A validated method for the quantification of curcumin in plasma and brain tissue by fast narrow-bore high-performance liquid chromatography with fluorescence detection

Abstract

Curcumin, a lipophilic polyphenol derived from the rhizome of the plant turmeric (Curcuma longa), might be useful in the prevention and treatment of a number of degenerative brain disorders, including glioma multiforma and Alzheimer's disease. Thus, there is growing interest in measuring curcumin concentrations in the brain and other target tissues in relevant animal models. We therefore developed and validated (according to the Food and Drug Administration guidelines for bioanalytical method validation), a simple, fast and reliable method for the quantification of curcumin in biological matrices by fast high-performance liquid chromatography with fluorescence detection. This method involves a simple extraction with 95% ethyl acetate and 5% methanol, rapid separation (<2 min if external standards and <4 min if the internal standard beta-estradiol 17-acetate is used) on a Jasco Reprosil-Pur Basic C(18) column (75 x 2 mm, 1.8 mum) with an eluent of acetonitrile, methanol, de-ionised water and acetic acid (49:20:30:1, v/v; flow rate, 0.4 mL/min) and fluorescence detection (excitation wavelength, 420 nm; emission wavelength, 470 nm). The method is selective, precise (<15% RSD at the lower limit of quantification), accurate (<15% of the coefficient of variation at the lower limit of quantification) and sensitive over a linear range of 0.05-10 microg/mL for curcumin. The developed method was used for the quantification of curcumin in the brains of mice force-fed (50 mg/kg bw) or i.p. injected (100 mg/kg bw) with curcumin. Brain curcumin concentrations of the mice were below the limit of detection at 30, 60 and 120 min after oral gavage and reached 4-5 microg/g brain 20-40 min after i.p. injection. In conclusion, the developed and validated method should be useful for the accurate and precise quantification of curcumin in target organs from relevant animal models of human diseases.