Preparation method of directional adsorption material

Abstract

The invention discloses a method for preparing macroporous adsorption resin. The method comprises the following steps: carrying out polymerization reaction on an oil phase containing styrene, divinylbenzene, a pore-foaming agent, an initiator and span 80 and a water phase containing water and a dispersing agent under the protection of nitrogen; the particle size of the polymer is controlled by the stirring speed, and the pore size of the resin is controlled by the addition amount of xylene so as to obtain the macroporous adsorption resin with a spherical structure.

Description

Technical Field

[0001] The present invention relates to the technical field of a method for manufacturing a directional adsorption material, and particularly to a method for manufacturing a directional adsorption material. Background Art

[0002] Currently, nucleic acids include DNA and RNA and are the main objects of molecular biology research. Therefore, nucleic acid purification is the most important and fundamental technique in molecular biology experimental techniques. Currently, nucleic acid purification mainly includes two methods: silica column method and magnetic bead method. In the silica column method, a plastic tube is filled with a silicon-based glass fiber membrane, and then a nucleic acid sample containing high salt and high alcohol is added to the plastic tube. After centrifugation by a centrifuge, the filtrate is at the bottom of the tube, and the nucleic acid is adsorbed on the silicon matrix. Then, the nucleic acid is eluted with water. This method can obtain high-purity DNA, but it has high costs, and at the same time, it cannot achieve automated operation and has very low efficiency.

[0003] In the magnetic bead method, Fe3O4 nanoparticles are coated with a layer of silica, thus having the property of adsorbing nucleic acids. Its principle is the same as that of the silica column method. However, the magnetic bead method can be compatible with automated equipment to achieve automation and improve efficiency. However, the magnetic bead method for purifying nucleic acids also faces some problems, such as multiple purification steps, long purification time (a purification process is greater than 30 minutes), the need to use a large amount of ethanol or isopropanol, low recovery efficiency, and after multiple-step operations, the final nucleic acid recovery rate is less than 50%. Therefore, a resin material is needed that can efficiently and directionally adsorb various impurities in a nucleic acid sample to achieve the purpose of nucleic acid cleaning. Summary of the Invention

[0004] In order to solve certain or some technical problems existing in the prior art, the purpose of the present application is to provide a method for manufacturing a directional adsorption material, which can efficiently and directionally adsorb various impurities in a nucleic acid sample to achieve the purpose of nucleic acid cleaning.

[0005] In order to solve the above-mentioned existing technical problems, one of the purposes of the present application is achieved by adopting the following technical solutions:

[0006] A method for manufacturing a directional adsorption material, which comprises the following steps:

[0007] S1, oil phase: After mixing 9 g of styrene, 8 g of divinylbenzene, 22 g of xylene, 0.23 g of benzoyl peroxide, and 1 g of span 80 and ultrasonic dissolving, it is added to the water phase (the water phase composition: 150 ml of water, 1 ml of methylene blue), and stirred at 1600 rpm and 45 degrees for half an hour under nitrogen protection to be fully emulsified;

[0008] S2, maintaining a rotation speed of 1600 rpm under nitrogen protection, heating to 85 degrees, and stirring for 5 hours;

[0009] S3. After stirring, the system is transferred to a separatory funnel to remove the liquid.

[0010] S4. The solid is loaded into a glass column and washed by passing methanol through the column to remove xylene.

[0011] S5. The obtained solid is the macroporous adsorption resin with a fixed pore size.

[0012] Further, in step S1: The aqueous phase composition is 150 ml of water and 1 ml of methylene blue.

[0013] Further, in step S1, the dispersant is Span 80, the initiator is benzoyl peroxide, the xylene is a pore-forming agent, and the volume ratio of the oil phase to the aqueous phase is 4:15.

[0014] Further, the resin particle size is controlled by the stirring speed, and the resin pore size is controlled by the amount of xylene added.

[0015] Further, the pore-forming agent is selected from xylene or toluene, and the initiator is selected from azobisisobutyronitrile or benzoyl peroxide.

[0016] Further, the pore-forming agent is toluene; the initiator is benzoyl peroxide.

[0017] Further, in step (1), the stirring speed is 1600 rpm.

[0018] Further, the oil droplet size is 200 - 600 μm.

[0019] Further, the mass percentage of xylene in the oil phase is 55% to obtain a suitable pore size.

[0020] Further, in step S1, during the stirring process, the mixture is heated, heated and raised to 85 °C in 1 - 2 hours, and the temperature is controlled at a constant temperature of 85 ± 2 °C to carry out the polymerization reaction.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] (1) The macroporous adsorption resin is a resin material. By controlling the size of the resin particles and the pore size on the resin surface, various impurities in the nucleic acid sample can be efficiently and directionally adsorbed to achieve the purpose of nucleic acid purification. At the same time, the resin pore size meets the requirements for nucleic acid purification. By using this method, spherical macroporous adsorption resin with a particle size of 80 - 200 μm can be effectively prepared. Description of the Drawings

[0023] Figure 1 It is the chemical formula of the macroporous adsorption resin in this application; Detailed Embodiments

[0024] Next, in combination with the accompanying drawings and specific embodiments, the present application will be further described. It should be noted that, on the premise of no conflict, the following-described embodiments or technical features can be arbitrarily combined to form new embodiments.

[0025] In the description of the present application, it should be understood that the orientation or positional relationship indicated by terms such as "upper", "lower", "left", "right", etc. is based on the orientation or positional relationship shown in the accompanying drawings. It is only for the convenience of describing the present application 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 therefore should not be construed as a limitation to the present application.

