Dynamic gradient mixing and conveying device of chromatographic column and chromatographic column analysis equipment

Abstract

The invention relates to the technical field of cell analysis equipment, and provides a dynamic gradient mixing and conveying device of a chromatographic column and chromatographic column analysis equipment. The dynamic gradient mixing and conveying device of the chromatographic column comprises a sample injection bottle, a B-phase input unit, a mixed phase output unit and a liquid flow velocity control unit. An A phase is contained in the sample injection bottle, and the B phase input unit is connected with the sample injection bottle so as to input a B phase into the sample injection bottle to be mixed with the A phase to form a mixed phase. And the mixed phase output unit is connected with the sample injection bottle and the chromatographic column so as to convey the mixed phase in the sample injection bottle into the chromatographic column. The liquid flow velocity control unit is used for adjusting the flow velocity of the mixed phase conveyed into the chromatographic column. In the ultra-narrow inner diameter chromatographic analysis process, the flow velocity of a mixed phase entering the ultra-narrow inner diameter chromatographic column can be controlled through the liquid flow velocity control unit, so that the flow velocity is more stable, the proportion of the mobile phase can be changed in real time, the mixed phase entering the ultra-narrow inner diameter chromatographic column is ensured to be uniform, and the stability and reproducibility of an analysis result are further improved.

Description

Technical Field

[0001] The present invention relates to the technical field of cell analysis equipment, and in particular, to a dynamic gradient mixing and conveying device for a chromatographic column and a chromatographic column analysis equipment. Background Art

[0002] Single-cell analysis is a technology with great potential, which can deeply reveal the molecular mechanisms of diseases, promote the early diagnosis of diseases, and provide important support for the development of precision treatment methods. Currently, common chromatographic separation technologies are used for single-cell analysis. Through chromatographic separation technology, analytes can be better separated, reducing the interference effects caused by co-elution, thus significantly reducing the possibility of ion suppression. Chromatographic separation enables each analyte to obtain a more uniform distribution before entering the mass spectrometry system, enabling the ion source to work in a more efficient state, greatly improving the detection sensitivity. In addition, chromatographic separation can also extend the acquisition time of the mass spectrometry, enabling the mass spectrometry analysis to accumulate signals for a longer time, thereby improving the accuracy and coverage depth of identification. Through the combination of chromatography and mass spectrometry, the precision and sensitivity of the analysis have been significantly improved, contributing to a more refined revelation of the molecular characteristics in single cells.

[0003] As an efficient separation tool, ultra-narrow inner diameter chromatographic columns are increasingly widely used. Due to their small inner diameter (generally below 5 μm), ultra-narrow inner diameter chromatographic columns can provide higher separation efficiency and lower sample consumption, and are suitable for analyzing trace samples and complex sample systems.

[0004] However, currently in the market during chromatographic column analysis, most use gradient pumps, whose flow rate is difficult to control. For ultra-narrow inner diameter chromatographic column analysis, gradient lag problems are more likely to occur, which will greatly affect the stability and reproducibility of the analysis results. Summary of the Invention

[0005] In view of the above technical problems, the present invention provides a dynamic gradient mixing and conveying device for a chromatographic column and a chromatographic column analysis equipment.

[0006] In a first aspect of the present invention, there is provided a dynamic gradient mixing and conveying device for a chromatographic column, including: a sample injection bottle, in which a phase A is contained; a phase B input unit, which is connected to the sample injection bottle to input phase B into the sample injection bottle; a mixed phase output unit, the sample injection bottle is connected to the mixed phase output unit, and the mixed phase output unit is used to be connected to a chromatographic column to convey the mixed phase in the sample injection bottle into the chromatographic column; a liquid flow rate control unit, which extends into the sample injection bottle and is used to adjust the flow rate of the mixed phase conveyed into the chromatographic column.

[0007] A dynamic gradient mixing and conveying device for a chromatographic column according to the present invention, wherein the liquid flow rate control unit includes a first capillary tube and a first pressure control valve. One end of the first capillary tube is connected to a gas source, and the other end of the first capillary tube extends into the sample injection bottle and is above the liquid level of the mixed phase in the sample injection bottle. The first pressure control valve is arranged on the first capillary tube.

