caa a22 4500 445802006 CHVBK 20180317145144.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s10337-010-1841-1 doi (NATIONALLICENCE)springer-10.1007/s10337-010-1841-1 Selective and Sensitive Determination of Pheomelanin in Biological Samples Using MEKC with Laser-Induced Fluorescence Detection Based on Intramolecular Excimer-Forming Fluorescence Derivatization [Elektronische Daten] [Xiaoling Zhang, Bin Yuan, Qiao Yang] A highly selective micellar electrokinetic capillary chromatography (MEKC) method with laser-induced fluorescence (LIF) detection for the sensitive determination of pheomelanin in diverse biological materials was originally described. The derivatization reagent, 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), allowed for the selective detection of the two aminohydroxyphenylalanines (AHPs) markers for pheomelanin monitored at 500nm. Multiple labeling of two AHPs with PSE allowed the formation of intramolecular excimers that emit at longer wavelengths (500nm) than the mono-labeled analytes (360-420nm) based on intramolecular excimer-forming fluorescence derivatization. Optimal separation of the labeled polyamines was achieved using a separation buffer consisting of 20mM phosphate pH 7.4, 30mM cholate, and 30% methanol. Using these conditions, the two AHPs were separated within 12min, and the relative standard deviations (RSDs) were less than 1.5 and 1.6% (intra-run), 3.8 and 4.6% (inter-run, for a 6-day period) for the migration times and peak areas (n=10), respectively. This method was successfully applied to the monitoring of pheomelanin in diverse biological samples with the spiked recoveries in the range of 94-101%. At a signal-to-noise ratio of 3, the detection limit for AHPs in the real samples was 31pM for 3-AHP and 35pM for 4-AHP, respectively, which are superior to those previously reported in the literature using fluorescence detection. Springer-Verlag, 2011 Micellar electrokinetic capillary chromatography nationallicence Pheomelanin nationallicence Aminohydroxyphenylalanine nationallicence Derivatization nationallicence 4-(1-Pyrene)butyric acid N -hydroxysuccinimide ester nationallicence Zhang Xiaoling East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 200090, Shanghai, China aut Yuan Bin Department of Biochemistry and Molecular Biology, Second Military Medical University, 200433, Shanghai, China aut Yang Qiao East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 200090, Shanghai, China aut Chromatographia Springer-Verlag 73/5-6(2011-03-01), 527-534 0009-5893 73:5-6<527 2011 73 10337 https://doi.org/10.1007/s10337-010-1841-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10337-010-1841-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Xiaoling East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 200090, Shanghai, China aut NATIONALLICENCE 700 1- Yuan Bin Department of Biochemistry and Molecular Biology, Second Military Medical University, 200433, Shanghai, China aut NATIONALLICENCE 700 1- Yang Qiao East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 200090, Shanghai, China aut NATIONALLICENCE 773 0- Chromatographia Springer-Verlag 73/5-6(2011-03-01), 527-534 0009-5893 73:5-6<527 2011 73 10337 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer