Self-Classified Topic Area(s): Assays Leveraging MS
An UPLC-MS/MS Method for Quantification of 8-OHdG in Urine
Chia-Ni Lin, Hsin-Lian Lin,Ya-Ching Huang Department of Laboratory Medicine, Chang Gung Memorial Hospital, Taoyuan, Taiwan
Chia-Ni Lin, PhD (Presenter) Chang Gung Memorial Hospital
Presenter Bio: Dr. Chia-Ni Lin joined Chang Gung Memorial Hospital in 2012. She received her Ph.D. degree in Clinical Laboratory Sciences and Medical Biotechnology from National Taiwan University in Taiwan followed by a fellowship in Clinical Chemistry at ARUP/University of Utah. She is board certificated by the American Board of Clinical Chemistry (ABCC) and a fellow of AACC academy. Her research interests are development, method improvement and validation of clinical laboratory assays, and mass spectrometry applications in clinical laboratory including metabolites in biological fluids, illicit drugs, trace elements and hormones.
Relevant Financial Disclosures
(within past 24 months)
No relevant financial relationship(s) to disclose.
Abstract
INTRODUCTION: 8-OHdG (8-hydroxy-2-deoxyguanosine) is generated after the repair of ROS-mediated DNA damages and has been widely used as a biomarker for oxidative stress. Oxidative damage to DNA has been associated with numerous pathological conditions, including cancer, diabetes and cardiovascular disease. This study aimed to develop a method to measure 8-OHdG in urine by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).
METHODS: Stable isotope-labelled 8-OHdG was used as internal standard. 8-OHdG was separated from the biological fluid using solid phase extraction (SPE). The elution solvent was injected into a LC-MS/MS (Waters Xevo TQ-XS). Samples were separated on a BEH Amide column followed by positive electrospray ionization and tandem mass spectrometric detection.
RESULTS: The linearity of 8-OHdG was between 0.2 to 20 ng/mL with a correlation coefficient of 0.997. The coefficient variation for inter- and intra-day imprecision was less than 4.8%. The accuracy was evaluated by spike recovery and the mean recovery was 106%-109%. No carryover and ion suppressant or enhancement was observed in this validated method.
CONCLUSION: In conclusion, we have developed a rapid, sensitive, and robust UPLC-MS/MS method for the quantification of 8OHdG in urine, which can provide results to assist in the assessment of oxidative stress.
= Discovery stage. (24.37%, 2023)
= Translation stage. (39.50%, 2023)
= Clinically available. (36.13%, 2023)
