Extraction And Purification Of Terpenoids Of Sargassum Fusiforme

Sargassum fusiforme, the edible brown algae, is one of the most widely consumed seaweeds in China, Japan and Korea, has been used as a source of food and as a traditional Chinese medicine and is officially listed in the Chinese Pharmacopoeia2010. Nowadays, the researchers of S. fusiforme forcuse on the study of polysaccharides because of its high content in S. fusiforme, and get a lot of exciting achievements. However, terpenoids of S. fusiforme often overlooked due to its low levels. Studies have shown that terpenes have gained more and more attention because it has significant acrivities of anti-tumor, Alzheimer prevention and so on. In this study, a method of microwave-assisted extraction coupled with high-speed counter-current chromatography was successfully developed for the separation and purification terpenoids of from S. fusiforme.1. A method of microwave-assisted extraction was successfully developed for terpenoids of from S. fusiforme. Compared with six kinds of commonly used extraction solvent, water has the highest extraction yield of three kinds of terpenoids. The main extraction conditions including microwave power, liquid/solid ratio, irradiation time and extraction temperature were optimized using orthogonal array design:800W microwave power,1:30liquid/solid ratio,15min irradiation time and80℃. Under the optimized conditions, the proposed approach has been evaluated in comparison with the Ultrasonic-assisted extraction (UAE) and conventional heat-reflux extraction (HRE). The reduction of the extraction time (from2h to15min) and remarkable higher efficiency (4.38-31.6%improved) supports the suitability of the proposed approach. This study reveals that MAE can effectively extract the three kinds of terpenoids from S.fusiforme.2. A method of microwave-assisted extraction with nature deep eutectic solvents (NADES) was successfully developed for terpenoids of from S. fusiforme. The results show that the terpenoids yield of25%NADES2.83%-36.92%,4.39%-9.51%and13.9%-57.7%is higher than water, respectively; The extraction yields of terpenoids is bigger when the content of water is50%or75%; NADES can accelerate the extraction rate, save about two-thirds of the time; Terpenoids were recovered from NADES with a yield between69%and75.5%. This study reveals the potential of NADES for applications involving the extraction of terpenoids from S. fusiforme.3. A method of High-speed countercurrent chromatography with a suitable solvents system consisting of n-hexane-ethyl acetate-methanol-water (1:2:1:2, v/v) was successfully developed for the separation and purification terpenoids of from S. fusiforme. A total of33.87mg mixture of Loliolide and Isololiolide,12.35mg (3R)-4-[(2R,4S)-4-acetoxy-2-hydroxy-2,6,6-trimethylcyclohexylidene]-3-buten-2-one were obtained from300mg S. fusiforme in240min,(3R)-4-[(2R,4S)-4-acetoxy-2-hydroxy-2,6,6-trimethylcyclohexylidene]-3-buten-2-one’purity is95.5%; A method of p-HPLC with a suitable mobile phase consisting of acetonitrile-water (20:80, v/v) was successfully developed for the separation and purification terpenoids from S. fusiforme. A total of4.24mg Loliolide and3.86mg Isololiolide were obtained from15mg mixture of Loliolide and Isololiolide in60min with purities of96.2%and95.8%, respectively. This study reveals that HSCCC is suitable for separation and enrichment of terpenoids and p-HPLC is suitable for separation of terpenoids isomers from S.fusiforme...