Substance separating and refining device based on conversion from linear chromatography to plane chromatography and application of substance separating and refining device

Abstract

The invention belongs to the technical field of sample separation and analysis, and particularly relates to a substance separation and refining device based on conversion from linear chromatography to plane chromatography and application of the substance separation and refining device. The device comprises a planar solid-phase chromatographic plate, a sealing bin, a chromatographic pipeline, a filter and a liquid collecting bin, the planar solid-phase chromatographic plate is arranged in the sealing bin, a flow guide groove is formed in the outer edge of the planar solid-phase chromatographic plate, and the filter is embedded in the planar solid-phase chromatographic plate; the chromatographic pipeline penetrates through the sealing bin and then is connected with the filter, and the chromatographic pipeline can drive the planar solid-phase chromatographic plate to rotate so as to realize chromatographic centrifugal separation; the liquid collecting bin is arranged below the plane solid phase chromatographic plate and is used for collecting centrifugal liquid. According to the invention, through conversion from linear chromatography to plane chromatography, under a pressure environment, through a coupling effect of mobile phase elution and centrifugal force separation, multi-dimensional separation and substance collection capabilities are realized.

Description

Technical Field

[0001] The present invention belongs to the technical field of sample separation and analysis, and in particular relates to a substance separation and purification device based on linear to planar chromatography conversion and an application thereof. Background Art

[0002] With the advancement of technology and exploration in fields such as life sciences, biological testing, food and drug testing, and active ingredient preparation, higher requirements are being placed on sample separation and analysis based on the baseline expression of relevant active substances. This requires analyzing substances at different concentrations or expression levels in the sample, particularly to accurately understand the properties of certain substances, in order to obtain the overall material distribution of a complex sample system and explore and understand the potential effects within the sample. Chromatographic analysis, as the primary method for sample testing and inspection, occupies the majority of application scenarios in the analytical testing field, with conventional linear separation chromatography accounting for a large proportion. The resulting multidimensional chromatography methods are generally performed using valve switching technology or independent parallel connections. Their separation efficiency and sample purification efficiency are not significantly improved due to the superposition of chromatographic efficiency. Conventional planar chromatography, operating at conventional pressures and driven by capillary action, also has significant limitations in its separation capabilities. To date, no chromatographic separation device has been developed that couples conventional linear chromatography with planar chromatography and operates under a specific pressure to enhance separation, analysis, and purification capabilities. Summary of the Invention

[0003] In view of the above problems, the purpose of the present invention is to provide a substance separation and purification device based on linear to planar chromatography conversion and its application, so as to solve the problem of poor chromatographic analysis ability of conventional technology.

[0004] In order to achieve the above object, the present invention adopts the following technical solutions:

[0005] On one hand, the present invention provides a substance separation and purification device based on linear to planar chromatography conversion, comprising a planar solid-phase chromatography plate, a sealed chamber, a chromatography pipeline, a filter, and a liquid collection chamber, wherein the planar solid-phase chromatography plate is arranged in the sealed chamber, a guide groove is provided along the circumference of the outer edge of the planar solid-phase chromatography plate, and the filter is embedded in the planar solid-phase chromatography plate;

[0006] The chromatographic pipeline passes through the sealed chamber and is connected to the filter. The chromatographic pipeline can drive the planar solid phase chromatographic plate to rotate to achieve chromatographic centrifugal separation;

[0007] The liquid collecting chamber is arranged below the planar solid phase chromatography plate and is connected to the guide groove on the outer edge of the planar solid phase chromatography plate. The liquid collecting chamber is used to collect liquid driven by centrifugation and pressure.

[0008] The bottom of the guide groove is provided with a plurality of through holes connected with the liquid collecting bin.

[0009] The liquid collecting bin is a conical structure, and a liquid guiding channel is provided at the bottom of the liquid collecting bin and extends to the outside of the sealing bin.

[0010] The filter is a cylindrical structure made by a sieve plate, and the sieve plate is a stainless steel filter sieve plate covered with 0.22 μm pores.

[0011] The upper end of the chromatographic pipeline is rotatably connected to the external pipeline through a converter.

[0012] The converter includes a downward guide rail and a rotor bearing, wherein the rotor bearing is arranged at the upper end of the chromatographic pipeline, the downward guide rail is arranged at the lower end of the external pipeline, and the rotor bearing and the downward guide rail are circumferentially slidably matched.

[0013] The chromatographic pipeline is connected to an external rotary drive motor, which provides power for the rotational movement of the chromatographic pipeline.

[0014] The inner edge of the guide groove is provided with a filtering type blocking ring along the circumferential direction to prevent the loss of filler.

