Method for detecting related substances of dronedarone hydrochloride tablets

Abstract

The invention discloses a method for detecting related substances of dronedarone hydrochloride tablets, and belongs to the technical field of medicines. According to the method, the related substances in the dronedarone tablet are detected by adopting HPLC (High Performance Liquid Chromatography), and under chromatographic conditions, a chromatographic column is a silica gel column which is prepared by adopting an ethylene bridge (BHE) hybrid particle technology and is bonded with a carbamate group on the surface; the mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is a buffer salt-organic phase, and the volume ratio of the buffer salt to the organic phase is (40-50): (60-50); a mobile phase B: a buffer salt-organic phase, wherein the volume ratio of the buffer salt to the organic phase is (25-35): (75-65); the buffer salt is monopotassium phosphate with the concentration of 10 to 20 mmol / L; the organic phase is a mixed solvent of methanol and acetonitrile in a volume ratio of (15-25): (85-75). The detection method provided by the invention has the characteristics of strong specificity, high accuracy, good precision, good linearity, high sensitivity, good stability, good durability and the like, and is simple to operate and easy to understand and master.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and in particular relates to a method for detecting related substances of dronedarone hydrochloride tablets. Background Art

[0002] Dronedarone Hydrochloride Tablets is a safe antiarrhythmic drug originally developed by Sanofi that can significantly reduce the morbidity and mortality of patients with atrial fibrillation / atrial flutter. The drug is suitable for patients with sinus rhythm who have a history of paroxysmal or persistent atrial fibrillation to reduce the risk of hospitalization due to atrial fibrillation (AF). The structural formula of its raw material is as follows:

[0003]

[0004] The related substances of preparations mainly study the interaction between APIs and excipients, and the degradation impurities of the APIs themselves. They can provide data support for excipient screening, prescription process research, stability research, and selection of packaging and storage methods. Therefore, the detection of related substances and degradation impurities is particularly important in the entire life cycle of preparations. Therefore, accurate and reliable detection methods are the premise and important guarantee for preparation quality research.

[0005] The related substances of Dronedarone Hydrochloride Tablets include impurities A, B, C and D, among which impurities A and B are process impurities, and impurities C and D are degradation impurities, and their structural formulas are as follows:

[0006]

[0007]

[0008] Dronedarone hydrochloride tablets have been approved for marketing in China and have import registration standards (standard number: JX201000086), USP and EP standards, as well as reports on their detection methods in patents CN109975434A and CN103901117B.

[0009] The related substance detection methods involved in the statutory standards all use cyano columns, and the sweeping agent triethylamine is used in the mobile phase (but the tailing factor of dronedarone is 4.1), which will lead to a short life of the cyano column, high analysis cost, and the tailing of the main peak, which will encapsulate the degradation impurity D. In the chromatographic analysis method of related substances of dronedarone hydrochloride in CN109975434A, the mobile phase system also uses the sweeping agent triethylamine, which not only cannot solve the tailing of the main component dronedarone, but also has a great risk of unknown impurities being encapsulated by the main peak, and the service life of the chromatographic column is short. And CN103901117B is a method for detecting dronedarone hydrochloride, which uses octadecylsilane bonded silica gel, and the mobile phase is a mixture of methanol, acetonitrile and phosphate buffer (a mixed solution of potassium dihydrogen phosphate and alkaline compounds), but its chromatographic column is ordinary C18, and there is no description of the detection of impurity D.

[0010] The above disclosed related material methods cannot take into account the detection of both the symmetry of the dronedarone peak and the oxidative degradation impurity nitrogen oxides (impurity D) at the same time.

[0011] Therefore, developing an accurate detection method for related substances in dronedarone hydrochloride tablets, including impurity D, is of great significance for providing medication safety and quality control of dronedarone hydrochloride tablets. Summary of the invention

[0012] The purpose of the present invention is to establish a detection method for dronedarone hydrochloride tablets which has good separation, high accuracy, short detection time, can stably and reliably detect impurities related to dronedarone hydrochloride tablets, and can better control product quality.

[0013] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0014] A method for detecting related substances in dronedarone hydrochloride tablets adopts high performance liquid chromatography. In the chromatographic conditions, the chromatographic column is a silica gel column with surface bonded carbamate groups made by ethylene bridge (BHE) hybrid particle technology; the mobile phase includes mobile phase A and mobile phase B; mobile phase A: buffer salt-organic phase, the volume ratio of buffer salt to organic phase is 40-50:60-50; mobile phase B: buffer salt-organic phase, the volume ratio of buffer salt to organic phase is 25-35:75-65;

[0015] The buffer salt is 10-20 mmol / L potassium dihydrogen phosphate;

[0016] The organic phase is a mixed solvent of methanol and acetonitrile in a volume ratio of 15-25:85-75;

[0017] The gradient elution method was adopted, and the elution procedure was as follows:

[0018]

[0019] The related substances include impurity A, impurity B, impurity C, and impurity D;

[0020] The chromatographic conditions also include: column temperature of 30-40° C., flow rate of 1.0-1.4 ml / min; and detection wavelength of 244-248 nm.

