Development of an HPLC diode array detector method for the determination of human plasma adenosine concentrations

Assays of adenosine and its derivatives in biological fluids involve fluori metric, radioimmunological or chromatographic analyses. Techniques currentl y used are tedious because they involve either an extraction using immunolo gical methods or one or two freeze drying cycles when using high performanc e liquid chromatography (HPLC). In this context, we describe a "quick" HPLC method using a diode array detector for the spectral analysis and quantita tion of adenosine and its derivatives in less than two hours following samp ling. We compared our results to those obtained with an HPLC technique coup led to UV detection. Although there is a good correlation between the two techniques, the "quick " method provide values that are on average 30% higher than those obtained with the classical method. The absence of a lyophilization step in the clas sical method could explain this difference. The utilization of an HPLC syst em coupled to a diode array detector leads to a rapid and reliable assay of adenosine and its derivatives. The possibility of having a rapid assay met hod available could be very useful in cases of certain cardiovascular or ne urological disorders.