Fluorimetric assay for ornithine decarboxylase by high-performance liquid chromatography.
A highly sensitive method for the assay of ornithine decarboxylase in sample solutions prepared from rat tissue homogenate is described which employs high-performance liquid chromatography with fluorescence detection. Putrescine formed from ornithine under the optimal conditions for the enzyme reaction is treated by Cellex P column chromatography for clean-up and converted into the fluorescamine derivative in the presence of cupric ion which inhibits the reaction of interfering amines with fluorescamine. The derivative is separated by reversed-phase chromatography on LiChrosorb RP-18 with linear gradient elution. The lower limit of detection for putrescine formed enzymatically is 5 pmol.
D000818:Animals; D002262:Carboxy-Lyases / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005753:Gastric Mucosa / Q000201:enzymology; D009955:Ornithine Decarboxylase / Q000302:isolation & purification*; D011700:Putrescine; D051381:Rats
D002262:Carboxy-Lyases; D009955:Ornithine Decarboxylase; D011700:Putrescine