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Application of liquid chromatography tandem mass spectrogram method in biological monitoring of 3-alkylation adenine in mass smokers

A technology of alkylated adenine and tandem mass spectrometry, applied in the field of 3-alkylated adenine (3-methyladenine 3-MeA

Active Publication Date: 2015-02-18
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT +1
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Problems solved by technology

[0003] At present, there are many reports on the application of DNA adduct analysis methods in urine, but large-scale collection of 3-alkylated adenine (3-methyladenine 3-MeA; 3-Hydroxyethyladenine 3-EtA; 3-Hydroxyethyladenine 3-HOEtA) has not been reported

Method used

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  • Application of liquid chromatography tandem mass spectrogram method in biological monitoring of 3-alkylation adenine in mass smokers
  • Application of liquid chromatography tandem mass spectrogram method in biological monitoring of 3-alkylation adenine in mass smokers
  • Application of liquid chromatography tandem mass spectrogram method in biological monitoring of 3-alkylation adenine in mass smokers

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example 1

[0047] 1. Instruments and reagents:

[0048] Agilent 1200 liquid chromatography (Agilent, USA), equipped with G1367D autosampler, G 1312B binary mixing pump and G1316B column thermostat; API 5500 triple quadrupole tandem mass spectrometer (Applied Biosystems, USA), equipped with Electrospray ion source (ESI) and Analyst 1.5 software data processing system.

[0049] Ammonia was analytically pure; ammonium formate, ethyl acetate, and methanol were all chromatographically pure (TEDIA, USA); Oasis MCX solid-phase extraction column (6 mL, Waters, USA).

[0050] 3-methyladenine, 3-ethyladenine, 3-hydroxyethyladenine were purchased from Canada TRC company, deuterated-3-methyladenine, deuterated-3-ethyladenine internal standard was purchased from American CIL laboratory.

[0051] 2. Volunteer Recruitment / Sample Collection

[0052] A total of 261 (193 smokers, 58 non-smokers) healthy volunteers were recruited to undergo physical examination and sign an informed consent form. All e...

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Abstract

The invention relates to the application of a liquid chromatography tandem mass spectrogram method in biological monitoring of 3- alkylation adenine in mass smokers. The application is characterized in that a testing process comprises the following steps: defrosting urine excreted for 24 h at a room temperature; mixing the urine evenly and conducting a centrifugal filtration; purifying and enriching the urine through an Oasis MCX solid phase extraction column; mixing well and adopting an LC-MS / MS system to analyze; accurately detecting the content levels of the 3- alkylation adenine (3- methyl adenine, 3-Me A, 3- ethyl adenine, 3-Et A, 3- hydroxyethyl adenine, 3-H O Et A) in the urine. According to the invention, deuterium generation standards are utilized as internal standard quantitative analyte, so that inaccuracies occurred during the pre-treatment process of samples are reduced; the tandem mass spectroscopy method better improves the selectivity, accuracy and the sensitivity of the method; due to the choice and optimization of the chromatographic columns and the elution conditions, the chromatographic fractionation process is better improved; the time of chromatographic analysis is shortened; the consumption amount of organic solvent is reduced.

Description

technical field [0001] The invention belongs to the technical application field of physical and chemical testing of urine samples, and mainly relates to 3-alkylated adenine (3-methyladenine 3-MeA; 3-ethyladenine 3-EtA; 3-hydroxyethyl adenine) in urine Adenine 3-HOEtA) measurement technology application field, specifically a liquid chromatography-electrospray ionization-tandem mass spectrometer (LC-ESI-MS / MS) to measure 3-alkylated adenine (3-HOEtA) in urine - Application of the method of methyladenine 3-MeA; 3-ethyladenine 3-EtA; 3-hydroxyethyladenine 3-HOEtA). Background technique [0002] 3-Alkylated adenine (3-methyladenine 3-MeA; 3-ethyladenine 3-EtA; 3-hydroxyethyladenine 3-HOEtA) is a free radical and The product of adenine conjugation. Because the background value of endogenous 3-alkylated adenine in urine is low and can more accurately reflect the characteristics of the prototype compound, it is widely used as a biological monitoring indicator for the biological mo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 田永峰杨进王安刘勇侯宏卫胡清源韩书磊吴帅宾陈欢刘彤
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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