[0008] A dynamic gradient mixing and conveying device for a chromatographic column according to the present invention, wherein the B-phase input unit includes a B-phase liquid storage tank and a second capillary tube. One end of the second capillary tube is connected to the B-phase liquid storage tank, and the other end of the second capillary tube extends into the sample injection bottle and is inserted into the A phase.

[0009] A dynamic gradient mixing and conveying device for a chromatographic column according to the present invention, wherein the B-phase input unit further includes a third capillary tube and a second pressure control valve. One end of the third capillary tube is connected to the gas source, and the other end of the third capillary tube extends into the B-phase liquid storage tank and is above the liquid level of the B-phase liquid storage tank. The second pressure control valve is arranged on the third capillary tube.

[0010] A dynamic gradient mixing and conveying device for a chromatographic column according to the present invention, wherein the dynamic gradient mixing and conveying device of the chromatographic column further includes a mixer, which is connected to the sample injection bottle and is used for mixing the mixed phase in the sample injection bottle.

[0011] A dynamic gradient mixing and conveying device for a chromatographic column according to the present invention, wherein the chromatographic column is used to extend into the sample injection bottle and is inserted into the mixed phase.

[0012] A dynamic gradient mixing and conveying device for a chromatographic column according to the present invention, wherein a first interface is arranged on the sample injection bottle, the first capillary tube passes through the first interface and extends into the sample injection bottle, and a first sealing device is arranged between the first capillary tube and the first interface.

[0013] A dynamic gradient mixing and conveying device for a chromatographic column according to the present invention, wherein a second interface is arranged on the sample injection bottle, the second capillary tube passes through the second interface and extends into the sample injection bottle, and a second sealing device is arranged between the second capillary tube and the second interface.

[0014] A dynamic gradient mixing and conveying device for a chromatographic column according to the present invention, wherein a third interface is arranged on the sample injection bottle, the chromatographic column passes through the third interface and extends into the sample injection bottle, and a third sealing device is arranged between the chromatographic column and the third interface.

[0015] According to a second aspect of the present invention, there is provided a chromatographic column analysis device, including a chromatographic column and a dynamic gradient mixing and conveying device for the chromatographic column as described above, and the chromatographic column is connected to the mixed phase output unit.

[0016] In the dynamic gradient mixing and conveying device for the chromatographic column provided by the present invention, it includes a sample injection bottle, a B-phase input unit, a mixed phase output unit, and a liquid flow rate control unit. The sample injection bottle contains the A-phase, the B-phase input unit is connected to the sample injection bottle, and it can input the B-phase into the sample injection bottle to mix with the A-phase to form a mixed phase. The mixed phase output unit is connected to the sample injection bottle and the chromatographic column, and it can convey the mixed phase in the sample injection bottle into the chromatographic column. The liquid flow rate control unit extends into the sample injection bottle and is used to adjust the flow rate of the mixed phase output sent into the chromatographic column.

[0017] Through this structural arrangement, during the ultra-narrow inner diameter chromatographic analysis process, the flow rate of the mixed phase entering the ultra-narrow inner diameter chromatographic column can be controlled by the liquid flow rate control unit, making its flow rate more stable, thereby improving the stability and reproducibility of the analysis results. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0019] Figure 1 It is a schematic structural diagram of the dynamic gradient mixing and conveying device for the chromatographic column provided by the present invention.

[0020] Reference numerals: 100, sample injection bottle; 110, first interface; 120, second interface; 130, third interface; 210, first capillary; 220, first pressure control valve; 310, B-phase liquid storage tank; 320, second capillary; 330, third capillary; 340, second pressure control valve. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0021] The following will further describe in detail the embodiments of the present invention in conjunction with the drawings and embodiments. The following embodiments are used to illustrate the present invention, but cannot be used to limit the scope of the present invention.

[0022] In the description of the embodiments of the present invention, it should be noted that the orientation or positional relationship indicated by the terms "center", "longitudinal", "transverse", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the embodiments of the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as a limitation on the embodiments of the present invention. In addition, the terms "first" and "second" are only used for descriptive purposes and should not be construed as indicating or implying relative importance.