[0015] A solid spherical cap is provided below the sealing chamber, and an annular gap for liquid to flow out is formed between the solid spherical cap and the inner wall of the liquid collecting chamber.

[0016] On the other hand, the present invention provides an application of the material separation and purification device based on linear to planar chromatography conversion as described above, in which the sample is injected through an external part, and the sample is separated and the main substances are purified and collected by utilizing the chromatographic separation of the chromatographic pipeline and the centrifugal separation of the planar solid-phase chromatography plate fixedly connected to the chromatographic pipeline and rotating therewith.

[0017] The advantages and beneficial effects of the present invention are: the present invention can realize single-dimensional chromatographic separation of samples by converting linear separation into planar separation by embedding the chromatographic system, and form multidimensional chromatographic separation by driving the planar centrifugal motion, thereby realizing sample separation, analysis and material purification operations, and can provide multidimensional chromatographic separation and analysis efficiency for life sciences, biological testing, food and drug testing, active ingredient preparation, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Schematic diagram of the structure of the substance separation and purification device based on the conversion from linear to planar chromatography of the present invention;

[0019] Figure 2 It is a structural schematic diagram of the pressing bin in the present invention;

[0020] Figure 3 Schematic diagram of the structure of the filter in the present invention;

[0021] Figure 4It is a structural schematic diagram of the chromatography pipeline in the present invention.

[0022] In the figure: 1. Planar solid-phase chromatography plate; 2. Sealing chamber; 3. External pipeline; 4. Chromatographic pipeline; 5. Filter; 6. Guide trough; 7. Liquid collecting chamber; 8. Liquid guide channel; 9. Lower plate; 10. Upper plate; 12. Converter; 13. Downward guide rail; 14. Rotor bearing. DETAILED DESCRIPTION

[0023] In order to make the objectives, technical solutions and advantages of the present invention more clear, the present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0024] See also Figure 1 and Figure 2 As shown, an embodiment of the present invention provides a material separation and purification device based on linear to planar chromatography conversion, comprising a planar solid-phase chromatography plate 1, a sealed chamber 2, a chromatography pipeline 4, a filter 5 and a liquid collection chamber 7, wherein the planar solid-phase chromatography plate 1 is arranged in the sealed chamber 2, and the outer edge of the planar solid-phase chromatography plate 1 is circumferentially provided with a circular low-dead-volume guide groove 6, and the filter 5 is embedded in the center of the planar solid-phase chromatography plate 1; the chromatography pipeline 4 passes through the sealed chamber 2 and is connected to the filter 5; the chromatography pipeline 4 can drive the planar solid-phase chromatography plate 1 to rotate to realize chromatographic centrifugal separation; the liquid collection chamber 7 is arranged below the planar solid-phase chromatography plate 1 and is connected to the guide groove 6 on the outer edge of the planar solid-phase chromatography plate 1, and the liquid collection chamber 7 is used to collect liquid driven by centrifugation and pressure.

[0025] Furthermore, a plurality of through holes communicating with the liquid collecting bin 7 are provided at the bottom of the guide groove 6 , and the liquid entering the guide groove 6 flows into the liquid collecting bin 7 through the through holes at the bottom.

[0026] Specifically, the liquid collecting bin 7 is a conical structure, and a liquid conducting channel 8 is provided at the bottom of the liquid collecting bin 7 and extends to the outside of the sealing bin 2 . The liquid conducting channel 8 is used to connect to a receiving bottle or an external detection device.

[0027] See also Figure 3 As shown, in the embodiment of the present invention, the filter 5 is a cylindrical structure made of a sieve plate, which is a stainless steel filter sieve plate covered with 0.22 μm pores, so that the liquid flowing out of the chromatographic pipeline 4 can be evenly injected into the planar solid phase chromatography plate 1 in a circular injection manner.

[0028] See also Figure 1 and Figure 4As shown, in the embodiment of the present application, the upper end of the chromatographic pipe 4 is connected with the external pipe 3 through the converter 12, and the external pipe 3 is used to connect with the chromatographic sample injection and mobile phase system. The converter 12 comprises a downward guide rail 13 and a rotor bearing 14, wherein the rotor bearing 14 is arranged at the upper end of the chromatographic pipe 4 and has upward rolling balls; the downward guide rail 13 is arranged at the lower end of the external pipe 3, and the rotor bearing 14 and the downward guide rail 13 are in circumferential sliding fit. The rotor bearing 14 and the downward guide rail 13 are connected by using a sealed chromatographic connector, and there is a small gap between the pipe heads of the chromatographic pipe 4 and the external pipe 3, while the rotor bearing 14 and the downward guide rail 13 can be smoothly contacted, the rolling balls of the rotor bearing 14 can slide on the downward guide rail 13, and the fixed part of the downward guide rail 13 can be in a stationary state.

