High-efficiency and high-capacity conical chromatographic column with segmented particle size damping and application of high-efficiency and high-capacity conical chromatographic column

Abstract

The invention discloses a segmented particle size damping high-efficiency large-capacity conical chromatographic column and application, and relates to the technical field of chromatographic equipment, the key points of the technical scheme are as follows: the segmented particle size damping high-efficiency large-capacity conical chromatographic column comprises a soft chromatographic column tube, the soft chromatographic column tube is tightly sleeved with a column tube jacket, the soft chromatographic column tube is internally filled with three kinds of chromatographic fillers with different particle sizes, and the column body is conical; an upper sieve plate, a first fixing connector, a first sealing connector and an inlet pipeline are arranged at the upper outlet, the inlet pipeline is connected with a back pressure device, the back pressure device is connected with an infusion pump connecting pipe, and a lower sieve plate, a second fixing connector, a second sealing connector and an outlet pipeline are arranged at the lower outlet. Under the use of particle size damping and back pressure increase, the device can display a better enrichment effect and maintain corresponding column efficiency, the special design provides a more effective way for sample online enrichment and volume concentration in life science, biological detection and food and drug detection, and the device is suitable for separation and enrichment of trace substances and has a wide application prospect. And the method is suitable for large-capacity substance analysis.

Description

Technical Field

[0001] This invention relates to the field of chromatography equipment technology, and more specifically, to a high-efficiency, high-capacity conical chromatographic column with segmented particle size damping and its application. Background Technology

[0002] Conventional liquid chromatography columns primarily utilize solid-liquid separation, separating different components through the dynamic equilibrium process of substance distribution or adsorption between the mobile and stationary phases. However, the separation efficiency of common linear chromatographic methods is limited by factors such as coexisting substances, the amount of substances adsorbed or distributed on the stationary phase, and the composition of the mobile phase. Furthermore, because the chromatographic process involves mass transfer, it is essentially a dilution process for component analysis, limiting the analysis of substances within a low dynamic range and thus restricting method sensitivity. Additionally, the column material itself can exhibit some activity, causing broadening of components and poor separation. Moreover, the high pressure required for the system also restricts the choice of suitable materials. Summary of the Invention

[0003] The purpose of this invention is to provide a high-efficiency, large-capacity conical chromatographic column with segmented particle size damping and its application. It employs three particle sizes for segmented packing. During chromatographic separation, the damping caused by the particle size and the change in the column's inner diameter create a kinetic process different from conventional chromatography. This results in better applicability for separating large-capacity material systems and provides a more effective method for online enrichment and volumetric concentration of samples in life sciences, biological detection, and food and drug testing. It is suitable for the separation and enrichment of trace substances and for large-capacity material analysis.

[0004] The above-mentioned technical objective of the present invention is achieved through the following technical solution: a segmented particle size damping high-efficiency large-capacity conical chromatographic column, comprising a flexible chromatographic column tube, a column tube jacket tightly fitted to the outside of the flexible chromatographic column tube, the interior of the flexible chromatographic column tube being filled with three different particle sizes of chromatographic packing material, the column body being conical, the upper outlet of the flexible chromatographic column tube being provided with an upper sieve plate, a fixed joint one, and a sealing joint one in sequence from bottom to top, the fixed joint one and the sealing joint one being provided with an inlet pipe penetrating through themselves at their center, the other end of the inlet pipe being connected to a back pressure device, the back pressure device being connected to an infusion pump connecting pipe, the lower outlet of the flexible chromatographic column tube being provided with a lower sieve plate, a fixed joint two, and a sealing joint two in sequence from top to bottom, the fixed joint two and the sealing joint two being provided with an outlet pipe penetrating through themselves at their center.

[0005] The present invention is further configured such that: the column tube jacket is woven from resin fiber combined with stainless steel wire, and the flexible chromatography column tube is made of polytetrafluoroethylene.

[0006] The present invention is further configured such that the soft chromatographic column is wider at the top and narrower at the bottom, and the inlet diameter is 5-10 times the outlet diameter.

[0007] The present invention is further configured such that the counter-pressure device is composed of large particle packing material, sintered stainless steel, or inert ceramic powder.

