Development and validation of a high-performance liquid chromatography assay and a capillary electrophoresis assay for the analysis of adenosine and the degradation product adenine in infusions.

A high-performance liquid chromatography (HPLC) method and a capillary electrophoresis (CE) method for the analysis of adenosine and the degradation product adenine in infusion solutions have been developed and validated. The HPLC separation of the analytes was achieved on a RP-18 column, using a mobile phase, consisting of 20mM ammonium acetate, pH 6.0, containing 5% of acetonitrile at a flow rate of 1ml/min. Thymidine was used as internal standard. The CE separation was performed in a fused-silica capillary with a 100mM sodium phosphate buffer, pH 2.7, at an applied voltage of 25kV, using cytidine as internal standard. The assays were validated with regard to linearity, range, limit of detection (LOD), limit of quantitation (LOQ), specificity, and precision. Both methods were specific allowing reliable quantification of the analytes. Compared to the CE method, HPLC analysis yielded a two- to five-fold lower LOD. With respect to analysis time, CE was faster than HPLC. The applicability of both methods for the determination of the purity and stability of adenosine in the infusion solutions is demonstrated.