Separation and purification device and separation and purification method

Abstract

The invention relates to a separation and purification device and a separation and purification method. The separation and purification device comprises a sample loading pump, a solvent pump, a chromatographic separation mechanism, a first multi-way valve, a second multi-way valve, a third multi-way valve, a storage mechanism and a fourth multi-way valve. The sample loading pump and the solvent pump are respectively communicated with the first multi-way valve; the first multi-way valve, the chromatographic separation mechanism, the second multi-way valve, the third multi-way valve, the storage mechanism and the fourth multi-way valve are communicated in sequence, and the fourth multi-way valve is communicated with the first multi-way valve to form a circulating liquid path. And the second multi-way valve is provided with a purified sample discharge port for discharging liquid out of the circulating liquid path and an impurity discharge port. According to the separation and purification device and the separation and purification method, the chromatographic separation mechanism can circularly perform separation and purification through the circulating liquid path with a specific structure, the number of the chromatographic separation mechanism can be reduced, the structure is simple, the equipment is compact, and the operation is easy.

Description

Technical Field

[0001] The invention relates to the technical field of liquid chromatography, and in particular to a separation and purification device and a separation and purification method. Background Art

[0002] In traditional chromatographic column separation and purification, the single-column process is usually operated intermittently, that is, it is necessary to continuously perform the steps of feeding, eluting, and collecting, which has the disadvantages of cumbersome operation, long time consumption, large amount of eluent, and high production cost. In addition, in the traditional single-column chromatographic separation process, only a part of the chromatographic column is involved in the separation, and most of the column is not effectively utilized.

[0003] Compared with the traditional single-column process, the simulated moving bed (SMB) achieves countercurrent between the stationary phase and the mobile phase through its special process equipment and automatic control technology, making the contact between the separated substance and the stationary phase in the system more complete, thereby effectively separating the different components in the mixture and obtaining higher purity samples with less eluent consumption; in addition, it can also realize the automatic and continuous operation of the system and reduce labor costs.

[0004] However, the traditional simulated moving bed (SMB) usually requires multiple chromatographic columns and more complex piping and valve systems to realize the functions of the simulated moving bed. The equipment has a complex structure, which brings disadvantages such as large footprint, high investment cost, difficult cleaning, and high maintenance cost. Summary of the invention

[0005] Based on this, it is necessary to provide a separation and purification device and a separation and purification method to solve the problem that the traditional simulated moving bed SMB usually requires multiple chromatographic columns and more complex piping and valve systems to realize the function of the simulated moving bed and the equipment structure is complex.

[0006] One of the objects of the present invention is to provide a separation and purification device, the scheme is as follows:

[0007] A separation and purification device, characterized in that it comprises a sample loading pump, a solvent pump, a chromatographic separation mechanism, a first multi-way valve, a second multi-way valve, a third multi-way valve, a storage mechanism and a fourth multi-way valve;

[0008] The loading pump and the solvent pump are respectively connected to the first multi-way valve; the first multi-way valve, the chromatographic separation mechanism, the second multi-way valve, the third multi-way valve, the storage mechanism and the fourth multi-way valve are connected in sequence, and the fourth multi-way valve is connected to the first multi-way valve to form a circulating liquid circuit; the third multi-way valve is connected to the solvent pump, and the second multi-way valve has a purified sample discharge port and an impurity discharge port for discharging liquid outside the circulating liquid circuit.

[0009] In one embodiment, the separation and purification device further includes a first pressure sensor, and the first pressure sensor is disposed on a pipeline between the sample loading pump and the first multi-way valve.

[0010] In one embodiment, the separation and purification device further includes a second pressure sensor, which is disposed on a pipeline between the solvent pump and the first multi-way valve.

[0011] In one embodiment, the separation and purification device further comprises a first detector, which is disposed on a pipeline between the chromatographic separation mechanism and the second multi-way valve, and is used to detect the physical and / or chemical properties of an effluent from the chromatographic separation mechanism.

[0012] In one embodiment, the separation and purification device further comprises a second detector, which is disposed on a pipeline between the storage mechanism and the fourth multi-way valve, and is used to detect the physical and / or chemical properties of the discharge of the storage mechanism.

[0013] In one embodiment, the first detector is one or more of an ultraviolet detector, an evaporative light detector, and a differential refractometer detector.

[0014] In one embodiment, the second detector is one or more of an ultraviolet detector, an evaporative light detector, and a differential refractometer detector.

[0015] In one embodiment, the third multi-way valve is connected to the first multi-way valve.

