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- Title
New validated LC-MS/MS method for the determination of three alkylated adenines in human urine and its application to the monitoring of alkylating agents in cigarette smoke.
- Authors
Tian, Yongfeng; Hou, Hongwei; Zhang, Xiaotao; Wang, An; Liu, Yong; Hu, Qingyuan
- Abstract
A highly specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of urinary N-methyladenine ( N-MeA), N-ethyladenine ( N-EtA), and N-(2-hydroxyethyl)adenine ( N-HOEtA). Chromatographic separation was achieved on a hydrophilic interaction liquid chromatography column, with a mobile phase gradient prepared from aqueous 10 mM ammonium formate-acetonitrile (5:95 v/v, pH 4.0). Quantification of the analytes was done by multiple reaction monitoring using a triple-quadrupole mass spectrometer in positive-ionization mode. The limits of quantification were 0.13, 0.02, and 0.03 ng/mL for N-MeA, N-EtA, and N-HOEtA, respectively. Intraday and interday variations (relative standard deviations) ranged from 0.6 to 1.3 % and from 3.7 to 7.5 %. The recovery ranges of N-MeA, N-EtA, and N-HOEtA in urine were 80.1-97.3 %, 83.3-90.0 %, and 100.0-110.0 %, respectively. The proposed method was successfully applied to urine samples from 251 volunteers including 193 regular smokers and 58 nonsmokers. The results showed that the levels of urinary N-MeA, N-EtA, and N-HOEtA in smokers were significantly higher than those in nonsmokers. Furthermore, the level of urinary N-MeA in smokers was found to be positively correlated with the level of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol ( r = 0.48, P < 0.001, N = 192). This method is appropriate for routine analysis and accurate quantification of N-MeA, N-EtA, and N-HOEtA. It is also a useful tool for the surveillance of alkylating agent exposure. [Figure not available: see fulltext.]
- Subjects
ADENINE; URINALYSIS; ALKYLATING agents; CIGARETTE smoke; LIQUID chromatography-mass spectrometry
- Publication
Analytical & Bioanalytical Chemistry, 2014, Vol 406, Issue 22, p5293
- ISSN
1618-2642
- Publication type
Article
- DOI
10.1007/s00216-014-7979-1