Determination of free intracellular amino acids in single mouse peritoneal macrophages after naphthalene-2,3-dicarboxaldehyde derivatization by capillary zone electrophoresis with electrochemical detection.
A method is described for the direct identification and quantification of amino acids in individual mouse peritoneal macrophages by capillary zone electrophoresis with electrochemical detection after on-column derivatization with naphthalene-2,3-dicarboxaldehyde (NDA) and CN-. In this method, individual macrophages and then the lysing/ derivatizing buffer are injected into the front end of the separation capillary by electromigration with the aid of an inverted microscope. The front end of the separation capillary is used as a chamber to lyse the macrophage and derivatize its contents, which minimizes dilution of amino acids of a single macrophage during derivatization. Six amino acids (serine, alanine, taurine, glycine, glutamic acid, and aspartic acid) in single mouse peritoneal macrophages have been identified. Quantitation has been accomplished through the use of calibration curves, where the concentration ratios of these standard amino acids are similar to the concentration ratios of amino acids in macrophages. Cellular levels of the amino acids in these cells range from 0.27 +/- 0.20 fmol/ cell for alanine to 6.4 +/- 4.6 fmol/cell for taurine.
Weng
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Q
Laboratory of Analytical Science, School of Chemistry, Shandong University, Jinan, China.
Jin
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W
D000409:Alanine / Q000032:analysis; D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D001224:Aspartic Acid / Q000032:analysis; D002457:Cell Extracts / Q000737:chemistry; D004563:Electrochemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D018698:Glutamic Acid / Q000032:analysis; D005998:Glycine / Q000032:analysis; D007424:Intracellular Fluid / Q000737:chemistry; D017737:Macrophages, Peritoneal / Q000737:chemistry* / Q000166:cytology; D051379:Mice; D009281:Naphthalenes / Q000737:chemistry*; D012694:Serine / Q000032:analysis; D013654:Taurine / Q000032:analysis
D000596:Amino Acids; D002457:Cell Extracts; D009281:Naphthalenes; D013654:Taurine; D001224:Aspartic Acid; D018698:Glutamic Acid; D012694:Serine; C027717:2,3-naphthalenedicarboxaldehyde; D000409:Alanine; D005998:Glycine