Rapid radioassay for metabolites of adenosine and deoxyadenosine in erythrocytes

• Published in: Journal of chromatography. B, Biomedical applications Vol. 674; no. 1; pp. 132 - 137
• Main Authors: Szabados, Eve, Christopherson, Richard I.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-12-1995
• Subjects: Adenosine - blood; Adenosine - metabolism; Blood Chemical Analysis - methods; Cellulose - analogs & derivatives; Chromatography, Thin Layer; Deoxyadenosines - blood; Deoxyadenosines - metabolism; Dibutyl Phthalate; Erythrocytes - chemistry; Erythrocytes - metabolism; Humans; Kinetics; Perchlorates; Polyethyleneimine - analogs & derivatives
• ISSN: 0378-4347; 1572-6495
• DOI: 10.1016/0378-4347(95)00288-X

A radioassay has been developed to quantify the uptake and initial metabolism of adenosine (Ado) or deoxyadenosine (dAdo) by human erythrocytes. Cell suspension and [
3H]Ado are mixed at 3-s intervals with a novel dual-syringe apparatus, and uptake and metabolism of Ado is stopped by centrifuging the cells through a dibutylphthalate layer into perchloric acid. The neutralized cell extract is analyzed by two-dimensional chromatography on poly(ethyleneimine)-cellulose plates by two procedures using combinations of solvents optimised for the separation of nucleosides and nucleobases, and for nucleotides derived from the exogenous [
3H]Ado.