4.5 Article

Quantification of carnosine-related peptides by microchip electrophoresis with chemiluminescence detection

Journal

ANALYTICAL BIOCHEMISTRY
Volume 393, Issue 1, Pages 105-110

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2009.06.012

Keywords

Microchip electrophoresis; Chemiluminescence detection; Peptide analysis; N-(4-Aminobutyl)-N-ethylisoluminol; Carnosine-related peptides; Human cerebrospinal fluid; Canine plasma

Funding

  1. National Natural Science Foundation of China [20665002, 20875019]
  2. U.S. National Institutes of Health [S06GM08047]

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A microchip electrophoresis (MCE) method with chemiluminescence (CL) detection was developed for the determination of carnosine-related peptides, including carnosine, homocarnosine, and anserine, in biological samples. A simple integrated MCE-CL system was built to perform the assays. The highly sensitive CL detection was achieved by means of the CL reaction between hydrogen peroxide and N-(4-aminobutyl)-N-ethylisoluminol-tagged peptides in the presence of adenine as a CL enhancer and Co2+ as a catalyst. Experimental conditions for analyte labeling, MCE separation, and CL detection were studied. MCE separation of the above-mentioned three peptides took less than 120 s. Detection limits (signal/noise ratio [S/N] = 3) of 3.0 x 10(-8), 2.8 x 10(-8), and 3.4 x 10(-8) M were obtained for carnosine, anserine, and homocarnosine, respectively. The current MCE-CL method was applied for the determination of carnosine, anserine, and homocarnosine in human cerebrospinal fluid (CSF) and canine plasma. Homocarnosine was detected at the micromolar (mu M) level in the CSF samples analyzed, whereas the levels of carnosine and anserine in these samples were below the detection limit of the assay. Interestingly, both carnosine and anserine were detected in the canine plasma samples, whereas homocarnosine was not. (C) 2009 Elsevier Inc. All rights reserved.

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