[0016] In one embodiment, the separation and purification device further comprises a diluent pump, and the diluent pump is connected to the pipeline between the fourth multi-way valve and the first multi-way valve through a diluent pipeline.

[0017] In one of the embodiments, the separation and purification device further includes a third pressure sensor, and the third pressure sensor is disposed on the diluent pipeline.

[0018] In one embodiment, the fourth multi-way valve has a drain port for discharging liquid out of the circulating liquid circuit.

[0019] In one embodiment, the chromatographic separation mechanism is a single chromatographic column.

[0020] Another object of the present invention is to provide a separation and purification method, the scheme is as follows:

[0021] A separation and purification method, using the separation and purification device described in any of the above embodiments, the separation and purification method comprises the following steps:

[0022] Loading the sample into the chromatographic separation mechanism via the first multi-way valve by the loading pump;

[0023] The solvent is input into the chromatographic separation mechanism through the first multi-way valve by the solvent pump to elute the chromatographic separation mechanism, so that the sample is spread in the chromatographic separation mechanism, and the front impurities, the front impurity contaminated sample, the purified sample, the rear impurity contaminated sample and the rear impurities are discharged in sequence;

[0024] Discharge the front impurities and the rear impurities from the impurity outlet of the second multi-way valve respectively; discharge the purified sample from the purified sample outlet of the second multi-way valve; discharge the front impurity contaminated sample and the rear impurity contaminated sample into the storage mechanism for storage via the second multi-way valve and the third multi-way valve respectively;

[0025] The front impurity contaminated sample and the rear impurity contaminated sample are input into the chromatographic separation mechanism via the fourth multi-way valve and the first multi-way valve to continue separation and purification.

[0026] Compared with the traditional solution, the above separation and purification device and separation and purification method have the following beneficial effects:

[0027] The separation and purification device and separation and purification method described above enable the chromatographic separation mechanism to circulate for separation and purification through a circulating liquid path of a specific structure, thereby reducing the number of chromatographic separation mechanisms, having a simple structure, compact equipment, and easy operation.

[0028] The separation and purification device has high purity and high yield of the separated product.

[0029] The separation and purification device can realize automated and continuous separation and purification. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is a schematic structural diagram of a separation and purification device according to an embodiment;

[0031] Figure 2 Liquid chromatography of the first substance obtained by separation and purification in Example 2;

[0032] Figure 3 This is the liquid chromatography of the second substance separated and purified in Example 2.

[0033] Description of reference numerals:

[0034] 100. Separation and purification device; 101. Sample loading pump; 102. Solvent pump; 103. Chromatographic separation mechanism; 104. First multi-way valve; 105. Second multi-way valve; 1051. Purified sample outlet; 1052. Front impurity outlet; 1053. Rear impurity outlet; 106. Third multi-way valve; 107. Storage mechanism; 108. Fourth multi-way valve; 1081. Liquid outlet; 109. First pressure sensor; 110. Second pressure sensor; 111. Third pressure sensor; 112. First detector; 113. Second detector; 114. Dilution pump. DETAILED DESCRIPTION

[0035] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

[0036] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as limiting the present invention.

[0037] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the present invention, the meaning of "plurality" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined.

[0038] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0040] like Figure 1 As shown, a separation and purification device 100 according to an embodiment of the present invention includes a loading pump 101, a solvent pump 102, a chromatographic separation mechanism 103, a first multi-way valve 104, a second multi-way valve 105, a third multi-way valve 106, a storage mechanism 107, and a fourth multi-way valve 108.

[0041] The sample pump 101 and the solvent pump 102 are respectively connected to the first multi-way valve 104 .

[0042] The first multi-way valve 104, the chromatographic separation mechanism 103, the second multi-way valve 105, the third multi-way valve 106, the storage mechanism 107 and the fourth multi-way valve 108 are connected in sequence, and the fourth multi-way valve 108 is connected to the first multi-way valve 104 to form a circulating liquid circuit.

[0043] The third multi-way valve 106 is connected to the solvent pump 102. The second multi-way valve 105 has a purified sample discharge port 1051 for discharging liquid out of the circulating liquid path and an impurity discharge port.

[0044] When using the separation and purification device 100 to perform separation and purification, the following steps can be followed:

[0045] Step S1 : The sample is loaded into the chromatographic separation mechanism 103 via the first multi-way valve 104 by the loading pump 101 .

[0046] Step S2, inputting the solvent into the chromatographic separation mechanism 103 through the first multi-way valve 104 by the solvent pump 102, eluting the chromatographic separation mechanism 103, allowing the sample to be spread in the chromatographic separation mechanism 103, and discharging the front impurities, the front impurity contaminated sample, the purified sample, the rear impurity contaminated sample and the rear impurities in sequence.

