RAPID AND SIMULTANEOUS HPLC ANALYSIS OF CURCUMIN AND ITS METABOLITE TETRAHYDROCURCUMIN FROM PLASMA AND LIVER HOMOGENATES

Abstract

A rapid and sensitive reverse phase high performance liquid chromatography (HPLC) method was developed for simultaneous determination of curcumin (Cur) and its active metabolite tetrahydrocurcumin (THC) from rat plasma and liver homogenates. 17β-estradiol was used as the internal standard (IS). A Waters Spherisorb RP C18 column (4.6 mm × 250 mm, 5 µm) and a mobile phase comprising acetonitrile-methanol-water adjusted to pH 3.0, at a flow rate of 1.0 mL min⁻¹ was optimized. THC and 17β-estradiol were detected at 280 nm and Cur at 425 nm. Cur revealed three peaks – diferuloylmethane, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC). All the analytes were well-separated with resolution >1.5. The calibration curves were linear over the range 100–5000 ngmL⁻¹ for diferuloylmethane and THC. Extraction of diferuloylmethane and THC was consistent and reproducible as indicated by a low coefficient of variation, and a constant analyte/IS ratio. A simple, reproducible HPLC method with good resolution and simultaneous detection of analytes was developed and successfully applied for pharmacokinetic evaluation and liver uptake in rats following oral administration of diferuloylmethane in single processing sample which makes this method a rapid and cost effective analysis.

Keywords:

• 17β-estradiol
• curcumin
• diferuloylmethane
• liver-homogenate
• plasma
• tetrahydrocurcumin

ACKNOWLEDGMENT

Authors are thankful to the University Grants Commission (UGC) Government of India for financial support and Institute of Chemical Technology, Mumbai, India for Prof. M.M. Sharma Fellowship to Anisha.

Notes

^a All values expressed in Mean ± S.D.