Flow injection measurements of S-nitrosothiols species in biological samples using amperometric nitric oxide sensor...

A novel flow injection analysis (FIA) system suitable for measurement of S-nitrosothiols (RSNOs) in blood plasma is described. In the proposed (FIA) system, samples and standards containing RSNO species are injected into a buffer carrier stream that is mixed with the reagent stream containing 3,3'-dipropionicdiselenide (SeDPA) and glutathione (GSH). SeDPA has been shown previously to catalytically decompose RSNOs in the presence of a reducing agent, such as GSH, to produce nitric oxide (NO). The liberated NO is then detected downstream by an amperometric NO sensor. This sensor is prepared using an electropolymerized m-phenylenediamine (m-PD)/resorcinol and Nafion composite films at the surface of a platinum electrode. Using optimized flow rates and reagent concentrations, detection of various RSNOs at levels in the range of 0.25-20 microM is possible. For plasma samples, detection of background sensor interference levels within the samples must first be carried out using an identical FIA arrangement, but without the added SeDPA and GSH reagents. Subtraction of this background sensor current response allows good analytical recovery of RSNOs spiked into animal plasma samples, with recoveries in the range of 90.4-101.0%.