PMID- 20143832
OWN - NLM
STAT- MEDLINE
DCOM- 20100614
LR  - 20131121
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 82
IP  - 5
DP  - 2010 Mar 1
TI  - Potent method for the simultaneous determination of glutathione and hydrogen 
      peroxide in mitochondrial compartments of apoptotic cells with microchip 
      electrophoresis-laser induced fluorescence.
PG  - 2006-12
LID - 10.1021/ac902741r [doi]
AB  - The first application of microchip electrophoresis with laser-induced 
      fluorescence (MCE-LIF) detection to simultaneously determine glutathione (GSH) 
      and hydrogen peroxide (H(2)O(2)) in mitochondria was described. Organoselenium 
      probe Rh-Se-2 and bis(p-methylbenzenesulfonate)dichlorofluorescein (FS) 
      synthesized in our laboratory were utilized as fluorescent probes for GSH and 
      H(2)O(2), respectively. Rh-Se-2, which is nonfluorescent, reacts with GSH to 
      produce rhodamine 110 (Rh110) with high quantum yield. Similarly, nonfluorescent 
      FS reacts with H(2)O(2) and produces dichlorofluorescein (DCF) accompanied by 
      drastic fluorescence enhancement. Both probes exhibit good sensitivity toward 
      their respective target molecule determination. Fast, simple, and sensitive 
      determination of GSH and H(2)O(2) was realized within 37 s using a running buffer 
      of 50 mM mannitol, 40 mM HEPES (pH 7.4), and an electric field of 360 V/cm for 
      separation. The linear ranges of the method were 3.3 x 10(-9)-1.0 x 10(-7) M/2.9 
      x 10(-7)-1.0 x 10(-4) M and 2.7 x 10(-9)-4.0 x 10(-7) M with detection limits 
      (signal-to-noise ratio = 3) of 1.3 nM (0.16 amol) and 1.0 nM (0.12 amol) for GSH 
      and H(2)O(2), respectively. The relative standard deviations (RSDs) of migration 
      time and peak area were less than 1.0% and 4.0%, respectively. The MCE-LIF assay 
      was utilized to investigate the levels of GSH and H(2)O(2) in mitochondria 
      isolated from HepG2 cells and were found to be 2.01 +/- 0.21 mM and 5.36 +/- 0.45 
      microM, respectively. The method was further extended to observe situations of 
      the two species in mitochondria of HepG2 cells experiencing cell apoptosis that 
      were induced by doxorubicin and photodynamic therapy (PDT).
FAU - Chen, Zhenzhen
AU  - Chen Z
AD  - College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory 
      of Molecular and Nano Probes, Ministry of Education, Shandong Normal University, 
      Jinan 250014, China.
FAU - Li, Qingling
AU  - Li Q
FAU - Wang, Xu
AU  - Wang X
FAU - Wang, Zhiyuan
AU  - Wang Z
FAU - Zhang, Ruirui
AU  - Zhang R
FAU - Yin, Miao
AU  - Yin M
FAU - Yin, Lingling
AU  - Yin L
FAU - Xu, Kehua
AU  - Xu K
FAU - Tang, Bo
AU  - Tang B
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - BBX060AN9V (Hydrogen Peroxide)
RN  - GAN16C9B8O (Glutathione)
SB  - IM
MH  - *Apoptosis
MH  - Electrophoresis, Microchip/*methods
MH  - Glutathione/*analysis
MH  - Hydrogen Peroxide/*analysis
MH  - Lasers
MH  - Mitochondria/*chemistry
EDAT- 2010/02/11 06:00
MHDA- 2010/06/15 06:00
CRDT- 2010/02/11 06:00
PHST- 2010/02/11 06:00 [entrez]
PHST- 2010/02/11 06:00 [pubmed]
PHST- 2010/06/15 06:00 [medline]
AID - 10.1021/ac902741r [doi]
PST - ppublish
SO  - Anal Chem. 2010 Mar 1;82(5):2006-12. doi: 10.1021/ac902741r.