Separation and purification method of chiral compound

Abstract

The invention belongs to the technical field of separation and analysis, and particularly relates to a separation and purification method of a chiral compound. The invention provides a separation and purification method of a chiral compound, which comprises the following steps: separating a chiral compound sample by using an axial electromagnetic field chromatography device to obtain a target chiral compound and racemization impurities; the axial electromagnetic field chromatography device comprises a separation system, wherein the separation system comprises a chromatographic column and an electromagnetic field system; the electromagnetic field system comprises a conductive spiral coil, the conductive spiral coil is arranged on the periphery of the chromatographic column in a surrounding manner, and the conductive spiral coil can generate an electromagnetic field along the axial direction of the chromatographic column. The method provided by the invention can realize effective separation of chiral biological macromolecular substances, and has the characteristics of high separation efficiency, good selectivity, high detection sensitivity, automatic operation and wide application.

Description

Technical Field

[0001] The invention belongs to the technical field of separation and analysis, and in particular relates to a method for separating and purifying chiral compounds. Background Art

[0002] At present, liquid chromatography uses liquid as the mobile phase and utilizes the differences in chemical forces of different substances on the stationary phase, including differences in distribution or adsorption and other forces, to form differences in elution on the chromatogram to achieve separation. Its essence is to utilize the chemical forces between substances. However, traditional liquid chromatography has a single separation mechanism, and it is difficult to achieve effective separation analysis and preparation amplification for different compounds, especially chiral compounds with similar structures and physicochemical properties.

[0003] Capillary electro chromatography (CEC) is a new and efficient separation and analysis technology that has emerged in recent years. It uses electroosmotic flow or electroosmotic flow combined with pressure flow to drive the mobile phase. It has the dual separation mechanisms of capillary electrophoresis and high performance liquid chromatography and can separate both charged and neutral substances.

[0004] However, the traditional electrochromatographic device consists of a high-voltage power supply, a capillary column with fillers (filling column) and a detection device. As the filler particle size in the filling column becomes smaller and smaller, the tiny particle size fillers are more likely to aggregate and clog the column when separating biochiral macromolecules, thus greatly reducing the service life of the analyzer. Summary of the invention

[0005] The purpose of the present invention is to provide a method for separating and purifying chiral compounds. The method for separating and purifying chiral compounds provided by the present invention can achieve effective separation of chiral biomacromolecules, and has the characteristics of high separation efficiency, good selectivity, high detection sensitivity, automated operation and wide application.

[0006] In order to achieve the above object, the present invention provides the following technical solutions: The present invention provides a method for separation and purification of chiral compounds, which uses an axial electromagnetic field chromatography device to separate chiral compound samples to obtain target chiral compounds and racemic impurities; the axial electromagnetic field chromatography device comprises a separation system, which comprises a chromatographic column and an electromagnetic field system; the electromagnetic field system comprises a conductive spiral coil, which is arranged around the outer periphery of the chromatographic column, and can generate an electromagnetic field along the axial direction of the chromatographic column.

[0007] Preferably, the south pole of the electromagnetic field generated by the conductive spiral coil is located at the inlet end of the chromatographic column, and the north pole of the electromagnetic field generated by the conductive spiral coil is located at the outlet end of the chromatographic column.

[0008] Preferably, the method comprises the following steps: The electromagnetic field system is started to generate an electromagnetic field along the axis of the chromatographic column in the chromatographic column; the mobile phase and the chiral compound sample enter the chromatographic column from the inlet end of the chromatographic column, and the target chiral compound and racemic impurity are respectively obtained from the outlet end of the chromatographic column.

[0009] Preferably, the chiral compound sample is a chiral polypeptide drug.

[0010] Preferably, the chemical structure of the target chiral compound is: .

[0011] Preferably, the mobile phase used for separation includes phase A and phase B; the phase A is an acetic acid aqueous solution, and the volume content of acetic acid in the acetic acid aqueous solution is 0.1%; the phase B is acetonitrile; The gradient elution procedure for the separation is: phase B 5% (v:v) to 5% (v:v) within 5 min; from 5.1 min to 25 min, phase B increases from 5% (v:v) to 35% (v:v), and phase B is maintained at 35% (v:v) for at least Tmin; from 25.1+Tmin to 36 min, phase B increases from 35% (v:v) to 95% (v:v), and phase B is maintained at 95% (v:v) for at least 6 min.

