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The photoluminescence-following electron-transfer (PFET) technique, developed in our laboratory, is a sensitive chromatographic detection method for oxidizable analytes. Because the oxidations are homogeneous, the technique avoids the problem of electrode fouling. A liquid-phase oxidant reacts with the electrochemically active analytes after separation, becoming capable of photoluminescence. Laser-induced photoluminescence is measured to quantitate the analytes. Thus, the electrochemical properties of the oxidant determine the detection selectivity, and the spectroscopic properties define the sensitivity. The properties of tris(2,2'-bipyridine)osmium (1) were investigated for use as the liquid-phase oxidant in the PFET system. The redox potential of the complex is less positive than that of tris(2,2'-bipyridine)ruthenium (2); thus, on-line generation of 1(3+) by reaction with PbO2, and selective oxidation of catechols by 1(3+), was possible. The mild oxidizing power of 1(3+) led to a lower background signal (compared to 2(3+)) when mixed with acidic mobile phases. Photoluminescence from 1(2+) was much weaker than that from 2(2+); nonetheless, the system achieved subnanomolar detection limits for dopamine, 3-methoxytyramine, and serotonin. Dopamine and 3-methoxytyramine in rat brain striatal dialysates were determined before and after the injection of nomifensine. The pH of the mobile phase can govern the detection selectivity, since oxidation of most organics is accompanied by proton transfer. Reaction of 1 with catechols showed pH-dependent sensitivity resulting from pH-dependent reaction rate changes. Since the reaction rate is also temperature dependent, increased temperature at the mixer resulted in higher sensitivity. However, the noise level also increased at elevated temperature; thus, the detection limit did not improve.

analytical chemistry

Use of Tris(2,2‘-bipyridine)osmium as a Photoluminescence-Following Electron-Transfer Reagent for Postcolumn Detection in Capillary High-Performance Liquid Chromatography