard samples of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2 and preparation impurity 3 and ibuprofen tablet fine powder respectively, accurately weigh them, add solvent to make a mixed solution containing about 1μg, 1μg, 1μg, 1μg, 1μg, 1μg, 1μg, 1μg, 2mg of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2, preparation impurity 3 and ibuprofen per 1ml of solution, respectively, as the low-concentration mixed test solution, and prepare 3 copies in parallel;

[0140] Take the standard samples of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2 and preparation impurity 3 and ibuprofen tablet fine powder respectively, accurately weigh them, add solvent to make a mixed solution containing about 4μg, 5μg, 3μg, 4μg, 3μg, 3μg, 3μg, 2mg of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2, preparation impurity 3 and ibuprofen per 1ml of solution, as the medium concentration mixed test solution, and prepare 3 copies in parallel;

[0141] Take the standard samples of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2 and preparation impurity 3 and ibuprofen tablet fine powder respectively, accurately weigh them, add solvent to make a mixed solution containing about 6μg, 7.5μg, 4.5μg, 6μg, 4.5μg, 4.5μg, 4.5μg, 2mg of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2, preparation impurity 3 and ibuprofen per 1ml of solution, respectively, as the high concentration mixed test solution, and prepare 3 copies in parallel;

[0142] Take standard samples of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2 and preparation impurity 3 respectively, weigh them accurately, add solvent to make reference solution containing about 4μg, 5μg, 3μg, 4μg, 3μg, 3μg, and 3μg of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2 and preparation impurity 3 per 1ml of solution, respectively, and prepare two parallel aliquots; and

[0143] Ibuprofen tablets were dissolved in a solvent (acetonitrile-water solution with a volume ratio of 1:1) to prepare an unspiked test solution containing approximately 2 mg of ibuprofen per 1 ml of solution, and two parallel aliquots were prepared.

[0144] Test results:

[0145] The above solution was used to perform detection according to the chromatographic conditions in Example 2, and the recovery rate was calculated by the external standard method based on the detection results. Recovery rate = (measured value of spiked sample - measured value of sample) / spiked amount × 100%.

[0146] The recovery rate of impurity J was 100.5% (RSD was 0.84%), the recovery rate of impurity E was 99.7% (RSD was 1.62%), the recovery rate of impurity N was 100.9% (RSD was 0.82%), the recovery rate of impurity B was 96.9% (RSD was 6.80%), the recovery rate of preparation impurity 1 was 100.9% (RSD was 1.92%), the recovery rate of preparation impurity 2 was 103.0% (RSD was 1.31%), and the recovery rate of preparation impurity 3 was 102.8% (RSD was 1.89%).

[0147] Example 6 Sensitivity

[0148] Take standard samples of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2, preparation impurity 3 and ibuprofen respectively, weigh them accurately, add solvent to make solutions containing 0.2μg, 0.2μg, 0.14μg, 0.22μg, 0.16μg, 0.52μg, 0.52μg and 0.2μg of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2, preparation impurity 3 and ibuprofen per 1ml of solution, as the quantitative limit solution; accurately measure 3ml of the quantitative limit solution, place it in a 10ml volumetric flask, and dilute to the scale with solvent as the detection limit solution.

[0149] The detection was carried out according to the chromatographic conditions of Example 2. The results are shown in Table 3 below, where the values ​​are S / N.

[0150] Table 3

[0151]

[0152] Comparative Example 1

[0153] The verification solution was tested using substantially the same chromatographic conditions as in Example 2, except that the elution procedure used in this comparative example was different from that in Example 2. The elution procedure used in this comparative example is specifically shown in Table 4 below:

[0154] Table 4

[0155]

[0156] Test results:

[0157] Each impurity in the verification solution can be detected individually, but the overall peak is at the front, which overlaps with the impurity of phenylephrine hydrochloride (but not overlapping), affecting the location of the impurity.

[0158] Comparative Example 2

[0159] The verification solution was tested using substantially the same chromatographic conditions as in Example 2, except that the chromatographic column used in this comparative example was different from that in Example 2. In this comparative example, a variety of different chromatographic columns were used to replace the C18 (Kromasil 4.6×250 mm, 5 μm) in Example 2 for testing, and the results are shown in Table 5 below.

[0160] Table 5

[0161]

[0162] Comparative Example 3

[0163] The verification solution was tested using substantially the same chromatographic conditions as in Example 2, with the only difference being that the column temperature used in this comparative example was different from that in Example 2. The test results are shown in Table 6 below.

[0164] Table 6

[0165]

[0166] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0167] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A method for detecting related substances of ibuprofen in ibuprofen renin tablets, characterized in that: The detection method adopts high performance liquid chromatography, and the chromatographic conditions include: the chromatographic column uses octadecylsilane bonded silica gel as a filler; the mobile phase consists of mobile phase A and mobile phase B and performs gradient elution; the mobile phase A is C 6-12 Sodium alkanesulfonate solution; the mobile phase B is acetonitrile; and the gradient elution procedure includes a first elution stage, a second elution stage, a third elution stage and a fourth elution stage performed continuously, The first elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 75% to 85% to 45% to 55%; the volume ratio of the mobile phase B changes from 15% to 25% to 45% to 55%; and the time of the first elution stage is 15 to 25 minutes, wherein the second elution stage is an isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the time of the second elution stage is 25 to 35 minutes, The third elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 45% to 55% to 75% to 85%, and the volume ratio of the mobile phase B changes from 45% to 55% to 15% to 25%; and the time of the third elution stage is 1.5 to 2.5 minutes, The fourth elution stage is isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the duration of the fourth elution stage is 6 to 10 minutes.

