Revista de bioanálisis y biomedicina

Determination of Curcumin in Rat Plasma by Liquid–liquid Extraction using LC–MS/MS with Electrospray Ionization: Assay Development, Validation and Application to a Pharmacokinetic Study

Abstract

Singh S. P, Wahajuddin and Jain G. K

A simple, speci fi c and rapid LC–MS/MS method has been developed and validated for the estimation of curcumin in rat plasma, using biochanin as internal standard (IS). The assay procedure involved liquid–liquid extraction of curcumin and IS from rat plasma. The recovery of curcumin and IS in rat plasma were 87.62 and 88.25%, respectively. The resolution of peaks was achieved with 0.01 M ammonium acetate (pH 5.5):acetonitrile (10:90, v/v) on a Supelco Discovery C18 column. The total chromatographic run time was 4 min. Speci fi city and matrix effect on ionization was determined and found that method was speci fi c and there was no signi fi cant matrix effect. The method was proved to be accurate and precise at linearity range of 10–2000 ng/mL with a correlation coef fi cient (r) of ≥ 0.995. The MS/MS ion transitions monitored were 367 → 217 for curcumin and 283 → 268 for IS. The intra- and inter-day assay precision ranged from 2.79 to 8.20% and 4.15 to 7.76%, respectively; and intra- and inter-day assay accuracy was between 92.83–107.83% and 93.92–104.26%, respectively. Practical utility of this LC–MS/MS method was demonstrated in a pilot pharmacokinetic study in male Sprague–Dawley rats following intravenous administration of curcumin.