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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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