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using ethanol as solvent (pa, Químis) for 24 hours followed by the measurement of the absorbency at 425 nm in a spectrophotometer (GBC, Model UV/VVIS 916). The concentration (wCurcumin) of the curcuminoids expressed in weight percent is computed using the equation as follows (Takahashi, 1987). w [wt.%] A f V[ml] (2) Curcumiin m[g]E1% 1cm Where A is the absorbency, V is the volume of solution in ml, m is the sample mass in mg, f s the dilution factor and E 1% is the absorptivity of curcumin ( E 1% = 1607). 1cm 1cm 2.5.2 Chemical Identification of the Curcuminoids by HPLC The chemical identification of curcumin and the two curcuminoids (desmethoxycurcumin and bis-desmethoxycurcumin) in the oilresin have been performed by high-pressure liquid chromatography using the procedure as follows (Taylor and McDowell, 1992). The samples (1.0- 9.0 mg) were diluted in a standard solution of acetic acid (1.0 mg/ml) with pH adjusted at 2.8, prepared in the proportion (55:45 vol/vol) with acetonitrile (chromatographyc grade, Science) and deionized water filtered using membranes of nylon with pores of 0.45 (Supelco). Afterward, the mobile phase at a rate of 1ml/mn have been injected in the HPLC (Shimadzu, Model: LC-10AD) equipped with an UV-Vis detector (Shimadzu, Model: SPD-10A), a manual splitting system (Shimadzu, Model: SV-10AM), a communication system (Shimadzu, Model: CBM-10A) coupled to a personal computer and a control software (Shimadzu, Class: LC10) for the data processing, a stationary phase column (Shimadzu, Model: Hypersil ODS-C18) of 250 mn x 4.5 nm. The detector has been adjusted at 425 nm and 298 K. The concentration of curcumin and the two curcuminoids (desmethoxycurcumin and bis-desmethoxycurcumin) in the oilresin expressed in g/ml has been computed using calibration curves, whose relates the concentration to the integration area (AI) of the three different peaks in the chromatogram as follows. 6 CCurcumin[μμg/ml0.167615.8635x10 *AI (3) 6 CBDMC[μμg/ml 0.00443 6.4176x10 * AI (4) 6 CDMC[μμg/ml0.024076.3673x10 *AI (5)PDF Image | SUPERCRITICAL CO2 EXTRACTION OF CURCUMINS
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