[0021] Furthermore, the pH value of the buffer salt is 7.6±0.2; the pH value of the buffer salt can be adjusted with sodium hydroxide solution.

[0022] Furthermore, the chromatographic column may be Waters XBridge Shield RP18; the specifications of the Waters XBridgeShield RP18 column are inner diameter×column length×particle diameter, 4.6 mm×150 mm×3.5 μm;

[0023] Furthermore, the volume ratio of buffer salt to organic phase in the mobile phase A is 45:55;

[0024] Furthermore, the volume ratio of the buffer salt to the organic phase in the mobile phase B is 30:70;

[0025] Further, the column temperature is preferably 35°C;

[0026] Further, the flow rate is preferably 1.2 ml / min;

[0027] Further, the detection wavelength is preferably 246 nm;

[0028] Furthermore, the injection volume is preferably 5 to 10 μl;

[0029] Furthermore, the mobile phase buffer salt is preferably 15mmol / L potassium dihydrogen phosphate;

[0030] Further, the pH value of the mobile phase A buffer is preferably 7.6;

[0031] Furthermore, the method for detecting related substances in dronedarone hydrochloride tablets specifically comprises the following steps:

[0032] (1) Solution preparation: Prepare the solvent, system suitability solution, impurity stock solution, control solution, and test sample solution separately.

[0033] (2) Detection method: Set the chromatographic conditions, wait until the system is stable, inject the solvent, system suitability solution, control solution, and test solution into the chromatograph, and record the chromatogram;

[0034] (3) Impurity calculation formula:

[0035]

[0036]

[0037] Total impurities (%) = sum of unknown impurities (%) + sum of known impurities (%)

[0038] Among them: A 杂质 —The peak area of ​​each known impurity in the test solution; A 单杂 —The peak area of ​​an unknown single impurity in the test solution; A 对照 —Peak area of ​​control solution.

[0039] In the step (1), the solvent is a mixture of water and acetonitrile, and the volume ratio of water to acetonitrile is 40-60:60-40; in some embodiments, the volume ratio of water to acetonitrile is 50:50.

[0040] The system suitability solution in step (1) is a mixture of dronedarone hydrochloride, impurity A, impurity B, impurity C, and impurity D reference substances dissolved in a solvent, wherein the concentrations of dronedarone per ml are about 0.8 mg to 1.2 mg, and impurities A, B, C, and D are 1.6 to 2.4 μg each.

[0041] The test solution in step (1) can be prepared according to the following method:

[0042] Grind dronedarone hydrochloride tablets, weigh accurately, place in a 100ml volumetric flask, add about 70ml of solvent, sonicate for 10 minutes, cool to room temperature, dilute to scale with solvent, shake well, filter with a 0.22μm PTFE filter, discard 3ml of the initial filtrate, and take the subsequent filtrate as the test solution, the solubility of which is about 0.8mg~1.2mg dronedarone per 1ml.

[0043] The control solution in step (1) is a diluted solution of the test solution, and its solubility is about 1.6 μg to 2.4 μg of dronedarone per 1 ml. In some embodiments, the solubility of dronedarone in the control solution is 2 μg / ml.

[0044] The impurity stock solution in step (1) is a mixture of impurity A, B, C, D reference substances dissolved in a solvent, and the solubility of each impurity is about 8 μg / ml to 12 μg / ml. In some embodiments, the solubility of each impurity in the impurity stock solution is 10 μg / ml.

[0045] Compared with the prior art, the present invention has the following beneficial effects:

[0046] 1. The present invention abandons the cyano column and adopts a silica gel column made of ethylene bridge (BHE) hybrid particle technology, improves the peak shape of dronedarone by potassium phosphate buffer and methanol, increases the retention of various impurities, has a good peak shape, a long chromatographic column life, and reduces the analysis cost; establishes a related substance detection method for detecting various impurities of dronedarone hydrochloride tablets and the rarely reported impurity D under the same method.

