Optimized method for the determination of phosphoarginine in abalone tissue by high-performance liquid chromatography

A rapid high-performance liquid chromatography method for the determination of phosphoarginine (PArg) in invertebrate tissue has been redeveloped and validated. The method employs a reversed-phase amino column and a KH2PO4-acetonitrile mobile phase. PArg peak identity was confirmed by comparison with a known standard and via enzymatic conversion. Additionally linear calibration data, low intra-assay variability (<4%), and a detection limit of 5 pmol were determined. The method was demonstrated using PArg extracted from red abalone (Haliotis rufescens) adductor muscle. Validation of the extraction procedure was also completed, including the measurement of a 100.2+/-0.9% extraction efficiency.