[0008] By adopting the above technical solution, in actual operation, the column is packed with three types of packing materials of small, medium and large particle sizes in sequence by pressure packing. Due to the packing of different particle sizes and the conical design, samples with larger volumes or concentrations can be injected than those in general chromatograms of the same type. Under the action of particle size damping and with the use of increased back pressure, better enrichment effect can be shown, and the corresponding column efficiency can be maintained. This special design provides a more effective way for online enrichment and volumetric concentration of samples in life sciences, biological detection, and food and drug detection. It is suitable for the separation and enrichment of trace substances and for large-volume substance analysis.

[0009] This invention further provides an application of a segmented particle size damped high-efficiency large-capacity conical chromatographic column in online sample enrichment and volumetric concentration in life sciences, biological detection, and food and drug detection.

[0010] The present invention is further configured such that the segmented particle size damped high-efficiency large-capacity conical chromatographic column is suitable for the separation and enrichment of trace substances, as well as large-capacity substance analysis.

[0011] In summary, the present invention has the following beneficial effects: The present invention attempts to redesign the chromatographic column system, deviating from the traditional approach of changing the stationary phase, mobile phase, column diameter, and packing particle size. Instead, it selects a soft, inert material to construct the column tube. While choosing a conical shape, an outer protective sleeve is added to the soft column tube to enhance its pressure resistance. In actual operation, the column is packed with packing material of small, medium, and large particle sizes in sequence using a pressure-loading method. Due to the different particle sizes and the conical design, a larger volume or concentration of sample can be injected compared to typical chromatographic methods. Under the effect of particle size damping and with increased back pressure, better enrichment is achieved while maintaining the corresponding column efficiency. This special design provides a more effective method for online enrichment and volumetric concentration of samples in life sciences, biological detection, and food and drug testing. It is suitable for the separation and enrichment of trace substances and for large-volume material analysis. Attached Figure Description

[0012] Figure 1 This is a schematic diagram of the structure of the high-efficiency, high-capacity conical chromatographic column with segmented particle size damping according to the present invention.

[0013] In the diagram: 1. Column tube jacket; 2. Fixed connector one; 3. Sealing connector one; 4. Inlet pipe; 5. Back pressure device; 6. Infusion pump connection pipe; 7. Upper sieve plate; 8. Soft chromatography column tube; 9. Lower sieve plate; 10. Outlet pipe; 11. Sealing connector two; 12. Fixed connector two. Detailed Implementation

[0014] The following is in conjunction with the appendix Figure 1 The present invention will be described in further detail below.

[0015] Example 1: A high-efficiency, high-capacity conical chromatographic column with segmented particle size damping, such as... Figure 1 As shown, the system includes a flexible chromatography column 8, which is tightly fitted with a column sleeve 1. The column sleeve 1 is woven from resin fiber and stainless steel wire, while the flexible chromatography column 8 is made of polytetrafluoroethylene (PTFE). The flexible chromatography column 8 is filled with three different particle sizes of chromatographic packing materials. Its column body is conical, wider at the top and narrower at the bottom, with the inlet diameter being 5-10 times the outlet diameter. At the outlet of the flexible chromatography column 8, from bottom to top, there is an upper sieve plate 7, a fixed connector 2, and a sealing connector 3. An inlet pipe 4 penetrates the center of each of the fixed connector 2 and sealing connector 3. The other end of the inlet pipe 4 is connected to a backpressure device 5, which is composed of large-particle packing, sintered stainless steel, or inert ceramic powder. The back pressure device 5 is connected to the infusion pump connecting pipe 6. The lower outlet of the soft chromatographic column tube 8 is provided with a lower sieve plate 9, a fixed joint 12 and a sealing joint 11 from top to bottom. An outlet pipe 10 is provided at the center of the fixed joint 12 and the sealing joint 11.

[0016] Example 2: Principle chromatographic column 1, packed with C18 10μ, 5μ, 3.5μ packing material in a 1:2:3 ratio, 5cm long, with an upper inlet inner diameter of 4.6mm and a lower outlet diameter of 2.1mm. Methanol with a small amount of isopropanol was used as a suspension, packed in the order of 3.5μ, 5μ, 10μ, and compacted at 300psi under pressure, ultimately reaching 500psi. The column remained intact. The retention solution was methanol-water (90:10). The column was then loaded into a liquid chromatography system with a mobile phase of methanol:water (80:20) and a sample of 0.1% caffeine in methanol. UV detection was performed.

