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Combination method of step-by-step focusing-axial mixing-high-performance liquid chromatographic separation system

A high-performance liquid chromatography and axial mixing technology, which is applied in the joint field of step-by-step focusing-axial mixing-high performance liquid chromatography separation system, can solve problems such as deformation, splitting, and broadening of chromatographic peaks, and achieve a simple and efficient system. Improved mixing effect, low cost effect

Active Publication Date: 2017-03-15
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method still has the following disadvantages: the inventors further in-depth research found that if thinner chromatographic columns and pipelines are used or compounds with weaker retention are analyzed, chromatographic peak broadening, splitting and deformation will still occur if the injection volume is further increased

Method used

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  • Combination method of step-by-step focusing-axial mixing-high-performance liquid chromatographic separation system
  • Combination method of step-by-step focusing-axial mixing-high-performance liquid chromatographic separation system
  • Combination method of step-by-step focusing-axial mixing-high-performance liquid chromatographic separation system

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0029] 1. Composition of step-by-step focusing-axial mixing-high performance liquid chromatography separation system

[0030] see figure 1 In Figure I, the step-by-step focusing-axial mixing-high performance liquid chromatography separation system in this example includes an interconnected step-by-step focusing sampling device B, a switchable axial mixing device C, and a high-performance liquid chromatography device A.

[0031] see figure 1 In Fig. I in the figure, the above-mentioned high-performance liquid chromatography device includes a high-pressure pump 5, an autosampler, a chromatographic column 6 and a detector 7; wherein the autosampler consists of a mechanical arm 1, an autosampler needle 2, a sample bottle 3 and an inlet The sample port 4 is composed of an automatic sample needle 2 that sucks the sample solution 14 into the sample port 4 under the control of the mechanical arm of the automatic sampler 1 .

[0032] see figure 1 In Figure I, the above-mentioned ste...

example 2

[0048] In order to compare the effect of the described method of the invention with the prior art, the inventor has done the following experiments:

[0049] Comparative experiment one

[0050] High performance liquid chromatography-tandem mass spectrometry (i.e. figure 1 The high-efficiency liquid chromatography device in A) detects tetrodotoxin standard solution by conventional sampling method, and mobile phase, eluent and elution gradient and mass spectrometry conditions are all identical with example 1, and the result is as follows figure 2 Shown in c figure.

[0051] Comparative experiment two (equivalent to the method described in the invention patent application with publication number CN103134889A)

[0052] 1. Experimental system: the figure 1 The pre-column 12, mixer 13 and second high-pressure six-way switching valve 11 in the figure I are removed, and the ⑤ port and the ⑥ port of the original second high-pressure six-way switching valve 11 are short-circuited wit...

example 3

[0057] In order to compare the sample introduction of the method described in the invention with the existing step-by-step focusing method (that is, the method described in the invention patent application with the publication number CN103134889A), the inventor has done the following experiments:

[0058] 1. Use figure 1 In the combined system shown, the injection is increased to 2mL, and the parameters of the combined method are adjusted according to the example, and the results are as follows image 3 Shown in e figure.

[0059] After adjusting the parameters, the specific method is as follows:

[0060] (1) First, according to the mixing ratio of the tetrodotoxin standard solution and the mobile phase acetonitrile, the time length of the first high-pressure six-way switching valve 8 in the sample loading position a is set to 2 min, and the time length in the sample injection position is set to 0.2 min; Set the flow rate of the mobile phase to 1mL / min, and the equilibration...

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Abstract

The invention relates to a combination method of a step-by-step focusing-axial mixing-high-performance liquid chromatographic separation system. The combination method includes the steps: (1) respectively setting duration of a first high-pressure six-way switching valve positioned at a sample carrying gear and a sample injection gear according to the mixing ratio of sample liquid and a flowing phase; (2) positioning the first high-pressure six-way switching valve at the sample carrying gear, positioning a second high-pressure six-way switching valve at a mixing gear, starting a high-pressure pump and an automatic sample injector; (3) starting a detector, positioning the first high-pressure six-way switching valve at the sample carrying gear, positioning a second high-pressure six-way switching valve at a non-mixing gear, and performing gradient elution, separation and detection. Timing is started after the automatic sample injector completely injects the sample liquid into a quantitative loop, the first high-pressure six-way switching valve is switched to the sample injection gear after reaching the set duration of the sample carrying gear, and then the first high-pressure six-way switching valve is switched back to the sample carrying gear after reaching the set duration of the sample injection gear. The process is repeated until sample injection is finished, and analyte is focused at a chromatographic column. By the method, sample injection volume can be further increased.

Description

technical field [0001] The present invention relates to the testing or analysis of materials by means of determining their chemical or physical properties, and in particular to the testing or analysis of materials involving the separation of materials into their individual components by chromatography. Background technique [0002] High performance liquid chromatography (HPLC) is a powerful analytical tool with strong separation ability and good versatility. However, the chromatographic injection volume severely limits the improvement of analytical sensitivity: the increase in injection volume (especially for samples rich in strong solvents) is likely to cause broadening and distortion of spectral peaks, resulting in a sharp drop in resolution. Rapid chromatography technology and ultra-high performance liquid chromatography technology have developed rapidly in recent years. The chromatographic column is shorter than the conventional liquid chromatographic column, and the pac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/20
CPCG01N30/20G01N2030/201
Inventor 潘加亮龙菲马安德
Owner SOUTHERN MEDICAL UNIVERSITY
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