Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1216, Issue 39, Pages 6746-6751Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2009.08.008
Keywords
Microchip electrophoresis; Chemiluminescence detection; Single cell analysis; Ascorbic acid; Tryptophan; Rat hepatocytes
Funding
- National Natural Science Foundations of China (NSFC) [20665002, 20875019]
- US National Institutes of Health [S06GM08047]
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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. (C) 2009 Elsevier B.V. All rights reserved.
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