[0047] 2. The detection method of the present invention has the characteristics of strong specificity, high accuracy, good precision, good linearity, high sensitivity, good stability, good durability of the method, etc. At the same time, it is simple to operate and easy to understand and use. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 The chromatogram of the solution for detecting the suitability of the system in Example 1;

[0049] Figure 2 The chromatogram of the test solution detected in Example 2;

[0050] Figure 3 This is the solution chromatogram of the detection system suitability in Comparative Example 1;

[0051] Figure 4 The chromatogram of the mixed solution detected in Comparative Example 2;

[0052] Figure 5 The chromatogram of the test solution detected in Comparative Example 2;

[0053] Figure 6 This is the solution chromatogram of the detection system suitability in Comparative Example 3;

[0054] Figure 7 This is the solution chromatogram of the detection system suitability in Comparative Example 4;

[0055] Figure 8 This is an enlarged chromatogram of the solution for testing the suitability of the system in Comparative Example 4. DETAILED DESCRIPTION

[0056] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention is described in detail below through specific embodiments, but the content and scope of the present invention are not limited by the following embodiments.

[0057] The reagents used in the present invention can be purchased from the market or prepared by the method described in the present invention. Among them, acetonitrile and methanol are HPLC grade, dronedarone hydrochloride tablets (specification: 400 mg) are from SANOFI AVENTISUS LLC; dronedarone hydrochloride and impurity A, B, C, D reference substances are purchased from Jiangsu Huiju Pharmaceutical Co., Ltd. Unless otherwise specified, the high performance liquid chromatograph used in the present invention is Thermo Fisher Ultimate 3000.

[0058] Example 1 System suitability test

[0059] (1) Chromatographic conditions

[0060] Chromatographic column: Waters XBridge Shield RP18 (150 mm × 4.6 mm, 3.5 μm)

[0061] Mobile phase A: buffer salt-organic phase (45:55, v / v)

[0062] Mobile phase B: buffer salt-organic phase (30:70, v / v)

[0063] Column temperature: 35°C

[0064] Flow rate: 1.2ml / min

[0065] Detection wavelength: 246nm

[0066] Injection volume: 10 μl

[0067] Gradient Program:

[0068]

[0069] (2) Solution preparation

[0070] Solvent: Take 500 ml of acetonitrile and 500 ml of water, mix well, and degas by ultrasonication.

[0071] Buffer salt: Take 2.04 g of potassium dihydrogen phosphate, add 1000 ml of water to dissolve it, adjust the pH value to 7.6 with 1 mol / L sodium hydroxide solution, filter, and degas by ultrasonication.

[0072] Organic phase: Measure 1600 ml of acetonitrile and 400 ml of methanol, mix well, and degas by ultrasonication.

[0073] Mobile phase A: Measure 450 ml of buffer salt, add 550 ml of organic phase, mix well, and degas by ultrasonication.

[0074] Mobile phase B: Measure 300 ml of buffer salt, add 700 ml of organic phase, mix well, and degas by ultrasonication.

[0075] Impurity stock solution ①: Take about 20 mg of each impurity A, B, C, and D reference substance, accurately weigh, place in a 100 ml volumetric flask, add solvent and ultrasonically dissolve, cool to room temperature, dilute to scale with solvent, shake well,

[0076] Impurity stock solution ②: Accurately measure 5 ml of impurity stock solution ①, place it in a 100 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0077] System suitability solution: Take about 26.6 mg of dronedarone hydrochloride reference substance, weigh accurately, place in a 25 ml volumetric flask, add appropriate amount of solvent and sonicate to dissolve, cool to room temperature, accurately add ②5 ml of impurity stock solution, dilute to scale with solvent, shake well, and obtain.

[0078] (3) Detection methods and test results

[0079] Set the chromatographic conditions, wait for the system to stabilize, inject the system suitability solution into the chromatograph, and record the chromatogram. Figure 1 , Table 1 is the results of system suitability solution determination.

[0080] Table 1 System suitability solution test results

[0081]

[0082]

[0083] The experimental results show that under this condition, the separation degree between the main peak and adjacent impurities in the system applicability solution is greater than 1.5, and the separation degree between each impurity is greater than 1; the dronedarone peak is symmetrical, and the tailing factor is 0.99, and the system applicability meets the requirements.

[0084] Example 2 Specificity Test

[0085] (1) Chromatographic conditions

[0086] Same as Example 1.

[0087] (2) Solution preparation

[0088] The preparation of solvent, mobile phase A and mobile phase B is the same as in Example 1.

[0089] Test solution: Take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, place in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, dilute to the scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, and take the subsequent filtrate as the test solution.

[0090] Acid destruction of the test sample: Take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, place in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, add 1 ml of 0.1 mol / L hydrochloric acid solution, leave at room temperature for 4 hours, neutralize with 1 ml of 0.1 mol / L sodium hydroxide solution, dilute to the scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, and take the subsequent filtrate to obtain.

