Novel method for producing and packing capillary chromatographic column sieve plate

A technology of capillary chromatographic column and capillary, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., and can solve the problems of difficult loading and unloading, increased experiment cost and difficulty, time-consuming and laborious, etc.

Inactive Publication Date: 2004-09-29
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Since the homogenate in the container requires electromagnetic stirring, a larger container is required to put the magnets in. For capillary tubes filled with only a few milligrams or even less than a milligram of chromatographic media, it is a waste of chromatographic media and makes loading and unloading difficult, time-consuming and laborious.
[0006] 3. When filling the capillary with a thinner homogenizing tube, ultrasonic waves are usually used to homogenize the chromatographic medium and solvent, which is not easy to achieve the purpose and complicates the column packing process
[0007] 4. When the traditional capillary column is packed, special devices and accessories are required, which increases the cost and difficulty of the experiment
[0008] 5. When using high-pressure nitrogen to provide pressure, because it is not easy to provide a large pressure, it is difficult to fill a long capillary chromatographic column, and it is easy to dry out the chromatographic medium in the capillary chromatographic column, reducing the chromatographic separation efficiency
[0009] 6. When packing the capillary column by vacuum extraction method, because the general vacuum pump is difficult to provide a high vacuum degree, it is impossible to fill a longer capillary column, and a special vacuum pump is required, which increases the cost of column packing

Method used

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  • Novel method for producing and packing capillary chromatographic column sieve plate
  • Novel method for producing and packing capillary chromatographic column sieve plate
  • Novel method for producing and packing capillary chromatographic column sieve plate

Examples

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Embodiment Construction

[0018] 1. Separation of trypsin lysate of myoglobin by reversed-phase capillary column

[0019] Instrument: Nano LC chromatographic system of Diane Company and Q-TOF Micro mass spectrometer of Micromass Company Capillary chromatographic column: 40cm×75μm i.d.; Chromatographic medium: C18 reverse phase chromatography filler, 3μm, 120 chromatographic conditions: mobile phase A, 2% acetonitrile aqueous solution +0.1% formic acid; mobile phase B, 20% acetonitrile aqueous solution +0.1% formic acid Flow rate, 0.2μl / min, detection wavelength, 214nm; linear gradient, 0-30min, 100%A-60%B; 30-35min, 60% B-100% B; 35-85min, 100%B-100%B, 85-90min, 100%B-100%A, 100%A extended to 100min.

[0020] Sample: trypsin hydrolyzate of myoglobin, the concentration is 5.6 pmol / μl; the loading volume, 5 μl. See attached chromatogram figure 2 .

[0021] 2. Separation of plasma whole protein trypsin lysate by reversed-phase capillary column

[0022] Instrument: Nano LC chromatographic system of Di...

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Abstract

The present invention relates to new technology of producing capillary chromatographic column sieve plate simply, fast and in low cost and reliable and efficient column filling method. Compared with available technology, the present invention has the following advantages: the utilization of common efficient liquid chromatographic connection tube suitable for filling milligram level stuffing as homogenating tube and vortex mixer with high efficiency and no stuffing lamination for vibration; the utilization of stainless steel or engineering plastic tube capable of being bent into U-shape before filling resulting in easy operation; and utilization of high pressure lab constant-flow pump for filling resulting in high column filling efficiency and low cost.

Description

technical field [0001] The present invention belongs to capillary chromatography and electrochromatographic separation in the field of separation science. Background technique [0002] The separation and analysis of a small amount of complex samples is a huge challenge and task faced by scientific and technological workers in this field, and it is also an important opportunity and driving force for the development of this field. In particular, the completion of genome projects such as human, mouse and rice and the gradual start of different organs, tissues, cells and subcellular proteome projects have put forward higher and higher requirements for separation and analysis. Generally, biological samples are characterized by small quantities and complex components, and people need to know the qualitative, quantitative and interaction information of tens of thousands of biological macromolecules, so the separation method with high separation efficiency and small sample volume—— ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/56G01N30/60
Inventor 钱小红张养军王京兰
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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