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A Urine Metabolomics Analysis Method Based on Osmolality Correction

A technology of metabolomics and analysis methods, which is applied in the field of urine metabolomics analysis based on osmotic pressure for normalization before data collection, which can solve the problem that creatinine is no longer applicable, and improve the repeatability and accuracy of results. Effect

Active Publication Date: 2022-06-28
SHANGHAI JIAOTONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] A. Take several urine samples to measure the osmotic pressure of each urine sample; as the gold standard for judging the status of urine in clinical practice, it is more comprehensive to judge the concentration of metabolites by using osmotic pressure as a benchmark to solve the problem of kidney disease Improves the applicability of the method by affecting creatinine metabolism such that creatinine-based corrections are no longer applicable;

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  • A Urine Metabolomics Analysis Method Based on Osmolality Correction
  • A Urine Metabolomics Analysis Method Based on Osmolality Correction
  • A Urine Metabolomics Analysis Method Based on Osmolality Correction

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Experimental program
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Effect test

Embodiment 1

[0073] like Figure 13 As shown, a urine metabolomics analysis method includes the following steps:

[0074] A. Take urine samples to determine the osmotic pressure of each urine sample

[0075] The urine samples of 15 patients with hydronephrosis (disease group) were taken as Examples 1-15, and the urine samples of 15 healthy people were taken as Examples 16-30.

[0076] B. Sample pretreatment

[0077] According to the osmotic pressure measurement results, the osmotic pressure span of the actual sample is about 10 times. In order to prevent the solvent effect caused by the excessive injection volume, the dilution ratio of each sample is determined according to the principle that the product of the osmotic pressure of all samples and the injection volume is equal. , taking the disease group as an example, the dilution factor is shown in Table 1:

[0078] Table 1 - Determination of dilution factor

[0079]

[0080]

[0081] According to the actual dilution factor dete...

Embodiment 2

[0106] A urine metabolomics analysis method, the difference from Example 1 is:

[0107] A. Simulation of the state of clinical urine samples

[0108] Three clinical urine samples were randomly selected and diluted by 2, 4, 8, and 12 times, respectively, to simulate the actual state of the dilution of endogenous metabolite concentrations in clinical urine samples due to the influence of physiological and pathological conditions.

[0109] Determination of osmotic pressure

[0110] Using YASN freezing point osmotic pressure (OM-819.C), use pure water and standard solution to perform three-point calibration at 0 mosm, 300 mosm and 900 mosm. After the calibration is completed, the sample can be measured. Take 100μL of sample into the sample tube, immerse the probe in the sample solution, press the probe, the cold trap starts to freeze the sample, read the reading after the reading is stable according to the instrument prompt, discard the sample tube, wipe the probe, and test the n...

Embodiment 3

[0127] A urine metabolomics analysis method, the difference from Example 1 is,

[0128] A. Take urine samples to determine the osmotic pressure of each urine sample

[0129] 10 urine samples from patients with hydronephrosis (disease group) and 10 urine samples from healthy people were taken, and the method was validated by a calibrated metabolomics experiment in RPLC(+) mode.

[0130] B. Sample pretreatment

[0131] According to the osmotic pressure measurement results, the osmotic pressure span of the actual sample is about 10 times. In order to prevent the solvent effect caused by the excessive injection volume, the dilution ratio of each sample is determined according to the principle that the product of the osmotic pressure of all samples and the injection volume is equal. . Add reagents for dilution according to the actual dilution factor determined for each sample. After the above-mentioned treatment of the sample, the sample was vortexed for 1 min to mix evenly, and...

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Abstract

The invention discloses a urine metabolomics analysis method based on osmotic pressure correction, which relates to the field of medical technology. The difficulty of diluting changes due to the influence of physiological and pathological conditions such as diseases. It is achieved through the following steps: (1) Determination of the osmotic pressure of the urine sample by the freezing point method; (2) Correct the injection volume or dilute the sample according to the osmotic pressure of the sample, so that the product of the osmotic pressure and the injection volume of all samples is at the same value (3) Use ultra-high performance liquid chromatography-high resolution mass spectrometry for detection and analysis; (4) Use software and databases to identify and statistically analyze compounds to find differential metabolites and metabolic pathways. The invention can be widely applied to various animal and clinical urine samples, especially for kidney diseases, and provides exemplary research for urine metabolomics analysis.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a urine metabolomics analysis method based on osmotic pressure correction, in particular to a urine metabolomics analysis method based on osmotic pressure for normalization before data collection. Background technique [0002] Metabolomics (metabonomic / metabolomics) is the qualitative and quantitative analysis of all low molecular weight (MW<1500) metabolites in a certain organism or cell, and the dynamic changes of metabolites produced under the action of internal and external factors are correlated with pathophysiology. associated disciplines. As the most downstream of systems biology, it reflects the actual state of organisms after various internal and external environmental disturbances, and the phenotypic differences between different individuals. Metabolomics has a wide range of applications in the biomedical field, such as finding biomarkers for clinical diagnosis and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/72G01N30/86
CPCG01N30/02G01N30/72G01N30/8634
Inventor 冯蕾何祉安
Owner SHANGHAI JIAOTONG UNIV