[0091] Alkali destruction of the test sample: take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, put into a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, add 1 ml of 0.1 mol / L sodium hydroxide solution, let it stand at room temperature for 4 hours, neutralize with 1 ml of 0.1 mol / L hydrochloric acid solution, dilute to the scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, and take the subsequent filtrate to obtain.

[0092] Oxidative destruction of the test sample: Take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, place in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, add 1 ml of 3% hydrogen peroxide solution, let stand at room temperature for 4 hours, dilute to the scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, and take the subsequent filtrate to obtain.

[0093] High-temperature destruction of solid test sample: Take an appropriate amount of fine powder of dronedarone hydrochloride tablets in a weighing bottle, spread it into a thin layer ≤5 mm thick, place it in an oven at 80°C for 4 hours, take it out, cool it to room temperature, take about 165 mg (equivalent to 100 mg of dronedarone), accurately weigh it, place it in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, dilute to the scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, take the subsequent filtrate, and obtain it.

[0094] High temperature destruction of the test solution: take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, put into a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, place in a water bath at 80°C for 4 hours, take out, cool to room temperature, dilute to the scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, and take the subsequent filtrate to obtain.

[0095] Solid photodestruction of the test sample: Take an appropriate amount of fine powder of dronedarone hydrochloride tablets in a weighing bottle, spread it into a thin layer ≤5mm thick, place it under an illumination of 4500lx±500lx, and leave it for 10 days. Take about 165mg (equivalent to 100mg of dronedarone), accurately weigh it, place it in a 100ml volumetric flask, add about 50ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, dilute to the scale with solvent, shake well, filter with a 0.22μm PTFE filter, discard 3ml of the initial filtrate, and take the subsequent filtrate to obtain.

[0096] Photodestruction of the test solution: Take about 165 mg of the fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, place in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, dilute to the scale with solvent, shake well, place under 4500lx±500lx illumination for 2 days, take out, filter with a 0.22μm PTFE filter, discard 3 ml of the initial filtrate, take the subsequent filtrate to obtain.

[0097] At the same time, take blank excipients and prepare blank excipient solution according to the test sample solution, and prepare blank excipient destruction solution in the same way as the acid destruction, alkali destruction, oxidation destruction, high temperature destruction and light destruction of the test sample.

[0098] (4) Detection methods and test results

[0099] Set the chromatographic conditions, wait for the system to stabilize, inject the solvent, test solution, blank excipient and destruction solution under each condition into the chromatograph, and record the chromatogram. Figure 2 Table 2 shows the test results of the test solution, and Table 3 shows the test results of the forced destruction test of the test sample. There was no change before and after the destruction of the blank auxiliary material.

[0100] Table 2 Test results of the test solution

[0101] name Retention time (min) Tailing Factor Dronedarone 23.126 0.98

[0102] Table 3 Forced degradation results

[0103]

[0104] According to Table 2 and Figure 2 The results show that the solvent does not interfere with the detection, and the main peak of dronedarone is symmetrical. According to the results in Table 3, under the various destruction conditions of the test sample, impurities C and D increase under high temperature, oxidation and solution light destruction, and there are no obvious unseparated impurities before and after the main peak of the test sample, and the separation of each impurity is greater than 1.5; the material balance under each condition is between 99.6% and 101.7%, and the purity of the main peak meets the requirements. This method has good specificity for the determination of related substances in dronedarone hydrochloride tablets.

[0105] Example 3 Detection Limit and Quantification Limit

[0106] (1) Chromatographic conditions

[0107] Same as Example 1.

[0108] (2) Solution preparation

[0109] The preparation of solvent, mobile phase A, mobile phase B and impurity stock solution ② is the same as that in Example 1.

[0110] Dronedarone hydrochloride reference substance stock solution (10μg / ml): Take about 21.3mg of dronedarone hydrochloride reference substance, accurately weigh it, place it in a 100ml volumetric flask, add solvent and ultrasonically dissolve it, cool it to room temperature, dilute it to the scale with solvent, shake it well, and you have it. Accurately measure 5ml of the solution, place it in a 100ml volumetric flask, dilute it to the scale with solvent, shake it well, and you have it.

[0111] Quantitative limit solution: Accurately measure 1 ml of impurity stock solution ② and dronedarone hydrochloride reference stock solution, put them into the same 25 ml volumetric flask, add solvent to dilute to the scale, shake well, and obtain.

[0112] Detection limit solution: Accurately measure 3 ml of the quantitative limit solution, place it in a 10 ml volumetric flask, add solvent to dilute to the scale, shake well, and obtain.

[0113] (3) Detection methods and test results

[0114] Set the chromatographic conditions, and after the system is stable, inject the detection limit solution and the quantification limit solution into the chromatograph, and record the chromatograms. Table 4 shows the results of the quantification limit and detection limit experiments.

