A SIMPLE AND RAPID MICROASSAY METHOD FOR ADENOSINE USING COMBINED EFFECTS OF ISOTOPE DILUTION AND ENZYMATIC CATALYSIS

Open Access
Publisher Website
Google Scholar
Abstract
To determine the applicability of this microassay method for measurement of tissue levels of adenosine [a coronary vasodilator], adenosine contents in normal and ischemic hearts from Wistar rats were measured. Under light ethylether anesthesia, intra-tracheal intubation was carried out and the chest opened after initiation of artificial respiration. Following application of artificial respiration for 30 min the control heart was removed and immediately frozen with dry ice ethanol. The ischemic heart was obtained by arresting artificial respiration for 5 min, after which the heart was excised and frozen. After extraction using 0.5 M perchloric acid, the extract was neutralized with KOH solution, lyophilized in vacuo and used for determination of adenosine. Adenosine content in the ischemic heart showed a significant increase as compared with that in controls; this level of increase was essentially in good parallel with that reported elsewhere. An advantage of this method is that commercially available adenosine deaminase [E.C. 3.5.4.4] can be used directly for the assay. This microassay method for adenosine is relatively simple, quite sensitive and reproducible, and should be useful for determining the content of adenosine in small amounts of tissue.