Detecting method for modified nucleoside in urine

A detection method and a technology for modifying nucleosides, which are applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as pollution, and achieve the effects of avoiding pollution, improving accuracy, and improving stability

Active Publication Date: 2016-05-04
HANGZHOU HEALTH BANK MEDICAL LAB CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

Aiming at the shortcoming that the test sample is susceptible to microbial contamination, the detection method provided by the present invention stores the urine on the filter paper sheet, which not only greatly improves its stability and avoids microbial contamination, but also has the advantages of simplicity and convenience

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  • Detecting method for modified nucleoside in urine
  • Detecting method for modified nucleoside in urine
  • Detecting method for modified nucleoside in urine

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Embodiment

[0044] Instruments and materials:

[0045] 6495 tandem mass spectrometer from Agilent Company of the United States, and Angilent 1290 liquid chromatograph, the chromatographic column is Agilent C18 column, the specification is 50mm×3.0mm, 2.7μm.

[0046] Drugs and reagents:

[0047] Standard: pseudouridine, cytidine, 1-methyladenosine, uridine, adenosine, inosine, guanosine, 1-methylhypoxanthine Glycoside, 5-methyluridine, xanthine nucleoside, 3-methyluridine, 1-methylguanosine, 6-methyladenosine, 2-methylguanosine, N4-acetylcytidine, N2 , N2-dimethylguanosine were purchased from Sigma-Aldrich Company.

[0048]Internal standard: 8-bromoguanosine (8-Br-G), purchased from Sigma-Aldrich Company.

[0049] Methanol: chromatographically pure, Merck Company.

[0050] Formic acid: chromatography pure, Merck company.

[0051] Ultrapure water: Thermo Corporation.

[0052] Urine sample: volunteer urine sample.

[0053] Accurately weigh the above-mentioned 16 kinds of standard subs...

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Abstract

The invention provides a detecting method for modified nucleoside in urine. The method specifically comprises the following steps: pretreating the urine and storing the pretreated urine on a piece of filter paper, then carrying out extraction to obtain a to-be-tested sample; carrying out liquid chromatography tandem mass spectrometry detection on the to-be-tested sample, so as to obtain the variety and the content of the modified nucleoside. In the detecting method provided by the invention, the modified nucleoside in the urine can be quickly and effectively detected qualitatively and quantitatively by storing the urine on the filter paper, then carrying out extraction and using the liquid chromatography tandem mass spectrometry method; as the urine is stored on the filter paper, and the urine sample is converted into a solid form to be stored, the stability is greatly improved, and the microbial pollution to the detected sample is avoided; furthermore, the method provided by the invention is easier and more convenient than the existing freezing method.

Description

technical field [0001] The invention relates to the technical field of liquid spectrometry and mass spectrometry, in particular to a method for detecting modified nucleosides in urine. Background technique [0002] Modified nucleosides mainly exist in RNA and are formed by specific methyltransferases and ligases acting on polynucleoside molecules after transcription. RNA metabolism requires specific enzymes to catalyze. Normal nucleosides can be degraded or phospholipidized and reused after enzymatic hydrolysis, while modified nucleosides are unique nucleosides formed by the modification or combination of glycosidic bonds, bases, and sugar hydroxyl groups of normal nucleosides. Due to its highly modified structure, the enzyme activity and specificity required to degrade the modified nucleosides are high. Therefore, the modified nucleosides are released from the cells and finally excreted through urine. Therefore, the content of modified nucleosides in urine can be used to e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02G01N2030/027
Inventor 童鸿斌
Owner HANGZHOU HEALTH BANK MEDICAL LAB CO LTD
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