[0115] Table 4 Results of detection limit and quantification limit

[0116]

[0117] The above test results show that the quantitative limit and detection limit of each impurity are low and the sensitivity of the method is high.

[0118] Example 4 Linearity and Correction Factor

[0119] (1) Chromatographic conditions

[0120] Same as Example 1.

[0121] (2) Solution preparation

[0122] The preparation of solvent, mobile phase A, mobile phase B and impurity stock solution ① is the same as that in Example 1.

[0123] Dronedarone hydrochloride reference substance stock solution (0.2 mg / ml): Take about 21.3 mg of dronedarone hydrochloride reference substance, accurately weigh, place in a 100 ml volumetric flask, add solvent and ultrasonically dissolve, cool to room temperature, dilute to the scale with solvent, shake well, and obtain.

[0124] Linear stock solution: Accurately measure 5 ml of dronedarone hydrochloride reference stock solution and impurity stock solution ①, place in a 100 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0125] Linear-200% concentration solution: Accurately measure 10 ml of linear stock solution, place in a 25 ml volumetric flask, dilute to the scale with solvent, and shake well.

[0126] Linear-150% concentration solution: Accurately measure 7.5 ml of linear stock solution, place in a 25 ml volumetric flask, dilute to the scale with solvent, and shake well.

[0127] Linear-100% concentration solution: Accurately measure 5 ml of linear stock solution, place in a 25 ml volumetric flask, dilute to the scale with solvent, and shake well.

[0128] Linear-80% concentration solution: Accurately measure 4 ml of linear stock solution, place in a 25 ml volumetric flask, dilute to the scale with solvent, and shake well.

[0129] Linear-50% concentration solution: Accurately measure 2.5 ml of linear stock solution, place in a 25 ml volumetric flask, dilute to the scale with solvent, and shake well.

[0130] Linearity-quantitation limit solution: Accurately measure 1 ml of linear stock solution, place in a 25 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0131] (3) Detection methods and test results

[0132] Set the chromatographic conditions, and after the system is stable, inject each linear solution into the chromatograph and record the chromatogram. Table 5 shows the linearity and correction factor results.

[0133] Table 5 Linear range and correction factor

[0134]

[0135]

[0136] The results showed that the linear relationship of each impurity was good within the range of 0.05% to 0.40%, and the correction factor was within the range of 0.9 to 1.1, and all of them could be calculated by the self-control method without adding the correction factor.

[0137] Example 5 Accuracy

[0138] (1) Chromatographic conditions

[0139] Same as Example 1.

[0140] (2) Solution preparation

[0141] The preparation of solvent, mobile phase A and mobile phase B is the same as in Example 1.

[0142] Accuracy impurity reference stock solution (40μg / ml): Take approximately 40mg of each impurity A, B, C, and D reference substance, accurately weigh them, place them in a 100ml volumetric flask, add solvent and sonicate to dissolve them, cool to room temperature, dilute to the mark with solvent, shake well, accurately transfer 10ml of the solution, place it in a 100ml volumetric flask, dilute to the mark with solvent, and shake well.

[0143] LOQ accuracy solution: Take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, place in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, accurately add 1 ml of accuracy impurity reference stock solution, dilute to scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, and take the subsequent filtrate. Prepare 3 portions in parallel (starting from weighing).

[0144] LOQ accuracy control solution: accurately measure 1 ml of LOQ accuracy solution, place it in a 50 ml volumetric flask, dilute it to the mark with solvent, and shake well; accurately measure 5 ml of the solution, place it in a 50 ml volumetric flask, dilute it to the mark with solvent, and shake well. Take LOQ accuracy solution and prepare control solution respectively.

[0145] 100% accuracy solution: Take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, place in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, accurately add 5 ml of accuracy impurity reference stock solution, dilute to scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, take the subsequent filtrate, and prepare 3 portions in parallel (starting from weighing).

[0146] 100% accuracy control solution: accurately measure 1 ml of 100% accuracy solution, place it in a 50 ml volumetric flask, dilute it to the mark with solvent, and shake well; accurately measure 5 ml of the solution, place it in a 50 ml volumetric flask, dilute it to the mark with solvent, and shake well. Take 100% accuracy solution and prepare control solution respectively.

[0147] 150% accuracy solution: Take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), accurately weigh, place in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, accurately add 7.5 ml of accuracy impurity reference stock solution, dilute to scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, take the subsequent filtrate, and prepare 3 portions in parallel (starting from weighing).

