Abstract
The efficiency of ion-pair reversed-phase HPLC on a Vydac C 18 column with 50 m M ammonium acetate (pH 4.75)—methanol—acetonitrile (88:9:3, v/v/v) as the mobile phase with isocratic separation and fluorescence detection for the determination of cAMP in cellular extracts was evaluated. This method was compared with a radioimmunoassay technique in terms of linearity, reproducibility and sensitivity. No interactions with other nucleotides such as AMP, ADP, ATP and cGMP were observed. Application to the measurement of cAMP modifications was studied in a neuroblastoma cell line: LA-N-2 cells stimulated by a neuropeptide, vasoactive intestinal peptide.
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