Optimization of Ethanol Extraction Conditions from Propolis (a Bee Product) Using Response Surface Methodology

A central composite design was used to optimize extraction of propolis materials using ethanol. The independent variables in extraction experiments were ethanol concentration (50, 60, 70, 80, 90%, v/v) and extraction time (1, 2, 3, 4, and 5 h). Higher ethanol concentration and shorter extraction time increased total polyphenol content, but total polyphenol concentration began to decrease when ethanol concentration was higher than 80% (v/v). Ethanol concentration was more important than extraction time in optimization of total polyphenol content in propolis extracts. Electron-donating ability increased with ethanol concentration and shorter extraction time, with ethanol concentration being of greater significance. Antioxidant ability in extracts was optimal at an ethanol concentration of 65 - 75% and with an extraction time of 2.2 - 3.6 h. Nitrite-scavenging ability was increased with use of higher ethanol concentration and shorter extraction time. Total flavonoid content was maximized with an ethanol concentration of 68 - 82% and an extraction time of 2.4 - 3.7 h. Total flavonoid content was affected by both ethanol concentration and extraction time. By superimposition of contour plots, an ethanol concentration of 72 - 82% and an extraction time of 2.2 - 3.3 h were optimal for preparation of propolis extracts.