PMID- 19206207
OWN - NLM
STAT- MEDLINE
DCOM- 20100901
LR  - 20131121
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 81
IP  - 6
DP  - 2009 Mar 15
TI  - Simultaneous determination of superoxide and hydrogen peroxide in macrophage RAW 
      264.7 cell extracts by microchip electrophoresis with laser-induced fluorescence 
      detection.
PG  - 2193-8
LID - 10.1021/ac801777c [doi]
AB  - A method for the first time to simultaneously determine superoxide and hydrogen 
      peroxide in macrophage RAW 264.7 cell extracts by microchip electrophoresis with 
      laser-induced fluorescence detection (MCE-LIF) was developed. 
      2-Chloro-1,3-dibenzothiazolinecyclohexene (DBZTC) and 
      bis(p-methylbenzenesulfonyl) dichlorofluorescein (FS), two probes that can be 
      specifically derivatized by superoxide and hydrogen peroxide, respectively, were 
      synthesized and used. Parameters influencing the derivatization and on-chip 
      separation were optimized. With the use of a HEPES (20 mM, pH 7.4) running 
      buffer, a 50 mm long separation channel, and a separation voltage of 1800 V, 
      baseline separation was achieved within 48 s for the two derivatization products, 
      DBZTC-oxide (DBO) and 2,7-dichlorofluorescein (DCF). The linearity ranges of the 
      method were 0.08-5.0 and 0.02-5.0 microM with detection limits (signal-to-noise 
      ratio = 3) of 10 nM (1.36 amol) and 5.6 nM (0.76 amol) for superoxide and 
      hydrogen peroxide, respectively. The relative standard deviations (RSDs) of 
      migration time and peak area were less than 2.0% and 5.0%, respectively. The 
      recoveries of the cell extract samples spiked with 1.0 microM standard solutions 
      were 96.1% and 93.0% for superoxide and hydrogen peroxide, respectively. With the 
      use of this method, superoxide and hydrogen peroxide in phorbol myristate acetate 
      (PMA)-stimulated macrophage RAW 264.7 cell extracts were found to be 0.78 and 
      1.14 microM, respectively. The method has paved a way for simultaneously 
      determining two or more reactive oxygen species (ROS) in a biological system with 
      high resolution.
FAU - Li, Hongmin
AU  - Li H
AD  - College of Chemistry, Chemical Engineering and Materials Science, Engineering 
      Research Center of Pesticide and Medicine Intermediate Clean Production, Ministry 
      of Education, Key Laboratory of Molecular and Nano Probes, Ministry of Education, 
      Shandong Normal University, Jinan 250014, People's Republic of China.
FAU - Li, Qingling
AU  - Li Q
FAU - Wang, Xu
AU  - Wang X
FAU - Xu, Kehua
AU  - Xu K
FAU - Chen, Zhenzhen
AU  - Chen Z
FAU - Gong, Xiaocong
AU  - Gong X
FAU - Liu, Xin
AU  - Liu X
FAU - Tong, Lili
AU  - Tong L
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  - 0 (Fluoresceins)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Reactive Oxygen Species)
RN  - 0 (bis(p-methylbenzenesulfonyl)dichlorofluorescein)
RN  - 11062-77-4 (Superoxides)
RN  - BBX060AN9V (Hydrogen Peroxide)
SB  - IM
MH  - Animals
MH  - Cell Line, Tumor
MH  - Electrophoresis, Microchip/*methods
MH  - Fluoresceins/chemistry
MH  - Fluorescent Dyes/*chemistry
MH  - Hydrogen Peroxide/*analysis
MH  - *Lasers
MH  - Macrophages/metabolism
MH  - Mice
MH  - Reactive Oxygen Species/metabolism
MH  - Superoxides/*analysis
EDAT- 2009/02/12 09:00
MHDA- 2010/09/02 06:00
CRDT- 2009/02/12 09:00
PHST- 2009/02/12 09:00 [entrez]
PHST- 2009/02/12 09:00 [pubmed]
PHST- 2010/09/02 06:00 [medline]
AID - 10.1021/ac801777c [pii]
AID - 10.1021/ac801777c [doi]
PST - ppublish
SO  - Anal Chem. 2009 Mar 15;81(6):2193-8. doi: 10.1021/ac801777c.