Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1217, Issue 36, Pages 5732-5736Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2010.07.012
Keywords
Microchip electrophoresis; Chemiluminescence detection; Single cell analysis; Sulphydryl compounds
Funding
- National Natural Science Foundations of China (NSFC) [20875019]
- US National Institutes of Health [SC1 GM089557]
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An analytical method based on microchip electrophoresis (MCE) and chemiluminescence detection (CL) was developed for the determination of intracellular sulphydryl compounds. Cell injection/loading, cytolysis, electrophoretic separation, and CL detection were integrated onto a simple cross-microfluidic chip. Selective CL detection of sulphydryl compounds was achieved by deploying the luminol-Na(2)S(2)O(8) reaction. Under the CL conditions selected, many endogenous compounds in biological systems such as amino acids, biogenic amines, peptides and proteins did not produce any CL signal, which further ensured a high selectivity of the proposed MCE-CL assays. Sulphydryl compounds including cysteine (Cys), glutathione (GSH), and hemoglobin (Hb) were selected as the test compounds. The MCE separation was completed within 120 s. The detection limits were estimated to be 7 amol for Cys, 32 amol for GSH and 69 amol for Hb, respectively. The present method was applied to analyze individual red blood cells collected from both healthy subjects and cancer patients. It was found that the average intracellular contents of Cys, GSH and Hb were in the ranges of 26-43 amol/cell,128-323 amol/cell and 522-667 amol/cell, respectively for cancer patients, compared to 579-609 amol Hb/cell and not detectable Cys and GSH for healthy subjects. (C) 2010 Elsevier B.V. All rights reserved.
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