Abstract
Curcumin (Cur) is a potent anti-ischemic drug. Recently, on the basis of in vitro and in vivo studies reports, Cur (found in Curcuma longa) has been found to be very effective in cerebral ischemia. The aim was to develop and validate a highly sensitive ultrahigh-pressure liquid chromatography (UHPLC-MS/MS) method for estimation of Cur in rat brain homogenate and plasma. The chromatographic separation was performed on Waters ACQUITY UPLC (TM) BEH C18 column (2.1 mm x 100 mm; 1.7 mu m) with mobile phase conditions as: isocratic mobile phase of acetonitrile: 2 mM ammonium acetate (85:15 v/v and flow rate of 0.25 mL min(-1)). The transitions occurred at m/z 367.08/217.03 for Cur and m/z 307.04/143.02 for the Internal Standard (IS) warfarin, respectively. The linear dynamic range established for Cur was at concentration of 0.50-1000.0 ng mL(-1) (r(2); 0.9984 +/- 0.0011). The lower limit of quantification (LLOQ) as well as lower limit of detection (LLOD) was 0.054 and 0.017 ng/mL, respectively. The peaks observed having good resolution. The degradation kinetics for curcumin were observed in the stability order as: acidic medium (t(1/2) 104.74/h; t(0.9), 16.59/h) > alkaline medium (t(1/2), 261.93/h; t(0.9), 41.09/h) > oxidative medium (t(1/2), 22.03/h; t(0.9), 4.21/h). The UHPLC-MS/MS method was developed and validated as well as successfully employed for degradation kinetics and pharmacokinetic studies in brain homogenate of Wistar rat with acceptable precision, adequate sensitivity and satisfied accuracy.