[0012] Preferably, the intensity of the electromagnetic field is 0~12mT and is not 0.

[0013] Preferably, the electromagnetic field system further includes a DC power supply, and the voltage of the DC power supply is 0~36V and is not 0.

[0014] Preferably, the axial electromagnetic field chromatography device also includes an infusion system, a quantitative injection system, a detection system and a data processing system; the output end of the infusion system is connected to the quantitative injection system, the downstream of the quantitative injection system is connected to the inlet end of the chromatographic column, the outlet end of the chromatographic column is connected to the detection system, and the detection system is connected to the data processing system; the quantitative injection system includes a quantitative injection valve.

[0015] Preferably, the column tube material of the chromatographic column includes glass, plastic or resin.

[0016] The present invention provides a method for separation and purification of chiral compounds, wherein an axial electromagnetic field chromatography device is used to separate a chiral compound sample to obtain a target chiral compound and a racemic impurity; the axial electromagnetic field chromatography device comprises a separation system, the separation system comprises a chromatographic column and an electromagnetic field system; the electromagnetic field system comprises a conductive spiral coil, the conductive spiral coil is arranged around the periphery of the chromatographic column, and the conductive spiral coil can generate an electromagnetic field along the axial direction of the chromatographic column. The axial electromagnetic field chromatography device provided by the present invention utilizes the large-scale and efficient application of liquid chromatography in analysis and separation, and on the basis of relying on the chemical forces between substances to achieve separation, utilizes the electromagnetic field formed by the conductive spiral coil to apply an electromagnetic field on the chromatographic column, so that the chiral compound sample is subjected to a magnetic field of a certain magnitude and direction during the chromatographic separation process, and due to the structural differences of the chiral compound sample itself and the polarity differences of the chiral compound sample, the electromagnetic field exerts different directions and magnitudes of the magnetic force on the target chiral compound and the racemic impurity in the chiral compound sample, thereby utilizing the difference in the moving speed of the chiral compound sample under the action of the electromagnetic field and the difference between the chemical forces of conventional liquid chromatography to achieve effective separation of the chiral compound sample. Moreover, the axial electromagnetic field chromatography device provided by the present invention adopts an external electromagnetic field system, and the conductive spiral coil is arranged around the outer periphery of the chromatographic column, and the electromagnetic field strength and direction can be adjusted and controlled, that is, the electromagnetic field direction and magnitude can be adjusted steplessly, so that the axial electromagnetic field chromatography device provided by the present invention superimposes the separation effect of the electromagnetic field force on the basis of a single chromatographic separation mechanism for different compounds, thereby showing a more efficient separation ability than a single chromatographic separation mechanism. A method for separating and purifying chiral compounds provided by the present invention has the following beneficial effects: The axial electromagnetic field chromatography device used in the present invention is simple, has good versatility, is suitable for industrialization, and has good compatibility with the process of ordinary preparative chromatography. Only slight adjustments are made to the chromatographic column and an external electromagnetic field system is added. The injection system, detection system, infusion system, etc. of the remaining chromatographic devices are all universal equipment.

[0017] In the present invention, since the electromagnetic field magnetic force acts on the chiral compound, the electromagnetic field driving force is superimposed on the fluid driving force, so that the chiral compound needs less fluid to flow out of the liquid chromatography than the conventional liquid chromatography, and the speed of achieving effective separation will be faster. That is, when treating the same mixed compound, due to the increase in the separation driving force and the increase in the dimension of the separation driving force, the material separation drive is simultaneously affected by multiple cross-dimensions under the premise of combining two separation mechanisms, and the separation effect is stronger than that of a single separation mechanism. At the same time, less solvents and reagents are required, and the time required is shorter, that is, it has better economy and higher work efficiency.

