A validated method for the quantification of curcumin in plasma and brain tissue by fast narrow-bore high-performance liquid chromatography with fluorescence detection
- 1.3k Downloads
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 β-estradiol 17-acetate is used) on a Jasco Reprosil-Pur Basic C18 column (75 × 2 mm, 1.8 μm) 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 μg/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 μg/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.
KeywordsCurcumin Fast HPLC Small particle size Validation Murine brain Plasma
GR acknowledges the financial support by the DFG Cluster of Excellence “Inflammation at Interfaces”.
- 3.Bengmark S, Mesa MD, Gil A (2009) Plant-derived health: the effects of turmeric and curcuminoids. Nutricion hospitalaria: organo oficial de la Sociedad Espanola de Nutricion Parenteral y Enteral 24:273–281Google Scholar
- 6.Food and Drug Administration (2001) Guidance for industry—bioanalytical method validation. Food and Drug Administration, Rockville, p 25Google Scholar
- 11.Pan MH, Huang TM, Lin JK (1999) Biotransformation of curcumin through reduction and glucuronidation in mice. Drug Metab Dispos 27:486–494Google Scholar
- 13.Ireson CR, Jones DJ, Orr S, Coughtrie MW, Boocock DJ, Williams ML, Farmer PB, Steward WP, Gescher AJ (2002) Metabolism of the cancer chemopreventive agent curcumin in human and rat intestine. Cancer Epidemiol Biomark Prev 11:105–111Google Scholar
- 19.Cheng AL, Hsu CH, Lin JK, Hsu MM, Ho YF, Shen TS, Ko JY, Lin JT, Lin BR, Ming-Shiang W, Yu HS, Jee SH, Chen GS, Chen TM, Chen CA, Lai MK, Pu YS, Pan MH, Wang YJ, Tsai CC, Hsieh CY (2001) Phase I clinical trial of curcumin, a chemopreventive agent, in patients with high-risk or pre-malignant lesions. Anticancer Res 21:2895–2900Google Scholar
- 20.Sharma RA, McLelland HR, Hill KA, Ireson CR, Euden SA, Manson MM, Pirmohamed M, Marnett LJ, Gescher AJ, Steward WP (2001) Pharmacodynamic and pharmacokinetic study of oral Curcuma extract in patients with colorectal cancer. Clin Cancer Res 7:1894–1900Google Scholar