[0023] In the description of the embodiments of the present invention, it should be noted that unless otherwise clearly specified and limited, the terms "connected" and "connected" 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 directly connected or indirectly connected through an intermediate medium. For those of ordinary skill in the art, the specific meanings of the above terms in the embodiments of the present invention can be understood according to specific circumstances.

[0024] In the embodiments of the present invention, unless otherwise clearly specified and limited, the first feature being "on" or "under" the second feature can be that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, the first feature being "above", "over" and "on top of" the second feature can be that the first feature is directly above or obliquely above the second feature, or merely indicates that the first feature has a higher horizontal height than the second feature. The first feature being "under", "below" and "beneath" the second feature can be that the first feature is directly below or obliquely below the second feature, or merely indicates that the first feature has a lower horizontal height than the second feature.

[0025] In the description of this specification, the description referring to terms such as "one embodiment", "some embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the embodiments of the present invention. In this specification, the schematic descriptions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples. To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0026] The following will be combined with Figure 1 A dynamic gradient mixing and conveying device for a chromatographic column and a chromatographic column analysis device provided by an embodiment of the present invention will be described. It should be understood that the following is only a schematic embodiment of the present invention and does not constitute any special limitation to the present invention.

[0027] An embodiment of the first aspect of the present invention provides a dynamic gradient mixing and conveying device for a chromatographic column, as Figure 1 shown. The dynamic gradient mixing and conveying device for the chromatographic column includes: a sample injection bottle 100, in which a phase A is contained; a phase B input unit, which is connected to the sample injection bottle 100 to input phase B into the sample injection bottle 100; a mixed phase output unit, the sample injection bottle 100 is connected to the mixed phase output unit, and the mixed phase output unit is used to be connected to the chromatographic column to convey the mixed phase in the sample injection bottle 100 into the chromatographic column; a liquid flow rate control unit, which extends into the sample injection bottle 100 and is used to adjust the flow rate of the mixed phase conveyed into the chromatographic column.

[0028] In the dynamic gradient mixing and conveying device of the chromatographic column provided by the present invention, it includes a sample injection bottle 100, a B-phase input unit, a mixed-phase output unit, and a liquid flow rate control unit. The sample injection bottle 100 contains the A-phase. The B-phase input unit is connected to the sample injection bottle 100 and can input the B-phase into the sample injection bottle 100 to mix with the A-phase to form a mixed phase. The mixed-phase output unit is connected to the sample injection bottle 100 and the chromatographic column, and can convey the mixed phase in the sample injection bottle 100 into the chromatographic column. The liquid flow rate control unit extends into the sample injection bottle 100 and is used to adjust the flow rate of the mixed phase conveyed into the chromatographic column. For example, the A-phase is a mobile phase such as water, and the B-phase is an organic phase.

[0029] Through this structural arrangement, during the ultra-narrow inner diameter chromatographic analysis process, the flow rate of the mixed phase entering the ultra-narrow inner diameter chromatographic column can be controlled by the liquid flow rate control unit, making its flow rate more stable, thereby improving the stability and reproducibility of the analysis results.

[0030] In an embodiment of the present invention, the liquid flow rate control unit includes a first capillary 210 and a first pressure control valve 220.

[0031] Wherein, one end of the first capillary 210 is connected to a gas source, the other end of the first capillary 210 extends into the sample injection bottle 100 and is above the liquid level of the mixed phase in the sample injection bottle 100, and the first pressure control valve 220 is arranged on the first capillary 210.

[0032] For example, the gas source is a nitrogen gas source. That is, one end of the first capillary 210 is connected to the nitrogen gas source, and the other end of the first capillary 210 extends above the liquid level of the mixed phase in the sample injection bottle 100. Nitrogen can enter the sample injection bottle 100 through the first capillary 210 to squeeze the mixed phase in the sample injection bottle 100 into the chromatographic column through the mixed-phase output unit. The first pressure control valve 220 is arranged on the first capillary 210. The first pressure control valve 220 can control the nitrogen pressure input from the nitrogen gas source into the sample injection bottle 100, and further, the flow rate of the mixed phase can be controlled.

[0033] In an embodiment of the present invention, the B-phase input unit includes a B-phase liquid storage tank 310 and a second capillary 320. One end of the second capillary 320 is connected to the B-phase liquid storage tank 310, and the other end of the second capillary 320 extends into the sample injection bottle 100 and is inserted into the A-phase.

