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Ultra-short chromatographic micro-column for rapid separation of biological sample, and preparation method thereof

A biological sample and rapid technology, applied in the field of chromatographic micro-columns, can solve the problems of difficulty in accurately controlling the length of the column bed and the inability to realize the production of column-bed chromatographic columns, and achieves the effects of simple operation, solving adsorption problems and convenient use.

Pending Publication Date: 2017-10-03
瑞谱信(厦门)科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the traditional preparation process of capillary liquid chromatography column, the production of the plunger often needs to be completed by sintering part of the filler, and it is difficult to precisely control the length of the column bed, so that the production of a shorter column bed chromatography column cannot be realized

Method used

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  • Ultra-short chromatographic micro-column for rapid separation of biological sample, and preparation method thereof
  • Ultra-short chromatographic micro-column for rapid separation of biological sample, and preparation method thereof
  • Ultra-short chromatographic micro-column for rapid separation of biological sample, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Preparation of ultra-short chromatographic micro-column of 1mm column bed

[0027] Cut the hollow quartz capillary 2 with an inner diameter of 100 μm and a hollow quartz capillary with an outer diameter of 365 μm (Hebei Yongnian Ruifeng Chromatography Device Co., Ltd.) of 5 cm, and place one end of the hollow quartz capillary 2 on a porous silica microsphere 1 with a diameter of 110 μm (U.K. In a centrifuge tube from X-tec Company), a single porous silica microsphere 1 is pressed into one end of the capillary as an outlet plunger.

[0028] Take a 20 μL pipette tip 7, insert the opening end of the above-mentioned capillary into the tip of the tip, and seal the joint and the opening of the pipette tip 7 with a sealing tape (such as figure 2 shown). Weigh 1 mg of C18 reverse-phase silica gel bonded filler (Waters Company) with a particle size of 5 μm as the chromatographic stationary phase 3 , and add the filler into the pipette tip 7 . Place the pipette tip ...

Embodiment 2

[0029] Embodiment 2: Fast chromatographic separation efficiency evaluation

[0030] Connect the ultra-short chromatographic micro-column obtained in Example 1 to a nano-flow liquid chromatograph, connect a section of capillary with an inner diameter of 20 μm as a connecting tube to guide the liquid flow into an ultraviolet detector, and test the chromatogram under the condition of 214nm ultraviolet light The ability of the column to separate benzene homologues (thiourea, benzene, toluene, ethylbenzene, propylbenzene). The mobile phase ratio is CAN / H 2 O=60 / 40, the volume flow rate is 200nL / min, the separation time is 60s, the obtained chromatogram is as follows image 3 As shown, the separation column efficiency of the last component propylbenzene is 33600 trays / m.

Embodiment 3

[0031] Embodiment 3: Ultrashort chromatographic microcolumn is used for the dot plate of matrix-assisted laser desorption ionization (MALDI) mass spectrometry

[0032] Such as Figure 4 As shown, five kinds of protein mixed solutions were taken as samples to be tested (Cytochrome C, Lysozyme, Ribonuclease A, Myoglobin, Albumin Bovine), and a 1mL syringe was used to draw the equilibrium solution (CAN / H 2 (0=5 / 95) stand-by, dip the head of the inlet end of the ultra-short chromatographic micro-column prepared in Example 1 to take a small amount of sample, connect it to a 1mL syringe through a PTFE sleeve, and then slowly push the plunger of the syringe so that the column tube Fully soaked inside. Take another 1mL syringe 4 and fill it up with the eluting solution (CAN / H 2 (0=60 / 40), remove the column tube from the first syringe and connect it to the syringe 4, then slowly push the plunger of the syringe so that the elution fraction droplet 6 falls, and the fraction droplet fal...

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Abstract

The invention provides an ultra-short chromatographic micro-column for rapid separation of a biological sample, and a preparation method thereof, and relates to a chromatographic micro-column. The ultra-short chromatographic micro-column contains a hollow quartz capillary, which is filled with fixed phase filler particles with a column bed length of less than 1 mm, wherein both ends of the micro-column are respectively provided with two porous silica microspheres as plungers. The preparation method comprises: intercepting a hollow quartz capillary, and pressing a porous silica microsphere into one end of the hollow quartz capillary so as to be used as an outlet plunger; weighing a filled chromatographic fixed phase, placing into a pipette tip, connecting the tip end portion of the pipette tip and the hollow quartz capillary, sealing with a glue strip, placing the obtained pipette tip into a centrifuge, and stacking the fixed phase particles into a column bed through the driving with the centrifugal force; and after completing the filling, pressing one end of the hollow quartz capillary into a porous silica microsphere as an inlet plunger, cutting the remaining hollow pipe with a ceramic cutter blade under a microscope along the inlet plunger position, such that the preparation of the ultra-short chromatographic micro-column for the rapid separation of the biological sample is completed.

Description

technical field [0001] The invention relates to a chromatographic microcolumn, in particular to an ultrashort chromatographic microcolumn for rapid separation of biological samples and a preparation method thereof. Background technique [0002] Miniaturization is a major trend in the development of science and technology today, and this trend also appears in the evolution and development of high performance liquid chromatography. The sample extraction process from living systems (local diseased tissues, single cells, etc.) is cumbersome and the amount available is extremely small. The high volume flow rate of conventional analytical columns and micro-diameter columns will cause serious dilution effects on trace samples. Reduce the sensitivity of analytical detection. In order to reduce the dilution effect of liquid chromatographic flow relative to trace samples, it is necessary to further reduce the diameter of the chromatographic column to the micron level, and the capilla...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/18
CPCG01N30/02G01N30/06G01N30/18G01N2030/065
Inventor 张博郭睿
Owner 瑞谱信(厦门)科技有限公司
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