Solid-phase extraction method for improving experimental recovery rate of immunoaffinity column

Abstract

The invention discloses a solid-phase extraction method for improving the experimental recovery rate of an immunoaffinity column, and relates to the technical field of solvent extraction, and the method comprises the following steps: S1, putting the immunoaffinity column in a feeding position of a solid-phase extraction instrument; s2, a reagent is pumped into the plunger, and the plunger is filled with the reagent, so that the plunger is pre-filled with fluid; s3, the plunger is moved downwards to be connected with the immunoaffinity column, and when the internal pressure of the immunoaffinity column reaches a threshold value, a lower cover of the immunoaffinity column is driven to be automatically separated; and S4, the plunger is moved upwards. A reagent is filled into the plunger in advance, so that air in the plunger is emptied. And then the plunger is jointed to enter the immunoaffinity column, so that the condition that the protection liquid is completely discharged by air is not easy to occur during subsequent sample loading. Besides, the lower cover at the bottom of the immunoaffinity column is automatically removed by pressing down the plunger, and air in the plunger is emptied, so that air compression is not easy to occur in the process of pressing down the plunger, and the condition that the protective liquid is pushed out by compressed air expansion at the moment of removing the cover is not easy to occur.

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 technology that uses solid adsorbents to selectively adsorb and elute to separate target substances for enrichment and purification. It is based on the principle of liquid-solid chromatography, is simple and efficient to operate, and is widely used in the fields of medicine, food, and environment. Compared with traditional liquid-liquid extraction, SPE saves solvent and has a higher recovery rate.

[0003] Solid phase extraction is used to separate and enrich target compounds. Different types of solid phase extraction columns are used for different substances. Conventional solid phase extraction columns are generally composed of column tubes, fillers and sieve plates, but there are some special solid phase extraction columns, such as immunoaffinity columns, which, in addition to column tubes, fillers and sieve plates, also have protective liquid in the column tube to protect the filler and keep it active.

[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 objective of the present invention is achieved through the following technical solutions: A solid phase extraction method for improving the recovery rate of immunoaffinity column experiments comprises the following steps: S1. Place the immunoaffinity column into the loading position of the solid phase extractor; S2. Pump the reagent into 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 a threshold, the lower cover is driven to automatically detach.

[0007] 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.

[0008] Preferably, in S2, a waste liquid collection tank is provided below the plunger.

[0009] 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.

[0010] Preferably, a solution collecting tank is arranged beside the waste liquid collecting tank, and the waste liquid collecting tank and the solution collecting tank are arranged on a transfer platform.

[0011] Preferably, in S2, the reagent is pumped into the plunger by a syringe pump.

[0012] Preferably, the plunger comprises a column, a pipeline is arranged in the column, and the bottom end of the pipeline has an inverted cone-shaped opening.

[0013] 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.

[0014] Preferably, a sealing ring is sleeved on the outer wall of the plunger.

[0015] The beneficial effects of the present invention are: 1. The air in the plunger is emptied by pre-filling the reagent into the plunger. The plunger is then engaged with the immunoaffinity column, making it less likely that the 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, it is less likely that the air will be compressed during the plunger pressing process, and it is even less likely that the compressed air will expand and push out the protective liquid when the cover is removed.

[0016] 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

[0017] Figure 1 is a schematic diagram of the structure of the plunger; Figure 2 Schematic diagram of the structure of the immunoaffinity column.

[0018] Figure numerals: 1. immunoaffinity column; 2. plunger; 3. lower cover; 4. waste liquid collecting tank; 5. lower cover collecting tank; 6. solution collecting tank; 7. injection pump; 8. column; 9. pipeline; 10. sealing ring; 11. three-way valve. DETAILED DESCRIPTION

[0019] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0020] A solid phase extraction method for improving the recovery rate of immunoaffinity column experiments comprises the following steps: S1, placing the immunoaffinity column 1 with the upper cover removed into the loading position of the solid phase extractor; S2, pumping the reagent into and filling the plunger 2, so that the plunger 2 is pre-filled with fluid; S3, the plunger 2 is moved downward to engage with the immunoaffinity column 1, and when the internal pressure of the immunoaffinity column 1 reaches a threshold value, the lower cover 3 is driven to automatically detach; S4. Move the plunger 2 upward to form negative pressure in the immunoaffinity column 1.

