Spectroscopic and chromatographic evidences of NADPH in human placental extract used as wound healer.

An aqueous extract of human placenta, which is used as wound healer, has been investigated in terms of fluorescence properties. When excited at 340 nm, it results in fluorescence emission having maxima around 436 nm, which is fairly specific for nicotinamide adenine dinucleotide, reduced form (NADH) and nicotinamide adenine dinucleotide phosphate, reduced form (NADPH). The excitation spectra, having emission at 440 nm, show patterns comparable to these nucleotides. Thin layer chromatography and reversed-phase (RP) HPLC confirm presence of only NADPH in the extract. The emission and excitation patterns of NADPH purified after HPLC resemble exactly with the reference compound. Its content has been estimated to be 0.018 +/- 0.003 mg/ml based on fluorescence emission with respect to a standard calibration curve (n=6). Biological functionality of NADPH in the extract has been confirmed by glutathione reductase assay (n=5).