Extraction, purification and identification of cytidine 3',5'-cyclic monophosphate from rat tissues.
The large-scale extraction and purification to homogeneity of cyclic CMP and its unequivocal identification are described. Rat liver, kidney, heart, spleen and lung tissues were subjected to a sequential purification procedure involving freeze-clamping, perchlorate extraction, alumina and boronate column chromatography, polyacrylamide-gel column electrophoresis and high-voltage paper electrophoresis. The purified sample co-chromatographed with authentic cyclic CMP on t.l.c. and high-pressure liquid chromatography and was positive in a cyclic CMP radio-immunoassay. The u.v., i.r. and p.m.r. spectra were each essentially identical with those of authentic cyclic CMP. Fast-atom bombardment of authentic cyclic CMP yielded a mass spectrum containing a molecular protonated ion: mass-ion-kinetic-energy scanning of this ion produced a spectrum unique to 3',5'-cyclic CMP. The extracted nucleotide produced an identical mass-ion-kinetic-energy spectrum.
D000818:Animals; D002845:Chromatography; D003563:Cyclic CMP / Q000302:isolation & purification; D003597:Cytosine Nucleotides / Q000302:isolation & purification; D004590:Electrophoresis, Paper; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D051381:Rats; D011919:Rats, Inbred Strains; D013053:Spectrophotometry
D003597:Cytosine Nucleotides; D003563:Cyclic CMP