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Journal of Analytical Chemistry

, Volume 63, Issue 1, pp 52–56 | Cite as

Quantitative determination of 8-hydroxy-2′-deoxyguanosine as a biomarker of oxidative stress in thalassemic patients using HPLC with an electrochemical detector

  • C. Matayatsuk
  • P. Wilairat
Articles

Abstract

A simple and robust HPLC method with electrochemical detection was developed for the quantitative determination of 8-hydroxy-2′-deoxyguanosine (8-OHdG), a DNA damage product excreted in urine. Sample cleanup was carried out using solid-phase extraction (SPE) prior to chromatographic separation. 8-OHdG was well separated on an Eclipse XDB®-C18 column (150 × 4.6 mm i.d., 5 μm) with an Eclipse XDB®-C18 guard column (12 × 4.6 mm i.d., 5 μm). Two mobile phases containing methanol and 10 mM sodium formate (pH 4.5) at a ratio of 10: 90 and 50: 50 v/v, respectively, were used. The retention time of 8-OHdG was 9.8 ± 0.5 min. The recovery of 8-OHdG was found to be 97.2 ± 3.3% (n = 6). Intraday and interday precisions of the method were 4.0 ± 2.9% (n = 6) and 6.6 ± 1.7% (n = 6), respectively. The detection limit was 5 ng/mL. Preliminary investigation showed that the mean value of 8-OHdG, normalized with the amount of creatinine in the sample, from the thalassemic group was significantly higher than that from healthy subjects (211 ± 214 ng/mg creatinine vs. 31.4 ± 32.2 ng/mg creatinine, respectively), indicating oxidative stress.

Keywords

Urine Sample Thalassemia Thalassemic Patient Sodium Formate Human Urine Sample 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Pleiades Publishing, Ltd. 2008

Authors and Affiliations

  • C. Matayatsuk
    • 1
  • P. Wilairat
    • 1
  1. 1.Department of Chemistry, Faculty of ScienceMahidol UniversityBangkokThailand

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