Abstract
4,5-Diaminophthalhydrazide (DPH) was found to be a highly sensitive chemiluminescence derivatization reagent for α-dicarbonyl compounds in high-performance liquid chromatography. The reagent reacts with α-dicarbonyl compounds in dilute hydrochloric acid in the presence of β-mercaptoethanol to give highly chemiluminescent quinoxaline derivatives which produce chemiluminescence by reaction with hydrogen peroxide and potassium hexacyanoferrate(III). The DPH derivatives of five α-dicarbonyl compounds were separated on a reversed-phase column, TSKgel ODS-120T, with a mixture of acetonitrile and 10 m M ammonium acetate, followed by chemiluminescence detection. The detection limits are in the range 1.1–300 fmol for a 20-μl injection volume (signal-to-noise ratio 3).
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