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Hollow Column Liquid Chromatography System and Separation of Substances Using the System

A liquid chromatography, hollow technology, applied in the direction of material separation, analysis materials, instruments, etc., can solve problems such as inability to analyze, achieve the effect of reducing the amount of use and realizing the environmental burden

Active Publication Date: 2021-03-02
SCI & HEALTH WITH CHEM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the size of the intermediate produced during protein aggregation is as large as several nm to tens of μm, it cannot pass through the gap between the fillers in the general-purpose column, therefore, it cannot be analyzed

Method used

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  • Hollow Column Liquid Chromatography System and Separation of Substances Using the System
  • Hollow Column Liquid Chromatography System and Separation of Substances Using the System
  • Hollow Column Liquid Chromatography System and Separation of Substances Using the System

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0115] Embodiment 1 has the construction of the liquid chromatography system of hollow column

[0116] In Examples, the following liquid chromatography systems were used.

[0117] System 1:

[0118] MP 711 pump (GL Science, Tokyo, Japan)

[0119] MI 709 Autosampler (GL Science)

[0120] Connection L from pump to autosampler: Hollow capillary (30 μm inner diameter, 0.73 m)

[0121] amicro 21UV-02 UV array detector (Jasco, Tokyo, Japan)

[0122] HPLC System Organizer (EZ Chrom Elite v.3.1.7J)

[0123] Capillary: Polymicro Technologies (Phoenix, AZ, USA)

[0124] Connector: GL Science

[0125] System 2:

[0126] MP 711 pump (GL Science, Tokyo, Japan)

[0127] C4 manual injector (VICI AG, Schenkon, Switzerland)

[0128] There is no connection between the pump and the manual injector, directly connected

[0129] Electrophoresis system (Agilent 7100 capillary electrophoresis system)

[0130] Connecting part: GL Science PTFE tube

[0131] System 3:

[0132]Configuration:...

Embodiment 2

[0158] Example 2 Research on the Influence of Column Internal Diameter and Column Length on Separation Capacity

[0159] (1) Influence of the inner diameter of the column on the separation ability

[0160] Separation experiments were performed using the liquid chromatography system of System 2. As a sample to be separated, DNA (salmon-derived DNA, 90 to 270 bp) and fluorescein were used.

[0161] Hollow Capillary Columns The following hollow capillary columns were used, namely: a hollow capillary column made of fused silica with an inner diameter of 50 μm and a length of 17.8 m; a hollow capillary tube made of fused silica with an inner diameter of 25 μm and a length of 47.6 m Column; a hollow capillary column made of fused silica with an internal diameter of 5 μm and a length of 5.0 m. As the mobile phase, pure water was used.

[0162] The result is as Figure 8 shown. Figure 8 A represents the result using a column with an inner diameter of 50 μm and a length of 17.8 m...

Embodiment 3

[0170] Embodiment 3 uses the separation that liquid chromatography system carries out

[0171] 1. Use pure water as mobile phase for analysis

[0172] (1) Separation of low molecular substances

[0173] Since the thiourea (0) of the neutral low-molecular-weight substance can ignore the influence of charge and size and can show the dissolution of the mobile phase, it can be used as a standard substance and can be used with the charged low-molecular-weight substance BCECF (2',7' -bis(carboxyethyl)-4 or 5-carboxyfluorescein (-4), fluorescein (-2), rhodamine (0), rhodamine 6G (+1) for the separation of standard substances. The number indicates the number of charges. As a chromatographic column, use a hollow capillary column made of fused silica with an inner diameter of 25 μm and a length of 9.0 m, and use pure water as the mobile phase. The linear velocity (flow rate) is 1.8 ± 0.1 mm / s , was detected by UV254nm.

[0174] The result of the separation is as Figure 11-1 shown.

...

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Abstract

The present invention is a liquid chromatography system having the objective of making it possible to separate substances that cannot be separated in a conventional liquid chromatography system because the separation effect is small. This liquid chromatography system is used to separate biologically active substances and is characterized by at least being provided with a hollow column. The hollow column is a capillary column of non-treated, hollow quartz silica or natural quartz that is not filled with a filler.

Description

technical field [0001] The invention relates to a material separation and analysis method using hollow column liquid chromatography. Background technique [0002] Chromatography, as a means of separation and analysis, is a general-purpose method that can be used in almost all fields such as medicine, engineering, pharmacy, and medicine. It is not an exaggeration to say that it is one of the basic technologies that support current scientific research and social life. With the development of society and technology, although higher sensitivity, high selectivity, high degradability, high speed, automation and miniaturization have been achieved, in the fields of life science and nanotechnology, better separation and analysis methods are expected. For example, in recent years, in the analysis of environmental samples and biological samples, in order to reduce environmental pressure and improve separation capabilities, especially for small molecular weight compounds, high-efficienc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/60G01N30/88
CPCG01N30/60G01N30/88
Inventor 冨永宪子
Owner SCI & HEALTH WITH CHEM CO LTD
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