[0047] Step S3, discharge the front impurities and the rear impurities from the impurity discharge port of the second multi-way valve 105. Discharge the purified sample from the purified sample discharge port 1051 of the second multi-way valve 105. Discharge the front impurity contaminated sample and the rear impurity contaminated sample into the storage mechanism 107 for storage via the second multi-way valve 105 and the third multi-way valve 106, respectively. The front impurity contaminated sample and the rear impurity contaminated sample constitute the internal circulation sample.

[0048] Step S4, inputting the inner circulation samples, including the front impurity contaminated samples and the rear impurity contaminated samples, into the chromatographic separation mechanism 103 via the fourth multi-way valve 108 and the first multi-way valve 104 to continue separation and purification.

[0049] In this way, the separation and purification device 100 can achieve continuous cyclic separation and purification.

[0050] After the impurities are discharged in each cycle, a regeneration liquid may be input through the solvent pump 102 to regenerate the chromatographic separation mechanism 103 , or a balance liquid may be input through the solvent pump 102 to balance the chromatographic separation mechanism 103 .

[0051] The separation and purification device 100 uses a circulating liquid path with a specific structure to enable the chromatographic separation mechanism 103 to circulate for separation and purification, which can reduce the number of chromatographic separation mechanisms 103 and has a simple structure, compact equipment, and easy operation.

[0052] The separation and purification device 100 has high purity and high yield of the separated product.

[0053] The separation and purification device 100 can realize automated and continuous separation and purification.

[0054] The storage mechanism 107 may be a pipeline or other container.

[0055] The first multi-way valve 104 , the second multi-way valve 105 , the third multi-way valve 106 and the fourth multi-way valve 108 are, for example, three-way valves, four-way valves, five-way valves or valves with multiple channels.

[0056] In one example, the separation and purification device 100 further includes a first pressure sensor 109, which is disposed on a pipeline between the sample loading pump 101 and the first multi-way valve 104. The first pressure sensor 109 is used to detect the pressure of the liquid inlet of the sample loading pump 101. The first pressure sensor 109 may be, but is not limited to, a PT pressure sensor.

[0057] In one example, the separation and purification device 100 further includes a second pressure sensor 110, which is disposed on a pipeline between the solvent pump 102 and the first multi-way valve 104. The second pressure sensor 110 is used to detect the pressure of the liquid entering the solvent pump 102. The second pressure sensor 110 may be, but is not limited to, a PT pressure sensor.

[0058] In one example, the separation and purification device 100 further includes a first detector 112 , which is disposed on a pipeline between the chromatographic separation mechanism 103 and the second multi-way valve 105 , and is used to detect the physical and / or chemical properties of the discharge of the chromatographic separation mechanism 103 .

[0059] Optionally, the first detector 112 may be, but is not limited to, one or more of an ultraviolet detector, an evaporative light detector, and a differential refractive index detector.

[0060] In one example, the separation and purification device 100 further includes a second detector 113 , which is disposed on a pipeline between the storage mechanism 107 and the fourth multi-way valve 108 , and is used to detect the physical and / or chemical properties of the discharge from the storage mechanism 107 .

[0061] Optionally, the second detector 113 may be, but is not limited to, one or more of an ultraviolet detector, an evaporative light detector, and a differential refractive index detector.

[0062] In one example, the third multi-way valve 106 is in communication with the first multi-way valve 102 .

[0063] In one example, the separation and purification device 100 further includes a diluent pump 114 , and the diluent pump 114 is connected to the pipeline between the fourth multi-way valve 108 and the first multi-way valve 104 through a diluent pipeline.

[0064] In step S4, during the separation and purification of the front impurity contaminated sample and the back impurity contaminated sample, the diluent pump 114 can be selectively used to input a diluent for dilution according to the actual application scenario. In addition, the loading pump 101 can be used to continue to replenish the sample to be separated and purified, and then the solvent pump 102 is used to elute the chromatographic separation mechanism 103, and the replenished sample and the front impurity contaminated sample and the back impurity contaminated sample of the previous cycle are simultaneously layered in the chromatographic separation mechanism 103.

[0065] In one example, the separation and purification device 100 further includes a third pressure sensor 111, which is disposed on the diluent pipeline. The third pressure sensor 111 is used to detect the pressure of the liquid transported in the diluent pipeline.

