PMID- 18692851
OWN - NLM
STAT- MEDLINE
DCOM- 20081114
LR  - 20131121
IS  - 0021-9673 (Print)
IS  - 0021-9673 (Linking)
VI  - 1206
IP  - 1
DP  - 2008 Oct 3
TI  - High-throughput assay of nitric oxide metabolites in human plasma without 
      deproteinization by lab-on-a-chip electrophoresis using a zwitterionic additive.
PG  - 41-4
LID - 10.1016/j.chroma.2008.07.065 [doi]
AB  - In order to develop a high-throughput assay for nitric oxide metabolites, nitrite 
      (NO2-) and nitrate (NO3-), in biological fluids, we have investigated the 
      simultaneous determination of them using an electrophoretic lab-on-a-chip 
      (microchip capillary electrophoresis, MCE) technique. In this study, in order to 
      establish an MCE assay process without deproteinization, the addition of a 
      zwitterionic additive into the running buffer to reduce the adsorption of protein 
      onto the surface of channel was investigated. Initially, some zwitterionic 
      additives were investigated by making a comparison of relative standard 
      deviations (RSDs) of the migration times for NO2(-) and NO3(-) on capillary 
      electrophoresis. From the results of our comparison of the RSD values, 2% (w/w) 
      N-cyclohexyl-2-aminoethanesulfonic acid (CHES) was selected. As a result of the 
      application of the running buffer with CHES to the MCE process, the complete 
      separation of NO2(-) and NO3(-) in human plasma without deproteinization was 
      achieved within 1 min. Since the RSD values of the positions of the peaks were 
      less than 2.3%, beneficial reduction effects on MCE were suggested. When we used 
      an internal standard method in order to correct the injection volume, the RSDs of 
      the peak heights and areas were less than 10%, and the correlation coefficients 
      of spiked calibration curves ranging from 0 to 350 microM were 0.999 and 0.997 
      for NO2(-) and NO3(-), respectively. The limits of detection (S/N=3) were 53 
      microM for NO2(-) and 41 microM for NO3(-). Moreover, the correlation 
      coefficients in excess of 0.99 between the MCE method and a conventional Griess 
      method were achieved for both NO2(-) and NO3(-). Consequently, the possibility of 
      establishing a high-throughput assay process was obtained by utilizing 2% (w/w) 
      CHES to reduce protein adsorption.
FAU - Miyado, Takashi
AU  - Miyado T
AD  - Health Technology Research Center (HTRC), National Institute of Advanced 
      Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 
      563-8577, Japan.
FAU - Wakida, Shin-ichi
AU  - Wakida S
FAU - Aizawa, Hitoshi
AU  - Aizawa H
FAU - Shibutani, Yasuhiko
AU  - Shibutani Y
FAU - Kanie, Tomohiko
AU  - Kanie T
FAU - Katayama, Makoto
AU  - Katayama M
FAU - Nose, Kazutoshi
AU  - Nose K
FAU - Shimouchi, Akito
AU  - Shimouchi A
LA  - eng
PT  - Journal Article
DEP - 20080726
PL  - Netherlands
TA  - J Chromatogr A
JT  - Journal of chromatography. A
JID - 9318488
RN  - 0 (Nitrates)
RN  - 0 (Nitrites)
RN  - 103-47-9 (2-(N-cyclohexylamino)ethanesulfonic acid)
RN  - 1EQV5MLY3D (Taurine)
RN  - 31C4KY9ESH (Nitric Oxide)
SB  - IM
MH  - Electrophoresis, Microchip/*methods
MH  - Humans
MH  - Microchip Analytical Procedures/*methods
MH  - Nitrates/*blood
MH  - Nitric Oxide/*metabolism
MH  - Nitrites/*blood
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - Taurine/analogs & derivatives
EDAT- 2008/08/12 09:00
MHDA- 2008/11/15 09:00
CRDT- 2008/08/12 09:00
PHST- 2008/04/14 00:00 [received]
PHST- 2008/07/18 00:00 [revised]
PHST- 2008/07/22 00:00 [accepted]
PHST- 2008/08/12 09:00 [pubmed]
PHST- 2008/11/15 09:00 [medline]
PHST- 2008/08/12 09:00 [entrez]
AID - S0021-9673(08)01265-X [pii]
AID - 10.1016/j.chroma.2008.07.065 [doi]
PST - ppublish
SO  - J Chromatogr A. 2008 Oct 3;1206(1):41-4. doi: 10.1016/j.chroma.2008.07.065. Epub 
      2008 Jul 26.