Analysis of 8-hydroxy-2′-deoxyguanosine in human urine using hydrophilic interaction chromatography with tandem mass spectrometry

Hosozumi, Chiemi; Toriba, Akira; Chuesaard, Thanyarat; Kameda, Takayuki; Tang, Ning; Hayakawa, Kazuichi

doi:10.1016/j.jchromb.2012.02.043

Cited by 18 publications

(16 citation statements)

References 33 publications

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“…The LOD was 1 fmol and the LOQ was 3 fmol, which is consistent with the result obtained by HILIC method53. This indicated that the sensitivity of the analytical method developed was excellent.…”

Section: Resultssupporting

confidence: 89%

Association between Oxidative DNA Damage and Risk of Colorectal Cancer: Sensitive Determination of Urinary 8-Hydroxy-2′-deoxyguanosine by UPLC-MS/MS Analysis

Guo

Wang

et al. 2016

Sci Rep

119

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Oxidative DNA damage plays crucial roles in the pathogenesis of numerous diseases including cancer. 8-hydroxy-2′-deoxyguanosine (8-OHdG) is the most representative product of oxidative modifications of DNA, and urinary 8-OHdG is potentially the best non-invasive biomarker of oxidative damage to DNA. Herein, we developed a sensitive, specific and accurate method for quantification of 8-OHdG in human urine. The urine samples were pretreated using off-line solid-phase extraction (SPE), followed by ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. By the use of acetic acid as an additive to the mobile phase, we improved the UPLC-MS/MS detection of 8-OHdG by 2.7−5.3 times. Using the developed strategy, we measured the contents of 8-OHdG in urine samples from 142 healthy volunteers and 84 patients with colorectal cancer (CRC). We observed increased levels of urinary 8-OHdG in patients with CRC and patients with tumor metastasis, compared to healthy controls and patients without tumor metastasis, respectively. Additionally, logistic regression analysis and receiver operator characteristic (ROC) curve analysis were performed. Our findings implicate that oxidative stress plays important roles in the development of CRC and the marked increase of urinary 8-OHdG may serve as a potential liquid biomarker for the risk estimation, early warning and detection of CRC.

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Section: Resultssupporting

confidence: 89%

Association between Oxidative DNA Damage and Risk of Colorectal Cancer: Sensitive Determination of Urinary 8-Hydroxy-2′-deoxyguanosine by UPLC-MS/MS Analysis

Guo

Wang

et al. 2016

Sci Rep

119

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“…In light of our findings, we recommend that thawing at 37 °C is a necessary part of sample preparation for the analysis of oxidized guanine lesions in urine. Previous reports in the literature have used a variety of sample workup steps e.g., thawing of urine at RT [36] , removal of precipitates by filtration or centrifugation [38] , [46] , and diluting the urine with deionized water [47] prior to analysis. We speculate these steps may contribute to some of the apparent assay variability between laboratories, as noted previously [48] , [49] .…”

Section: Discussionmentioning

confidence: 99%

Clinical relevance of guanine-derived urinary biomarkers of oxidative stress, determined by LC-MS/MS

et al. 2019

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A reliable and fast liquid chromatography-tandem mass spectrometry method has been developed for the simultaneous determination of three oxidized nucleic acid damage products in urine, 8-oxoguanine (8-oxoGua), 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodGuo) and 8-oxo-7,8-dihydroguanosine (8-oxoGuo). We applied this method to assess the effect of various urine workup procedures on the urinary concentrations of the oxidized nucleic acid products. Our results showed that frozen urine samples must be warmed (i.e., to 37 °C) to re-dissolve any precipitates prior to analysis. We showed that common workup procedures, such as thawing at room temperature or dilution with deionized water, are not capable of releasing fully the oxidized nucleic acid products from the precipitates, and result in significant underestimation (up to ~ 100% for 8-oxoGua, ~ 86% for both 8-oxodGuo and 8-oxoGuo).With this method, we further assessed and compared the ability of the three oxidized nucleic acid products, as well as malondialdehyde (MDA, a product of lipid peroxidation), to biomonitor oxidative stress in vivo. We measured a total of 315 urine samples from subjects with burdens of oxidative stress from low to high, including healthy subjects, patients with chronic obstructive pulmonary disease (COPD), and patients on mechanical ventilation (MV). The results showed that both the MV and COPD patients had significantly higher urinary levels of 8-oxoGua, 8-oxodGuo, and 8-oxoGuo (P < 0.001), but lower MDA levels, compared to healthy controls. Receiver operating characteristic curve analysis revealed that urinary 8-oxoGuo is the most sensitive biomarker for oxidative stress with area under the curve (AUC) of 0.91, followed by 8-oxodGuo (AUC: 0.80) and 8-oxoGua (AUC: 0.76). Interestingly, MDA with AUC of 0.34 failed to discriminate the patients from healthy controls. Emerging evidence suggests a potential clinical utility for the measurement of urinary 8-oxoGuo, and to a lesser extent 8-oxodGuo, which is strongly supported by our findings.

