Development of adenosine sensor: effect of physiological buffers on activity and sensitivity in adenosine determinations by fast scan voltammetry.

Abstract

A new fast scan voltammetry (FSV) method was tested in the determinations of adenosine in physiological buffers at pH 7.4. The buffers can be used in the determinations of adenosine in vivo and include 7 x 10(-2) M phosphate, Krebs-Henseleit (K-H) and Hanks' Balanced Salts (HBSS). A new method of fabrication of carbon fiber electrodes (CFEs) by polishing, followed by electrochemical pretreatment (ECP) was developed for the determination. After the ECP of CFE, CFE background current was stable in FSV determinations even though an increase in the background current was observed after the ECP in the buffers at pH 7.4. The sensitivity in FSV determinations of adenosine at the pretreated electrodes was tested in the physiological buffers at the potential scan rate of 500 V s(-1) at pH 7.4. Buffer composition and pH was the same during the ECP of CFE and in the FSV determinations. The sensitivity in the FSV determinations of adenosine at the new CFEs was high, compared to that previously reported at CFEs prepared by other methods, and showed a limited dependence on buffer composition. However, a small increase in buffer pH above 7.4 resulted in a decrease in sensitivity in the determinations of adenosine. The decrease in sensitivity was associated with an additional increase in CFE background current at pH > 7.4. At pH 7.4 best sensitivity and limit of detection was obtained in 7 x 10(-2) M phosphate buffer, where the background current was lowest. The sensitivity was half that in K-H and HBSS. Standard deviation of measurements was ca. 1%. The results demonstrate the feasibility of sensitive FSV determinations of adenosine in physiological buffers at pH 7.4 at CFEs.