Premium
Discontinuous electrolyte systems for improved detection of biologically active amines and acids by capillary electrophoresis with laser‐induced native fluorescence detection
Author(s) -
Hsieh MingMu,
Chang HuanTsung
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200406123
Subject(s) - chemistry , capillary electrophoresis , chromatography , electrolyte , electrophoresis , tris , citric acid , propanoic acid , detection limit , acetic acid , capillary action , acetonitrile , laser induced fluorescence , fluorescence , analytical chemistry (journal) , electrode , biochemistry , materials science , physics , organic chemistry , quantum mechanics , composite material , stereochemistry
On‐line concentration and separation of biologically active amines and acids by capillary electrophoresis (CE) in conjunction with laser‐induced fluorescence using an Nd:YAG laser at 266 nm under discontinuous conditions is presented. The suitable conditions for simultaneous analysis of amines and acids were: samples were prepared in a solution (pH* 3.1) consisting of 10 m M citric acid, 89% acetonitrile (ACN), and water; a capillary was filled with 1.5 M Tris‐borate (TB) buffer (pH 10.0); and the anodic vial contained PTG10 buffer (pH* 9.0) that consists of 50 m M propanoic acid, Tris, 10% glycerol, and water. After injecting a large‐volume sample, amines and acids were separately stacked at the front (cathodic side) and back (anodic side) of the acidic sample zone, mainly because of changes in their electrophoretic mobilities as a result of changes in pH, viscosity, and electric field when high voltage was applied. When the sample was injected at 15 kV for 360 s, the concentration limits of detection (LODs) for 5‐hydroxytryptamine (5‐HT) and 5‐hydroxyindole‐3‐acetic acid (5‐HIAA) were 0.27 and 0.31 n M , respectively, which are about 400‐ and 800‐fold sensitivity improvements when compared to those injected at 1 kV for 10 s. For the analysis of amines, samples were prepared in 100 m M citric acid (pH* 1.8) containing 89% ACN and both the capillary and anodic vial were filled with 400 m M PTG20 (propanoic acid, Tris, 20% glycerol, and water) at pH* 4.5. Using a large injection volume (15 kV for 360 s), we achieved concentration LODs of 17 p M and 0.3 n M for tryptamine and epinephrine, which are about 5200‐ and 14 000‐fold sensitivity improvements, respectively, in comparison with those injected at 1 kV for 10 s. The features of simplicity (no sample pretreatment), rapidity (12 min), and sensitivity for identification of amines and acids of interest in urine samples show diagnostic potential of the two approaches developed in this study.