[0066] In one example, the fourth multi-way valve 108 has a liquid discharge port 1081 for discharging liquid out of the circulating liquid path, so that the front contaminated sample and the rear contaminated sample in the storage mechanism 107 can be discharged to the outside.

[0067] In one example, the impurity discharge outlet of the second multi-way valve 105 includes a front impurity discharge outlet 1052 and a rear impurity discharge outlet 1053. The front impurity discharge outlet 1052 is used for discharging front impurities, and the rear impurity discharge outlet 1053 is used for discharging rear impurities.

[0068] In one example, the chromatographic separation mechanism 103 is a chromatographic column. In one example, the chromatographic separation mechanism 103 is a single chromatographic column.

[0069] The above-mentioned separation and purification device 100 can reduce the number of chromatographic columns and also utilizes the countercurrent operation of the simulated moving bed SMB to realize the continuous and automated circulation operation of a single-column chromatography. The main advantage is that a single chromatographic column is used to realize the countercurrent operation and continuous operation functions of the simulated moving bed, thereby realizing continuous separation and purification of a multi-component mixture at a relatively low investment cost.

[0070] Furthermore, the present invention also provides a separation and purification method using any of the separation and purification devices 100 described above.

[0071] The separation and purification method of one embodiment comprises the following steps:

[0072] Step S1 : The sample is loaded into the chromatographic separation mechanism 103 via the first multi-way valve 104 by the loading pump 101 .

[0073] Step S2, inputting the solvent into the chromatographic separation mechanism 103 through the first multi-way valve 104 by the solvent pump 102, eluting the chromatographic separation mechanism 103, allowing the sample to be spread in the chromatographic separation mechanism 103, and discharging the front impurities, the front impurity contaminated sample, the purified sample, the rear impurity contaminated sample and the rear impurities in sequence.

[0074] Step S3, discharge the front impurities and the rear impurities from the impurity outlet of the second multi-way valve 105. Discharge the purified sample from the purified sample outlet 1051 of the second multi-way valve 105. Discharge the front impurity contaminated sample and the rear impurity contaminated sample into the storage mechanism 107 through the second multi-way valve 105 and the third multi-way valve 106 for storage.

[0075] Step S4, inputting the front impurity contaminated sample and the rear impurity contaminated sample into the chromatographic separation mechanism 103 via the fourth multi-way valve 108 and the first multi-way valve 104 to continue separation and purification.

[0076] During the loading and feeding process, the chromatographic separation mechanism 103 and the storage mechanism 107 are in an independent state, that is, no liquid flows through them. The loading flow rate controlled by the loading pump 101 can be adjusted according to the actual application scenario, and the loading time can be adjusted according to the actual application scenario.

[0077] During the development process, the chromatographic separation mechanism 103 and the storage mechanism 107 are in an independent state, that is, no liquid is circulated, and the development reagent is consistent with the mobile phase of the single column circulation chromatography method and the system SCRC chromatography. The flow rate of the development reagent can be adjusted according to the actual application scenario. The development time can be adjusted according to the actual application scenario.

[0078] In the process of discharging the front impurities, the chromatographic separation mechanism 103 and the storage mechanism 107 are in an independent state, that is, no liquid is circulated. The flow rate of the solvent pump 102 is consistent with the flow rate of the developing reagent. The elution time of the front impurities can be adjusted according to the actual application scenario. In the actual application scenario, the front impurities can be collected and stored separately when they are valuable, and can be directly discharged into the waste liquid for treatment if they are not valuable.

[0079] In the process of discharging the front impurity contaminated sample, the chromatographic separation mechanism 103 and the storage mechanism 107 are connected in series. The flow rate of the solvent pump 102 is consistent with the flow rate of the developing reagent. The transfer time of the front impurity contaminated sample can be adjusted according to the actual application scenario.

[0080] During the process of discharging the purified sample, the chromatographic separation mechanism 103 and the storage mechanism 107 are in an independent state, that is, no liquid is circulated. The flow rate of the solvent pump 102 is consistent with the flow rate of the developing reagent. The collection time of the purified sample can be adjusted according to the actual application scenario.

[0081] In the process of discharging the contaminated samples, the chromatographic separation mechanism 103 and the storage mechanism 107 are connected in series. The flow rate of the solvent pump 102 is consistent with the flow rate of the developing reagent. The transfer time of the contaminated samples can be adjusted according to the actual application scenario.

