文献汇报高速逆流色谱和制备高效液相色谱法-分离贯叶金丝桃中的儿茶素和黄酮类化合物.pptVIP

Accept by Journal of Chromatography A Auther：Wei Y, Xie Q, Dong W, Ito Y. May 8，2009 2014 IF :4.169 5-year IF:4.298 Introduction 2. Experimental 3. Results and discussion content 4. Conclusions Hypericum perforatum L.贯叶金丝桃 Introduction Naphthodianthrones :hypericin金丝桃素 Phloroglucinols :hyperforin贯叶金丝桃素 Flavonoids 黄酮类:quercetin槲皮素, quercitrin槲皮苷 Phenolic acids:chlorogenic acid绿原酸 Epigallocatechin表焙儿茶素 etc. Isolation and purification——Chromatography Introduction High Performance Liquid Chromatography（HPLC）高效液相色谱法 Preparative liquid chromatography separation(Prep-LC)制备型高效液相 Introduction High-Speed Countercurrent Chromatography (HSCCC)高速逆流色谱技术 a support-free liquid–liquid partition method 液-液色谱分离技术 eliminates irreversible adsorption of sample onto the solid support 无不可逆吸附 HSCCC Lower phase Peak fraction Experimental Crude extract Prep-LC HPLC HSCCC: Preparation of two-phase solvent Experimental Separation procedure HSCCC： The lower phase：mobile phase flow rate:2.0 ml/min；revolution speed:800 rpm The upper phase：stationary phase UV detector at 254 nm Preparation of two-phase solvent and sample solution for HSCCC HSCCC中两相溶剂和样品的制备 n-hexane-ethyl acetate-methanol-water 正己烷-乙酸乙酯-甲醇-水(1:1:1:1, v/v) solvent system ethyl acetate-methanol-water 乙酸乙酯-甲醇-水 (5:1:5 or 10:1:10 or 50:1:50, v/v) sample solution dissolving the crude extract in the lower phase Experimental Experimental Separation procedure Prep-LC reversed phase C18 column (19×300 mm, 7 μm, Symmetry Prep TM) eluent A:methanol—eluent B:water(45:55, v/v) flow rate:5 ml/min 30min inject 2ml sample solution Experimental HPLC analyses and identification of HSCCC peak fractions YMCPack ODS-A, 150×4.6 mm I.D. ODS column; eluent A: MeOH—eluent B: 0.08% H3PO4 in water gradient elution Time(min) B(%) 0 95 5 70 25 55 30 50 35 50 45 95 flow rate:0.8 ml/min sample quantity:10μl Flavonoids黄酮类 UV detection wavelength：254nm Results and discussion Fig.1 Peak 1: epigallocatechin. Peak