Optimization of Extraction Conditions for Maximal Phenolic, Flavonoid and Antioxidant Activity from Melaleuca bracteata Leaves Using the Response Surface Methodology

Melaleuca bracteata is a yellow-leaved tree belonging to the Melaleuca genus. Species from this genus are known to be good sources of natural antioxidants, for example, the "tea tree oil" derived from M. alternifolia is used in food processing to extend the shelf life of products. In order to determine whether M. bracteata contains novel natural antioxidants, the components of M. bracteata ethanol extracts were analyzed by gas chromatography-mass spectrometry. Total phenolic and flavonoid contents were extracted and the antioxidant activities of the extracts evaluated. Single-factor experiments, central composite rotatable design (CCRD) and response surface methodology (RSM) were used to optimize the extraction conditions for total phenolic content (TPC) and total flavonoid content (TFC). Ferric reducing power (FRP) and 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH·) scavenging capacity were used as the evaluation indices of antioxidant activity. The results showed that the main components of M. bracteata ethanol extracts are methyl eugenol (86.86%) and trans-cinnamic acid methyl ester (6.41%). The single-factor experiments revealed that the ethanol concentration is the key factor determining the TPC, TFC, FRP and DPPH·scavenging capacity. RSM results indicated that the optimal condition of all four evaluation indices was achieved by extracting for 3.65 days at 53.26°C in 34.81% ethanol. Under these conditions, the TPC, TFC, FRP and DPPH·scavenging capacity reached values of 88.6 ± 1.3 mg GAE/g DW, 19.4 ± 0.2 mg RE/g DW, 2.37 ± 0.01 mM Fe2+/g DW and 86.0 ± 0.3%, respectively, which were higher than those of the positive control, methyl eugenol (FRP 0.97 ± 0.02 mM, DPPH·scavenging capacity 58.6 ± 0.7%) at comparable concentrations. Therefore, the extracts of M. bracteata leaves have higher antioxidant activity, which did not only attributed to the methyl eugenol. Further research could lead to the development of a potent new natural antioxidant.