[0082] In the process of discharging the rear impurities, the chromatographic separation mechanism 103 and the storage mechanism 107 are in an independent state, that is, no liquid is circulated. The flow rate of the solvent pump 102 is consistent with the flow rate of the developing reagent. The elution time of the rear impurities can be adjusted according to the actual application scenario. In the actual application scenario, the rear impurities can be collected and stored separately when they are valuable, and can be directly discharged into the waste liquid for treatment if they are not valuable.

[0083] After the impurities are discharged in each cycle, a regeneration liquid may be input through the solvent pump 102 to regenerate the chromatographic separation mechanism 103 ; or a balance liquid may be input through the solvent pump 102 to balance the chromatographic separation mechanism 103 .

[0084] During the regeneration process, the chromatographic separation mechanism 103 and the storage mechanism 107 are in an independent state, that is, no liquid is circulated. The regeneration liquid enters the chromatographic separation mechanism 103 through the solvent pump 102. The flow rate of the solvent pump 102 is consistent with the flow rate of the developing reagent. The regeneration time can be adjusted according to the actual application scenario.

[0085] During the balancing process, the chromatographic separation mechanism 103 and the storage mechanism 107 are in an independent state, that is, no liquid is circulated. The balancing liquid enters the chromatographic separation mechanism 103 through the solvent pump 102. The flow rate of the solvent pump 102 is consistent with the flow rate of the developing reagent. The balancing time can be adjusted according to the actual application scenario.

[0086] In step S4, during the process of circulating separation and purification of the inner circulation sample, the diluent pump 114 can be selectively used to input diluent for dilution according to the actual application scenario. And the loading pump 101 can also be used to continue to replenish the sample to be separated and purified.

[0087] During the transfer of the inner circulation sample, the chromatographic separation mechanism 103 and the storage mechanism 107 are in a serial connection state. The outlet of the storage mechanism 107 is connected to the inlet of the chromatographic separation mechanism 103. The solvent pump 102 switches to input the mobile phase into the storage mechanism 107 through the third multi-way valve 106. During the transfer of the inner circulation sample, the diluent pump 114 can be selectively used to input the diluent according to the actual application scenario to dilute the inner circulation sample online. The flow rate of the solvent pump 102 and the diluent pump 114 can be adjusted according to the actual application scenario.

[0088] During the process of supplementing the sample and feeding, the chromatographic separation mechanism 103 and the storage mechanism 107 are in an independent state, that is, no liquid is circulated. The time period for supplementing the sample and feeding can be selected before, within or after the time period for transferring the inner circulation sample, preferably within the time period for transferring the inner circulation sample, and more preferably in the middle of the time period for transferring the inner circulation sample. The sample supplement flow rate controlled by the sample loading pump 101 is consistent with the flow rate of the spreading reagent. The time for supplementing the sample and feeding can be adjusted according to the actual application scenario.

[0089] During the process of sample replenishment and feeding, the step of transferring the inner circulation sample should be suspended, and the storage mechanism 107 and the chromatographic separation mechanism 103 should be disconnected. After the sample replenishment and feeding are completed, the storage mechanism 107 and the chromatographic separation mechanism 103 are connected again to continue the transfer of the inner circulation sample.

[0090] Compared with the traditional single column process and simulated moving bed SMB process, the advantages of the separation and purification device 100 of the present invention include: simpler equipment structure, flexible and convenient operation, reduced eluent and filler usage, higher separation efficiency and yield, automated continuous operation, saving time and labor costs, low investment cost, and easy maintenance and cleaning.

[0091] The following specific examples are provided to further illustrate the present invention. The present invention provides the following specific examples for a better understanding of the present invention, but is not limited to the specific implementation methods and does not limit the protection scope of the present invention.

[0092] Example 1

[0093] The present embodiment provides a separation and purification device 100, including a sample pump 101, a solvent pump 102, a chromatographic separation mechanism 103, a first multi-way valve 104, a second multi-way valve 105, a third multi-way valve 106, a storage mechanism 107, a fourth multi-way valve 108, a first pressure sensor 109, a second pressure sensor 110, a third pressure sensor 111, a first detector 112, a second detector 113 and a diluent pump 114.

[0094] The chromatographic separation mechanism 103 is a single chromatographic column. The size of the chromatographic column is 250 mm×10 mm, and the filler is a chiral filler with a particle size of 10 μm (Jiangsu Hanbang Technology Co., Ltd.). The storage mechanism 107 is a pipeline with a volume of 18 ml.

[0095] The sample loading pump 101 and the solvent pump 102 are respectively connected to the first multi-way valve 104. The first multi-way valve 104, the chromatographic separation mechanism 103, the second multi-way valve 105, the third multi-way valve 106, the storage mechanism 107 and the fourth multi-way valve 108 are connected in sequence, and the fourth multi-way valve 108 is connected to the first multi-way valve 104 to form a circulating liquid path. The third multi-way valve 106 is connected to the solvent pump 102.