[0026] The terms "first", "second", etc. in the present application are used to distinguish similar objects, rather than to describe a specific order or sequence. It should be understood that such data can be interchanged under appropriate circumstances so that the embodiments of the present application can be implemented in an order other than those illustrated or described herein, and the objects distinguished by "first", "second", etc. are usually of the same category, and the number of objects is not limited. For example, the first object can be one or multiple. In addition, "and / or" in the specification and claims means at least one of the connected objects, and the character " / ", generally represents an "or" relationship between the front and rear associated objects, which is a method for making a directional adsorption material.

[0027] Embodiment 1:

[0028] As Figure 1 shown, this embodiment provides a method for making a directional adsorption material, which is characterized by including the following steps:

[0029] S1, oil phase: After mixing 9 g of styrene, 8 g of divinylbenzene, 22 g of xylene, 0.23 g of benzoyl peroxide, and 1 g of Span 80 and ultrasonic dissolution, it is added to the water phase (water phase composition: 150 ml of water, 1 ml of methylene blue). Under nitrogen protection, it is stirred at 1600 rpm and 45 °C for half an hour to be fully emulsified;

[0030] S2, Under nitrogen protection, maintain a rotation speed of 1600 rpm, heat up to 85 °C, and stir for 5 hours;

[0031] S3, After the stirring ends, the system is transferred to a separating funnel to remove the liquid;

[0032] S4, The solid is loaded into a glass column and washed through the column with methanol to remove xylene;

[0033] In S5, the obtained solid is the macroporous adsorption resin with fixed pore size. This macroporous adsorption resin is a resin material. By controlling the size of resin particles and the pore size on the resin surface, various impurities in the nucleic acid sample can be efficiently and directionally adsorbed to achieve the purpose of nucleic acid purification. At the same time, the resin pore size meets the requirements for nucleic acid purification. By using this method, spherical macroporous adsorption resin with a particle size of 80 - 200 μm can be effectively prepared.

[0034] Furthermore, in step S1: the water phase composition is 150 ml of water and 1 ml of methylene blue.

[0035] Furthermore, in step S1, the dispersant is Span 80, the initiator is benzoyl peroxide, xylene is the pore-forming agent, and the volume ratio of the oil phase to the water phase is 4:15.

[0036] Furthermore, the resin particle size is controlled by the stirring speed, and the resin pore size is controlled by the addition amount of xylene.

[0037] Furthermore, the pore-forming agent is selected from xylene or toluene, and the initiator is selected from azobisisobutyronitrile or benzoyl peroxide.

[0038] Furthermore, the pore-forming agent is toluene; the initiator is benzoyl peroxide.

[0039] Furthermore, in step (1), the stirring speed is 1600 rpm.

[0040] Furthermore, the oil droplet size is 200 - 600 μm.

[0041] Furthermore, the mass ratio of xylene in the oil phase is 55%, so as to obtain a suitable pore size.

[0042] Furthermore, in step S1, during the stirring process, the mixture is heated, heated and raised to 85 °C in 1 - 2 hours, and the temperature is controlled at a constant temperature of 85 ± 2 °C to carry out the polymerization reaction.

[0043] The above embodiments are only the preferred embodiments of the present application, and the scope of protection of the present application cannot be limited thereby. Any non-substantive changes and substitutions made by those skilled in the art based on the present application belong to the scope of protection required by the present application.

Claims

1. A method for manufacturing a directional adsorption material, characterized in that, It includes the following steps: S1, oil phase: After mixing 9 g of styrene, 8 g of divinylbenzene, 22 g of xylene, 0.23 g of benzoyl peroxide, and 1 g of Span 80 and dissolving them by ultrasonic treatment, add them to the aqueous phase (aqueous phase composition: 150 ml of water, 1 ml of methylene blue). Under nitrogen protection, stir at 1600 rpm and 45 °C for half an hour to fully emulsify. S2, Under nitrogen protection, maintain a rotation speed of 1600 rpm and heat up to 85 °C, and stir for 5 hours. S3, After the stirring ends, transfer the system to a separatory funnel to remove the liquid. S4, Load the solid into a glass column and wash it through the column with methanol to remove xylene. S5, The obtained solid is the macroporous adsorption resin with a fixed pore size.

2. The manufacturing method of a directional adsorption material according to claim 1, characterized in that, In step S1: The aqueous phase composition is 150 ml of water and 1 ml of methylene blue.

3. The manufacturing method of a directional adsorption material according to claim 2, characterized in that: In step S1, the dispersant is Span 80, the initiator is benzoyl peroxide, xylene is the pore-forming agent, and the volume ratio of the oil phase to the aqueous phase is 4:

15.

4. The manufacturing method of a directional adsorption material according to claim 3, characterized in that: Control the resin particle size by the stirring speed and control the resin pore size by the amount of xylene added.

5. The manufacturing method of a directional adsorption material according to claim 4, characterized in that: The pore-forming agent is selected from xylene or toluene, and the initiator is selected from azobisisobutyronitrile or benzoyl peroxide.

6. The manufacturing method of a directional adsorption material according to claim 5, characterized in that: The pore-forming agent is toluene; the initiator is benzoyl peroxide.

7. The manufacturing method of a directional adsorption material according to claim 6, characterized in that: In step (1), the stirring speed is 1600 rpm.

8. The manufacturing method of a directional adsorption material according to claim 7, characterized in that: The oil droplet size is 200 - 600 μm.

9. The manufacturing method of a directional adsorption material according to claim 8, wherein: The mass ratio of xylene in the oil phase is 55%, so as to obtain a suitable pore size.

10. The manufacturing method of a directional adsorption material according to claim 9, characterized in that: In step S1, during the stirring process, heat the mixture, heat up for 1 - 2 hours to 85 °C, and control the constant temperature at 85 ± 2 °C to carry out the polymerization reaction.

Images