| As a carotenoid compounds, fucoxanthin is involved in photosyntheses and widely exists in kinds of algae, phytoplankton and shellfish animals, especially in brown algae, golden algae and yellow-green algae. Fucoxanthin has many biological activities, such as anti-cancer, weight loss and antioxidant effects. We mainly studied the separation conditions of fucoxanthin from Sargassum thunbergii by HPLC, and the optimal process parameters of extracting fucoxanthin by ultrasonic assisted extraction and accelerated solvent extraction. We discussed factors during the extracting process, determining the optimal condition through response surface method. This paper also studied the Sargassum thunbergii extract and grape seed extract combined use on reducing serum lipid of hyperlipidemia rats.The main research contents and results are as follows:(1) The topic mainly studied the separation of fucoxanthin by HPLC, analyzing the different mobile phase and mobile phase ratio. The optimal chromatographic condition was carried out on Welch C18(4.6mm×250mm, 5μm) column, using water-methanol(1:9, v/v) as mobile phase A, ethyl acetate as mobile phase B. And during 20 min, the rate of mobile phase B was from 0 to 100%. The flow rate was 1m L·min-1. The detection wavelength was set at 450 nm. The sample volume was 20μL.(2) The topic mainly studied the optimal parameters of ultrasonic assisted extraction, analyzing the ratio of liquid to material, the extraction time and temperature on fucoxanthin extraction rate. Then determined the optimal condition through response surface method. The experimental results showed that, with the extraction time 44.41 min, the temperature 67.84°C and the ratio 42.85 m L/g, the fucoxanthin extraction rate was 3.72μg/g.(3) The topic mainly studied the optimal parameters of accelerated solvent extraction, analyzing the extraction time, the extraction temperature and cycle times on fucoxanthin extraction rate. Then determined the optimal condition through response surface method. The experimental results showed that, with the extraction time 12.61 min, the temperature 62.60°C and extracting for 2.42 times, the fucoxanthin extraction rate was 6.97μg/g. It is better than ultrasonic assisted extraction.(4) The topic mainly studied the Sargassum thunbergii extract and grape seed extract combined use on reducing serum lipid. 80 male SD rats(150±10g) were divided into 8 groups as control group, model group, grape seed group(grape seed extract 33.3mg/Kg/d), fucoxanthin group(Sargassum thunbergii extract 33.3mg/Kg/d), low dose group(grape seed extract 16.7mg/Kg/d + Sargassum thunbergii extract 16.7mg/Kg/d), medium dose group(grape seed extract 33.3mg/Kg/d + Sargassum thunbergii extract 33.3mg/Kg/d), high dose group(grape seed extract 100mg/Kg/d + Sargassum thunbergii extract 100mg/Kg/d), simvastatin positive drug group(20mg/kg/d). Control group and Model group were given 0.01 m L/g water. All groups were given high fat diet excepted Control group which was fed with normal diet. The groups were continuously gavaged administration for 30 days. The weight, four serum lipid indexes including cholesterol(TC), triglyeerides(TG), high- density lipoprotein(HDL), low-density lipoprotein(LDL), the wet weight of fat and the weight of liver were measured and recorded during the experiment. The pathological structural variation of liver cell and adipose cell were observed by HE staining technique. The results showed that compared with the control group, the body weight, TC, TG, and LDL of the model group were increased. The sizes of fat cells were significantly bigger and the liver cells were appeared pathological structure. Compared with the model group, the Sargassum thunbergii extract and grape seed extract in three dose levels had an effect on lowing the serum lipid lever, ameliorating the structure of fat and liver cell and decreasing the size of the fat cells. |
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