阅读说明：本技术 一种利伐沙班中有关物质的分离方法 (Method for separating related substances in rivaroxaban ) 是由 邹金球 张国丽 何莉 于 2019-11-25 设计创作，主要内容包括：本发明涉及一种利伐沙班中有关物质的分离方法,该方法采用高效液相色谱法,色谱柱为C18柱,使用磷酸溶液和乙腈的混合溶液为流动相进行梯度洗脱。该方法可以同时对利伐沙班片中利伐沙班及其有关物质进行分离检测,所述有关物质包括原料合成当中带入的工艺杂质,制剂制备工艺当中产生的工艺降解杂质。与现有技术相比,本发明杂质研究更加全面,且分离方法操作简单、方便、杂质分离度好、灵敏度高、环境友好。(The invention relates to a method for separating related substances in rivaroxaban, which adopts high performance liquid chromatography, adopts a C18 column as a chromatographic column, and adopts a mixed solution of phosphoric acid solution and acetonitrile as a mobile phase for gradient elution. The method can simultaneously separate and detect rivaroxaban and related substances thereof in the rivaroxaban tablets, wherein the related substances comprise process impurities brought in the raw material synthesis and process degradation impurities generated in the preparation process of the preparation. Compared with the prior art, the method has the advantages of more comprehensive impurity research, simple and convenient separation method operation, good impurity separation degree, high sensitivity and environmental friendliness.)

1. A method for separating related substances in rivaroxaban is high performance liquid chromatography, wherein a chromatographic column is a C18 chromatographic column, and is characterized in that gradient elution is carried out by taking 0.01mol/L phosphoric acid solution as a mobile phase A and acetonitrile as a mobile phase B, and the pH value of the mobile phase A is 1.5-2.0.

2. The separation method according to claim 1, wherein the procedure of the gradient elution is as shown in the following table:

3. the separation method according to claim 2, wherein the procedure of the gradient elution is as shown in the following table:

4. the separation process of claim 1, wherein the mobile phase a has a pH of 1.8.

5. The separation method according to any one of claims 1 to 4, wherein the analysis conditions of the liquid chromatography are one or more of the following (i) to (viii):

the column has a size of 4.0 x 55mm, 3um,

(ii) the chromatography column is of the type Purospher Star RP-18 Endcapped,

(iii) the column temperature is 45. + -. 5 ℃ C,

(iv) variation range of mobile phase ratio: plus or minus 2 percent of the total weight of the mixture,

(v) a flow rate of 0.9 to 1.1ml/min,

(vi) the sample injection amount is 3-8 mul,

(vii) an elution time of 30 to 60min,

(viii) a wavelength range of 230 to 270 nm.

6. Use of the separation method according to any one of claims 1 to 4 for the separation of pharmaceutical impurities.

Technical Field

The invention belongs to the field of medical analysis, and particularly relates to a method for separating and detecting related substances in rivaroxaban by using a high performance liquid chromatography.

Background

Rivaroxaban (rivaroxban) the first orally bioavailable factor Xa inhibitor worldwide, developed by the cooperation of hadron and bayer. The trade name is Bylermon, which is approved to be on the market by more than 120 countries in the world. Can be used for preventing and treating venous thrombosis. Compared with the common warfarin medicine, the rivaroxaban has the advantages of good absorption, stable blood concentration, less interference by medicines and food, quick response and quick excretion, does not need to go to a hospital regularly to test the blood coagulation function, does not need to adjust the dosage generally, and is convenient for patients to use. Rivaroxaban is chemically named: 5-chloro-nitrogen- ({ (5S) -2-oxo-3- [ -4- (3-oxo-4-morpholinyl) phenyl ] -1, 3-oxazolidin-5-yl } methyl) -2-thiophene-carboxamide, structural formula:

research on related substances is one of key problems in the research on the quality of the preparation, and many reports are made on the research on related substances of rivaroxaban at home and abroad. The difficulty of detecting related substances of rivaroxaban is that technological impurities are generated in the rivaroxaban bulk drug synthesis process, preparation degradation impurities are generated in the preparation production process, degradation impurities are also generated in the drug storage and transportation process, and the publicly determined rivaroxaban impurities comprise more than ten kinds of acetyl oxamide, oxalamide urea, dechlorinated compounds, oxophthaloyl, 4, 5-dichloro compounds, diamide, triamide, oxophthalimide and the like. Many impurities are difficult to separate, and some impurities have large polarity and some impurities have small polarity and are difficult to detect in the same chromatographic system.

The CN108152412A patent researches a method for separating and determining rivaroxaban and related substances thereof by using a liquid chromatography, the method adopts a high performance liquid chromatography to separate and detect rivaroxaban and 13 impurities thereof, a chromatographic column is an octadecyl silane bonded silica gel chromatographic column, and a mobile phase is acetonitrile, methanol, buffer salt, an ion pair reagent and a phosphoric acid aqueous solution. The impurities detected by the method are many process impurities, no degradation impurities are detected, the method is complex in mobile phase components and troublesome in preparation, and a surfactant ion pair reagent is required to be used, so that the method is environment-friendly.