[0034] Furthermore, in an embodiment of the present invention, the B-phase input unit further includes a third capillary 330 and a second pressure control valve 340.

[0035] One end of the third capillary 330 is connected to a gas source, and the other end of the third capillary 330 extends into the B-phase liquid storage tank 310 and is above the liquid level of the B-phase liquid storage tank 310. A second pressure control valve 340 is provided on the third capillary 330.

[0036] Specifically, both ends of the second capillary 320 are respectively connected to the B-phase liquid storage tank 310 and the sample injection vial 100. In particular, the second capillary 320 is inserted into the interior of the A-phase to improve the mixing uniformity of the A-phase and the B-phase. The B-phase in the B-phase liquid storage tank 310 can enter the sample injection vial 100 through the second capillary 320 and mix with the A-phase to form a mixed phase. A third capillary 330 is provided above the liquid level of the B-phase liquid storage tank 310, and the third capillary 330 can communicate with a nitrogen gas source. Nitrogen gas can enter the B-phase liquid storage tank 310 through the third capillary 330 to squeeze the B-phase in the B-phase liquid storage tank 310 into the sample injection vial 100 through the second capillary 320. A second pressure control valve 340 is provided on the third capillary 330. By adjusting the second pressure control valve 340, the flow rate of the B-phase entering the sample injection vial 100 can be controlled to flexibly adjust the mixing ratio of the mixed phase in real time, ensuring the uniformity of the mixed phase entering the ultra-narrow inner diameter chromatographic column.

[0037] In addition, by adjusting the first pressure control valve 220 and the second pressure control valve 340, the mixed phase after real-time mixing in the sample injection vial 100 can be transported to the chromatographic column to avoid the problem of gradient lag.

[0038] For example, the first pressure control valve 220 and the second pressure control valve 340 are pressure reducing valves provided at the outlet of the nitrogen gas cylinder.

[0039] In an embodiment of the present invention, the dynamic gradient mixing and transporting device of the chromatographic column further includes a mixer, which is connected to the sample injection vial 100 and is used to mix the mixed phase in the sample injection vial 100 to improve the mixing effect of the mixed phase, thereby improving the accuracy of the analysis result.

[0040] In an embodiment of the present invention, the chromatographic column is used to extend into the sample injection vial 100 and be inserted into the interior of the mixed phase. During the analysis process, nitrogen gas enters the sample injection vial 100 through the first capillary 210, causing the mixed phase in the sample injection vial 100 to be squeezed into the chromatographic column.

[0041] In an embodiment of the present invention, a first interface 110 is provided on the sample injection vial 100. The first capillary 210 passes through the first interface 110 and extends into the sample injection vial 100. A first sealing device is provided between the first capillary 210 and the first interface 110.

[0042] In an embodiment of the present invention, a second interface 120 is provided on the sample vial 100. The second capillary 320 passes through the second interface 120 and extends into the sample vial 100. A second sealing device is provided between the second capillary 320 and the second interface 120.

[0043] Furthermore, in an embodiment of the present invention, a third interface 130 is provided on the sample vial 100. The chromatographic column passes through the third interface 130 and extends into the sample vial 100. A third sealing device is provided between the chromatographic column and the third interface 130.

[0044] For example, the first interface 110, the second interface 120, and the third interface 130 on the sample vial 100 are all peek connectors. The first capillary 210, the second capillary 320, and the chromatographic column respectively pass through the corresponding peek connectors and extend into the sample vial 100. A first sealing device, a second sealing device, and a third sealing device are respectively provided between the first capillary 210, the second capillary 320, and the chromatographic column and the corresponding peek connectors to ensure the sealing of the sample vial 100. For example, the first sealing device, the second sealing device, and the third sealing device are all rubber sealing rings.

[0045] According to the embodiments described above, the dynamic gradient mixing and conveying device of the chromatographic column can achieve precise and uniform mixing, ensure the continuity of gradient elution, ensure the stable supply of phase A, improve the system operation efficiency, can control the dynamic ratio of the mixed phase in real time, significantly improve the separation effect and analysis resolution, has strong adaptability, is particularly suitable for ultra-narrow inner diameter chromatographic columns, and can meet the high-precision analysis requirements of trace samples and complex samples.