[0096] The first pressure sensor 109 is arranged on the pipeline between the sample pump 101 and the first multi-way valve 104, and is used to detect the pressure of the liquid inlet of the sample pump 101. The second pressure sensor 110 is arranged on the pipeline between the solvent pump 102 and the first multi-way valve 104, and is used to detect the pressure of the liquid inlet of the solvent pump 102. The first detector 112 is arranged on the pipeline between the chromatographic separation mechanism 103 and the second multi-way valve 105. The first detector 112 is an ultraviolet detector. The second detector 113 is arranged on the pipeline between the storage mechanism 107 and the fourth multi-way valve 108. The second detector 113 is an ultraviolet detector. The diluent pump 114 is connected to the pipeline between the fourth multi-way valve 108 and the first multi-way valve 104 through the diluent pipeline. The third pressure sensor 111 is arranged on the diluent pipeline, and is used to detect the pressure of the liquid transported in the diluent pipeline.

[0097] The second multi-way valve 105 has a purified sample outlet 1051 for discharging liquid outside the circulating liquid path and an impurity outlet. The impurity outlet includes a front impurity outlet 1052 and a rear impurity outlet 1053. The front impurity outlet 1052 is used for discharging front impurities. The rear impurity outlet 1053 is used for discharging rear impurities.

[0098] Example 2

[0099] This embodiment provides a separation and purification method, using the separation and purification device 100 provided in Example 1 to separate and purify a chiral sample raw material. The separation and purification method includes the following steps:

[0100] The chiral sample raw material to be separated and purified is dissolved in anhydrous ethanol to prepare a sample solution with a concentration of 25 mg / ml. The chiral sample raw material includes 50.1% by mass of the first substance and 49.9% by mass of the second substance.

[0101] The sample solution is input into the separation and purification device 100 provided in Example 1 for separation and purification. The separation and purification process includes sequentially performing sample loading, layer development, qualified sample collection of the first substance (pre-impurity removal stage), transfer of the first substance contaminated sample (pre-impurity contaminated sample transfer stage), transfer of the first substance and the second substance contaminated sample (qualified sample collection stage), transfer of the second substance contaminated sample (post-impurity contaminated sample transfer stage), collection of qualified sample of the second substance (post-impurity removal stage), regeneration, equilibrium, first cycle, transfer of the first substance and the second substance contaminated sample (inner cycle sample transfer stage), supplementary sample feeding, layer development, qualified sample collection of the first substance (pre-impurity removal stage), transfer of the first substance contaminated sample (pre-impurity contaminated sample transfer stage), transfer of the first substance and the second substance contaminated sample (qualified sample collection stage), transfer of the second substance contaminated sample (post-impurity contaminated sample transfer stage), collection of qualified sample of the second substance (post-impurity removal stage), regeneration, equilibrium and the i-th (i≥2) cycle.

[0102] The loading parameters include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are in an independent state, the flow rate of the loading pump 101 is 6 ml / min, and the time is 0.13 min.

[0103] The developing parameters include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are in an independent state, the developing reagents are 35.9% by volume of isopropanol, 63.9% by volume of n-hexane and 0.2% by volume of trifluoroacetic acid, the flow rate of the developing reagents pumped by the solvent pump 102 is 6 ml / min, and the developing time is 3 min.

[0104] The parameters for collecting the qualified samples of the first substance include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are independent, the eluent and the developing reagent are the same, the flow rate of the solvent pump 102 pumping the eluent is 6 ml / min, and the collection time of the qualified samples of the first substance is 1.3 min.

[0105] The parameters for transferring samples contaminated by the first substance, the parameters for transferring samples contaminated by the first substance and the second substance, and the parameters for transferring samples contaminated by the second substance all include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are connected in series, the eluent and the developing reagent are the same, the flow rate of the eluent pumped by the solvent pump 102 is 6 ml / min; the total time for the three stages of transferring samples contaminated by the first substance, transferring samples contaminated by the first substance and the second substance, and transferring samples contaminated by the second substance is 3 min.

[0106] The parameters for collecting the qualified samples of the second substance include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are independent, the eluent and the developing reagent are the same, the flow rate of the solvent pump 102 pumping the eluent is 6 ml / min, and the collection time of the qualified samples of the second substance is 2 min.

