HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF CYCLIC 3',5'-AMP WITH FLUORESCENCE DETECTION - VASOACTIVE INTESTINAL PEPTIDE-INDUCED MODIFICATION OF ITS CONCENTRATION IN NEUROBLASTOMA-CELLS

The efficiency of ion-pair reversed-phase HPLC on a Vydac C-18 column with 50 mM ammonium acetate (pH 4.75)-methanol-acetonitrile (88:9:3, v /v/v) as the mobile phase with isocratic separation and fluorescence d etection for the determination of cAMP in cellular extracts was evalua ted. This method was compared with a radioimmunoassay technique in ter ms of Linearity, reproducibility and sensitivity, No interactions with other nucleotides such as AMP, ADP, ATP and cGMP were observed. Appli cation to the measurement of cAMP modifications was studied in a neuro blastoma cell line: LA-N-2 cells stimulated by a neuropeptide, vasoact ive intestinal peptide.