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Sensitive detection of near-infrared fluorescent dyes using high-performance liquid chromatography with peroxyoxalate
K Kimoto1, R Gohda, K Murayama
1Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Abstract:
The highly sensitive detection of four near-infrared (near-IR) fluorescent dyes, methylene blue, pyridine 1, oxazine 1 and 3,3'-diethylthiadicarbocyanine iodide (DTDCI) by high-performance liquid chromatography (HPLC) with post-column peroxyoxalate chemiluminescence (PO-CL) detection utilizing bis(4-nitro-2-(3, 6,9-trioxadecyloxycarbonyl)phenyl) oxalate (TDPO) and hydrogen peroxide was examined. The detection limits for methylene blue, pyridine 1, oxazine 1 and DTDCI were 120, 27, 31, 0.19 fmol on-column at a signal-to-noise ratio of 2, respectively. The sensitivity for DTDCI was 250 times that obtained by HPLC with the conventional fluorescent detection.