A solid-phase extraction method for improving the experimental recovery rate of an immunoaffinity column

Abstract

The present invention discloses a solid phase extraction method for improving the recovery rate of immunoaffinity column experiments, which relates to the field of solvent extraction technology and comprises the following steps: S1, placing the immunoaffinity column into the loading position of a solid phase extractor; S2, pumping a reagent into and filling the plunger so that the plunger is pre-filled with fluid; S3, moving the plunger downward to engage with the immunoaffinity column, and when the internal pressure of the immunoaffinity column reaches a threshold value, driving the lower cover thereof to automatically detach; S4, moving the plunger upward. By pre-filling the plunger with reagent, the air in the plunger is emptied. The plunger is then engaged with the immunoaffinity column, making it less likely that air will drain the protective liquid during subsequent sample loading. In addition, the lower cover at the bottom of the immunoaffinity column is automatically removed by pressing down the plunger. Since the air in the plunger is emptied, air compression is less likely to occur during the process of pressing down the plunger, and it is even less likely that the compressed air will expand and push out the protective liquid when the cover is removed.

Description

Technical Field

[0001] The invention relates to the technical field of solvent extraction, in particular to a solid phase extraction method for improving the recovery rate of an immunoaffinity column experiment. Background Art

[0002] Solid-phase extraction (SPE) is a sample pretreatment technique that uses a solid adsorbent for selective adsorption and elution to separate target compounds for enrichment and purification. Based on the principles of liquid-solid chromatography, SPE is simple and efficient, and widely used in the pharmaceutical, food, and environmental fields. Compared to traditional liquid-liquid extraction, SPE uses less solvent and offers higher recovery rates.

[0003] Solid-phase extraction (SPE) instruments are used to separate and enrich target compounds. Different types of SPE cartridges are used for different substances. Conventional SPE cartridges generally consist of a column tube, filler, and sieve plate. However, some specialized SPE cartridges, such as immunoaffinity columns, contain a protective fluid in addition to the column tube, filler, and sieve plate to maintain the filler's activity.

[0004] When conventional solid-phase extraction methods are used to operate immunoaffinity columns, since the conventional small columns themselves are dry, there is no special design to avoid drying, which will eventually cause the immunoaffinity columns to be blown dry, resulting in a decrease in their recovery rate. Summary of the Invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a solid phase extraction method for improving the recovery rate of immunoaffinity column experiments.

[0006] The object of the present invention is achieved through the following technical solutions:

[0007] A solid phase extraction method for improving the recovery rate of immunoaffinity column experiments comprises the following steps:

[0008] S1. Place the immunoaffinity column in the loading position of the solid phase extractor; S2. Pump the reagent in and fill the plunger so that the plunger is pre-filled with fluid; S3. Move the plunger downward to engage with the immunoaffinity column. When the internal pressure of the immunoaffinity column reaches the threshold, the lower cover is automatically detached.

[0009] Preferably, the method further comprises: S4, moving the plunger upward to form a negative pressure in the immunoaffinity column; wherein, under the action of the negative pressure, the protective liquid in the immunoaffinity column moves upward and contacts the fluid inside the plunger.

[0010] Preferably, in said S2, a waste liquid collection tank is provided below the plunger.

[0011] Preferably, a partition net is provided in the waste liquid collecting tank to define the upper portion of the waste liquid collecting tank as a lower cover collecting tank.

[0012] Preferably, a solution collecting tank is provided beside the waste liquid collecting tank, and the waste liquid collecting tank and the solution collecting tank are provided on a transfer platform.

[0013] Preferably, in S2 , the reagent is pumped into the plunger by a syringe pump.

[0014] Preferably, the plunger comprises a column, a pipeline is provided in the column, and the bottom end of the pipeline has an inverted cone-shaped opening.

[0015] Preferably, the pumping of the reagent in S2 includes the following stages performed in sequence: a liquid filling stage and a liquid adding stage, wherein the fluid pumping speed in the liquid adding stage is lower than the fluid pumping speed in the liquid filling stage.

[0016] Preferably, a sealing ring is sleeved on the outer wall of the plunger.