[0046] An embodiment of the second aspect of the present invention provides a chromatographic column analysis device, including a chromatographic column and the dynamic gradient mixing and conveying device of the chromatographic column as described above. The chromatographic column is connected to the mixed phase output unit.

[0047] In particular, the chromatographic column is an ultra-narrow inner diameter chromatographic column.

[0048] In the chromatographic column analysis device provided by the present invention, since it includes the dynamic gradient mixing and conveying device of the chromatographic column as described above, it also has the above-mentioned various advantages.

[0049] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A dynamic gradient mixing and delivery device for a chromatographic column, characterized in that: include: A sampling bottle (100), wherein the sampling bottle (100) contains phase A; A phase B input unit, the phase B input unit being connected to the injection bottle (100) so as to input the phase B into the injection bottle (100); a mixed phase output unit, the injection bottle (100) being connected to the mixed phase output unit, the mixed phase output unit being used to be connected to a chromatographic column so as to transport the mixed phase in the injection bottle (100) into the chromatographic column; A liquid flow rate control unit, the liquid flow rate control unit extends into the injection bottle (100) and is used to adjust the flow rate of the mixed phase delivered to the chromatographic column.

2. The dynamic gradient mixing and delivery device of a chromatographic column according to claim 1, characterized in that: The liquid flow rate control unit comprises a first capillary tube (210) and a first pressure control valve (220). One end of the first capillary tube (210) is connected to a gas source, the other end of the first capillary tube (210) extends into the injection bottle (100) and is located above the liquid level of the mixed phase in the injection bottle (100), and the first pressure control valve (220) is provided on the first capillary tube (210).

3. The dynamic gradient mixing and delivery device of the chromatographic column according to claim 2, characterized in that: The B phase input unit comprises a B phase liquid reservoir (310) and a second capillary (320), one end of the second capillary (320) is connected to the B phase liquid reservoir (310), and the other end of the second capillary (320) extends into the injection bottle (100) and is inserted into the interior of the A phase.

4. The dynamic gradient mixing and delivery device of the chromatographic column according to claim 3, characterized in that: The B-phase input unit further includes a third capillary tube (330) and a second pressure control valve (340). One end of the third capillary tube (330) is connected to the gas source, the other end of the third capillary tube (330) extends into the B-phase liquid reservoir (310) and is located above the liquid surface of the B-phase liquid reservoir (310), and the second pressure control valve (340) is arranged on the third capillary tube (330).

5. The dynamic gradient mixing and transporting device for a chromatographic column according to any one of claims 1 to 4, characterized in that: The dynamic gradient mixing and conveying device of the chromatographic column further comprises a mixer, which is connected to the injection bottle (100) and is used to mix the mixed phase in the injection bottle (100).

6. The dynamic gradient mixing and delivery device for a chromatographic column according to claim 4, characterized in that: The chromatographic column is used to extend into the injection bottle (100) and be inserted into the mixed phase.

7. The dynamic gradient mixing and delivery device for a chromatographic column according to claim 2, characterized in that: The injection bottle (100) is provided with a first interface (110), the first capillary (210) passes through the first interface (110) and extends into the injection bottle (100), and a first sealing device is provided between the first capillary (210) and the first interface (110).

8. The dynamic gradient mixing and delivery device for a chromatographic column according to claim 3, characterized in that: The injection bottle (100) is provided with a second interface (120), the second capillary tube (320) passes through the second interface (120) and extends into the injection bottle (100), and a second sealing device is provided between the second capillary tube (320) and the second interface (120).

9. The dynamic gradient mixing and delivery device for a chromatographic column according to claim 6, characterized in that: The injection bottle (100) is provided with a third interface (130), the chromatographic column is used to pass through the third interface (130) and extend into the injection bottle (100), and a third sealing device is provided between the chromatographic column and the third interface (130).

10. A chromatographic column analysis device, characterized in that: A dynamic gradient mixing and transporting device comprising a chromatographic column and the chromatographic column according to any one of claims 1 to 9, wherein the chromatographic column is connected to the mixed phase output unit.

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