Total contact fluorescence detection pool for capillary electrophoresis

A capillary electrophoresis and fluorescence detection technology, applied in measurement devices, material analysis by optical means, instruments, etc., can solve the problems of high background signal and noise level, aggravate the optical distortion of the excitation beam, and reduce the sensitivity of the detector. Troubleshooting, eliminating stray light interference, reducing the effect of fluorescence intensity loss

Inactive Publication Date: 2014-09-17
徐静 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But it has the following disadvantages: 1. Before the excitation light reaches the sample, it has to pass through two optical interfaces: air-quartz capillary wall-sample solution; the significant difference in refractive index (RI) (1.0-1.47-1.33), making the light Reflection, refraction and total reflection inside the tube wall occur here, and the cylindrical shape of the capillary exacerbates the optical distortion of the excitation beam, and the reflected excitation light is received by the collection fiber, resulting in higher background signal and noise levels
2. Before the fluorescence reaches the collection fiber, it passes through three optical interfaces: sample solution-quartz capillary wall-air-quartz fiber; the refractive index (RI) is 1.33-1.47-1.0-1.47 in sequence; according to the Fresnel equation, in each optical There will be a corresponding reflection loss at the interface, and its size depends on the two RI values ​​of the optical interface. The greater the difference, the greater the loss; especially from the optically sparse medium (lower refractive index) into the optically dense medium ( When the refractive index is higher), when the angle of the incident light is greater than the critical value, total reflection will occur; in the on-column detection, the propagation of fluorescence has experienced two optically sparse-optical dense interfaces, that is, the sample solution-quartz capillary wall And air-silica fiber, resulting in a large loss of fluorescence intensity, reducing the sensitivity of the detector

Method used

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  • Total contact fluorescence detection pool for capillary electrophoresis
  • Total contact fluorescence detection pool for capillary electrophoresis
  • Total contact fluorescence detection pool for capillary electrophoresis

Examples

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Effect test

Embodiment 1

[0056] A full-contact fluorescent detection cell for capillary electrophoresis, assembled according to the following steps and parameters:

[0057] 1. Assemble the detection cell body assembly 100, as attached figure 1 and 2 Shown:

[0058] A transparent T-shaped tee pipe 110 made of quartz glass has tapered diameter-reducing sections 112, 122 and 132 on the inner walls of its main pipes 111, 121 and branch pipes 131, respectively. The center of the through pipe 110; the outer diameter of the T-shaped tee pipe 110 is 3 millimeters, the inner diameter is 500 microns, and the diameter is 100 microns after passing through the tapered section with an included angle of 45 degrees. Apply cured glue on the outer wall of the first port of the capillary with an outer diameter of 365 microns and an inner diameter of 100 microns, and then insert them from the open ends of the main pipes 111 and 121 until the ends reach the tapered diameter reducing sections 112 and 122; the outer wall...

Embodiment 2

[0065] According to Example 1, a full-contact fluorescence detection cell was set up, and the detection cell was used to assemble a light-emitting diode-induced fluorescence detector and applied to capillary electrophoresis to detect the vitamin B2 content in commercially available pork.

[0066] Assembly scheme: use a light-emitting diode with a central wavelength of 470nM, a band-pass interference filter BP470nm to filter the light source, a 5V constant voltage source connected in series with a 100Ω resistor to power the light-emitting diode; the excitation light is irradiated to the detection area through the optical fiber I. Use fiber II as the collection fiber to collect fluorescence in the direction perpendicular to the excitation light, conduct it through fiber II, pass through the fluorescence filter (BP530nm) to eliminate background interference light, and finally enter the photomultiplier tube to convert it into an electrical signal and output it to the workstation.

...

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Abstract

The invention discloses a total contact fluorescence detection pool for capillary electrophoresis. The detection pool comprises a detection pool body assembly, a positioning assembly and a fixing assembly. The fluorescence detection pool adopts a quadrature optical path structure, and makes an excited fiber, a fiber II and a liquid optical path vertically conjugate crossed in a pairwise conjugate manner; a simple structure is adopted to realize the physical contact of a liquid flow path, the excited fiber and a reception fiber, and a detection zone adopts a quartz material, guarantees the continuous running of an electroosmotic flow, and is suitable for capillary electrophoresis-fluorescence detection; and additionally, the positioning element and the fixing assembly matched with the detection zone are designed to make the installation and use of the detection pool convenient.

Description

technical field [0001] The invention relates to a contact type fluorescent detection cell for capillary electrophoresis. Background technique [0002] Fluorescence detection is one of the detection technologies with the highest sensitivity at present, and has high selectivity and response stability; capillary electrophoresis uses capillary as the separation channel, and ions or charged particles are distributed in the capillary according to their mobility or / and High-efficiency and rapid separation technology with different coefficients has the characteristics of high separation efficiency, short analysis time, and small injection volume. Capillary electrophoresis-fluorescence detection, which combines the advantages of the above two technologies, has been widely used in life sciences, bioengineering, environmental protection, food and drug monitoring and other related fields. [0003] In microfluidic analysis systems (including flow injection, microcolumn liquid chromatogr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/01
Inventor 徐静郑秋月曹冬梅曹际娟
Owner 徐静
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