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“…To date, the degree of DNA damage has been assessed by the measurement of 8-OH-dGuo in biological fluids by LC-MS or ELISA methods [6,14] Scarce data are available on other products of the oxidation of nucleic acids.…”

Section: Discussionmentioning

confidence: 99%

“…Our method consists of sample pre-treatment with solid phase extraction (SPE) followed by chromatographic separation, with a 14 HILIC column, by UHPLC-MS/MS. In recent years, hydrophilic interaction chromatography (HILIC) has been reported as a good alternative to reversephase liquid chromatography (RPLC) for the separation of highly polar compounds such as nucleotides and their derivatives [6,14]. In our experience, SPE is essential to facilitate the subsequent detection and quantification of metabolites.…”

Section: Discussionmentioning

confidence: 99%

“…Different analytical methods have been used to determine these compounds, mostly based on chromatographic and immunoassay techniques. Some authors applied ELISA immunoassay techniques to identify metabolites but encountered problems derived from the 4 lack of selectivity of the antibody in the urine matrix [6]. The method employed most widely to measure urinary 8-OH-dGuo is HPLC-MS/MS, due to its high sensitivity, specificity, and time saving characteristics [7].…”

Section: Introductionmentioning

confidence: 99%

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Effect of elite physical exercise by triathletes on seven catabolites of DNA oxidation

Villaño

Vilaplana

Medina

et al. 2015

Free Radical Research

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The oxidized nucleoside 8-hydroxy-2'-deoxyguanosine has been widely studied as a marker of DNA oxidation; however, data on the occurrence of other metabolites in plasma that are related to DNA damage are scarce. We have applied an improved, sensitive, robust, and reliable method, involving solid phase extraction and ultrahigh-performance liquid chromatography (UHPLC)-tandem mass spectrometry (MS/MS), to the precise quantitation of seven metabolites in the plasma of 15 elite triathletes after a 2-week training program. All compounds were eluted in the first 1.6 min, with limits of detection and quantification ranging between 0.001 and 0.3 ng.mL(-1) and 0.009 and 0.6 ng.mL(-1), respectively. Four compounds were detected in plasma: guanosine-3'-5'-cyclic monophosphate, 8-hydroxyguanine, 8-hydroxy-2'-deoxyguanosine, and 8-nitroguanosine. After two weeks of training, 8-hydroxyguanine exhibited the highest increase (from 0.031 ± 0.008 nM to 0.036 ± 0.012 nM) (p < 0.05), which could be related to the enhanced activity of DNA-repairing enzymes that excise this oxidized base. Increased levels of guanosine-3'-5'-cyclic monophosphate and 8-hydroxy-2'-deoxyguanosine were also observed. In contrast, levels of 8-nitroguanosine (p < 0.05) were significantly reduced, which might be a protective measure as this compound strongly stimulates the generation of superoxide radicals, and its excess is related to pathologies such as microbial (viral) infections and other inflammatory and degenerative disorders. The results obtained indicate an induced adaptive response to the increased oxidative stress related to elite training, and point to the benefits associated with regular exercise.

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Analysis of 8-hydroxy-2′-deoxyguanosine in human urine using hydrophilic interaction chromatography with tandem mass spectrometry

Cited by 18 publications

References 33 publications

Association between Oxidative DNA Damage and Risk of Colorectal Cancer: Sensitive Determination of Urinary 8-Hydroxy-2′-deoxyguanosine by UPLC-MS/MS Analysis

Association between Oxidative DNA Damage and Risk of Colorectal Cancer: Sensitive Determination of Urinary 8-Hydroxy-2′-deoxyguanosine by UPLC-MS/MS Analysis

Clinical relevance of guanine-derived urinary biomarkers of oxidative stress, determined by LC-MS/MS

Effect of elite physical exercise by triathletes on seven catabolites of DNA oxidation

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