Development and Validation of a Reversed Phase HPLC Method for Simultaneous Determination of Curcumin and Piperine in Human Plasma for Application in Clinical Pharmacological Studies

A sensitive and specific HPLC method was developed for the simultaneous determination of curcumin and piperine in human plasma in view of the potential therapeutic application of curcumin and piperine in various diseases. The HPLC method consisted of isocratic elution with acetonitrile-methanol-trifluoroacetic acid-water (17.6:35.3:0.1:47.0, v/v/v/v) with a flow rate of 2.5mL min-1 on a Chromolith (R) SpeedROD RP-18 (50x4.6mm) column at an ambient temperature. Ultraviolet detection was performed in programe mode at 415nm for curcumin, 335nm for piperine, and 280nm for -17-estradiol acetate (internal standard). Curcumin, piperine, and internal standard were extracted from plasma using ethyl acetate-propanol (9:1 v/v). Mean extraction recoveries for curcumin, piperine, and internal standard were 91.3, 91.4, and 92.9%, respectively. The assay was linear over the therapeutic concentration range (10-500ng mL-1) for both drugs with correlation coefficients of r20.99. Limit of detection and limit of quantification were 1ng mL-1 and 10ng mL-1 for both curcumin and piperine. The method was applied to the determination of the concentrations of curcumin and piperine in healthy volunteers after treatment with 1500mg curcumin and 500mg piperine, and should find an applications in pharmacokinetic studies of these compounds.