[0018] Furthermore, in the present invention, the electromagnetic field system further comprises a DC power supply, and the voltage of the DC power supply is 0-36V, and is not 0. The axial electromagnetic field chromatography device provided by the present invention has high safety, and the voltage of the DC power supply of the electromagnetic field system does not exceed 36V, does not involve high voltage conditions, and is within the safe voltage range for the human body.

[0019] In summary, the separation and purification method of chiral compounds provided by the present invention has high separation efficiency, good selectivity, high detection sensitivity, automated operation, wide application, and also has the characteristics of electrochromatography, and has significant discrimination, separation, and enrichment effects on chiral compounds due to the difference in the size of the electromagnetic field. At the same time, the present invention realizes the change of the direction and size of the electromagnetic field in the axial direction of the fluid through an external electromagnetic field system. For different compounds, especially compounds with similar structures and physicochemical properties, due to the difference in the forward and reverse magnetic fields of the compounds themselves, efficient separation is achieved under the dual action mechanism of magnetic field drive and chromatographic separation mechanism drive. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Analyze and detect the crude peptide (EYH008-002-0801); Figure 2 This is the conventional liquid chromatography purification spectrum in Comparative Example 1; Figure 3 This is the analysis and detection spectrum after conventional liquid chromatography purification in Comparative Example 1; Figure 4 The purification spectrum of axial electromagnetic field chromatography in Example 1; Figure 5 This is the analysis and detection spectrum after axial electromagnetic field chromatography purification in Example 1; Figure 6 A schematic diagram of the structure of the axial electromagnetic field chromatography device provided by the present invention; Figure 7 A schematic diagram of a partially enlarged structure of an axial electromagnetic field chromatography device provided by the present invention; In the figure: 1 is a liquid reservoir, 2 is an infusion pump, 3 is a sample bottle, 4 is a quantitative injection system, 5 is a separation system, 6 is a detection system, 7 is a waste liquid bottle, and 8 is a data processing system. DETAILED DESCRIPTION

[0021] The present invention provides a method for separation and purification of chiral compounds, which uses an axial electromagnetic field chromatography device to separate chiral compound samples to obtain target chiral compounds and racemic impurities; the axial electromagnetic field chromatography device comprises a separation system, which comprises a chromatographic column and an electromagnetic field system; the electromagnetic field system comprises a conductive spiral coil, which is arranged around the outer periphery of the chromatographic column, and can generate an electromagnetic field along the axial direction of the chromatographic column.

[0022] Figure 6 A schematic diagram of the structure of the axial electromagnetic field chromatography device provided by the present invention; Figure 7 This is a partial enlarged structural diagram of the axial electromagnetic field chromatography device provided by the present invention. Figure 6 and Figure 7 , the axial electromagnetic field chromatography device provided by the present invention is described in detail.

[0023] The axial electromagnetic field chromatography device provided by the present invention comprises a separation system 5. The separation system comprises a chromatography column and an electromagnetic field system; the electromagnetic field system comprises a conductive spiral coil, which is arranged around the periphery of the chromatography column and can generate an electromagnetic field along the axial direction of the chromatography column.

[0024] As one or more embodiments of the present invention, the south pole of the electromagnetic field generated by the conductive spiral coil is located at the inlet end of the chromatographic column, and the north pole of the electromagnetic field generated by the conductive spiral coil is located at the outlet end of the chromatographic column.

[0025] As one or more embodiments of the present invention, the intensity of the electromagnetic field is 0~12mT and is not 0.

[0026] As one or more embodiments of the present invention, the electromagnetic field system further includes a DC power supply, and a voltage of the DC power supply is 0-36V and is not 0.

[0027] As one or more embodiments of the present invention, the column tube material of the chromatographic column includes glass, plastic or resin.

[0028] As one or more embodiments of the present invention, the specifications of the chromatographic column are 250mm×10mm, 250mm×20mm, 250mm×21.2mm, 250mm×30mm or 250mm×50mm, and the filler is C18.

[0029] As one or more embodiments of the present invention, the particle size of the C18 is 7-20 μm, specifically 7 μm, 8 μm, 10 μm or 20 μm.

