Determination of the oxidative stress biomarker urinary 8-hydroxy-2'-deoxyguanosine by automated on-line in-tube solid-phase microextraction coupled with liquid chromatography-tandem mass spectrometry

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Hiroyuki KataokaAkihiro Saito

Abstract

A simple and sensitive method for the determination of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a marker of oxidative DNA damage in human urine, was developed using automated on-line in-tube solid-phase microextraction (SPME) coupled with stable isotope-dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS). Creatinine was also analyzed simultaneously to normalize urine volume by the in-tube SPME LC-MS/MS method, and 8-OHdG and creatinine were separated within 3min using a Zorbax Eclipse XDB-C8 column. Electrospray MS/MS for these compounds was performed on an API 4000 triple quadruple mass spectrometer in the positive ion mode by multiple reaction monitoring. The optimum in-tube SPME conditions were 20 draw/eject cycles of 40μL of sample at a flow rate of 200μL/min using a Carboxen 1006 PLOT capillary column as an extraction device. The extracted compounds were easily desorbed from the capillary by passage of the mobile phase, and no carryover was observed. The calibration curve for 8-OHdG using its stable isotope-labeled internal standard was linear in the range of 0.05-10ng/mL, and the detection limit was 8.3pg/mL. The intra-day and inter-day precision (relative standard deviations) were below 3.1% and 9.6% (n=5), r...Continue Reading

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Citations

Oct 21, 2016·Journal of Separation Science·Bruno Henrique FumesFernando Mauro Lanças
Feb 23, 2018·Journal of Environmental Science and Health. Part. B, Pesticides, Food Contaminants, and Agricultural Wastes·Ana L De ToffoliFernando M Lanças
May 6, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Natalia ManousiConstantinos K Zacharis
Dec 7, 2020·Electrophoresis·Asmin AndriesAnn Van Schepdael
Dec 29, 2020·Journal of Chromatography. a·Hiroyuki Kataoka
Feb 3, 2021·Journal of Separation Science·Caroline Fernandes GreccoMaria Eugênia Costa Queiroz
Sep 14, 2020·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Ying SunXuejun Kang

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