[0029] Further, the chromatographic pipe 4 is connected with an external rotary drive motor, and the external rotary drive motor provides power for the rotary motion of the chromatographic pipe 4. The planar solid-phase chromatographic plate 1 is pressed between two flat plates and is in a sealed state, so that the rotary separation under a certain pressure environment is realized. The external rotary drive motor drives the chromatographic pipe 4 to rotate for a certain number of revolutions, and the rotation of the chromatographic pipe 4 drives the planar solid-phase chromatographic plate 1 to rotate in the same direction. The mobile phase contained in the chromatographic pipe 4 can drive the related substances to be thrown into the flow guide groove 6 under the joint action of a certain pressure and centrifugal force, so that the mobile phase fluid is converted from linear flow to planar flow combined with the action of centrifugal force, and further diffusion separation is realized. The flow guide groove 6 receives the liquid fractions driven by centrifugal force and pressure, and the liquid fractions flow out through the fixed liquid collection bin 7 and the liquid guide channel 8.

[0030] Further, the inner side edge of the flow guide groove 6 is provided with a filtering type blocking ring for preventing the loss of filler along the circumference, and the side wall of the filtering type blocking ring is provided with filtering holes.

[0031] In the embodiment, the sealed bin 2 is formed by pressing the upper layer plate 10 and the lower layer plate 9 in parallel. The upper layer plate 10 and the lower layer plate 9 are both circular flat plates, and the planar solid-phase chromatographic plate 1 pressed between the upper layer plate 10 and the lower layer plate 9 is also circular. The planar chromatogram is sealed and pressed by the upper and lower layer plates, the filter 5 is embedded in the middle circular hole of the planar solid-phase chromatographic plate 1, and the height of the filter 5 is consistent with the thickness of the planar solid-phase chromatographic plate 1. In actual operation, the chromatographic pipe 4 can be filled with chromatographic filler or not filled, and the sample can be injected through the external part. The chromatographic separation of the chromatographic pipe 4 and the centrifugal separation of the planar solid-phase chromatographic plate 1 which can rotate synchronously can be used to separate and refine and collect the main substances of the sample, and can provide multi-dimensional chromatographic separation and analysis efficiency for life science, biological detection, food and drug detection, active ingredient preparation, etc.

[0032] Furthermore, the upper plate 10 has a circular etched groove in the middle, which contacts the top of the planar solid-phase chromatography plate 1. The lower plate 9 also has a circular etched groove in the middle, which contacts the bottom of the planar solid-phase chromatography plate 1. The upper and lower plates press the planar solid-phase chromatography plate 1 to form an integrated planar chromatography. The planar solid-phase chromatography plate 1 can be rotated by the chromatography pipe 4. The upper end of the chromatography pipe 4 is connected to other external devices such as the injection, mobile phase pump, collector, and detection and analysis equipment through the converter 12.

[0033] Furthermore, to reduce dead volume and minimize chromatographic diffusion, a solid spherical cap can be fixedly attached to the lower plate 9. An annular gap for liquid outflow is formed between the cap and the inner wall of the liquid collection chamber 7. After entering the flow channel 6, liquid flows out along the annular gap inside the liquid collection chamber 7. The coupling of linear and circular centrifugal separations under pressure enables continuous online multidimensional chromatography coupling, combining multiple separation driving forces to achieve separation and purification of substances.

[0034] Furthermore, the system can achieve both circular chromatography separation and analysis under closed conditions, and multidimensional chromatography separation and analysis coupled with linear and planar chromatography, for sample separation, analysis, and active substance purification. Sample separation and preparation are not limited to step-by-step operations and modular applications; they can all be performed through automated coupling and integration, utilizing static planar chromatography, rotating planar chromatography, and linear planar switching chromatography to meet requirements and enhance the efficiency of multidimensional sample concentration and preparation.

[0035] Example 1

[0036] Chromatographic conduit 4 is filled with C18, 10μ packing. The column is 15 cm long and 4.6 mm in diameter, including the lower filter 5, which is 0.2 cm high and has the same diameter as the column. The flat plate chromatographic packing is C18, 30μ packing with a radius of 2.5 cm. To prevent packing loss, a filter-type barrier ring is placed between the conduit 6 and the flow channel. It is uniformly distributed with 0.45μm pores and has a thickness of 1mm. A rotary motor is connected, capable of achieving speeds of 5,000-12,000 rpm. The upper portion of the chromatographic conduit 4 is connected to an external liquid chromatography system.