[0030] As one or more embodiments of the present invention, the pore size of the filler is preferably 100-200Å, and the embodiments may be 100Å, 120Å, 150Å or 200Å.

[0031] The axial electromagnetic field chromatography device provided by the present invention preferably also includes an infusion system. The infusion system includes a liquid reservoir 1 and an infusion pump 2. The liquid reservoir 1 is used to store the mobile phase.

[0032] The axial electromagnetic field chromatography device provided by the present invention preferably further comprises a quantitative injection system 4. The output end of the infusion system is connected to the quantitative injection system 4, and the downstream of the quantitative injection system 4 is connected to the inlet end of the chromatography column.

[0033] As one or more embodiments of the present invention, the quantitative injection system 4 includes a quantitative injection valve.

[0034] The axial electromagnetic field chromatography device provided by the present invention preferably further comprises a detection system 6. The outlet end of the chromatography column is connected to the detection system 6, and the detection system 6 is connected to the data processing system.

[0035] As one or more embodiments of the present invention, the detection system 6 includes a detector.

[0036] As one or more embodiments of the present invention, the detection system 6 is connected to a waste liquid bottle 7 .

[0037] The axial electromagnetic field chromatography device provided by the present invention preferably also includes a data processing system 8 .

[0038] The axial electromagnetic field chromatography device provided by the present invention utilizes a conductive spiral coil to form an electromagnetic field, and applies an electromagnetic field to a chromatographic column, so that the compound is subjected to a magnetic field of a certain magnitude and direction during the chromatographic separation process. Due to the difference in the structure of the chiral compound itself and the difference in the polarity of the chiral compound, under the conditions of a certain direction and a certain magnetic field strength, due to the difference in the forward and reverse magnetism with the electromagnetic field, the compound has a difference in the moving speed of different chiral compounds under the action of the electromagnetic field, and the difference in the chemical action force of the liquid chromatography is superimposed to achieve the separation of different compounds. Since different chiral compounds have a certain spin polarization direction, they themselves will form a small magnetic field at the microscopic level, with a magnetic field direction and magnitude, and under the action of the external magnetic field, the chiral compound Due to their own magnetic field characteristics, chiral compounds will show obvious paramagnetism or antimagnetism. For a pair of chiral compounds, their molecular configurations are chiral symmetric and their own magnetic field directions are opposite. Therefore, the racemic compound is in a racemic state in terms of optical rotation. In the magnetic field, due to the opposite symmetry, they cancel each other out and are also in a demagnetized state. However, under the external electromagnetic field device, due to the external magnetic field, the racemic mixed chiral compound is subjected to uneven force due to the paramagnetism and antimagnetism of its own magnetic field and the external magnetic field. The chiral compound with the same direction as the external magnetic field is subjected to the force in the direction of the magnetic field, while its enantiomer is subjected to the force in the direction of the antimagnetic field due to the opposite magnetic field direction. Therefore, after the racemic mixture (i.e., the sample) passes through the magnetic field, the force between the pair of enantiomers differs by at least 2∆F 磁, when subjected to other chromatographic forces at the same time, the racemic enantiomers can be effectively separated after passing through this special electromagnetic field liquid chromatography, and this separation difference can be achieved by adjusting the chromatographic parameters or adjusting the magnetic field parameters. For example, adjusting the size of the external magnetic field can significantly change and does not require special design of the chromatographic column, and the electromagnetic field intensity and direction can be controlled, that is, the electromagnetic field direction and electromagnetic field intensity can be adjusted by steplessly adjusting the electromagnetic field direction and size, so that different parameters can be adjusted for different compounds to achieve effective separation.

[0039] In the present invention, unless otherwise specified, all preparation raw materials / components are commercially available products well known to those skilled in the art.

[0040] In the present invention, the chiral compound sample is a racemic chiral compound.

[0041] In the present invention, the method for separating and purifying the chiral compound preferably comprises the following steps: The electromagnetic field system is started to generate an electromagnetic field along the axis of the chromatographic column in the chromatographic column; the mobile phase and the chiral compound sample enter the chromatographic column from the inlet end of the chromatographic column, and the target chiral compound and racemic impurity are respectively obtained from the outlet end of the chromatographic column.

