Abstract
Fluorescence reaction of pyridoxal (PAL) and pyridoxal-5-phosphate (PALP) with hydrazides (Table I) was examined. PAL and PALP reacted with iso-butyric acid hydrazide (IBH) to form hydrazones in an acetate buffer solution which gave green fluorescence in an alkaline medium. IBH was suitably used for the determination of PAL and PALP. By the proposed method, 0.011.0 μg per ml of PAL hydrochloride and 0.11.0 μg per ml of PALP could be determined. Ten fold amount of pyridoxine hydrochloride and equal amount of pyridoxamine hydrochloride did not interfere.
Separatory determination of PAL and PALP has been done by the combination of the proposed method with an enzymatic method using acid phosphatase.
