Determination of Spironolactone and Canrenone in Human Plasma by High-performance Liquid Chromatography with

Abstract. A new simple, sensitive and LC-MS/MS method for quantification of spironolactone and its me-tabolite, canrenone, in human plasma is proposed. The analytes were analysed on a C18 column at 48 ºC, by using a mobile phase of 58 % methanol, 42 % 10 mmol dm−3 ammonim acetate in water and a flow rate of 1 mL/min. The detection of both analytes in plasma was performed as follows: ESI+, EIC (m/z 169;187;283;305) from m/z 341, after protein precipitation with methanol. Calibration curves were gener-ated over the ranges 2.77–184.50 ng/mL for spironolactone and 2.69–179.20 ng/mL for canrenone by us-ing a weighted (1/y) linear regression. The absolute values of within- and between-run precision and accu-racy for both analytes ranged between 3.1 and 13.9 %, and the mean recovery was 99.7 %. The analytes demonstrated good stability in various conditions. The validated method has been applied to a bioequiva-lence study of 50 mg spironolactone tablets on healthy volunteers. (doi: 10.5562/cca1761