[0148] 150% accuracy control solution: accurately measure 1 ml of 150% accuracy solution, place it in a 50 ml volumetric flask, dilute it to the mark with solvent, and shake well; accurately measure 5 ml of the solution, place it in a 50 ml volumetric flask, dilute it to the mark with solvent, and shake well. Take 150% accuracy solution and prepare control solution respectively.

[0149] (3) Detection methods and test results

[0150] Set the chromatographic conditions, wait for the system to stabilize, inject each accuracy solution into the chromatograph, and record the chromatogram. Table 6 shows the accuracy test results of each impurity.

[0151] Table 6 Accuracy results

[0152]

[0153] The results showed that the recoveries of each impurity at different concentration levels were all between 95.4% and 104.8%, and the RSDs of the recoveries were all less than 5%, indicating that the method had high accuracy.

[0154] Example 6 Repeatability and Intermediate Precision Test

[0155] (1) Chromatographic conditions

[0156] Same as Example 1

[0157] (2) Solution preparation

[0158] The preparation of solvent, mobile phase A and mobile phase B is the same as in Example 1.

[0159] Repeatability impurity reference stock solution (40μg / ml): Take about 40mg of impurity A, B, C, D reference substances, accurately weigh, place in a 100ml volumetric flask, add solvent and sonicate to dissolve, cool to room temperature, dilute to the scale with solvent, shake well, accurately transfer 10ml of the solution, place in a 100ml volumetric flask, dilute to the scale with solvent, and shake well.

[0160] Repeatability solution: Take about 165 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 100 mg of dronedarone), weigh accurately, put in a 100 ml volumetric flask, add about 70 ml of solvent, ultrasonicate for 10 minutes, cool to room temperature, accurately add 5 ml of repeatability impurity reference stock solution, dilute to scale with solvent, shake well, filter with a 0.22 μm PTFE filter, discard 3 ml of the initial filtrate, take the subsequent filtrate, and prepare 6 portions in parallel (starting from weighing).

[0161] Repeatability control solution: accurately measure 1 ml of repeatability solution, place it in a 50 ml volumetric flask, dilute it to the mark with solvent, shake well, then accurately measure 5 ml of the solution, place it in a 50 ml volumetric flask, dilute it to the mark with solvent, shake well, and you have it. Take 6 portions of repeatability solution and prepare control solutions respectively.

[0162] Intermediate precision solution and intermediate precision control solution: prepared by different test personnel together with the above repeatability solution and repeatability control solution.

[0163] (3) Detection methods and test results

[0164] Set the chromatographic conditions, and after the system is stable, inject each repetitive solution into the chromatograph and record the chromatogram.

[0165] Intermediate precision: The above operation was repeated on different days with different test personnel and different HPLC instruments (Agilent 1260 chromatograph was used). Table 7 shows the repeatability and intermediate precision results.

[0166] Table 7 Repeatability and intermediate precision results

[0167]

[0168]

[0169] The results showed that the RSD of the repeatability and intermediate precision of each impurity was less than 5%, and the detection precision of this method was high.

[0170] Example 7 Solution stability test

[0171] (1) Chromatographic conditions

[0172] Same as Example 1

[0173] (2) Solution preparation

[0174] The preparation of solvent, mobile phase A and mobile phase B is the same as in Example 1; the preparation of repeatability solution and repeatability control solution is the same as in Example 6.

[0175] (3) Detection methods and test results

[0176] Set the chromatographic conditions, and after the system is stable, inject the repetitive solutions (0h, 8h, 26h, 33h, 42h, 52h, 62h, 72h) and repetitive control solutions (0h, 11h, 29h, 65h, 77h) placed for different times into the chromatograph, and record the chromatograms. Tables 8 and 9 are the stability test results of the repetitive solution and repetitive control solution, respectively.

[0177] Table 8 Solution stability results (reproducible solution)

[0178] Time (h) Impurity A(%) Impurity B(%) Impurity C(%) Impurity D(%) 0 0.20 0.19 0.19 0.19 8 0.20 0.19 0.19 0.19 26 0.20 0.19 0.20 0.20 33 0.21 0.20 0.20 0.20 42 0.20 0.20 0.19 0.19 52 0.20 0.19 0.20 0.20 62 0.20 0.19 0.20 0.20 72 0.21 0.20 0.20 0.20 Difference % 0.01 0.01 0.01 0.01

[0179] Table 9 Solution stability test results - control solution (peak area)

[0180] Time (h) Main peak area Ratio to 0 hours 0 0.429 N / A 11 0.430 1.00 29 0.426 0.99 65 0.424 0.99 77 0.426 0.99

[0181] The results showed that the control solution was stable within 77 hours at room temperature, and the test solution was stable within 72 hours at room temperature. This method has good stability of the solution.