[0042] In the present invention, the electromagnetic field system is started and at the same time, the mobile phase and the chiral compound sample enter the chromatographic column from the inlet end of the chromatographic column.

[0043] In an embodiment of the present invention, the method for separating and purifying the chiral compound specifically includes the following steps: starting the electromagnetic field system to generate an electromagnetic field along the axis of the chromatographic column in the chromatographic column; using an infusion system to transport the mobile phase to the quantitative injection system 4, and entering the chromatographic column together with the sample from the inlet end of the chromatographic column to start adsorption, elution, and separation, and obtaining the target chiral compound and racemic impurities from the outlet end of the chromatographic column.

[0044] In the present invention, the chiral compound sample is preferably a chiral polypeptide drug.

[0045] In an embodiment of the present invention, the chemical structure of the target chiral compound is: .

[0046] In an embodiment of the present invention, the chemical structure of the racemic impurity is: .

[0047] In the present invention, the mobile phase includes phase A and phase B. The phase A is preferably an acetic acid aqueous solution, and the volume content of acetic acid in the acetic acid aqueous solution is preferably 0.1%. The phase B is preferably acetonitrile. The injection flow rate is preferably 15 mL / min.

[0048] In the present invention, before the elution, the present invention preferably uses a mobile phase to balance the chromatographic column. The volume ratio of phase A and phase B used to balance the chromatographic column is preferably 95:5. The time for balancing the chromatographic column is preferably 10-15 minutes.

[0049] In the present invention, the elution is preferably performed by gradient elution. The elution procedure is preferably: 5% (v:v) to 5% (v:v) of phase B within 5 min; from 5.1 min to 25 min, phase B increases from 5% (v:v) to 35% (v:v), and phase B is maintained at 35% (v:v) for at least Tmin; from 25.1+Tmin to 36 min, phase B increases from 35% (v:v) to 95% (v:v), and phase B is maintained at 95% (v:v) for at least 6 min. The Tmin is preferably 3 min, and the 25.1+Tmin is preferably 28.1 min.

[0050] In the present invention, the wavelength of the detector is preferably dual wavelength, and the dual wavelength is preferably 220 nm and 254 nm.

[0051] In the present invention, the voltage of the DC power supply is preferably 20V, and the intensity of the electromagnetic field is preferably 10mT. In the present invention, the sample moves from the inlet end of the chromatographic column to the outlet end as the fluid is eluted, and is affected by the conventional chromatographic force and the electromagnetic field. Due to the difference in the charged properties of different compounds in the sample, the external magnetic field parallel to the fluid direction applies different magnetic field forces to different compounds in the chromatographic column, and the mixed substances in the sample gradually change from being mixed into a mass to moving differently, and finally the elution of each compound in the sample is achieved in sequence.

[0052] In order to further illustrate the present invention, the technical solutions provided by the present invention are described in detail below in conjunction with embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0053] The raw materials in the following Example 1 and Comparative Example 1 are crude polypeptides; Among them, the chemical structure of the target product in the crude peptide is: .

[0054] The chemical structure of the main impurities (racemic impurities) in the crude peptide is: .

[0055] Comparative Example 1 (conventional liquid chromatography cannot effectively separate): (1) Sample processing: The crude peptide product obtained by solid phase synthesis has a purity (area normalization method) of 40.97%. The crude chiral peptide drug product (EYH008-002-0801) is about 300 mg. The analytical detection spectrum of the crude peptide product (EYH008-002-0801) is as follows: Figure 1 The analysis and detection methods of the crude polypeptide (EYH008-002-0801) are shown in Table 1.

[0056] Table 1 Analysis and detection methods of crude peptide (EYH008-002-0801)

[0057] The crude polypeptide was dissolved in methanol at a concentration of 2 g / L, and then filtered through a 0.45 μm organic filter membrane to obtain a crude polypeptide filtrate.

