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Procedure For conventional extraction, turmeric powder (0.1 g) was ac curately weighed and mixed with petroleum ether (b.p. 40-60°C) (10 mL) and left to stand overnight. The solvent was carefully re moved, and the residual powder was extracted overnight with methanol (10 mL) at room temperature. The yellowish methanol extract was then removed from the powder. A stock solution of 1000 ppm curcumin in methanol was pre pared. It was diluted 10-fold to give a 100-ppm solution. From this, serial dilutions in acetonitrile-water mixture (50:50) were made. Standard curves from 10 to 30 ppm were prepared for the total curcumin content determination. 20 μL of the extract was injected in triplicate onto the ODS-Hy- persil column and separated using acetonitrile-acetate buffer (pH 4.0, 0.05M), (45:55). The curcumin content was determined by comparison with a standard sample of curcumin. For supercritical fluid extraction, turmeric (100 mg) was placed in the 1-mL extraction vessel, which was placed in the sample loop position with the valve in the load position. Carbon dioxide (or CO2 with methanol) wasdelivered from thepump tothe de tector to equilibrate the system. Theextraction temperature was controlled by the oven. After equilibration, the valve was switched to the inject position to start the extraction. The extrac tion profile was monitored with an on-line variable-wavelength UV detector set at 254 nm. The analyte was collected at the outlet of the back-pressure regulator in a vial. On-line SFE-SFC using 20% methanol-modified carbon dioxide as the extractant at a flow rate of 2 mL/min was per formed at an extraction pressure of 280 bar and a temperature of 60°C, and the extract was passed through the pressure-relief valve to the separation column, which washeld at 100 bar. The analytewasdetectedat254nmandwascollectedattheoutletfor quantitative curcumin determination by HPLC. Results and Discussion Determination of curcumin by HPLC In the selection of a suitable eluent for the determination of curcumin by reversed-phase HPLC, a phosphate buffer (0.01M, pH 7.0) was initially used as the aqueous eluent, but inadequate separation was obtained. Acetonitrile-acetate buffer (0.05M, pH 4.0) (45:55) was found to give satisfactory separations at a flow rate of 1 mL/min, and was therefore used in subsequent HPLC analysis. HPLC separation of a standard curcumin sample showed the presence of all the three curcuminoids (Figure 2), and the total peak area of all three was used in the calculation of the curcumin content. SFE of turmeric SFE of turmeric was initially performed using pure carbon 22 Figure 2. HPLC separation of curcumin and minor curcuminoids using acetonitrile-acetate buffer (0.05M, pH 4.0), (45:55) as the mobile phase. Detection at 254 nm and 0.16 AUFS. Peaks: (1) bisdemethoxycurcumin; (2) demethoxycurcumin; (3) curcumin. Figure 3. Extraction profile of turmeric with supercritical carbon dioxide modified with 20% methanol at 60°C and 250 bar. Journal of Chromatographic Science, Vol. 31, January 1993PDF Image | Supercritical Fluid Extraction Turmeric
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