Nanospray capillary high pressure liquid chromatographic column and its preparation method

A high-pressure liquid chromatography and capillary column technology, applied in the field of analytical chemistry, can solve the problems of large back pressure, low flow rate, and the separation efficiency of difficult peptide samples

Inactive Publication Date: 2011-11-16
徐平
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When separating samples, the back pressure of this column is large and the flow rate is low, it is d

Method used

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  • Nanospray capillary high pressure liquid chromatographic column and its preparation method
  • Nanospray capillary high pressure liquid chromatographic column and its preparation method
  • Nanospray capillary high pressure liquid chromatographic column and its preparation method

Examples

Experimental program
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Effect test

Example Embodiment

[0016] Example 1

[0017] Put 0.5 ml of 100% methanol, a magnetic stir bar and 50 μg of 10-15 μm glass microspheres (Michrom Bioresources, Auburn, CA) into a 2 ml ampere flask into a pressure injection cell (pressure injection cell, Next Advance) Inc. 24Prospect Avenue, Averill Park, NY 12018), placed on a magnetic stirrer and suspended. A quartz tube (Polymicro Technologies, Phoenix, Ar) coated with an outer diameter of 360μm, an inner diameter of 200μm and a length of 25cm polyimide (Polymicro Technologies, Phoenix, Ar) is fitted with an M-130 end-fitting with SST filter. , IDEXhealth & Science, 619 Oak Street, Oak Harbor, WA) The filter is fixed, and the other end is inserted into the turbid liquid containing glass microspheres and fixed on the pressure vessel. Press to fill the glass microspheres into the quartz tube to a length of 15cm ( figure 2 ).

[0018] The 1 cm wide part in the middle of the quartz tube filled with glass microspheres was burnt on a fire for about 30 s...

Example Embodiment

[0020] Example 2

[0021] The method is the same as in Example 1, except that a polyimide-coated quartz tube (Polymicro Technologies, Phoenix, Ar) with an inner diameter of 150 μm and an outer diameter of 360 μm is used; the heat adjustment of the laser needle puller is set at 380 degrees Celsius, and the speed is set at 40. An empty nanojet capillary high-pressure liquid chromatography empty column with an inner diameter of 150 μm, low back pressure and high flow rate, was prepared.

Example Embodiment

[0022] Example 3

[0023] The method is the same as Example 1, except that a polyimide-coated quartz tube (Polymicro Technologies, Phoenix, Ar) with an inner diameter of 100 μm and an outer diameter of 360 μm (Polymicro Technologies, Phoenix, Ar) is used; the heat adjustment of the laser needle puller is set at 390 degrees, and the speed is set at 40. Repeat twice. Thus, an empty column with an inner diameter of 100 μm, a low back pressure and a high flow rate, was prepared by a nanojet capillary high pressure liquid chromatography.

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Abstract

The invention belongs to the field of analytical chemistry. More specifically, the invention discloses a nanospray capillary high pressure liquid chromatographic column and its preparation method. The preparation method of the liquid chromatographic hollow column comprises three steps of: filling a polyimide-coated quartz capillary with a large-grained material which can be melted and solidified, removing the polyimide coating outside the capillary and drawing the capillary chromatographic column. The particle size of the meltable and solidifiable large-grained material in the chromatographic column is 10-15 microns, and a hollow gap can be formed after the large-grained material is heated and melted; in comparison with regular chromatographic columns, the chromatographic column provided by the invention has advantages of small back pressure on the tip as well as fast flow velocity and fast column-filling speed under the same pressure; when separating samples, the column has relatively small back pressure and high flow velocity. Therefore, a longer chromatographic column can be used on a regular liquid chromatogram under the same pressure so as to effectively raise the resolution and target protein identification efficiency of the chromatographic column.

Description

technical field [0001] The invention belongs to the field of analytical chemistry. More specifically, the invention discloses a nanospray capillary high-pressure liquid chromatography column and a preparation method thereof. Background technique [0002] The tandem use of liquid chromatography and mass spectrometry has become a common technology platform in the fields of analytical chemistry and proteomics research. Quartz capillary liquid chromatography microcolumns have been widely used due to their high resolution, high sensitivity, and high precision, and have become an essential component and basis of high-throughput proteomics technology platforms (Link, A.J.; Eng, J.; Schieltz, D.M.; Carmack, E.; Mize, G.J.; Morris, D.R.; -82.). Quartz material capillary liquid chromatography empty column with low resistance, low back pressure and high flow rate is the prerequisite for the preparation of high resolution, high sensitivity and high precision liquid chromatography micr...

Claims

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Application Information

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IPC IPC(8): G01N30/60G01N30/56B01D15/22C03C23/00C03B37/025
Inventor 徐平杨大福
Owner 徐平
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