Simultaneous and Ultrarapid Determination of Reactive Oxygen Species and Reduced Glutathione by Micr

• 作 者: JianhuaQin;NannanYe;LinfenYu;DayuLiu;BingchengLin;
• 机 构: Department of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dali
• 出 处: 2005 World DNA and Genome Day(2005中国(大连)国际DNA和基因组节大会)
• 发表日期: 2005年期

　　In this study,a simultaneous and ultrarapid microchip electrophoresis method coupled with laser-induced fluorescence (LIF) detection was established for the simultaneous determination of two important intracellular signaling molecules (reactive oxygen species,ROS,and reduced glutathione,GSH) related to apoptosis and oxidative stress.The selected fluorogenic probes were dihydrorhodamine-123 (DHR-123),which can be converted intracellularly by ROS to the fluorescent rhodamine-123 (Rh-123).Probe naphthalene-2,3-dicarboxaldehyde (NDA) can react quickly with GSH in cell extracts to produce a fluorescent adduct.Rapid separation ofRh-123,GSH-NDA adduct was completed on a glass microchip within 30 s with a 20mM borate buffer (pH 9.2).The method gives a linear calibration curve for tested Rh123 and GSH within the range of 2.6 × 10-12-1.3 × 10-10 M and 5 × 10-7-5 × 10-5 M.The limit of detection (LOD) for Rh-123 and GSH was 1.88 pM (0.21 zmol) and 0.46 μM (5.43 amol),respectively.