HPLC-TOF-MS/MS method for simultaneous determination of 10 nucleosides and application

A technology of HPLC-TOF-MS and determination method, which is applied in the field of analysis of such substances in urine, to achieve the effects of stable properties, simple mobile phase ratio, and convenient storage

Inactive Publication Date: 2016-10-12
CHINA PHARM UNIV
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Problems solved by technology

[0004] In the existing liquid chromatography-mass spectrometry method, there is no simple and general analysis method for multiple nucleoside substances. In view of the importance of modified nucleosides, the present invention attempts to simultaneously detect multiple nucleosides and modified nucleoside substances. Finally, the secondary mass spectrogram and standard curve are listed for qualitative and quantitative research respectively, and the analysis results are statistically analyzed to find out the relationship between the substances to be analyzed and the occurrence of tumors

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  • HPLC-TOF-MS/MS method for simultaneous determination of 10 nucleosides and application
  • HPLC-TOF-MS/MS method for simultaneous determination of 10 nucleosides and application
  • HPLC-TOF-MS/MS method for simultaneous determination of 10 nucleosides and application

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[0024] An HPLC-TOF-MS / MS method and application for simultaneous determination of 10 nucleoside substances include the following steps.

[0025] 1. Experimental part

[0026] 1. Reagents and instruments:

[0027] Nucleoside substances mixed standard (including cytidine, inosine, guanosine, uridine, 1-methyladenosine, pseudouridine, 5-methylcytidine, 5-methyluridine, pseudouridine, N -4-acetylcytidine); acetonitrile; ultrapure water; formic acid.

[0028] Agilent 1250 high performance liquid chromatography (Agilent, USA), equipped with G1315DDAD detector, G1312A binary pump, G1322A vacuum degasser, G1367B autosampler, G1316A column thermostat; 6520Q-TOF mass spectrometer, ESI ion source; The chromatographic column is an Inertsil ODS-3 C18 column, its size is 250mm×4.6mm, i.d.5μm; electronic balance; orbital oscillator.

[0029] 2. Liquid quality conditions:

[0030] Mobile phase: A=0.1% formic acid water, B=0.1% formic acid acetonitrile, elution conditions: 0-6min, 5-10% B;...

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Abstract

Belonging to the technical field of small molecule metabolite analysis, the invention relates to an HPLC-TOF-MS/MS method for simultaneous determination of 10 nucleosides. The method specifically includes the steps of: mixing of 10 nucleosides to prepare solutions with different concentrations; HPLC-TOF-MS/MS analysis and identification of the10 nucleosides; HPLC-TOF-MS analysis and stability, precision, detection limit and quantification limit analysis of nucleoside mixed solutions with different solutions; drawing of a standard curve of the 10 nucleosides; HPLC-TOF-MS and HPLC-TOF-MS/MS analysis of biological samples; and qualitative and quantitative analysis of several specific nucleosides in the biological samples according to the HPLC-TOF-MS/MS data of each nucleoside and the standard curve. The method has the characteristics of rapid and simple analysis process, reliable detection result and high sensitivity, and is suitable for qualitative and quantitative analysis of nucleosides in blood samples, urine samples, saliva and other biological samples.

Description

technical field [0001] The invention relates to the fields of analytical chemistry and life sciences, and specifically belongs to the technical field of analysis of small molecular metabolites. It involves a qualitative analysis of 10 nucleoside substances based on HPLC-TOF-MS / MS, and a method for quantitative analysis based on HPLC-TOF-MS results, which is finally applied to such substances in urine analysis. Background technique [0002] The modified nucleosides in human urine and serum are mainly transfer RNA (Transfer Ribonucleic Acid, tRNA) metabolites, and its content reflects the metabolic level of the human body to a certain extent. Many reports have revealed the relationship between nucleosides and tumors. and some pathological inflammatory responses. When tumors occur, tumor cells need more nutrients to ensure their continuous reproduction. At this time, the body synthesizes a large amount of protein, and tRNA, as an important medium in the process from DNA to p...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02G01N2030/027
Inventor 顾月清吕丽伟张万存韩智豪张奇
Owner CHINA PHARM UNIV
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