[0107] The parameters of regeneration and balancing include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are independent, the regeneration liquid, the balancing liquid and the developing agent are the same, the flow rate of the solvent pump 102 pumping the eluent is 6 ml / min, the regeneration and balancing time is 3 min; the first cycle is completed.

[0108] According to the actual application scenario of chiral samples, this embodiment chooses not to use a dilution pump and a diluent to perform online dilution on the transferred first substance and second substance contaminated samples (inner circulation samples).

[0109] The parameters for transferring the samples contaminated by the first substance and the second substance (the first half) include: the storage mechanism 107 and the chromatographic separation and purification mechanism 1 are connected in series, the outlet of the storage mechanism 107 is connected to the inlet of the chromatographic separation mechanism 103, the solvent pump 102 inputs the mobile phase to the pipeline inlet through valve switching to push the samples contaminated by the first substance and the second substance (the first half) to be transferred to the chromatographic separation mechanism 103, the flow rate of the solvent pump 102 is 6 ml / min, and the transfer time is 1.5 min.

[0110] The parameters of sample replenishment feeding include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are in an independent state, the sample replenishment flow rate controlled by the sample loading pump 101 is consistent with the flow rate of the solvent pump 102 pumping the spreading reagent, the flow rate of the sample loading pump 101 is 6ml / min, and the sample replenishment feeding time is 0.13min.

[0111] The parameters for transferring the samples contaminated by the first substance and the second substance (the second half) include: the storage mechanism 107 and the chromatographic separation and purification mechanism 1 are connected in series, the outlet of the storage mechanism 107 is connected to the inlet of the chromatographic separation mechanism 103, the solvent pump 102 inputs the mobile phase to the pipeline inlet through valve switching to push the samples contaminated by the first substance and the second substance (the second half) to be transferred to the chromatographic separation mechanism 103, the flow rate of the solvent pump 102 is 6 ml / min, and the transfer time is 1.5 min.

[0112] The developing parameters include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are in an independent state, the developing reagents are 35.9% by volume of isopropanol, 63.9% by volume of n-hexane and 0.2% by volume of trifluoroacetic acid, the flow rate of the developing reagents pumped by the solvent pump 102 is 6 ml / min, and the developing time is 3 min.

[0113] The parameters for collecting qualified samples of the first substance include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are independent, the eluent and the developing reagent are the same, the flow rate of the eluent pumped by the solvent pump 102 is 6 ml / min, and the collection time of qualified samples of the first substance is 1.3 min.

[0114] The parameters for transferring samples contaminated by the first substance, the parameters for transferring samples contaminated by the first substance and the second substance, and the parameters for transferring samples contaminated by the second substance all include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are connected in series, the eluent and the developing reagent are the same, the flow rate of the eluent pumped by the solvent pump 102 is 6 ml / min; the total time for the three stages of transferring samples contaminated by the first substance, transferring samples contaminated by the first substance and the second substance, and transferring samples contaminated by the second substance is 3 min.

[0115] The parameters for collecting qualified samples of the second substance include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are independent, the eluent and the developing reagent are the same, the flow rate of the eluent pumped by the solvent pump 102 is 6 ml / min, and the collection time of qualified samples of the second substance is 2 min.

[0116] The parameters of regeneration and equilibrium include: the chromatographic separation and purification mechanism 1 and the storage mechanism 107 are in an independent state, the regeneration liquid, the equilibrium liquid and the developing agent are the same, the flow rate of the solvent pump 102 pumping the eluent is 6 ml / min, and the regeneration and equilibrium time is 3 minutes in total; the i-th (i≥2) cycle ends.

[0117] The subsequent steps are the same as the above i-th (i≥2) cycle steps, and so on.

[0118] During the separation and purification of the chiral sample repeated three times, the single column circulation chromatography SCRC system reached equilibrium.

[0119] The qualified samples of the first substance and the qualified samples of the second substance collected during the third cycle were analyzed using high performance liquid chromatography.

[0120] The content of the first substance and the content of the second substance are determined by high performance liquid chromatography, and the liquid phase conditions include:

[0121] NP7000-NU3000 high performance liquid chromatography (Jiangsu Hanbang Technology Co., Ltd.); the size of the chromatographic column is 250mm×4.6mm, and the filler is a chiral filler with a particle size of 10μm (Jiangsu Hanbang Technology Co., Ltd.); the mobile phase is isopropanol-n-hexane-trifluoroacetic acid (35.9:63.9:0.2); the flow rate is 1ml / min; the column temperature is 25℃; the injection volume is 20μL; the content of the first substance and the content of the second substance are determined by the external standard method.