[0182] Example 8 Durability Test

[0183] (1) Chromatographic conditions

[0184] The chromatographic conditions of Example 1 were used as standard conditions, and single variable minor adjustments were made to the column temperature, flow rate, mobile phase pH, mobile phase ratio, etc. of the chromatographic column. The specific durability change parameters are as follows:

[0185]

[0186]

[0187] (2) Solution preparation

[0188] The preparation of solvent and system suitability solution is the same as that in Example 1.

[0189] (3) Test results

[0190] Table 10 Durability-Separation Results

[0191]

[0192] Table 11 Durability-Test Results

[0193]

[0194]

[0195] The results showed that the method had good durability in detecting related substances in dronedarone hydrochloride tablets with slight changes in different chromatographic parameters.

[0196] Comparative Example 1

[0197] The comparative example used a chromatographic column of Waters SunFire C18 4.6 mm×150 mm, 3.5 μm, and other conditions were consistent with those of Example 1.

[0198] System suitability chromatograms such as Figure 3 , Table 12 shows the results of system suitability solution determination.

[0199] Table 12 System suitability solution test results

[0200] name Retention time (min) Separation Tailing Factor Impurity D 6.776 - - Impurity C 7.585 3.8 - Dronedarone 20.746 25.5 0.68 Impurity A 22.493 2.2 - Impurity B 23.057 Unseparated -

[0201] Under this condition, impurity B and impurity A could not be baseline separated, and the peak of the main component dronedarone was extended, making it impossible to accurately determine the related substances of dronedarone hydrochloride tablets.

[0202] Comparative Example 2: Legal reporting method

[0203] This comparison refers to the imported drug registration standard JX20100086.

[0204] (1) Chromatographic conditions

[0205] Chromatographic column: ZORBAX SB-CN 4.6mm×250mm, 5μm

[0206] Mobile phase A: aqueous solution (2 ml triethylamine to 950 ml water, adjust pH to 3.0 with phosphoric acid, add water to 1000 ml)

[0207] Mobile phase B: acetonitrile

[0208] Mobile phase: Mobile phase A-Mobile phase B (50:50)

[0209] Column temperature: 30°C

[0210] Flow rate: 0.8ml / min

[0211] Detection wavelength: 246nm

[0212] Injection volume: 20 μl

[0213] Run time: 15 minutes

[0214] (2) Solution preparation

[0215] Mixed solution: Take appropriate amount of dronedarone hydrochloride reference substance and impurity A, B, C, D reference substances respectively, dissolve them in methanol, and dilute with mobile phase to make a solution containing 10 μg / ml of dronedarone and impurity A, B, C, D per 1 ml.

[0216] Test solution: Take about 660 mg of fine powder of dronedarone hydrochloride tablets (equivalent to about 400 mg of dronedarone), accurately weigh, place in a 100 ml volumetric flask, add about 70 ml of methanol, ultrasonicate for 5 minutes, cool to room temperature, dilute to the scale with methanol, shake well, let stand for 10 minutes, accurately measure 5 ml of supernatant, place in a 20 ml volumetric flask, dilute to the scale with mobile phase, shake well, filter with a filter membrane, and take the filtrate.

[0217] (3) Detection methods and test results

[0218] Set the chromatographic conditions, wait until the system is stable, inject the mixed solution and the test solution into the chromatograph respectively, and record the chromatogram. Figure 4 is the mixed solution spectrum, Figure 5 Table 13 is the test result of the mixed solution, and Table 14 is the test result of the test solution.

[0219] Table 13 Mixed solution measurement results

[0220] name Retention time (min) Separation Impurity C 5.126 - Impurity A 5.973 4.7 Dronedarone 7.721 12.0 Impurity D 7.870 Unseparated Impurity B 9.874 12.1

[0221] Table 14 Test results of the test solution

[0222] name Retention time (min) Tailing Factor Dronedarone 7.114 4.1

[0223] Under this condition, impurity D and the main component dronedarone are not separated, and the content of impurity D cannot be accurately determined. Triethylamine in the mobile phase will reduce the service life of the cyano column and cause the chromatographic column to be easily worn.

[0224] Comparative Example 3

[0225] This comparative example is set up with reference to the detection method of Example 13 in CN103901117B.

[0226] (1) Chromatographic conditions

[0227] Chromatographic column: Agilent XDB-C18 4.6mm×150mm, 5μm

[0228] Mobile phase: methanol-acetonitrile-phosphate (pH 7.5) (32:50:18)

[0229] Column temperature: 40°C

[0230] Flow rate: 1.5ml / min

[0231] Detection wavelength: 220nm

[0232] Injection volume: 20 μl

[0233] Run time: 30 minutes

[0234] (2) Solution preparation

[0235] The preparation of the system suitability solution is the same as in Example 1.

