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

Abstract

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 2.5 mL min⁻¹ on a Chromolith® SpeedROD RP-18 (50 × 4.6 mm) column at an ambient temperature. Ultraviolet detection was performed in programe mode at 415 nm for curcumin, 335 nm for piperine, and 280 nm for β-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 (10–500 ng mL⁻¹) for both drugs with correlation coefficients of r² > 0.99. Limit of detection and limit of quantification were 1 ng mL⁻¹ and 10 ng 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 1500 mg curcumin and 500 mg piperine, and should find an applications in pharmacokinetic studies of these compounds.

Keywords:

• Curcumin
• Healthy volunteers
• HPLC
• Piperine

ACKNOWLEDGMENTS

We are grateful to Prof. G. Padamanaban, Indian Institute of Sciences, Banglore, for valuable discussion and guidance. Thanks are also due to Dr. N. C. Gupta for technical support. The authors sincerely acknowledge the financial support from Department of Biotechnology, New Delhi, to perform this study.

Notes

∗Numbers in the [] represents reference.

∗All QC samples were analysed in triplicate. Mean values are reported.

Obs. = observed; Dev. = deviation.