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Liquid chromatogram retention time database correction method based on SCAC-RI

A retention time and liquid chromatography technology, applied in the field of calibration of liquid chromatography compound retention time database, can solve problems such as database retention time offset, and achieve the effect of eliminating adverse effects, stable properties and not easy to degrade

Active Publication Date: 2021-01-22
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method solves the problem of shifting the retention time of the original database when method transplantation or method fine-tuning is required, and avoids the trouble of rebuilding the database

Method used

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  • Liquid chromatogram retention time database correction method based on SCAC-RI
  • Liquid chromatogram retention time database correction method based on SCAC-RI
  • Liquid chromatogram retention time database correction method based on SCAC-RI

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Retention Time Database Correction Based on Metabolite Mixed Standards

[0042] 1. Metabolite Mixed Standard Configuration

[0043] In this example, 75 metabolites detectable in the positive mode of ESI were selected as research objects (including SCACs). The selected metabolites have a large polarity range and can completely cover each retention time of reversed-phase chromatography. Use 50% methanol aqueous solution to prepare mixed standards with the concentration of each metabolite on the order of μM, and store them in a -20°C refrigerator for later use.

[0044] 2. Chromatographic conditions and mass spectrometry conditions

[0045] 2.1 Chromatographic conditions

[0046] In this embodiment, the initial chromatographic condition is the M0 method (Table 2), and another 10 different chromatographic methods (M1~M10) are also combined according to different gradients, flow rates, column temperatures and chromatographic columns, and are used to verify the r...

Embodiment 2

[0081] Embodiment 2: Retention time database correction based on biological samples

[0082] 1. Pretreatment of serum and tissue samples

[0083] Transfer the human serum mixed QC sample from the -80°C refrigerator to the 4°C refrigerator to thaw for 30 minutes, then place the sample on ice, and do not touch the sample area of ​​the EP tube with your hands during the operation. Use a pipette gun to accurately draw 50 μL of serum samples into a 1.5mL EP tube, add 225 μL of pre-cooled methanol solution (refrigerated at -20°C overnight), vortex for 30 sec, incubate in a refrigerator at 4°C for 30 min to fully precipitate the protein, and then place in Centrifuge at 14000g for 10min in a low-temperature centrifuge (4°C). Take 225 μL of supernatant and add 750 μL of pre-cooled MTBE solution (refrigerated overnight at -20°C), vortex for 30 sec, add 188 μL of pure water, and vortex for 30 sec. Place a low-temperature centrifuge (4° C.), and centrifuge at 14000 g for 5 min. Finally...

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Abstract

The invention belongs to the technical field of biological analysis, and particularly relates to a liquid chromatogram compound retention time database calibration method based on SCAC-RI. The methodcomprises the following steps: establishing an initial compound retention time database; calculating a compound retention index; and establishing a correction retention time database under a new method by taking SCAC as an index. According to the method, the situation that the database is unavailable due to retention time deviation caused by chromatographic condition change is improved to a greatextent, so that migration of the retention time database among different methods becomes possible, and adverse effects of unstable factors of other liquid phase systems on the retention time in an experiment can be eliminated so as to greatly improve the matching degree with a database during compound identification. The method is suitable for the field of non-targeted metabonomics and other chemical analysis in which a universal and flexible retention time database needs to be established to qualitatively analyze complex multi-component samples.

Description

technical field [0001] The invention belongs to the technical field of bioanalysis, and in particular relates to a method for calibrating a liquid chromatography compound retention time database based on straight-chain acylcarnitine retention index (SCAC-RI). This method is suitable for non-targeted metabolomics and other chemical analysis fields that need to establish a general and flexible retention time database to qualitatively complex and multi-component samples. Background technique [0002] Metabolomics can be divided into target metabolomics and non-target metabolomics technologies according to different research strategies, among which non-target metabolite detection technology based on high-resolution mass spectrometry can collect biological samples with high coverage without any prior knowledge Therefore, it is widely used in the discovery-type metabolomics research process. However, its high coverage also brings the problem that many compounds cannot be accurate...

Claims

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Application Information

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IPC IPC(8): G01N30/86G16C20/64G16C20/90
CPCG01N30/8668G16C20/64G16C20/90
Inventor 唐惠儒芦勤玮
Owner FUDAN UNIV
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