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A Mass Spectrometry Method for Quantifying Endogenous Metabolites in Tissue by Molecular Image

A metabolite and tissue technology, applied in the field of medical detection and mass spectrometry detection, can solve the problems of low throughput, high cost, unable to meet clinical needs, etc., and achieve the effect of high sensitivity

Active Publication Date: 2022-07-05
融智生物科技(青岛)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Biochemical methods are basically based on colorimetric methods or enzyme analysis methods, which require tedious and time-consuming sample pretreatment processes such as tissue homogenization; chromatography-mass spectrometry methods have low throughput, and can only analyze 2- HG content
Although magnetic resonance spectroscopy can non-invasively assess the content of 2-HG in patients before surgery, it is costly and requires complex data post-processing processes, such as baseline correction algorithms, etc.
Although atmospheric pressure mass spectrometry can quickly detect 2-HG qualitatively, it cannot meet the clinical needs in the quantitative detection of 2-HG

Method used

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  • A Mass Spectrometry Method for Quantifying Endogenous Metabolites in Tissue by Molecular Image
  • A Mass Spectrometry Method for Quantifying Endogenous Metabolites in Tissue by Molecular Image

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1 Determination of parameters and elements in the detection process

[0031] The choice of matrix is ​​critical for mass spectrometers equipped with a matrix-assisted laser desorption ionization source. In order to ensure and improve the ionization efficiency of 2-HG in glioma tissue, the choice of matrix is ​​the first step. The matrix needs to have low interference from background peaks in the low molecular weight region and be able to ionize the analyte in negative ion mode. This patent innovatively proposes N-phenylnaphthylamine compounds including N-(1-naphthyl)ethylenediamine dihydrochloride (NEDC) matrix for use with The mass spectrometer with matrix-assisted laser desorption ionization source was used to detect 2-HG ionization in glioma tissue (NEDC has no interference peak near 147.0 of 2-HG, and the signal is very strong). Although the detection limit, sensitivity and other indicators are different, other matrices such as 1-5-DAN, 9-AA, DHB verificat...

Embodiment 2

[0034] Example 2 Quantitative analysis of 2-HG in human glioma tissue

[0035]Mouse liver tissue and multiple human glioma tissues were cut into 12 μm thick sections at -17°C, transferred to the same ITO conductive slide, and dried in a vacuum desiccator for 20 minutes.

[0036] Prepare multiple gradient concentrations of 2-hydroxyglutaric acid disodium salt in the aqueous solution, mix them evenly, and use a pipette to pipette 0.2 μL of the standard solution of the above concentration gradient onto the surface of the mouse liver tissue, each 0.2 μL of the standard solution solution Covers about 0.785mm 2 The tissue area, the density of mouse liver tissue is 1g / cm 3 Calculation, then 0.2μL standard solution is located on about 9.42μg tissue, then the standard solution is converted into the content in the tissue, which are 0.1, 0.5, 1.0, 2.0, 5.0, 10.0, 20.0, 40.0, 80.0pmol / μg , the slides were placed in a vacuum desiccator for 10 minutes immediately after spotting the standa...

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Abstract

The present application provides a mass spectrometry method for quantifying endogenous metabolites in tissue by means of molecular images, characterized in that the method comprises the following steps: (1) selection of a simulated tissue model; (2) addition of standard substances; (3) Prepare matrix and add internal standard. The method of the present application uses N-(1-naphthyl)ethylenediamine dihydrochloride as a substrate, 13 C as an internal standard for the detection of 2-hydroxyglutarate in glioma tissue has achieved good results in terms of sensitivity, specificity and detection limit.

Description

technical field [0001] The invention belongs to the field of medical detection and mass spectrometry detection. Specifically, the present application provides a mass spectrometry method for quantifying endogenous metabolites in tissues, comprising the following steps: (1) selection of a simulated tissue model; (2) selection of a standard substance Add; (3) prepare matrix and add internal standard; it uses N-(1-naphthyl)ethylenediamine dihydrochloride as matrix, 13 C was used as an internal standard to quantitatively detect 2-hydroxyglutarate in glioma tissue. Background technique [0002] Mutations in the isocitrate dehydrogenase (IDH) gene are found in a variety of cancers, including glioma, acute myeloid leukemia, prostate cancer, thyroid cancer, colon cancer, chondrosarcoma, cholangiocarcinoma, melanoma and para Gangliomas, etc. IDH gene mutations are known to reduce α-Ketoglutarate (α-KG) to 2-Hydroxyglutarate (2-HG), resulting in the oncogenic metabolite 2-HG in tissu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/64
CPCG01N27/64
Inventor 兰春燕王磊张晶李海南蔡林波马旭
Owner 融智生物科技(青岛)有限公司
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