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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 893-894; pp. 173 - 176
• Main Authors: Hosozumi, Chiemi, Toriba, Akira, Chuesaard, Thanyarat, Kameda, Takayuki, Tang, Ning, Hayakawa, Kazuichi
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.04.2012
• Subjects: 8-Hydroxy-2′-deoxyguanosine; acetonitrile; Acetonitriles; ammonium acetate; biomarkers; Chromatography; Chromatography, Liquid - methods; Chromatography, Reverse-Phase; Deoxyguanosine - analogs & derivatives; Deoxyguanosine - urine; detection limit; Human; Humans; Hydrophilic interaction chromatography; hydrophilicity; Hydrophobic and Hydrophilic Interactions; Ionization; isotope labeling; Limit of Detection; Mass spectrometry; Monitoring; Monitors; Oxidative stress; rapid methods; Reproducibility of Results; Signal-To-Noise Ratio; Tandem mass spectrometry; Tandem Mass Spectrometry - methods; Urine; Urine; Tandem mass spectrometry; Oxidative stress; 8-Hydroxy-2′-deoxyguanosine; Hydrophilic interaction chromatography
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2012.02.043

► Improved sensitivity of mass spectrometric detection by HILIC column. ► A sensitive and rapid analytical method for urinary 8-hydroxy-2′-deoxyguanosine. ► Strong chromatographic retention of polar compounds under highly organic mobile phase. Urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) is a widely used noninvasive biomarker of oxidative stress. A selective, sensitive and rapid method for determining 8-OHdG in human urine was developed using hydrophilic interaction chromatography–tandem mass spectrometry (HILIC–MS/MS) with electrospray ionization. 8-OHdG and isotopically labeled 8-OHdG (internal standard) were separated on a HILIC column with a mobile phase of 10mM ammonium acetate: acetonitrile (1:9, v/v) within 10min and detected by using a positive electrospray ionization interface under the selected reaction monitoring mode. The detection limits of 8-OHdG (corresponding to a signal-to-noise ratio of 3) for the HILIC-MS/MS system and the conventional method using a reversed-phase column with MS/MS were 1.0 and 26.0fmol/injection, respectively. The proposed method makes it possible to monitor the basal level of urinary 8-OHdG from non-exposed healthy subjects and can be used for large-scale human studies.