The CN105651871A patent researches a method for separating and detecting rivaroxaban related substances by using high performance liquid chromatography, and the method adopts a buffer saline solution and acetonitrile as mobile phases to carry out gradient elution to separate the related substances. The method has less related substances separated and no degradation impurity.

The CN105259282A patent researches a method for separating and searching related substances of rivaroxaban by using a high performance liquid chromatography, and the method adopts a buffer salt solution and an organic phase ion pair as a mobile phase to carry out gradient elution and separation on the related substances. The method has the advantages of less related substances for separation and detection, no degradation impurity detection, use of surfactant ion pair reagents, and no environmental pollution.

The CN110057942A patent researches a method for detecting rivaroxaban and related substances of its preparation by high performance liquid chromatography, which uses buffered saline solution and acetonitrile as mobile phase to perform gradient elution to separate related substances. The method detects more related substances, but only detects one alkali degradation impurity, and the gradient program design is complex.

Therefore, it is necessary to develop a rivaroxaban related substance detection method which is simple and convenient in operation method, good in impurity separation degree, high in sensitivity and environment-friendly, especially to research on degraded impurities, guidance can be provided for the quality of the rivaroxaban preparation and further formulation of reasonable quality standards, a basis is provided for conditions of various links such as storage, transportation and use of the rivaroxaban preparation, and the quality and stability of the preparation are ensured.

Disclosure of Invention

The invention aims to provide a method for separating rivaroxaban tablet related substances, which can accurately and efficiently detect and separate related substances in rivaroxaban tablets, particularly acid degradation impurities and alkali degradation impurities, and is simple to operate and environment-friendly.

The technical scheme of the invention is as follows: and detecting and separating rivaroxaban and related substances in the rivaroxaban tablets by adopting a high performance liquid chromatography. The structural formula of rivaroxaban and related substances is as follows:

chromatographic conditions are as follows: the chromatographic column is a C18 column, 0.01mol/L phosphoric acid solution is a mobile phase A, the pH value of the mobile phase A is 1.5-2.0, and acetonitrile is a mobile phase B.

The particle size of octadecyl bonded silica gel particles is 4 mu m, the column length is 55mm, the temperature of a chromatographic column is 45 +/-5 ℃, the flow rate is 0.9-1.1 ml/min, the sample injection amount is 3-8 mu l, the elution time is 30-60 min, the wavelength range is 230-270 nm, and the elution procedure is gradient elution.

The procedure for gradient elution was set as follows:

time (min)	A	B
			0	90%~98%	10%~2%
23	49%~82%	51%~18%
			30	90%~98%	10%~2%

Further, it is preferable that the procedure of gradient elution is set as follows:

time (min)	A	B
			0	92%	8%
23	49%	51%
			25	92%	8%
30	92%	8%

Further, it is preferable that the conditions of chromatography are such that the pH of the mobile phase A is 1.8, the column temperature is 45 ℃, the flow rate is 1.0ml/min, the amount of sample is 5. mu.l, the elution time is 30min, and the wavelength is 250 nm.

The advantages of detecting and separating related substances in the rivaroxaban tablets by adopting the method are as follows:

1. the method is used for separating related substances in rivaroxaban, and the rivaroxaban and the related substances thereof have the advantages of good separation degree of each component, good peak shape, high sensitivity, strong reproducibility and detection result closer to a real result;

2. by adopting the method for separating rivaroxaban related substances, not only can intermediate impurities and process impurities be separated, but also acid and alkali degradation impurities which have great influence on the quality standard of the preparation can be separated.

3. Compared with the prior art, the method has the advantages of simple and convenient operation, simple mobile phase components and environmental protection.

Drawings

FIG. 1 is a chromatogram of a test sample of example 1;

FIG. 2 is a chromatogram of the mixed solution of example 1;

FIG. 3 is a chromatogram of a sample of example 2

FIG. 4 is a chromatogram of the mixed solution of example 2;

FIG. 5 is a chromatogram of a test sample of example 3;

FIG. 6 is a chromatogram of the mixed solution of example 3;

FIG. 7 is a chromatogram of the test sample of example 4;

FIG. 8 is a chromatogram of the mixed solution of example 4.

Detailed Description

The present application is described in further detail below with reference to specific embodiments and the attached drawings. The following examples are intended to be illustrative of the present application only and should not be construed as limiting the present application.

The reagents and starting materials used in the present invention are commercially available.

Unless otherwise specified, the following examples are set forth with reference to a high performance liquid chromatograph: agilent (1260 Infinity II), column was Purospher Star RP-18 Endcapsed (55 × 4.0mm, 3 μm).