Quantitative detecting method for various metabolites in biological sample, and metabolic chip

A quantitative detection method and detection method technology, applied in the direction of biological testing, biological material analysis, chemical instruments and methods, etc., can solve the problems of unsolved quantitative detection of multiple indicators, reduce the difficulty of detection, improve sensitivity, and avoid system errors Effect

Active Publication Date: 2019-02-01
麦特绘谱生物科技(上海)有限公司
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Problems solved by technology

However, the problem of quantitative detection of multiple indicators at the same time has not been solved

Method used

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  • Quantitative detecting method for various metabolites in biological sample, and metabolic chip
  • Quantitative detecting method for various metabolites in biological sample, and metabolic chip
  • Quantitative detecting method for various metabolites in biological sample, and metabolic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0041] Metabolic Chip Determination of Multiple Index Components in 10 Human Blood and Feces Samples

[0042] 1. Instrument

[0043] Liquid chromatography-tandem mass spectrometer (LC-MS / MS), equipped with electrospray ionization source (ESI).

[0044] 2. Sample preparation

[0045] Serum samples: collect venous whole blood samples, place them in anticoagulant tubes, mix them upside down 5-6 times immediately after blood collection, and centrifuge within 30 minutes to separate the plasma and place them in a centrifuge tube. Use cold methanol: chloroform: water in the centrifuge tube Extract with mixed solvent (volume ratio 3:1:1), shake for a few seconds, centrifuge the sample at 10,000-20,000 rpm and 4°C for 5-15 minutes to obtain the supernatant, and transfer the supernatant to an autosampler glass vial Medium; all aqueous serum or urine sample extracts were used for subsequent derivatization treatments.

[0046] Stool sample: Freeze-dry the stool sample; homogenize the a...

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Abstract

The invention discloses a quantitative detecting method for various metabolic components in a biological sample, and a metabolic chip used for the quantitative detecting method. The quantitative detecting method is characterized in that the biological sample is subjected to derivatization treatment, and the biological sample subjected to derivatization is detected through liquid chromatography andmass spectrometry combination; and during derivatization treatment, 3-nitrophenylhydrazine serves as a derivatization reagent, and 1-(3-dimethylaminopropyl)-3-ethly carbodiimide serves as a derivatization reaction catalyst. According to the quantitative detecting method, high-sensitivity detection can be achieved, and a plurality of level-crossing metabolic components can be covered during detection, and the quantitative detecting method is simple, fast and suitable for being applied to clinical and scientific research testing. The metabolic chip comprises a chip carrier microtitration plateand related reagents, and thus the multiple level-crossing metabolites such as amino acid, phenols, phenyl or benzyl derivatives, indole, organic acid, fatty acid, sugar and bile acid in the biological samples can be subjected to quantitative detection on the same microtitration plate.

Description

Technical field [0001] The present invention relates to the field of biological sample detection, specifically to methods for the quantitative determination of multiple metabolic components in biological samples, such as amino acids, phenols, phenyl or benzyl derivatives, indole, organic acids, fatty acids, sugars and bile acids. In particular, it relates to a detection method for measuring biological samples by chemical derivation method tandem mass spectrometry and a metabolic chip used in the method. Background technique [0002] Metabolomics involves the unbiased analysis of the complete set of metabolites (metabolome) present in cells, body fluids and tissues. Currently, nuclear magnetic resonance (NMR) or mass spectrometry (MS)-based metabolomics platforms detect many small molecule (MW<1500) metabolites but only in biological fluids (serum / plasma or urine) and in subjects with metabolic diseases. Relative (not absolute) concentrations are provided in the tissues o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/02
CPCG01N30/02G01N30/06G01N2030/067G01N2030/8813G01N30/7233G01N2560/00G01N2030/045G01N2800/7066G01N33/50B01L3/5085B01L2300/0681B01L2300/0819B01L2300/0829
Inventor 贾伟谢国祥王鹭
Owner 麦特绘谱生物科技(上海)有限公司
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