Abstract
A simple, speci fi c and rapid LC–MS/MS method has been developed and validated for the estimation of curcumin in rat plasma, using biochanin as internal standard (IS). The assay procedure involved liquid–liquid extraction of curcumin and IS from rat plasma. The recovery of curcumin and IS in rat plasma were 87.62 and 88.25%, respectively. The resolution of peaks was achieved with 0.01 M ammonium acetate (pH 5.5):acetonitrile (10:90, v/v) on a Supelco Discovery C18 column. The total chromatographic run time was 4 min. Speci fi city and matrix effect on ionization was determined and found that method was speci fi c and there was no signi fi cant matrix effect. The method was proved to be accurate and precise at linearity range of 10–2000 ng/mL with a correlation coef fi cient (r) of ≥ 0.995. The MS/MS ion transitions monitored were 367 → 217 for curcumin and 283 → 268 for IS. The intra- and inter-day assay precision ranged from 2.79 to 8.20% and 4.15 to 7.76%, respectively; and intra- and inter-day assay accuracy was between 92.83–107.83% and 93.92–104.26%, respectively. Practical utility of this LC–MS/MS method was demonstrated in a pilot pharmacokinetic study in male Sprague–Dawley rats following intravenous administration of curcumin.
Highlights
Curcuma spp. contain turmerin, essential oils, and curcuminoids, including curcumin [1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6heptadiene-3,5-dione] which is a phenolic substance derived from dietary spice herb Curcuma longa
Mobile phase comprising of 0.01 M ammonium acetate:acetonitrile (10:90, v/v) was delivered at a flow rate of 0.5 mL/min was found to be suitable during LC optimization and enabled the determination of electrospray response for curcumin and internal standard (IS)
Experiments were performed with different C18 columns and found that chromatographic resolution, selectivity and sensitivity were good with Supelco Discovery C18 column (4.6 × 50 mm, 5.0 m)
Summary
Curcuma spp. contain turmerin, essential oils, and curcuminoids, including curcumin [1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6heptadiene-3,5-dione] which is a phenolic substance derived from dietary spice herb Curcuma longa. Appropriate dilutions were made in methanol for curcumin to produce working stock solution of 20, 10, 5, 2, 1, 0.5, 0.2, 0.1 g/mL and on the day of analysis this set of stocks was used to prepare standards for the calibration curve. Calibration standards and QC samples were prepared by spiking 90 L of control pooled rat plasma with the appropriate working solution of curcumin (10 L) and IS (10 L) on the day of analysis. Samples for the determination of precision and accuracy were prepared by individually spiking control rat plasma at four concentration levels [10 ng/mL (lower limit of quantitation, LLOQ), 40 ng/mL (QC low), 800 ng/mL (QC medium) and 1600 ng/mL (QC high)] and stored at –80 ± 10°C until analysis. The residue was reconstituted in 200 L of the mobile phase and 10 L was injected onto analytical column
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
