Rapid automated high-performance liquid chromatographic analysis of cyclic adenosine 3′,5′-monophosphate : Synthesis in brain tissues

• Published in: Journal of Chromatography A 1985-01, Vol.341 (1), p.43-51
• Main Authors: Lin, Lillian, Saller, Charles F., Salama, Andre I.
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Nucleic acids; Nucleic bases, nucleotides
• ISSN: 0378-4347; 0021-9673
• DOI: 10.1016/S0378-4347(00)84008-6

A rapid and sensitive automated system for measuring cyclic adenosine 3′,5′-monophosphate (cAMP) synthesized from either radiolabelled adenine or adenosine 5′-triphosphate (ATP) in intact and broken cell tissue preparations, respectively, is described. After incubation with radiolabelled precursor, tissue samples are deproteinized and then injected directly onto a reversed-phase high-performance liquid chromatographic column. The column effluent fraction which contains cAMP is collected into scintillation vials and assayed for tritium by liquid scintillation spectrometry. Since the high-performance liquid chromatographic and fraction collection procedures are automated, over fifty samples can be analyzed in duplicate in a single day. The utility of this assay is illustrated by investigations of the effects of β-adrenergic receptor stimulation on cAMP synthesis in tissue slices prepared from rat cerebral cortex and dopamine on cAMP synthesis in striatal membrane preparations.