2. The detection method according to claim 1, characterized in that: The ibuprofen-related substances in the ibuprofen renin tablets include one or more of impurity J, impurity E, impurity N, impurity B, preparation impurity 1, preparation impurity 2 and preparation impurity 3, wherein the impurity J is (2RS)-2-[4-(2-methylpropyl)phenyl]propionic acid; the impurity E is 1-[4-(2-methylpropyl)phenyl]acetone; the impurity N is (2RS)-2-[4-(2-ethylpropyl)phenyl]propionic acid; the impurity B is (2RS)-2-[4-(2-butylpropyl)phenyl]propionic acid; the formulation impurity 1 is 3-((R)-1-hydroxy-2-(methylamino)ethyl)phenyl-2-(4-isobutylphenyl)propanoate; the formulation impurity 2 is (R)-1-(3-hydroxyphenyl)-2-(methylamino)ethyl-2-(4-isobutylphenyl)propionic acid ethyl ester; and the formulation impurity 3 is N-((R)-2-hydroxy-2-(3-hydroxyphenyl)ethyl)-2-(4-isobutylphenyl)-N-methylpropionamide.

3. The detection method according to claim 1 or 2, characterized in that: The first elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 78% to 82% to 48% to 52%; the volume ratio of the mobile phase B changes from 18% to 22% to 48% to 52%; and the time of the first elution stage is 18 to 22 minutes; Optionally, the second elution stage is an isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the time of the second elution stage is 28 to 32 minutes; Optionally, the third elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 48% to 52% to 78% to 82%, and the volume ratio of the mobile phase B changes from 48% to 52% to 18% to 22%; and the time of the third elution stage is 1.8 to 2.2 minutes; Optionally, the fourth elution stage is isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the duration of the fourth elution stage is 7 to 9 minutes.

4. The detection method according to any one of claims 1 to 3, characterized in that: The first elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 80% to 50%; the volume ratio of the mobile phase B changes from 20% to 50%; and the time of the first elution stage is 20 minutes; Optionally, the second elution stage is an isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the time of the second elution stage is 30 minutes; Optionally, the third elution stage is a linear gradient elution, wherein the volume ratio of the mobile phase A changes from 50% to 80%, and the volume ratio of the mobile phase B changes from 50% to 20%; and the time of the third elution stage is 2 minutes; Optionally, the fourth elution stage is isocratic elution, wherein the volume ratio of the mobile phase A to the mobile phase B remains unchanged; and the duration of the fourth elution stage is 8 minutes.

5. The detection method according to any one of claims 1 to 4, characterized in that: The mobile phase A is selected from an aqueous solution of sodium octane sulfonate, an aqueous solution of sodium hexane sulfonate or an aqueous solution of sodium decane sulfonate; Preferably, the concentration of the mobile phase A is 3 to 3.5 g / L; Preferably, the pH of the mobile phase A is 2.2 to 3.

6. The detection method according to any one of claims 1 to 5, characterized in that: The specification of the chromatographic column is 250×4.6 mm, and the filler particle size is 5 μm.

7. The detection method according to any one of claims 1 to 6, characterized in that: The column temperature of the chromatographic column is 30 to 40°C; Optionally, the flow rate of the mobile phase is 0.8 to 1.2 mL / min; Optionally, the chromatographic conditions also include a detection wavelength of 210 to 220 nm.

8. The detection method according to any one of claims 1 to 7, characterized in that: The concentration of the mobile phase A is 3.25 g / L; Optionally, the pH of the mobile phase A is 2.6; Optionally, the column temperature of the chromatographic column is 35°C; Optionally, the flow rate of the mobile phase is 1 mL / min; Optionally, the chromatographic conditions also include a detection wavelength of 214 nm.

9. The detection method according to any one of claims 1 to 8, characterized in that: The detection method comprises the following steps: S1. dissolving the bromelain tablets to be tested in a solvent to prepare a sample solution to be tested; S2. The sample solution to be tested was diluted 400 to 600 times to obtain a control solution; S3. using the high performance liquid chromatography to detect the sample solution and the control solution; and S4. Calculate the weight percentage of each of the ibuprofen-related substances relative to ibuprofen in the ibuprofen tablet to be tested based on the peak area of ​​the ibuprofen-related substances in the sample solution to be tested and the peak area of ​​ibuprofen in the control solution.

10. The detection method according to claim 9, characterized in that: The solvent comprises acetonitrile and water in a volume ratio of 1:1; Optionally, the test sample solution contains ibuprofen at a concentration of 1 to 3 mg / mL.

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