Microchip electrophoresis with chemiluminescence detection for assaying ascorbic acid and amino acids in single cells.
A method based on microchip electrophoresis (MCE) with chemiluminescence (CL) detection was developed for the determination of ascorbic acid (AA) and amino acids including tryptophan (Trp), glycine (Gly) and alanine (Ala) present in single cells. Cell injection, loading, lysing, electrophoretic separation and CL detection were integrated onto a simple cross microfluidic chip. A single cell was loaded in the cross intersection by electrophoretic means through applying a set of potentials at the reservoirs. The docked cell was lysed rapidly under a direct electric field. The intracellular contents were MCE separated within 130 s. CL detection was based on the enhancing effects of AA and amino acids on the CL reaction of luminol with K(3)[Fe(CN)(6)]. Rat hepatocytes were prepared and analyzed as the test cellular model. The average intracellular contents of AA, Trp, Gly and Ala in single rat hepatocytes were found to be 38.3, 5.15, 3.78 and 3.84 fmol (n=12), respectively.
Zhao
Shulin
S
College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 51004, China.
Huang
Yong
Y
Liu
Yi-Ming
YM
D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D001205:Ascorbic Acid / Q000032:analysis*; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D022781:Hepatocytes / Q000737:chemistry*; D008163:Luminescent Measurements / Q000379:methods*; D051381:Rats
D000596:Amino Acids; D001205:Ascorbic Acid
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