High-Performance Liquid Chromatographic Determination of Hydrogen Peroxide with Peroxyoxalate Chemiluminescence Detection

A high performance liquid chromatographic method with peroxyoxalate chemiluminescence (PO-CL) detection was developed for the determination of hydrogen peroxide. Hydrogen peroxide separated by a reversed phase column was detected with the CL produced by the PO-CL reaction using a mixture of 2,4,6,8-tetrathiomorpholi-nopyrimido[5,4-d]pyrimidine and bis[2-(3,6,9-trioxade-cyloxycarbonyl)-4-nitrophenyl]oxalate or bis(2,4,6-trichlorophenyl)oxalate in acetonitrile as a post column reagent. Hydrogen peroxide could be determined over the range of 5–100 pmol on column and the detection limit was 188 fmol at a signal-to-noise ratio of 3. Relative standard deviation of 10-replicate measurements of peak heights for 50 pmol was 2.36%. The detection system applied to the assay of hydrogen peroxide in cola drinks. Trace amounts of hydrogen peroxide could be determined in six cola drinks. The levels were 68.4 ± 2.6–174.2 ± 8.4 nmol/dl. Hydrogen peroxide contaminated in commercially available hydroperoxides was also determined by the proposed method. The amounts obtained were 0.061% for t-butyl hydroper-oxide and 0.003% for cumene hydroperoxide.