Spectrophotometric determination of acadesine (AICA-riboside) in plasma using a diazotization coupling technique with N-(1-naphthyl)ethylenediamine.

A simple spectrophotometric assay has been developed for determining concentrations of the new cardioprotective agent, acadesine (AICA-riboside), in human plasma. The method is adapted from the Bratton-Marshall (BMA) procedure for detection of primary aromatic amines. The assay was developed to measure concentrations of the drug in plasma during i.v. infusion to patients undergoing coronary artery bypass graft (CABG) surgery. The limit of quantitation of the assay is 0.25 microgram/ml using a 0.25 ml sample of plasma. Use of 96-well titer plates and reader can rapidly process many samples at one time. The colorimetric assay correlates well (r = 0.98) with a previously described high performance liquid chromatographic (HPLC) procedure in terms of range of quantitation, limit of quantitation, and precision.