Bioactive Compound Extraction of Hemp (Cannabis sativa L.) Leaves through Response Surface Methodology Optimization

Hemp, commonly known as Cannabis sativa L., is a medicinal plant species of the Cannabaceae family. For the efficient extraction of C. sativa leaves using the conventional stirring process with water as the solvent, three crucial extraction parameters (i.e., extraction duration, liquid–solid ratio, and temperature) were investigated through the response surface methodology (RSM). The concentrations of the extracted bioactive compounds (polyphenols, ascorbic acid, and carotenoids) showed significant variations in the RSM design points, suggesting the importance of finding the optimal extraction conditions in which liquid–solid ratio and extraction temperature were found to have the highest impact. Further analysis was conducted on the optimal extract employing several assays to determine their polyphenol content, total carotenoid content, color evaluation, anti-inflammatory activity, and antioxidant capacity through FRAP, DPPH, and H2O2 assays. Α low extraction time (30 min) at 50 °C and a high liquid–solid ratio (50:1) were required for the highest possible yield of polyphenols. The total polyphenol content was determined to be 9.76 mg gallic acid equivalents/g under optimum conditions, with pelargonin being the most abundant polyphenol (1.51 mg/g) in C. sativa extracts. Ascorbic acid was measured at 282.23 μg/g and total carotenoids at 356.98 μg/g. Correlation analyses revealed that anti-inflammatory activity was negatively correlated with specific polyphenols. As determined by DPPH (27.43 μmol ascorbic acid equivalents (AAE)/g), FRAP (49.79 μmol AAE/g), and H2O2 (230.95 μmol AAE/g) assays, the optimized aqueous extract showed a high antioxidant capacity. Furthermore, it demonstrated considerable anti-inflammatory activity at 17.89%, with the potential to increase to 75.12% under particular extraction conditions. Given the high added-value of the aqueous extracts, the results of this study highlight the potential utility of C. sativa leaves as a source of health-improving antioxidant compounds in the pharmaceutical and food industries.