Study On The Preparation Of Spinach Extracts

Spinacia oleracea L.belongs to Chenopodiaceae.Previous studies have reported that S.oleracea contains 20-hydroxyecdysone and show potential anti-muscle atrophy activity.The aim of this study is to develop new products:spinach extracts which contains 20-hydroxyecdysone in it.Single factor and response surface design tests were applied to investigate optimum conditions for extracting of 20-hydroxyecdysone from spinach.The optimum extraction conditions are as follow:extraction solvent,75%ethanol,liquid to solid ratio 11:1,extraction time 67 min and refluxing twice;The optimal purification conditions are as follow:concentration of sample solution 165 mg/mL,velocity of sample solution 2.0 mL/min,pH3.5,concentration of ethanol on desorption 87%,velocity 2.0 mL/min,and volume of 87%ethanol 8 BV.After the purification,the purity of20-hydroxyecdysone had reached a level of 0.92%.We have further explored a simple method to purify 20-hydroxyecdysone by using silica gel column chromatography and sephadex column chromatography,and got the 20-hydroxyecdysone with a purity of 83.9%.In the last chapter,the LC-MS~ntechnique was used to analyze the chemical constituents of spinach extracts.In addition to 20-hydroxyecdysone,it contains 8 major flavonoids:patuletin-3-O-β-D-glucopyranosyl-(1→6)-[β-D-apiofuranosyl-(1→2)]-β-D-glucopyranoside;patuletin-3-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside;spinacetin-3-O-β-D-glucopyranosyl-(1→6)-[β-D-apiofuranosyl-(1→2)]-β-D-glucopyranoside;spinacetin-3-O-β-D-glucopy ranosyl-(1→6)-β-D-glucopyranosie;3,6-dimethyl-quercetagetin-4’-O-glu curonide;5,7,4’-trihydroxy-3,6,3’-trimethoxyflavone-4’-O-glucuronide;5,3’,4’-trihydroxy-3-methoxy-6:7-methylenedtoxyflavone-4’-β-D-glucur onide;5,4’-dihydroxy-3,3’-dimethoxy-6:7-methylenedioxyflavone-4’-β-D-glucuronide...