[0122] The liquid chromatography of the first substance obtained is as follows: Figure 2 The specific chromatographic information is shown in Table 1. The liquid chromatogram of the obtained second substance is shown in Figure 3 The specific chromatographic information is shown in Table 2.

[0123] Table 1 Chromatographic information of the first substance

[0124]

[0125] Table 2 Chromatographic information of the second substance

[0126]

[0127] from Figure 2 It can be concluded from Table 1 that according to the collection of the first substance and the input raw materials, the purity of the first substance in this experiment was 99.4% and the yield was 99.7%.

[0128] from Figure 3 From Table 2, it can be concluded that based on the collected second substance and the input raw materials, the purity of the second substance in this experiment was 98.6% and the yield was 96.5%.

[0129] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0130] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A separation and purification device, characterized in that: It includes a sample loading pump, a solvent pump, a chromatographic separation mechanism, a first multi-way valve, a second multi-way valve, a third multi-way valve, a storage mechanism and a fourth multi-way valve; The loading pump and the solvent pump are respectively connected to the first multi-way valve; the first multi-way valve, the chromatographic separation mechanism, the second multi-way valve, the third multi-way valve, the storage mechanism and the fourth multi-way valve are connected in sequence, and the fourth multi-way valve is connected to the first multi-way valve to form a circulating liquid circuit; the third multi-way valve is connected to the solvent pump, and the second multi-way valve has a purified sample discharge port and an impurity discharge port for discharging liquid outside the circulating liquid circuit.

2. The separation and purification device according to claim 1, characterized in that: The separation and purification device meets at least one of the following characteristics (1) to (2): (1) The separation and purification device further includes a first pressure sensor, which is disposed on a pipeline between the sample loading pump and the first multi-way valve; (2) The separation and purification device further includes a second pressure sensor, which is disposed on a pipeline between the solvent pump and the first multi-way valve.

3. The separation and purification device according to claim 1, characterized in that: The separation and purification device meets at least one of the following characteristics (1) to (2): (1) The separation and purification device further comprises a first detector, which is disposed on a pipeline between the chromatographic separation mechanism and the second multi-way valve, and is used to detect the physical and / or chemical properties of the discharge of the chromatographic separation mechanism; (2) The separation and purification device further includes a second detector, which is disposed on a pipeline between the storage mechanism and the fourth multi-way valve, and is used to detect the physical properties and / or chemical properties of the discharge from the storage mechanism.

4. The separation and purification device according to claim 3, characterized in that: The separation and purification device meets at least one of the following characteristics (1) to (2): (1) The first detector is one or more of an ultraviolet detector, an evaporative light detector and a differential refractive index detector; (2) The second detector is one or more of an ultraviolet detector, an evaporative light detector and a differential refractometer detector.

5. The separation and purification device according to claim 3, characterized in that: The third multi-way valve is in communication with the first multi-way valve.

6. The separation and purification device according to any one of claims 1 to 5, characterized in that The separation and purification device further comprises a diluent pump, and the diluent pump is connected to the pipeline between the fourth multi-way valve and the first multi-way valve through a diluent pipeline.

7. The separation and purification device according to claim 6, characterized in that: The separation and purification device further comprises a third pressure sensor, and the third pressure sensor is arranged on the diluent pipeline.

8. The separation and purification device according to any one of claims 1 to 5, characterized in that The fourth multi-way valve has a liquid discharge port for discharging liquid out of the circulating liquid path.

9. The separation and purification device according to any one of claims 1 to 5, characterized in that The chromatographic separation mechanism is a single chromatographic column.

10. A separation and purification method, characterized in that: Using the separation and purification device according to any one of claims 1 to 9, the separation and purification method comprises the following steps: Loading the sample into the chromatographic separation mechanism via the first multi-way valve by the loading pump; The solvent is input into the chromatographic separation mechanism through the first multi-way valve by the solvent pump to elute the chromatographic separation mechanism, so that the sample is spread in the chromatographic separation mechanism, and the front impurities, the front impurity contaminated sample, the purified sample, the rear impurity contaminated sample and the rear impurities are discharged in sequence; Discharge the front impurities and the rear impurities from the impurity outlet of the second multi-way valve respectively; discharge the purified sample from the purified sample outlet of the second multi-way valve; discharge the front impurity contaminated sample and the rear impurity contaminated sample into the storage mechanism for storage via the second multi-way valve and the third multi-way valve respectively; The front impurity contaminated sample and the rear impurity contaminated sample are input into the chromatographic separation mechanism via the fourth multi-way valve and the first multi-way valve to continue separation and purification.

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