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Serum metabolic spectrum molecular weight calibrator kit, preparation method and use method thereof

A technology of molecular weight and calibrator, which is applied in the field of mass spectrometry, can solve the problems of interference spectrum peaks, spectrum stability cannot be guaranteed, and metabolic molecular region cannot be calibrated at the same time, achieving low cost, convenient preparation, and convenient use Effect

Active Publication Date: 2021-03-19
HANGZHOU WELL HEALTHCARE TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can cover the molecular weight range of 300-1000Da, but the peak of the silver cluster seriously interferes with the spectral peaks of complex biological samples, and at the same time changes the ion summation form of metabolic molecules. The spectral shape of metabolic molecules is related to the distribution of nanoparticles on the substrate. , the stability of the spectrum cannot be guaranteed
And the silver clusters can only form a list of molecules that can calculate the precise molecular weight in the positive ion mode, and cannot be used for the calibration of the metabolic molecular area in the negative ion mode at the same time
At the same time, due to the unavoidable ion suppression effect in direct ionization mass spectrometry detection technology, other internal standard methods are usually not conducive to the accurate detection of complex biological samples.

Method used

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  • Serum metabolic spectrum molecular weight calibrator kit, preparation method and use method thereof
  • Serum metabolic spectrum molecular weight calibrator kit, preparation method and use method thereof
  • Serum metabolic spectrum molecular weight calibrator kit, preparation method and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1 Extracting and preparing metabolic molecular calibrator from serum samples

[0072] Step 1: Obtain a blood sample, centrifuge it after coagulation, and obtain a clear light yellow serum supernatant;

[0073] Step 2: Take 10-50 μL of serum supernatant, add 5-10 times the volume of mixed extraction solvent, and go through shaking incubation and centrifugation steps to separate the serum samples;

[0074] In step 3, a solution containing metabolic molecules is obtained by separating the serum stratified sample in step 2, and its solvent is evaporated to dryness to obtain a metabolic molecular calibrator.

[0075] Step 4, redissolving the metabolic molecular calibrator with a dissolving solution to prepare a metabolic molecular calibrator solution. As a preference, the dissolving solution of the metabolic molecular calibrator extracted from the serum sample is a mixed solvent of isopropanol and water, and the ratio of the metabolic molecular calibrator to the dis...

Embodiment 2

[0078] Molecular Weight Determination of Metabolic Molecular Weight Calibrators and Obtaining a Molecular Weight Calibration List

[0079] Step 1: Add 100-1000 μL of the dissolving solution to the metabolic molecular calibrator, shake and redissolve to prepare the calibrator solution, aliquot 10 μL / tube in a single tube, and store at -80°C when not in use;

[0080] Step 2, add pure product mixed metabolism composed of lauric acid, palmitic acid, stearic acid, arachidic acid, lignoceric acid, 1,2-dipalmitic acid-sn-glycero-3-phosphoethanolamine to the calibrator solution Molecule, which constitutes a molecular weight calibrator in negative ion mode with an internal standard;

[0081] Step 3, add pure product mixed metabolic molecules composed of 1,2-distearoyl-sn-glycero-3-phosphocholine and 1,2,3-tripalmitin to the calibrator solution, the composition contains Molecular weight calibrators in positive ion mode for internal standards;

[0082] Step 4: Spot the metabolic molecu...

Embodiment 3

[0090] Calibration Effect Evaluation of Molecular Weight Internal Standard Calibration Method of Metabolic Molecular Weight Calibrator

[0091] Step 1, the molecular weight of the serum biological sample to be tested is calibrated:

[0092] 1) collecting, extracting or enriching metabolic molecules in serum;

[0093] 2) Dissolving 1 part of the metabolic molecule calibrator extracted from serum with the dissolving solution to obtain the metabolic molecule calibrator solution;

[0094] 3) Transfer the metabolic samples collected, extracted or enriched in the serum to the sample loading platform, and at the same time spot the metabolic calibrator solution on the sample loading platform, and the serum sample points to be tested are distributed around or adjacent to the metabolic molecular calibrator points Location;

[0095] 4) Start the mass spectrometry instrument to collect data, use the metabolic molecular calibrator to calibrate the instrument in the two detection modes, a...

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Abstract

The invention relates to a serum metabolic spectrum molecular weight calibrator kit, a preparation method and a use method thereof. The serum metabolic spectrum molecular weight calibrator kit comprises a metabolic molecule calibrator, a dissolving solution and a molecular weight calibration list, wherein the ratio of the metabolic molecule calibrator to the dissolving solution is as follows: 1 part of the metabolic molecule calibrator corresponds to 1-5 parts of the dissolving solution. According to the invention, the molecular weight calibrator is taken from a biological sample, and is low in cost and convenient to prepare; the kit can cover the range of 100-1000 Da of metabolic molecule detection, is also suitable for positive ion and negative ion detection modes, and can select a molecular weight calibrator and a calibration list as required; the kit is convenient to use and can be suitable for detection and molecular weight calibration of metabolic molecular regions of various ionsource mass spectrometers and various biological samples; and the kit is suitable for accurate molecular weight determination of the metabolic small molecules, structure identification and analysis of the metabolic small molecules, and precise molecular mass spectrometry imaging.

Description

technical field [0001] The invention relates to the technical field of mass spectrometry, in particular to a serum metabolic spectrum molecular weight calibrator kit, a preparation method and a use method thereof. Background technique [0002] Because metabolic molecules can reflect the phenotypic changes, physiological and pathological conditions of organisms in real time, their composition changes in complex biological samples such as blood, saliva, urine, tissues, and cells are closely related to disease monitoring. MALDI-MS matrix-assisted laser desorption ionization mass spectrometry is a high-throughput mass spectrometry technology. highly valued. However, due to the wide variety of metabolic molecular components, taking lipids as an example, more than 20,000 species have been discovered and named. When using MALDI and other direct ionization mass spectrometry technologies as platforms for direct detection and analysis of complex biological samples, molecular weight c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/08G01N30/72G01N30/86
CPCG01N30/06G01N30/08G01N30/72G01N30/8665
Inventor 邬建敏陈晓明余捷凯栾春燕
Owner HANGZHOU WELL HEALTHCARE TECH CO LTD