Determination of free intracellular amino acids in single mouse peritonealmacrophages after naphthalene-2,3-dicarboxaldehyde derivatization by capillary zone electrophoresis with electrochemical detection

Weng, QF Jin, WR

Qf. Weng et Wr. Jin, Determination of free intracellular amino acids in single mouse peritonealmacrophages after naphthalene-2,3-dicarboxaldehyde derivatization by capillary zone electrophoresis with electrochemical detection, ELECTROPHOR, 22(13), 2001, pp. 2797-2803

21

INGLESE

Article

Chemistry & Analysis

ELECTROPHORESIS

0173-0835 → ACNP

22

13

2001

2797 - 2803

ISI

0173-0835(200108)22:13<2797:DOFIAA>2.0.ZU;2-U

A method is described for the direct identification and quantification of a mino acids in individual mouse peritoneal macrophages by capillary zone ele ctrophoresis with electrochemical detection after on-column derivatization with naphthalene-2,3-dicarboxaldehyde (NDA) and CN-. In this method, indivi dual 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 use d as a chamber to lyse the macrophage and derivatize its contents, which mi nimizes dilution of amino acids of a single macrophage during derivatizatio n. Six amino acids (serine, alanine, taurine, glycine, glutamic acid, and a spartic acid) in single mouse peritoneal macrophages have been identified. Quantitation has been accomplished through the use of calibration curves, w here 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 alanin e to 6.4 +/-4.6 fmol/cell for taurine.