[0058] (2) Chromatographic separation: About 60 mL (about 120 mg) of the crude polypeptide solution was purified and separated by a liquid chromatography system. The chromatographic column specifications were 250 mm × 20 mm, the filler was C18, 20 μm, 100 Å, and the mobile phase A was: 0.1% (v:v) acetic acid aqueous solution; the mobile phase B was: acetonitrile; the mobile phase volume flow rate was: 15 mL / min; the chromatographic column was equilibrated with 5% (v:v) phase B and 95% (v:v) phase A for 10 min. Elution gradient: phase B 5% (v:v) ~ 5% (v:v) within 5 min, phase B increased from 5% (v:v) to 45% (v:v) gradient from 5.1 min to 30 min, and maintained at 45% (v:v) gradient for 5 min; from 35.1 min to 40 min, phase B increased from 45% (v:v) to 95% (v:v) gradient, and maintained at 95% (v:v) gradient for 8 min, detection system wavelength (dual wavelength): 220 nm, 254 nm.

[0059] Figure 2 This is the purification spectrum of the conventional liquid chromatography preparation process in Comparative Example 1. Figure 3 For Comparative Example 1, conventional liquid chromatography was used for purification and analysis (the analysis and detection method is shown in Table 1). The analytical instrument shows that the polypeptide compound and the impurity are relatively close in chromatographic retention behavior. Figure 2 In the above figure, we can see that the main component and impurities overlap, and there is a lack of good separation in analytical detection. During purification and preparation, due to the increase in sample processing volume, the separation performance of the chromatographic column is not as good as the analytical level. In the preparative chromatogram, it can be clearly observed that the main peak and its impurity peaks are greatly broadened and overlap with each other ( Figure 2 ), which is difficult to separate effectively. Figure 3 It can be seen that after conventional liquid chromatography separation in Comparative Example 1, the content of the target polypeptide compound in the obtained product is only 55.79%.

[0060] Embodiment 1 (Electromagnetic liquid chromatography separation): This embodiment adopts Figure 6 and Figure 7 The axial electromagnetic field chromatography device shown is used for separation and purification.

[0061] (1) Sample: Use the crude product solution (about 90 mL) dissolved in Example 1.

[0062] (2) Chromatographic separation: About 90 mL (about 180 mg) of the above crude product solution was purified and separated by an axial electromagnetic field chromatography device. The chromatographic column (high-pressure resin column) had a specification of 250 mm × 20 mm and a filler of C18, 20 μm, 100 Å. The mobile phase A was 0.1% (v:v) acetic acid aqueous solution; the mobile phase B was acetonitrile; the mobile phase volume flow rate was 15 mL / min; the external electromagnetic field system was set to a DC voltage of 20 V and a magnetic field strength of 10 mT. The magnetic field direction was the axial direction of the fluid flow. The south pole of the magnetic field was at the inlet end of the chromatographic column, and the north pole of the magnetic field was at the outlet end of the chromatographic column. The electromagnetic field was turned on at the beginning of elution. The mobile phase used 5% (v:v) phase B and 95% (v:v) phase A to equilibrate the chromatographic column for 10 min. Elution gradient: phase B 5% (v:v) ~ 5% (v:v) within 5 min, phase B increased from 5% (v:v) to 35% (v:v) gradient from 5.1 min to 25 min, and maintained at 35% (v:v) gradient for 3 min, phase B increased from 35% (v:v) to 95% (v:v) gradient from 28.1 min to 36 min, and maintained at 95% (v:v) gradient for 6 min, detection system wavelength (dual wavelength): 220 nm, 254 nm.

[0063] Due to the addition of an external electromagnetic field, the overall separation time of the polypeptide compound is shortened. Due to the introduction of the electromagnetic field strength, the original chromatographic elution gradient is overall reduced (using less organic phase) to achieve a similar elution effect, and the overall separation degree in purification preparation is significantly improved.