[0017] The beneficial effects of the present invention are:

[0018] 1. By pre-filling the plunger with reagent, the air inside the plunger is purged. The plunger is then engaged with the immunoaffinity column, making it less likely that air will drain the protective fluid during subsequent sample loading. Furthermore, the downward pressure of the plunger automatically removes the lower cap at the bottom of the immunoaffinity column. Since the air inside the plunger is purged, air compression is less likely to occur during the downward pressure, and the compressed air is less likely to expand and push out the protective fluid when the cap is removed.

[0019] 2. By moving the plunger upward, the protective liquid rises under the action of negative pressure and contacts the fluid inside the plunger, which further eliminates the gap between the protective liquid and the liquid inside the plunger. Under the action of liquid tension, the protective liquid is less likely to flow out due to gravity. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Schematic diagram of the structure of the plunger;

[0021] Figure 2 Schematic diagram of the structure of the immunoaffinity column.

[0022] Figure numerals: 1. Immunoaffinity column; 2. Plunger; 3. Lower cover; 4. Waste liquid collection tank; 5. Lower cover collection tank; 6. Solution collection tank; 7. Injection pump; 8. Column; 9. Pipeline; 10. Sealing ring; 11. Three-way valve. DETAILED DESCRIPTION

[0023] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the embodiments. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work shall fall within the scope of protection of the present invention.

[0024] A solid phase extraction method for improving the recovery rate of immunoaffinity column experiments comprises the following steps:

[0025] S1. Place the immunoaffinity column 1 with the upper cover removed into the loading position of the solid phase extraction instrument;

[0026] S2. Pump the reagent into and fill the plunger 2 so that the plunger 2 is pre-filled with fluid;

[0027] S3, the plunger 2 is moved downward to engage with the immunoaffinity column 1. When the internal pressure of the immunoaffinity column 1 reaches a threshold, the lower cover 3 is driven to automatically detach;

[0028] S4. Move the plunger 2 upward to form negative pressure in the immunoaffinity column 1.

[0029] See also Figure 1 、 Figure 2 , wherein, in S2, a waste liquid collection tank 4 is provided below the plunger 2. For example, the reagent can be pumped into the plunger 2 by the injection pump 7 of the solid phase extractor. Figure 1 , preferably, a syringe pump 7 with 11 interfaces is shown, through which any one of the eight reagents connected to the syringe pump 7 can be pumped into the plunger 2 through the three-way valve 11.

[0030] A waste liquid collection tank 4 is preferably provided below the plunger 2. In the above-described step S2, the reagent pumped into the plunger 2 will fill the internal pipeline 9 of the plunger 2, and the overflowed reagent will be discharged downward into the waste liquid collection tank 4. At this time, the air in the plunger 2 is evacuated, and the plunger 2 is then engaged with the immunoaffinity column 1, making it less likely that air will drain the protective liquid during subsequent sample loading.

[0031] In addition, see Figure 2 In S3, the downward pressure of the plunger 2 creates a positive pressure within the immunoaffinity column 1. When the pressure exceeds a threshold, the lower cover 3 at the bottom of the immunoaffinity column 1 automatically detaches. For example, the waste liquid collection tank 4 may also be provided with a screen that defines the upper portion of the waste liquid collection tank 4 as a lower cover collection tank 5. The automatically detached lower cover 3 then falls into the lower cover collection tank 5.

[0032] It will be appreciated that, prior to step S3, the lower cover 3 remains attached to the bottom of the immunoaffinity column 1, preventing the protective fluid from being expelled due to gravity. Furthermore, step S3 allows for automatic detachment of the lower cover 3, rather than manual removal. Furthermore, because the air within the plunger 2 is evacuated in step S2, when the plunger 2 is depressed, compressed air is not formed within the immunoaffinity column 1, as in the prior art. Consequently, until the lower cover 3 is removed, the compressed air within the immunoaffinity column 1 is less likely to return to atmospheric pressure and expand, pushing out the protective fluid.

[0033] In S4, plunger 2 is moved upward again, creating a negative pressure inside immunoaffinity column 1. Under this negative pressure, the protective liquid moves upward and contacts the fluid inside plunger 2, creating a zero gap between the protective liquid and the solvent. Furthermore, due to the liquid tension, the protective liquid is less likely to drain due to gravity.

[0034] After S4, subsequent solid phase extraction operation steps can be performed. Since the plunger 2 and the pipeline 9 in the immunoaffinity column 1 are in a full state, the immunoaffinity column 1 will always be in an infiltrated state, thereby further improving the recovery rate.

[0035] In a preferred example, the pumping speed of the reagent in S2 can be segmented, for example, it can specifically include the following stages in sequence: a filling stage, a liquid adding stage, and the fluid pumping speed in the liquid adding stage is lower than the fluid pumping speed in the filling stage.

[0036] Furthermore, the plunger 2 may preferably include a cylindrical body 8, in which a tube 9 is disposed, and the bottom end of the tube 9 has an inverted tapered opening. During the aforementioned liquid addition stage, as the fluid is injected into the plunger 2, the fluid gradually increases from the tube 9 to the opening, thereby helping the fluid to be subjected to greater liquid tension and remain in the tube 9, thereby preventing it from dripping.

[0037] Furthermore, a sealing ring 10 can be sleeved on the outer wall of the plunger 2 to fill the gap between the plunger 2 and the inner wall of the immunoaffinity column 1 , thereby making it easier to form a negative pressure in S3 to drive the protective liquid upward.

[0038] A solution collection tank 6 may be provided next to the waste liquid collection tank 4, into which the solid phase extraction solution may be discharged. For example, the waste liquid collection tank 4 and the solution collection tank 6 may be provided on a transfer platform (not shown), and the different collection tanks may be aligned with the plunger 2 by moving the transfer platform.

[0039] The downward pressing and upward movement of the above-mentioned plunger 2 can be achieved through an ejection mechanism, which includes but is not limited to a pneumatic cylinder, an electric cylinder, a screw motor or an oil cylinder. As long as it can meet the up and down movement of the plunger 2, it is not limited in the present invention.

[0040] The foregoing description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the form disclosed herein and should not be construed as excluding other embodiments. Rather, the present invention can be used in various other combinations, modifications, and environments and can be modified within the scope of the concept described herein through the above teachings or techniques or knowledge in the relevant field. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention are intended to be protected by the appended claims.

Claims

1. A solid phase extraction method for improving the recovery rate of immunoaffinity column experiments, characterized by: The steps include: S1. Place the immunoaffinity column (1) into the loading position of the solid phase extraction instrument; S2. Pumping the reagent into and filling the plunger (2) so that the plunger (2) is pre-filled with fluid; S3, moving the plunger (2) downward to engage with the immunoaffinity column (1), and when the internal pressure of the immunoaffinity column (1) reaches a threshold, driving the lower cover (3) thereof to automatically detach; Also includes: S4, moving the plunger (2) upward to form negative pressure in the immunoaffinity column (1); Under the action of negative pressure, the protective liquid in the immunoaffinity column (1) moves upward and contacts the fluid inside the plunger (2).

2. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiment according to claim 1, characterized in that: In the S2, a waste liquid collection tank (4) is provided below the plunger (2).

3. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiment according to claim 2, characterized in that: A partition net is provided in the waste liquid collection tank (4) to define the upper portion of the waste liquid collection tank (4) as a lower cover collection tank (5).

4. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiment according to claim 3, characterized in that: A solution collecting tank (6) is provided beside the waste liquid collecting tank (4), and the waste liquid collecting tank (4) and the solution collecting tank (6) are provided on a transfer platform.

5. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiment according to claim 1, characterized in that: In S2, the reagent is pumped into the plunger (2) by the syringe pump (7).

6. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiments according to claim 1, wherein: The plunger (2) comprises a column (8), a pipe (9) is arranged in the column (8), and the bottom end of the pipe (9) has an inverted cone-shaped opening.

7. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiment according to claim 1 or 5, characterized in that: The pumping of the reagent in S2 includes the following stages that are carried out in sequence: A liquid filling stage and a liquid adding stage, wherein the fluid pumping speed in the liquid adding stage is lower than the fluid pumping speed in the liquid filling stage.

8. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiments according to claim 1, wherein: A sealing ring (10) is sleeved on the outer wall of the plunger (2).

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