Capillary liquid-phase chromatographic collumn and making method

A liquid chromatography column and capillary technology, which is applied in the field of capillary liquid chromatography column and its preparation, can solve the problems of not being as durable as stainless steel column, high technical content, high price, etc., so as to improve the degree of modification and improve the separation efficiency , the effect of high mechanical strength

Inactive Publication Date: 2006-08-09
TSINGHUA UNIV
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Problems solved by technology

The column packing technology of this method is very important, which determines the performance of the column to a large extent, but this technology is difficult to master, so most researchers use the method of directly purchasing commercial capillary ion exchange chromatographic columns
However, due to the high technical content in the preparation of capillary liquid chromatography columns, it is expensive, and the price of a commercial capillary liquid chromatography column is almost twice or even higher than that of ordinary liquid chromatography columns.
At the same time, compared with the traditional liquid chromatography column with an inner diameter of 4.6mm, the service life of the capillary liquid chromatography column is very low. This is because the capillary column usually adopts an elastic quartz column tube, which has poor mechanical strength and is fragile, and is far less durable than the stainless steel column tube.
In addition, the composition of biological samples is complex, and the capacity of the capillary column is small, so it is easy to be contaminated
At present, the cost of proteomics research is very high, and one of the important factors is the need to purchase a large number of expensive capillary liquid chromatography columns
In addition, the separation efficiency of biomacromolecules on traditional particle-packed ion-exchange chromatography columns is low, mainly due to the slow diffusion process of solutes in the pores of particle packing, which affects the mass transfer process of large and irregularly shaped biomolecules

Method used

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Embodiment 1

[0019] The present embodiment capillary liquid chromatography column (such as figure 1 Shown) the preparation method comprises the following steps:

[0020] 1. Pretreatment of the capillary column: take an elastic quartz capillary 1 with an inner diameter of 0.32mm and a length of 20cm, use the negative pressure of a vacuum pump to pass a 0.1M sodium hydroxide solution through the capillary for 2 hours, then rinse the capillary with water until the pH is 7, Acetone that will be 50 times the capillary volume passes through the capillary, blow dry with nitrogen for 2 minutes, take 30% γ-methacrylic acid oxypropyltrimethoxysilane acetone solution to fill the capillary column, seal both ends with silicone rubber for 12 hours, and use Rinse with acetone and dry with nitrogen gas.

[0021] 2. The preparation of the ion exchange monolithic column is carried out in two steps:

[0022] (1) Glyceryl methacrylate and ethyl dimethacrylate are mixed in a mass ratio of 3:1 to form a monom...

Embodiment 2

[0027] The present embodiment capillary liquid chromatography column (such as figure 1 Shown) the preparation method comprises the following steps:

[0028] 1. Pretreatment of the capillary column: same as in Example 1.

[0029]2. The preparation of the ion exchange monolithic column is carried out in two steps:

[0030] (1) Glyceryl methacrylate and ethyl dimethacrylate are mixed in a mass ratio of 4:1 to form a monomer mixture, and the porogen n-propanol, 1,4-butanediol and water are mixed in a mass ratio of 4 : 3:1 mixing to form a porogen mixture, the monomer mixture and the porogen mixture are mixed together at a volume ratio of 3:4, and then add azobisisobutyronitrile whose mass is 0.8% of the polymerization monomer mass As an initiator, ultrasonically mix evenly, pass nitrogen into the mixture for 2 minutes to remove the air in the mixture, then fill the polymer mixed solution into the capillary, the amount of the mixed solution is determined according to the needs of...

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Abstract

Present invention relates to a liquid phase chromatography column and preparation method, which includes one elastic quartz capillary without adhesive joining. It features middle of said capillary set with integral stationary phase formed by organic polymer in situ polymerization and modified by weak acid cation, lower reaches of said stationary phase set with detect window removing coated surface polyimide. Said invented includes 1, to proceed general pretreatment to elastic quartz inner wall, 2, making integral stationary phase basal body using monomer mixture, porogenic agent mixture and initiator, 3, modifying stationary phase basal body by modifier to obtain subacidity cation exchange integral stationary phase, 4, making detect window to obtain subacidity cation exchange capillary liquid phase chromatography column. Said invention has los price, easy to control preparing condition, fine reproduction quality, fine consistency with biological sample, suitable for biomacromolecule sample isolation analysis.

Description

technical field [0001] The invention relates to a capillary liquid chromatography column and a preparation method thereof, in particular to a capillary liquid chromatography column capable of weakly acidic cation exchange and a preparation method thereof. Background technique [0002] Capillary liquid chromatography (μ-HPLC) is an efficient and rapid separation and analysis method, usually using a capillary column with an inner diameter of 50 μm to 530 μm. Compared with the traditional liquid chromatography column with an inner diameter of 4.6 mm, it can greatly save mobile phase, Due to its unique advantages, the consumption of stationary phase and sample, and the ease of on-line connection with mass spectrometers and other detectors, it has been increasingly widely used in biochemical analysis, environment, medicine and other fields in recent years. Especially in the late 1990s, with the development of proteomics, more and more attention has been paid to capillary liquid c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/96G01N30/60
Inventor 丁明玉郑睿马继平
Owner TSINGHUA UNIV
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