Abstract
Oxidative nucleic acid modifications have attracted increasing attention in recent years since they have been found to be related to a number of diseases including cancer. 8-Hydroxy-2′-deoxyguanosine (8-OHdG) and 8-hydroxyguanosine (8-OHG) are the typical markers of oxidative modification of DNA and RNA, respectively, and they are emerging biomarkers for the early detection of diseases. Urine is a favored biofluid for biomarker discovery due to its noninvasiveness to patients. Accurate quantification of these oxidative nucleic acid modifications still has challenges because their amounts in urine are very low and the interferences in urine samples are complicated. Herein, we developed and validated an accurate and robust solid-phase extraction (SPE) coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous quantification of these oxidative nucleic acid modifications in human urine. Stable isotope dilution strategy was utilized and the method shows good precision on intraday and interday measurements. Meanwhile, recovery was satisfactory by utilizing the Oasis hydrophilic–lipophilic balance (HLB) cartridge for sample pretreatment at three spiked levels. We successfully quantified urinary 8-OHdG and 8-OHG from 60 gastric cancer patients and 70 healthy controls by using this method. The measured contents of 8-OHdG and 8-OHG in urine from gastric cancer patients are both increased, compared with those in urine from healthy controls, indicating these oxidative nucleic acid modifications could act as potential non-invasive markers for early diagnosis of gastric cancer. Moreover, the present study will stimulate investigations of the effects of oxidative stress and nucleic acid modifications on the initiation and progression of gastric cancer.
Highlights
Reviewed by: Rebecca Whelan, University of Notre Dame, United States Daoyang Chen, Michigan State University, Specialty section: This article was submitted to Analytical Chemistry, a section of the journal Frontiers in Chemistry
We developed and validated an accurate and robust solid-phase extraction (SPE) coupled with stable isotope dilution ultra-performance liquid chromatography (UPLC)-MS/MS method for the simultaneous determination of urinary 8-OHdG and 8-OHG from gastric cancer patients
It has been demonstrated that acetic acid could improve the sensitivity of 8-OHdG in UPLC-MS/MS, whereas formic acid could improve the detection sensitivity of 8-OHG (Guo et al, 2016, 2020)
Summary
Reviewed by: Rebecca Whelan, University of Notre Dame, United States Daoyang Chen, Michigan State University, Specialty section: This article was submitted to Analytical Chemistry, a section of the journal Frontiers in Chemistry. The measured contents of 8-OHdG and 8-OHG in urine from gastric cancer patients are both increased, compared with those in urine from healthy controls, indicating these oxidative nucleic acid modifications could act as potential non-invasive markers for early diagnosis of gastric cancer. As the typical oxidative modifications of nucleic acids, 8-hydroxy-2′deoxyguanosine (8-OHdG) and 8-hydroxyguanosine (8-OHG) have drawn increasing attention since previous studies indicated that oxidative nucleic acid damage was associated with a variety of diseases, including neurodegenerative diseases, type 2 diabetes, cardiovascular diseases, aging, and cancer (Nunomura et al, 2012; Gao et al, 2019; Liang et al, 2020; Urbaniak et al, 2020). Many types of sorbents could be used for packing the cartridges, and it was selected on the basis of the property of the target analytes
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