[0037] After connecting the device, the flow conditions were methanol and water (50:50) as the mobile phase, a liquid phase display flow rate of 1.0 ml / min, a sample injection volume of 100 μL methanol, and a detector connected at the bottom. The experimental parameters are as follows:

[0038]

[0039]

[0040] The present invention realizes a mode of conversion from linear chromatography to rotating planar chromatography. Under common liquid phase conditions, due to the high-speed rotation, it has a better flow effect than linear chromatography. As the number of revolutions increases, the dead volume and dead time are significantly reduced, indicating that under the auxiliary drive of centrifugal force, the elution efficiency of the liquid phase is enhanced, and it has a better effect than the linear chromatography series mode.

[0041] Compared with the same double columns in series, the dead time and dead volume of the existing device are half of that of the conventional circular chromatography, and the reagents are saved by 2 times.

[0042] Example 2

[0043] The chromatographic pipeline 4 (chromatographic column) is filled with C18, 10μ filler. The column length of the chromatographic pipeline 4 is 15 cm, the column diameter is 4.6 mm, and the column length includes the lower filter 5. The filter 5 is 0.2 cm high and the diameter is consistent with the column body; the flat plate chromatographic filler is C18, 30μ filler with a radius of 2.5 cm. In order to prevent the loss of filler, there is a filter-type barrier ring between it and the guide groove 6, and the sieve holes with a pore size of 0.45 μm are evenly distributed on it, and the pressing thickness is 1 mm; it is connected to a rotating motor to achieve a rotation speed of 5000-12000 rpm. The upper part of the chromatographic pipeline 4 is externally connected to a liquid chromatography.

[0044] After connecting the device, the mobile phase was methanol and water (70:30), the liquid phase flow rate was 1.0 ml / min, the injection volume was 100 μL of a 0.05% methanol solution of benzene and toluene (1:1), and the detector was connected at the bottom. The experimental parameters are shown below:

[0045]

[0046]

[0047] Compared with conventional circular planar chromatography, its theoretical plate number should be more than 10 times that of the conventional one.

[0048] Example 3

[0049] The chromatographic pipe 4 (chromatographic column) is filled with C18, 10μ filler. The column length of the chromatographic pipe 4 is 15 cm, the column diameter is 4.6 mm, and the column length includes the lower filter 5. The height of the filter 5 is 0.2 cm, and the diameter is consistent with the column body; the flat plate chromatographic filler is C18, 30μ filler with a radius of 2.5 cm. In order to prevent the loss of filler, there is a filter-type barrier ring between it and the guide groove 6, and the sieve holes with a pore size of 0.45 μm are evenly distributed on it, and the pressing thickness is 1 mm; it is connected to a rotating motor to achieve a rotation speed of 5000-12000 rpm. The upper part of the chromatographic pipe 4 is externally connected to a liquid chromatography.

[0050] After connecting the device, the mobile phase was methanol and water (70:30), with a liquid chromatography flow rate of 1.0 ml / min. The injection volume was 100 μL of 0.05% rhubarb extract, with an emodin concentration of approximately 67 mg / kg. The detector was connected at the bottom. The experimental parameters are as follows:

[0051]

[0052] Due to the presence of impurities, the above recovery rates are converted to peak purity.

[0053] The above data show that the relevant separation and preparation efficiency has been greatly improved compared with the conventional method. It can be expected that it will be further improved based on the addition of temperature control and speed change.

[0054] In response to the problems that arise in chromatographic testing, the present invention analyzes the effectiveness of various existing technologies, proposes a multidimensional mode for coupling linear chromatography and planar chromatography, and further introduces the driving force of high-speed centrifugation, so that the overall separation system is sealed in a system that can provide enhanced standby pressure, thereby making up for the low separation efficiency of planar chromatography itself on the one hand, and on the other hand, multidimensional separation can be achieved at one time. Under pressure conditions, in a state of high-speed rotation, the separation ability of planar chromatography is greatly changed, and its theoretical plate number is increased by more than ten times compared with low-speed rotation under normal pressure. The mobile phase system is also expanded and can be converted to a conventional chromatographic mobile phase, which changes the common separation mechanism of planar chromatography as a whole. Moreover, due to the characteristics of planar chromatography, the material load of the sample can be increased in the linear part, thereby realizing the transformation from analysis to preparation and improving the refining effect of the substance.

[0055] This invention achieves a transition from linear chromatography to planar chromatography, achieving multidimensional separation and material collection capabilities through the coupling of mobile phase elution and centrifugal separation under pressure. A circular planar chromatography plate is compressed between two flat plates and sealed, enabling rotational separation under a certain pressure.

[0056] Based on the above design concept, another embodiment of the present invention provides an application of a substance separation and purification device based on the conversion from linear to planar chromatography. Samples are injected through an external portion, and the sample is separated and the main substances are purified and collected using chromatographic separation in a chromatographic channel 4 and centrifugal separation on a planar solid-phase chromatography plate 1 that is fixedly connected to and rotates with the chromatographic channel 4. By embedding the chromatographic system, single- and multi-dimensional chromatographic separation of samples can be performed, enabling sample separation, analysis, and substance purification operations. This provides multi-dimensional chromatographic separation and analysis capabilities for applications in life sciences, biological testing, food and drug testing, and active ingredient preparation.

[0057] The above description is only an embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Any modification, equivalent replacement, improvement, expansion, etc. made within the spirit and principle of the present invention are included in the scope of protection of the present invention.

Claims

1. A substance separation and purification device based on linear to planar chromatography conversion, characterized in that: The invention comprises a planar solid-phase chromatography plate (1), a sealing chamber (2), a chromatography pipeline (4), a filter (5) and a liquid collection chamber (7), wherein the planar solid-phase chromatography plate (1) is arranged in the sealing chamber (2), a guide groove (6) is provided on the outer edge of the planar solid-phase chromatography plate (1) along the circumference, and the filter (5) is embedded in the planar solid-phase chromatography plate (1); The chromatographic pipeline (4) passes through the sealed chamber (2) and is connected to the filter (5). The chromatographic pipeline (4) can drive the planar solid phase chromatographic plate (1) to rotate to achieve chromatographic centrifugal separation; The liquid collecting chamber (7) is arranged below the planar solid phase chromatography plate (1) and is connected to the guide groove (6) on the outer edge of the planar solid phase chromatography plate (1). The liquid collecting chamber (7) is used to collect liquid driven by centrifugation and pressure.

2. The substance separation and purification device based on linear to planar chromatography conversion according to claim 1, characterized in that: The bottom of the guide groove (6) is provided with a plurality of through holes communicating with the liquid collecting bin (7).

3. The substance separation and purification device based on linear to planar chromatography conversion according to claim 1, characterized in that: The liquid collecting bin (7) is a conical structure, and a liquid guiding channel (8) is provided at the bottom of the liquid collecting bin (7) and extends to the outside of the sealing bin (2).

4. The substance separation and purification device based on linear to planar chromatography conversion according to claim 1, characterized in that: The filter (5) is a cylindrical structure made by a sieve plate, and the sieve plate is a stainless steel filter sieve plate covered with 0.22 μm pores.

5. The substance separation and purification device based on linear to planar chromatography conversion according to claim 1, characterized in that: The upper end of the chromatographic pipeline (4) is rotatably connected to the external pipeline (3) through a converter (12).

6. The substance separation and purification device based on linear to planar chromatography conversion according to claim 5, characterized in that: The converter (12) includes a downward guide rail (13) and a rotor bearing (14), wherein the rotor bearing (14) is arranged at the upper end of the chromatographic pipeline (4), and the downward guide rail (13) is arranged at the lower end of the external pipeline (3), and the rotor bearing (14) and the downward guide rail (13) are circumferentially slidably matched.

7. The substance separation and purification device based on linear to planar chromatography conversion according to claim 1, characterized in that: The chromatographic pipeline (4) is connected to an external rotary drive motor, and the external rotary drive motor provides power for the rotational movement of the chromatographic pipeline (4).

8. The substance separation and purification device based on linear to planar chromatography conversion according to claim 1, characterized in that: The inner edge of the guide groove (6) is provided with a filtering type blocking ring along the circumferential direction to prevent the loss of filler.

9. The substance separation and purification device based on linear to planar chromatography conversion according to claim 1, characterized in that: A solid spherical cap is provided below the sealing chamber (2), and an annular gap for liquid outflow is formed between the solid spherical cap and the inner wall of the liquid collecting chamber (7).

10. An application of a substance separation and purification device based on linear to planar chromatography conversion according to any one of claims 1 to 9, characterized in that: The sample is injected through the external part, and the sample is separated and the main substances are purified and collected by utilizing the chromatographic separation of the chromatographic pipeline (4) and the centrifugal separation of the planar solid phase chromatographic plate (1) fixedly connected to the chromatographic pipeline (4) and rotating therewith.

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