[0064] Figure 4 This is the purification spectrum during the preparation process using the axial electromagnetic field chromatography device in Example 1, Figure 5 This is the analysis and detection spectrum after purification using the axial electromagnetic field chromatography device in Example 1 (the analysis and detection method is shown in Table 1). On the analytical instrument, it can be seen that the polypeptide compound and the impurity are still relatively close in chromatographic retention behavior, but during the electromagnetic chromatography purification preparation, due to the improvement of elution separation, it can be observed on the preparative chromatography that the overlap between the main peak and its impurity peaks is reduced significantly, the compound is better separated and purified, and the content of the main impurity peak has been effectively controlled. Figure 5It can be seen that after the electromagnetic field chromatography separation in Example 1, the target polypeptide compound with a total impurity of no more than 2.0% and a main peak purity of more than 98% is obtained.

[0065] After calculation, the maximum impurity of the finished product of this embodiment is 1.91%, the purity is 98.09%, and the purification yield of the main component is 58.2%.

[0066] From the above embodiments, it can be seen that the axial electromagnetic field chromatography method and device provided by the present invention can be used in all fields of liquid chromatography application, and the ability of electromagnetic chromatography can also be used to achieve certain applications and separation of compounds in fields where liquid chromatography has certain limitations. The axial electromagnetic field chromatography device provided by the present invention has a simple structure, wide versatility, and is easy to implement analysis, preparation and industrial application. In addition, an external electromagnetic field is used, and the strength and direction of the electromagnetic field can be arbitrarily adjusted according to the characteristics of the compound.

[0067] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. Other embodiments can be obtained based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for separating and purifying a chiral compound, characterized in that: An axial electromagnetic field chromatography device is used to separate a chiral compound sample to obtain a target chiral compound and a racemic impurity; the axial electromagnetic field chromatography device comprises a separation system, which comprises a chromatographic column and an electromagnetic field system; the electromagnetic field system comprises a conductive spiral coil, which is arranged around the periphery of the chromatographic column and can generate an electromagnetic field along the axial direction of the chromatographic column.

2. The separation and purification method according to claim 1, characterized in that The south pole of the electromagnetic field generated by the conductive spiral coil is located at the inlet end of the chromatographic column, and the north pole of the electromagnetic field generated by the conductive spiral coil is located at the outlet end of the chromatographic column.

3. The separation and purification method according to claim 1 or 2, characterized in that: The following steps are involved: Starting the electromagnetic field system to generate an electromagnetic field along the axial direction of the chromatographic column in the chromatographic column; The mobile phase and the chiral compound sample enter the chromatographic column from the inlet end of the chromatographic column, and the target chiral compound and the racemic impurity are respectively obtained from the outlet end of the chromatographic column.

4. The separation and purification method according to claim 1, characterized in that: The chiral compound sample is a chiral polypeptide drug.

5. The separation and purification method according to claim 1 or 4, characterized in that: The chemical structure of the target chiral compound is: 。 6. The separation and purification method according to claim 5, characterized in that: The mobile phase used for separation includes phase A and phase B; the phase A is an acetic acid aqueous solution, and the volume content of acetic acid in the acetic acid aqueous solution is 0.1%; the phase B is acetonitrile; The gradient elution procedure for the separation is: phase B 5% (v:v) to 5% (v:v) within 5 min; from 5.1 min to 25 min, phase B increases from 5% (v:v) to 35% (v:v), and phase B is maintained at 35% (v:v) for at least Tmin; from 25.1+Tmin to 36 min, phase B increases from 35% (v:v) to 95% (v:v), and phase B is maintained at 95% (v:v) for at least 6 min.

7. The separation and purification method according to claim 1 or 2, characterized in that: The intensity of the electromagnetic field is 0~12mT and is not 0.

8. The separation and purification method according to claim 1 or 2, characterized in that: The electromagnetic field system also includes a DC power supply, and the voltage of the DC power supply is 0~36V and is not 0.

9. The separation and purification method according to claim 1, characterized in that: The axial electromagnetic field chromatography device also includes an infusion system, a quantitative injection system, a detection system and a data processing system; the output end of the infusion system is connected to the quantitative injection system, the downstream of the quantitative injection system is connected to the inlet end of the chromatographic column, the outlet end of the chromatographic column is connected to the detection system, and the detection system is connected to the data processing system; the quantitative injection system includes a quantitative injection valve.

10. The separation and purification method according to claim 1, characterized in that: The column tube material of the chromatographic column includes glass, plastic or resin.

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