Determination of higenamine in plasma and urine by high-performance liquid chromatography with electrochemical detection

A method for the determination of higenamine in plasma and urine based on high-performance liquid chromatography (HPLC) with electrochemical detection was developed. The plasma or urine sample was treated by adsorption with acidic alumina and then higenamine was released by acid treatment. HPLC was performed on an ODS column with a mobile phase of acetonitrile-0.1%c phosphoric acid (9:91) and an electrochemical detector at an oxidation potential of 0.75 V. The lower limits of the assay for higenamine in plasma and urine were 2.645 and 10.58 ng / ml, respectively. The recoveries of higenamine after alumina treatment in rabbit plasma and urine were ca. 77.5 and 84.4%, respectively. Intra- and inter-day precision and accuracy reported as coefficients of variation in plasma and urine were less than 7%. The application of the assay was demonstrated successfully by the determination of the concentration of higenamine in rabbit plasma and urine samples after doses of higenamine hydrochloride at 20 mg/kg intravenously and 50 mg/akg orally.