T has been used to decrease dampness [8]. In several studies, phenolic acids, polyketides, diterpenes, triterpenes, glucosides, proteins, and sterols have been isolated from various plant parts, and phenols comprise the major constituents of the plant [9,10]. C. cyrtophyllum may be an excellent source of antioxidants: its methanolic Title Loaded From File extract has strong DPPH radical-scavenging activity [11]. However, the economic feasibility of industrial processing of C. cyrtophyllum requires more investigation to optimize the extraction process to increase the yield of extracted active substances. Ultrasonic-assisted extraction (UAE) has been used to extract functional components from different matrixes; it is more rapid than conventional extraction techniques. UAE creates shear forces that break cell walls mechanically, simultaneously facilitating theExtraction of Antioxidants from C. cyrtophyllumrelease of cellular constituents of plant material 11967625 into the extraction solvent without chemical degradation [12]. However, the efficiency of the extraction process is less than desireable [13]. An appropriate experimental design is necessary for any optimization study, and the two most common designs are single-factor experiments and response-surface methodology (RSM). Single-factor experiments were used here to provide data regarding extraction factors with significant effects on phenolic antioxidants from C. cyrtophyllum leaves. Next, these factors were analyzed by RSM for central composite rotatable design (CCRD) to more precisely determine optimal extraction conditions. Continuing our ongoing research into natural plant antioxidants, we report a method of optimal antioxidant extraction [14,15]. Our extraction parameters were ethanol concentration, extraction time, and extraction temperature. Phenolic compounds contain many hydroxyl groups and phenyls, so ethanol, a binary solvent, and water were critical extraction components. Our goal was to extract useful components from the leaves of C. cyrtophyllum while retaining optimal total phenolic content (TPC), total flavonoid content (TFC), and scavenging activity on 2,29diphenyl-1-picrylhydrazyl (DPPH) and 2,29-azino-bis (3-ethylbenzothiazoline- 6-sulphonicacid) (ABTS).yellow. The volume of the reaction mixture obtained was increased to 100 ml (q.s., deionized water) before usage. Then, 2 mL of diluted extracts were mixed with 2 mL of FC reagent. After 3 min, 750 mL of sodium Title Loaded From File carbonate anhydrous solution (7.5 , w/ v) was added and the sample was vortexed. The absorbance at 765 nm versus a blank control was measured with a UV light spectrophotometer (Shimadzu UV2754) after a 2-h incubation in the dark at room temperature. Measurements were calibrated to a standard curve of prepared gallic acid solution ranging from 0?00 mg/mL with y = 0.0480x ?0.0071 (R2 = 0.9991) and TPC was then expressed as mg of gallic acid equivalents (GAE) per g of dry weight (DW).TFC measurementEstimation of TFC in extracts was performed according to colorimetric method [19] with some modifications. The reaction mixture contained 1.0 mL of extract, 4 mL of 60 ethanol and 0.3 mL of 5 sodium nitrite. Six minutes later, 0.3 mL of 10 aluminium nitrite was added. In the next six minutes, 4 mL of 1 M sodium hydroxide solution were added and the volume was increased to 10 mL (q.s. 60 ethanol). Immediately, the reaction mixture absorbance was measured by a spectrophotometer at 510 nm against a blank (control) and used to calculate TFC using.T has been used to decrease dampness [8]. In several studies, phenolic acids, polyketides, diterpenes, triterpenes, glucosides, proteins, and sterols have been isolated from various plant parts, and phenols comprise the major constituents of the plant [9,10]. C. cyrtophyllum may be an excellent source of antioxidants: its methanolic extract has strong DPPH radical-scavenging activity [11]. However, the economic feasibility of industrial processing of C. cyrtophyllum requires more investigation to optimize the extraction process to increase the yield of extracted active substances. Ultrasonic-assisted extraction (UAE) has been used to extract functional components from different matrixes; it is more rapid than conventional extraction techniques. UAE creates shear forces that break cell walls mechanically, simultaneously facilitating theExtraction of Antioxidants from C. cyrtophyllumrelease of cellular constituents of plant material 11967625 into the extraction solvent without chemical degradation [12]. However, the efficiency of the extraction process is less than desireable [13]. An appropriate experimental design is necessary for any optimization study, and the two most common designs are single-factor experiments and response-surface methodology (RSM). Single-factor experiments were used here to provide data regarding extraction factors with significant effects on phenolic antioxidants from C. cyrtophyllum leaves. Next, these factors were analyzed by RSM for central composite rotatable design (CCRD) to more precisely determine optimal extraction conditions. Continuing our ongoing research into natural plant antioxidants, we report a method of optimal antioxidant extraction [14,15]. Our extraction parameters were ethanol concentration, extraction time, and extraction temperature. Phenolic compounds contain many hydroxyl groups and phenyls, so ethanol, a binary solvent, and water were critical extraction components. Our goal was to extract useful components from the leaves of C. cyrtophyllum while retaining optimal total phenolic content (TPC), total flavonoid content (TFC), and scavenging activity on 2,29diphenyl-1-picrylhydrazyl (DPPH) and 2,29-azino-bis (3-ethylbenzothiazoline- 6-sulphonicacid) (ABTS).yellow. The volume of the reaction mixture obtained was increased to 100 ml (q.s., deionized water) before usage. Then, 2 mL of diluted extracts were mixed with 2 mL of FC reagent. After 3 min, 750 mL of sodium carbonate anhydrous solution (7.5 , w/ v) was added and the sample was vortexed. The absorbance at 765 nm versus a blank control was measured with a UV light spectrophotometer (Shimadzu UV2754) after a 2-h incubation in the dark at room temperature. Measurements were calibrated to a standard curve of prepared gallic acid solution ranging from 0?00 mg/mL with y = 0.0480x ?0.0071 (R2 = 0.9991) and TPC was then expressed as mg of gallic acid equivalents (GAE) per g of dry weight (DW).TFC measurementEstimation of TFC in extracts was performed according to colorimetric method [19] with some modifications. The reaction mixture contained 1.0 mL of extract, 4 mL of 60 ethanol and 0.3 mL of 5 sodium nitrite. Six minutes later, 0.3 mL of 10 aluminium nitrite was added. In the next six minutes, 4 mL of 1 M sodium hydroxide solution were added and the volume was increased to 10 mL (q.s. 60 ethanol). Immediately, the reaction mixture absorbance was measured by a spectrophotometer at 510 nm against a blank (control) and used to calculate TFC using.