[0236] (3) Detection methods and test results

[0237] Set the chromatographic conditions, wait for the system to stabilize, inject the system suitability solution into the chromatograph, and record the chromatogram. Figure 6 , Table 15 is the system suitability solution test results.

[0238] Table 15 System suitability solution test results

[0239] name Retention time (min) Separation Impurity D 2.130 - Impurity C 2.679 10.3 Dronedarone 15.294 30.9 Impurity B 18.513 4.8 Impurity A 20.023 2.0

[0240] Under this condition, impurity D is weakly retained and the dead volume segment is eluted, making it impossible to stably and accurately determine the content of impurity D.

[0241] Comparative Example 4

[0242] This comparative example is set up with reference to the detection method of Example 1 in CN109975434A.

[0243] (1) Chromatographic conditions

[0244] Chromatographic column: Ultimate XB-C18 4.6mm×250mm, 5μm

[0245] Mobile phase A: 0.5% triethylamine aqueous solution (pH 4.0 adjusted with phosphoric acid)

[0246] Mobile phase B: acetonitrile

[0247] Column temperature: 30°C

[0248] Flow rate: 0.8ml / min

[0249] Detection wavelength: 246nm

[0250] Injection volume: 20 μl

[0251] Gradient Program:

[0252]

[0253] (2) Solution preparation

[0254] The preparation of the system suitability solution is the same as in Example 1.

[0255] (3) Test results

[0256] Set the chromatographic conditions, wait for the system to stabilize, inject the system suitability solution into the chromatograph, and record the chromatogram. Figure 7 , and its enlarged chromatogram is as follows Figure 8 , Table 16 is the system suitability solution test results.

[0257] Table 16 System suitability solution test results

[0258] name Retention time (min) Separation Tailing Factor Impurity C 11.780 - - Impurity A 19.352 25.3 - Dronedarone 20.437 2.7 3.6 Impurity D 21.894 3.8 - Unknown impurities 22.775 3.5 - Impurity B 23.034 1.0 -

[0259] Under this condition, the chromatographic column and gradient program were optimized on the basis of the statutory method, but the main peak tailing factor was 3.6, with no obvious improvement, far from meeting the main peak tailing requirement of 0.9-1.1 for the chromatographic system. There is a high possibility that unknown impurities are contained in the main peak, and the related substances of dronedarone tablets cannot be accurately determined.

Claims

1. A method for detecting related substances in dronedarone hydrochloride tablets, characterized in that: The method adopts high performance liquid chromatography, wherein the chromatography column is a silica gel column with surface bonded carbamate groups made by ethylene bridge hybrid particle technology; the mobile phase includes mobile phase A and mobile phase B; mobile phase A: buffer salt-organic phase, the volume ratio of buffer salt to organic phase is 40-50:60-50; mobile phase B: buffer salt-organic phase, the volume ratio of buffer salt to organic phase is 25-35:75-65; The buffer salt is 10-20 mmol / L potassium dihydrogen phosphate; The organic phase is a mixed solvent of methanol and acetonitrile in a volume ratio of 15-25:85-75; A gradient elution method was used.

2. The detection method according to claim 1, characterized in that: The elution procedure of the gradient elution is as follows:

3. The detection method according to claim 1 or 2, characterized in that: The related substances of the dronedarone hydrochloride tablets include impurity A, impurity B, impurity C, and impurity D, and their structural formulas are as follows:

4. The detection method according to claim 1, characterized in that: The chromatographic conditions also include: the column temperature of the chromatographic column is 30-40° C., the flow rate is 1.0-1.4 ml / min, and the detection wavelength is 244-248 nm.

5. The detection method according to claim 1 or 4, characterized in that: The chromatographic conditions are: column temperature: 35°C; flow rate: 1.2 ml / min; detection wavelength: 246 nm.

6. The detection method according to claim 1, characterized in that: The chromatographic column is a Waters XBridgeShield RP18 column.

7. The detection method according to claim 6, characterized in that: The specifications of the Waters XBridge Shield RP18 column are inner diameter×column length×particle diameter: 4.6 mm×150 mm×3.5 μm.

8. The detection method according to claim 1, characterized in that: The volume ratio of the buffer salt to the organic phase in the mobile phase A is 45:55, and the volume ratio of the buffer salt to the organic phase in the mobile phase B is 30:

70.

9. The detection method according to claim 1, characterized in that: The pH value of mobile phase A buffer was 7.6 ± 0.

2.

10. The detection method according to claim 1, characterized in that: The chromatographic conditions also include: the injection volume is 5 to 10 μl.

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