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Array multi-dimensional liquid-phase chromatographic column system

A multi-dimensional liquid chromatography and column system technology, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as limited sample loading, time-consuming and labor-consuming, and platform stability to be considered

Active Publication Date: 2016-01-13
XIAMEN CHROMATOGRAPHIC ANALYSIS INSTR CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Two-dimensional gel electrophoresis has high resolution, but there are still some inherent defects, such as low degree of automation, poor reproducibility, time-consuming and labor-intensive, and limited sample loading, which makes it difficult to meet the needs of large-scale proteomics research
At present, the high-efficiency separation of protein samples at home and abroad is mostly limited to the construction of single-channel multi-dimensional chromatography systems, and the construction of multi-channel multi-dimensional chromatography is relatively rare, and the stability of the platform needs to be considered.

Method used

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  • Array multi-dimensional liquid-phase chromatographic column system
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  • Array multi-dimensional liquid-phase chromatographic column system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Cytochrome C hydrolyzate was digested with trypsin (purchased from Sangon Bioengineering (Shanghai) Co., Ltd.), with a concentration of 5 mg / mL. When operating, first follow the instructions of the device figure 1 Connection, the two-channel array multidimensional liquid chromatography separation column system of the present invention is provided with a high-pressure nanoflow pump 1, a sample injector 2, a single capillary column 3, a tee 4, the first array capillary column 5, and the second array capillary column 6 , a multi-channel ultraviolet detector 7, a first empty capillary with a detection window 8 and a second empty capillary with a detection window 9.

[0028] The outlet of the high-pressure nanoflow pump 1 is connected to the injection port of the single capillary column 3 through the injector 2, the liquid flow outlet of the single capillary column 3 is connected to the liquid flow inlet of the tee 4, and the liquid flow outlet of the tee 4 is connected to t...

Embodiment 2

[0031] Similar to Example 1, the difference is that through a multi-channel connection device (3 PEEK tees, zero dead volume, 1 / 16 outer diameter, IDEX, the United States), a single chromatographic column (SCX, 100 μm inner diameter, 15 cm long ), 6 array chromatographic columns (RP, 100μm inner diameter, 7cm long) to establish a four-channel array multidimensional liquid chromatography separation column system, according to the schematic diagram of the device image 3 connect. Prepare the mobile phase, the sample volume used and the operation process as above, the flow rate used is 400nL / min, and the reverse-phase gradient elution program is extended as follows: 0-5% B, 10min; 5%-50% B, 15min; 50%-100% B, 15min. Finally, a multi-channel ultraviolet detector was used to obtain the separation and detection results. Experimental results such as Figure 4 .

[0032]This embodiment is provided with high-pressure nanoflow pump 1, injector 2, single capillary column 3, the first...

Embodiment 3

[0034] Similar to Example 1, the difference is that through a multi-channel connection device (1 PEEK four-way and 3 PEEK three-way, zero dead volume, 1 / 16 outer diameter, IDEX, the U.S.), a single chromatographic column (SCX , 100μm inner diameter, 15cm long), 9 array chromatographic columns (RP, 100μm inner diameter, 7cm long) to establish a six-channel array multidimensional liquid chromatography separation column system, according to the schematic diagram of the device Figure 5 connect. Prepare the mobile phase, the sample volume used and the operation process as above, the flow rate used is 550nL / min, and the gradient elution procedure is the same as in Example 2. Finally, a multi-channel ultraviolet detector was used to obtain the separation and detection results. Experimental results such as Figure 6 .

[0035] This embodiment is provided with high-pressure nanoflow pump 1, injector 2, single capillary column 3, four-way 18, the first three-way 4, the second three-...

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Abstract

The invention relates to a liquid-phase chromatographic column, in particular to an array multi-dimensional liquid-phase chromatographic column system. The system is provided with a high-pressure nanofluidic pump, a single chromatographic column, a multi-channel chromatography connection device and parallel array chromatographic columns. The high-pressure nanofluidic pump, the single chromatographic column, the multi-channel chromatography connection device and the parallel array chromatographic columns are sequentially connected in series, outlets of the parallel array chromatographic columns are connected with an inlet of a multi-channel ultraviolet detector, and an outlet of the multi-channel ultraviolet detector is connected with at least two empty capillary tubes with detection windows. First-dimension chromatographic separation is achieved through ion exchange chromatography, and after first salt gradient separation is completed, at the tails of the columns, eluate is equally divided and gathered to multiple reversed phase chromatographic column heads with the same separation effect; then, the multiple elution components are separated at the same time in a multi-column parallel mode at the second dimension. The operation is repeated until all salt gradients are run, and then all two-dimensional chromatographic separation processes are completed. Through the array two-dimensional chromatography system, a high-resolution two-dimensional separation effect can be achieved, and multiple sample flows can be obtained through one-off separation running.

Description

technical field [0001] The invention relates to a liquid chromatography column, in particular to an array type multidimensional liquid chromatography column system. Background technique [0002] In recent years, proteomics has developed rapidly. Due to the diversity of proteins expressed in the genome and the large abundance differences between proteins, the proteomic system is highly complex. Taking the human plasma proteome as an example, there are tens of thousands of proteins in it, which brings great challenges to the analysis of proteins. Therefore, high-resolution separation technology is required to reduce the complexity of samples. [0003] Two-dimensional gel electrophoresis is a classic method for protein separation in proteomics research. Two-dimensional gel electrophoresis has high resolution, but there are still some inherent defects, such as low degree of automation, poor reproducibility, time-consuming and labor-intensive, and limited sample loading, which m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/60
Inventor 张博丁芳芳
Owner XIAMEN CHROMATOGRAPHIC ANALYSIS INSTR CO LTD
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