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Detecting method for 3-ethyl adenine (3-HOEtA) in urine

A technology of ethyl adenine and a determination method, which is applied in the field of analyzing the urine of smoking and non-smoking people to achieve the effects of improving selectivity, accuracy and sensitivity, improving chromatographic separation process and reducing errors

Active Publication Date: 2014-06-11
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, there are many reports on the analysis and detection methods of DNA adducts in urine, but the extraction, purification, and detection of various targets in urine matrix are high The flux analysis method is not yet perfect

Method used

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  • Detecting method for 3-ethyl adenine (3-HOEtA) in urine
  • Detecting method for 3-ethyl adenine (3-HOEtA) in urine
  • Detecting method for 3-ethyl adenine (3-HOEtA) in urine

Examples

Experimental program
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Effect test

example 1

[0048] 1. Instruments and reagents:

[0049] 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.

[0050] 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).

[0051] 3-ethyladenine was purchased from Canadian TRC Company, and the deuterated-3-ethyladenine internal standard was purchased from CIL Laboratory in the United States.

[0052] 2. Sample handling:

[0053] The urine was thawed at room temperature and mixed evenly. Take 4 mL of urine, add 4 mL of ultrapure water, add 100 μL of internal standard (100 ng / mL), and adjust the temperature to 80 ° C with a water bath. The urine dil...

example 2

[0055] Example 2: As described in Example 1, the method established in this study was used to detect the content of 3-ethyladenine (3-EtA) in 20 urines, 3-ethyladenine (3-EtA) in urine - The content of EtA) is shown in Table 4.

[0056]

[0057] Table 4 Content of 3-EtA in urine

[0058]

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Abstract

The invention provides a detecting method for 3-ethyl adenine (3-HOEtA) in urine. The detecting method is characterized by comprising the steps of unfreezing 24-hour urine at room temperature, uniformly mixing the urine and centrifugally filtering the urine; purifying and enriching the urine through an OasisMCX solid-phase extraction column; introducing the urine to an LC-MS (liquid Chromatograph Mass Spectrometer) / MS system for analyzing after uniformly mixing to accurately detect the content of 3-EtA in the urine. The detecting method is a brand new detecting method for 3-EtA in the urine; a deuterated standard is used as a quantitative analyte of an internal standard, so that errors caused in a sample pretreatment process can be reduced; moreover, a tandem mass spectrometry is adopted so that the selectivity, accuracy and sensitivity of the method are better improved. Through selecting and optimizing a chromatographic column and an elution condition, the chromatographic separation process is well improved, the chromatographic analysis time is shortened, and the consumption of an organic solvent is reduced.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of physical and chemical testing of urine samples, and mainly relates to the technical field of determination of 3-ethyladenine (3-EtA) in urine, specifically a liquid chromatography-electrospray ionization-tandem mass spectrometer (LC-ESI-MS / MS) method for the determination of 3-ethyladenine (3-EtA) in urine. [0003] Background technique: [0004] 3-Ethyladenine (3-EtA) is the product of the combination of free radicals generated by the metabolism of exogenous compounds in vivo and adenine. Because the background value of endogenous 3-ethyladenine in urine is low and can reflect the characteristics of the prototype compound more accurately, it is widely used as a biological monitoring indicator for the biological monitoring of environmental alkylation pollutants. According to reports, 3-ethyladenine (N3-ethyladenine, 3-EtA) in urine has a strong correlation with smoking levels, and can be us...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 田永峰侯宏卫胡清源王安刘勇杨进陈欢韩书磊刘彤张小涛石龙凯刘洋
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT