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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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When separating samples, the back pressure of this column is large and the flow rate is low, it is difficult to improve the separation efficiency of polypeptide samples by increasing the column length

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
Comparison scheme
Effect test

Embodiment 1

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

[0018] Burn the 1 cm wide part of the quartz tube filled with glass microspheres on fire for about 30 seconds to remove t...

Embodiment 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 thermal adjustment of the laser needle pulling instrument is set at 380 degrees Celsius, and the speed is set at 40. In this way, an empty nanospray capillary high-pressure liquid chromatography column with an inner diameter of 150 μm and a low back pressure and high flow rate was prepared.

Embodiment 3

[0023] 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 100 μm and an outer diameter of 360 μm is used; the thermal adjustment of the laser needle pulling instrument is set at 390 degrees, and the speed is set at 40, repeat twice. In this way, an empty nanospray capillary high-pressure liquid chromatography column with an inner diameter of 100 μm and low back pressure and high flow rate was prepared.

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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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