[0017]

[0018] The results showed that the final chromatographic display was good within the flow rate range of 0.5-1.5 ml / min. The optimal flow rate was close to that of a conventional chromatographic column with an inner diameter of 4.6 mm, and no abnormal peak broadening was observed.

[0019] Example 3: Principle chromatographic column 2, packed with C18 packing material of 5μ, 3.5μ, and 1.7μ in a 1:2:3 ratio. The column is 5cm long, with an upper inlet inner diameter of 2.1mm and a lower outlet diameter of 1.2mm. Methanol with a small amount of isopropanol was used as a suspension, packed in the order of 1.7μ, 3.5μ, and 5μ, initially at 200psi and then compacted to 500psi. The column remained intact. The retention solution was methanol-water (90:10). The column was then loaded into a liquid chromatography system with a mobile phase of methanol:water (80:20) and a sample of 0.01% benzene and toluene-methanol solution. UV detection was performed.

[0020]

[0021]

[0022] Under good separation conditions, the loading (injection volume) does not show a significant effect, indicating that it can handle a high elution loading, with good separation effect and peak shape, and has a high theoretical plate number.

[0023] Example 4: Principle chromatographic column 2, packed with C18 packing material of 5μ, 3.5μ, and 1.7μ in a ratio of 1:2:3. The column length was 5cm, with an upper inlet inner diameter of 2.1mm and a lower outlet diameter of 1.2mm. Methanol with a small amount of isopropanol was used as a suspension, packed in the order of 1.7μ, 3.5μ, and 5μ, initially at 200psi and then compacted to 500psi. The column remained intact. The retention solution was methanol-water (90:10). The column was then loaded into a liquid chromatography system with a mobile phase of methanol:water (80:20) and a sample of 0.01% Coptis chinensis methanol solution. UV detection was performed.

[0024]

[0025] The mixture exhibits good separation performance, with no abnormal peak shapes for the target substances, demonstrating separation efficiency at high capacity.

[0026] This specific embodiment is merely an explanation of the present invention and is not intended to limit the invention. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they are within the scope of the claims of the present invention.

Claims

1. A high-efficiency, high-capacity conical chromatographic column with segmented particle size damping, characterized in that: The system includes a soft chromatography column (8), which is tightly fitted with a column sleeve (1). The soft chromatography column (8) is filled with three different particle sizes of chromatographic packing material. The column body is conical. At the upper outlet of the soft chromatography column (8), from bottom to top, there are an upper sieve plate (7), a fixed connector one (2), and a sealing connector one (3). At the center of the fixed connector one (2) and the sealing connector one (3), there is an inlet pipe (4) that penetrates through them. The other end of the inlet pipe (4) is connected to a back pressure device (5). The back pressure device (5) is connected to an infusion pump connecting pipe (6). At the lower outlet of the soft chromatography column (8), from top to bottom, there are a lower sieve plate (9), a fixed connector two (12), and a sealing connector two (11). At the center of the fixed connector two (12) and the sealing connector two (11), there is an outlet pipe (10) that penetrates through them.

2. The high-efficiency, high-capacity conical chromatographic column with segmented particle size damping according to claim 1, characterized in that: The column sleeve (1) is woven from resin fiber and stainless steel wire, and the soft chromatography column (8) is made of polytetrafluoroethylene.

3. The high-efficiency, high-capacity conical column with segmented particle size damping according to claim 1, characterized in that: The soft chromatographic column (8) is wider at the top and narrower at the bottom, with the inlet diameter being 5-10 times the outlet diameter.

4. The high-efficiency, high-capacity conical column with segmented particle size damping according to claim 1, characterized in that: The counterpressure device (5) is composed of large particle filler, sintered stainless steel or inert ceramic powder.

5. The application of the segmented particle size damped high-efficiency large-capacity conical chromatographic column according to claim 1 in online enrichment and volume concentration of samples in life sciences, biological detection, and food and drug detection.

6. The application according to claim 5, characterized in that: The segmented particle size damped high-efficiency, high-capacity conical column is suitable for the separation and enrichment of trace substances, as well as large-volume substance analysis.

Images