Optimization of the Reaction Conditions for the Peroxyoxalate Chemiluminescence Detection System of Fluorescent Compounds in a High-Performance Liquid Chromatography

Abstract

Using an instrument of automatic fluid injection (batch method), we optimized such reaction conditions as pH, concentration and kinds of buffer for the peroxyoxalate chemiluminscence (PO-CL) detection of fluorescent compounds for high-performance liquid chromatography (HPLC). The results were compared with those obtained by HPLC. Consequently, the usefulness of the batch method for investigating the reaction conditions for PO-CL detection in HPLC was demonstrated. With the optimized condition using a phosphate buffer for the eluent in HPLC, the detection limits for Dns-Val and dipyridamole were 70 amol and 20 amol on column (S/N=2), respectively.

Determination of C-21 Ketosteroids in Serum Using Trifluoromethanesulfonic Acid Catalyzed Precolumn Dansylation and 1,1‘-Oxalyldiimidazole Postcolumn Peroxyoxalate Chemiluminescence Detection

“…Although the imidazole buffer yielded the highest chemiluminescence intensity, the signal-to-noise ratio was better with phosphate buffer. This has also been shown by Gohda et al and is explained by the high background generated by imidazole 3 Time-resolved chemiluminescence emission profiles recorded in the stopped-flow system.…”

HighlightTrends in Chemiluminescence Detection for Liquid Separations

“…The efficacy of this approach has been demonstrated with the peroxyoxalate chemiluminescence (PO-CL) reaction as applied to the determination of fluorescent compounds. 4 The effects of pH and phosphate buffer concentration on the CL emission intensity of a fluorescent drug, dipyridamole (an anti-platelet aggregation drug), and dansylated valine (Dns-Val) were investigated using a batch luminometer and the optimum conditions were then used for PO-CL detection following LC separation on a TSKgel ODS-80TM column (4.6 mm id 3 150 mm, 5 mm) with a 60/40 buffer/acetonitrile mobile phase. Detection limits for Dns-Val and dipyridamole of 70 amol and 20 amol on column, respectively, were achieved.…”

Capillary Electrophoresis With a Chemiluminescence Detector Using the Two Reactions of Luminol and Peroxyoxalate

“…[8][9][10] Synthetic compounds that induce CL, including luminol, peroxyoxalate ester, ruthenium complex, lophine derivatives, acridinium ester, and adamantyldioxetane compounds, are well known in the field of analytical chemistry, and many applications have been reported. [11,12] The CL emitted by luminol is remarkably enhanced by catalysts, such as metal ions. This has been used for the determination of small amounts of metal ions and their complexes.…”