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@article{586321,
author = {Vrábel, Patrik and Táborský, Petr and Ryvolová, Markéta and Havel, Josef and Preisler, Jan},
article_location = {Holland},
article_number = {2},
keywords = {Electrophoresis; Fluorescence; Derivatization; RBITC; Proteins; Rhodamine; 532 nm Laser.},
language = {eng},
issn = {0022-2313},
journal = {Journal of Luminescence},
title = {Sensitive Detection and Separation of Fluorescent Derivatives using Capillary Electrophoresis with Laser-Induced Fluorescence Detection with 532 nm Nd:YAG Laser},
url = {http://www.elsevier.com/wps/find/journaldescription.cws_home/505700/description?navopenmenu=-2},
volume = {118},
year = {2006}
}
TY - JOUR
ID - 586321
AU - Vrábel, Patrik - Táborský, Petr - Ryvolová, Markéta - Havel, Josef - Preisler, Jan
PY - 2006
TI - Sensitive Detection and Separation of Fluorescent Derivatives using Capillary Electrophoresis with Laser-Induced Fluorescence Detection with 532 nm Nd:YAG Laser
JF - Journal of Luminescence
VL - 118
IS - 2
SP - 283-292
EP - 283-292
PB - Elsevier
SN - 00222313
KW - Electrophoresis
KW - Fluorescence
KW - Derivatization
KW - RBITC
KW - Proteins
KW - Rhodamine
KW - 532 nm Laser.
UR - http://www.elsevier.com/wps/find/journaldescription.cws_home/505700/description?navopenmenu=-2
N2 - Capillary Electrophoresis with Laser Induced Fluorescence detection (CELIF) is a powerful tool for separation and sensitive determination of fluorescent compounds. Biologically active compounds, such as amino acids, peptides and proteins may exhibit native fluorescence, which is however often low and/or an expensive laser is required for excitation in UV. Therefore, labeling of the analytes with a fluorescent dye is usually necessary. In this work, home-built CELIF instrument was developed and tuned up using rhodamine-based dyes with diode pumped frequency-doubled continuous wave Nd:YAG excitation laser with feedback power regulation (532 nm). First, the suitability and performance of this type of laser for LIF detection in a separation method was evaluated. Detection limit of 2E-13 M was achieved with rhodamine B, which is comparable to previously described CELIF with Ar+ laser. An exemplary separation of rhodamine dyes is given. Rhodamine based derivatizing agent was chosen for labeling of aminoacids and proteins with derivatization techniques suggested by several groups for fluorescein-based dyes. Derivatization conditions such as solvent, pH, time and temperature of derivatization as well as separation conditions were optimized and examples of electropherograms are presented.
ER -
VRÁBEL, Patrik, Petr TÁBORSKÝ, Markéta RYVOLOVÁ, Josef HAVEL and Jan PREISLER. Sensitive Detection and Separation of Fluorescent Derivatives using Capillary Electrophoresis with Laser-Induced Fluorescence Detection with 532 nm Nd:YAG Laser. \textit{Journal of Luminescence}. Holland: Elsevier, 2006, vol.~118, No~2, p.~283-292. ISSN~0022-2313.
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