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Method for detecting 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in urine by high-performance liquid chromatography tandem mass spectrometry technology

A hydroxydeoxyguanosine and high performance liquid chromatography technology, applied in the field of biochemical detection, can solve the problems of poor specificity, complex method, low sensitivity, etc., and achieve the effect of good recovery rate and stability

Inactive Publication Date: 2016-04-27
SHANGHAI DIAN CLINICAL TESTING CENT
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  • Application Information

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Problems solved by technology

[0003] The technical problem to be solved in the present invention is to provide a method for detecting 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in urine by high-performance liquid chromatography tandem secondary mass spectrometry, so as to solve the complicated and specific method existing in the prior art. Problems with poor performance and low sensitivity

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  • Method for detecting 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in urine by high-performance liquid chromatography tandem mass spectrometry technology
  • Method for detecting 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in urine by high-performance liquid chromatography tandem mass spectrometry technology
  • Method for detecting 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in urine by high-performance liquid chromatography tandem mass spectrometry technology

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Embodiment 1

[0044] 1. Materials

[0045] The samples of the methodological research experiment came from the urine samples of the employees of Shanghai Dean Medical Laboratory.

[0046] (1) Instrument: LC-20AD liquid chromatograph (Shimadzu, Japan), API4000 +Triple quadrupole mass spectrometer (ABSciex, the United States), Ultrapure water machine (ultrapure, Chengdu), multi-tube vortex mixer (Hangzhou Aosheng Instrument Co., Ltd.); rapid mixer (Xinkang, Jiangsu) , high-speed refrigerated centrifuge (Baiyang, Beijing), chromatographic separation column UltimatePolar-RP3μm2.1×100, adjustable pipette (ThermoFisher, 0.5-10μL, 10-100μL, 100-1000μL) and glassware, beaker, Graduated cylinder, etc.

[0047] (2) Reagent consumables: Methanol and acetonitrile are chromatographically pure, purchased from Tiandi Company of the United States; formic acid is chromatographically pure, purchased from Tianjin Zhiyuan Chemical Reagent Co., Ltd.; 1.5mL centrifuge tube (AXYGEN, USA); calcium chloride dihyd...

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Abstract

The invention discloses a method for detecting 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in urine by a high-performance liquid chromatography tandem mass spectrometry technology. According to the method, 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in a standard solution and a pretreated urine sample are separately detected by adopting the high-performance liquid chromatography tandem mass spectrometry technology, 8-hydroxydeoxyguanosine and 8-hydroxyguanosine are separated firstly by using a high-performance liquid chromatography technology, and then, a calibration curve is established by using a tandem mass spectrometry isotopic internal-standard quantitative method in a manner of taking a concentration ratio of a standard to an internal standard as an X axis and taking a peak area ratio of the standard to the internal standard as a Y axis, so as to figure out the content of 8-hydroxydeoxyguanosine and the content of 8-hydroxyguanosine. Compared with the prior art, the method disclosed by the invention is efficient and stable, is good in recovery ratio and stability and has the advantages of accuracy, simplicity, convenience, economical efficiency, high efficiency and the like, thereby being applicable to popularization and application in clinical detection.

Description

technical field [0001] The invention belongs to the field of biochemical detection, and in particular relates to a method for detecting 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in urine by high performance liquid chromatography tandem secondary mass spectrometry. Background technique [0002] The potential use and value of DNA and RNA oxidative damage stress products as biomarkers is an important aspect of biomedical research. As two important oxidation markers, 8-hydroxydeoxyguanosine and 8-hydroxyguanosine are produced in tissue cells, transported by blood, and excreted by the kidneys with urine. Since the two substances are relatively stable in the body, and urine samples are easy to obtain, non-invasive detection can be realized. The monitoring of the two substances in urine can provide help for the effective evaluation of oxidative damage caused by various factors in the body. In addition, the influence of potential factors such as gender, age and smoking can be ...

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/88G01N30/06
Inventor 张卫卫于嘉屏
Owner SHANGHAI DIAN CLINICAL TESTING CENT
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