[0021] 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 illustrated, 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.

[0022] A waste liquid collection tank 4 is preferably provided below the plunger 2. In the above 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 emptied, and then the plunger 2 is engaged into the immunoaffinity column 1, so that it is not easy for air to drain the protective liquid during subsequent sample loading.

[0023] In addition, see Figure 2 In S3, a positive pressure is formed in the immunoaffinity column 1 by pressing down the plunger 2, and after the pressure exceeds the threshold, the lower cover 3 at the bottom of the immunoaffinity column 1 will automatically detach. For example, the above-mentioned waste liquid collection tank 4 may also be provided with a partition net, and the upper part of the waste liquid collection tank 4 is defined as the lower cover collection tank 5 by the partition net, so that the automatically detached lower cover 3 will fall into the lower cover collection tank 5.

[0024] It can be understood that before S3, the bottom of the immunoaffinity column 1 always has the lower cover 3, so that the protective liquid inside is not easily discharged due to gravity, and the lower cover 3 is automatically detached through S3 instead of being manually performed. In addition, since the air inside the plunger 2 is evacuated in S2, when the plunger 2 is pressed down, there is no compressed air inside the immunoaffinity column 1 as in the prior art, and until the lower cover 3 is removed, it is not easy for the compressed air inside the immunoaffinity column 1 to restore the atmospheric pressure and expand to push out the protective liquid.

[0025] In S4, the plunger 2 is moved upward again, which forms a negative pressure inside the immunoaffinity column 1. Under the negative pressure, the protective liquid moves upward and contacts the fluid inside the plunger 2, forming a zero gap between the protective liquid and the solvent. And under the action of liquid tension, the protective liquid is less likely to flow out due to gravity.

[0026] 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 both in a full state, the immunoaffinity column 1 will always be in an infiltrated state, thereby achieving a further increase in the recovery rate.

[0027] In a preferred example, the pumping speed of the reagent in S2 may be segmented, for example, it may 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.

[0028] In addition, the plunger 2 may preferably include a column 8, and a pipe 9 is disposed in the column 8, and the bottom end of the pipe 9 has an inverted cone-shaped opening. In the above-mentioned liquid addition stage, as the fluid is injected into the plunger 2, the fluid gradually increases from the pipe 9 to the opening, thereby helping the fluid to be subjected to greater liquid tension and remain in the pipe 9, and not easy to drip.

[0029] Furthermore, a sealing ring 10 may 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 to move upward.

[0030] A solution collection tank 6 may be provided beside the waste liquid collection tank 4, and the solid phase extraction solution may be discharged downward into the solution collection tank 6. 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 relative alignment of different collection tanks and the plunger 2 may be achieved by moving the transfer platform.

[0031] The downward pressing and upward moving of the above-mentioned plunger 2 can be achieved by 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.

[0032] The above is only 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 regarded as excluding other embodiments, but 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 the technology or knowledge of the relevant field. The changes and modifications made by those skilled in the art shall not deviate from the spirit and scope of the present invention, and shall be within the scope of protection of the claims attached to the present invention.

Claims

1. A solid phase extraction method for improving the recovery rate of immunoaffinity column experiments, characterized in that: The steps include: S1. Place the immunoaffinity column (1) into the loading position of the solid phase extractor; S2, pumping the reagent into and filling the plunger (2), so that the plunger (2) is pre-filled with fluid; S3, the plunger (2) is moved downward to engage with the immunoaffinity column (1), and when the internal pressure of the immunoaffinity column (1) reaches a threshold value, the lower cover (3) is driven to automatically detach.

2. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiment according to claim 1, characterized in that: include: S4, moving the plunger (2) upward to form a 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).

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

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

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

6. 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 a syringe pump (7).

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

8. The solid phase extraction method for improving the recovery rate of immunoaffinity column experiment according to claim 1 or 6, characterized in that: The pumping of reagents in S2 includes the following stages that are 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.

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

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