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

1、Separation of epigallocatechin and flavonoids from Hypericum perforatum L. by high-speed counter-current chromatography and followed by preparative liquid chromatography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,Accept by Journal of Chromatography A Auther：Wei Y, Xie Q, Dong W, Ito Y. May 8，2009,2014 IF :

2、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 Epigallocate

3、chin表焙儿茶素 etc.,Isolation and purificationChromatography,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 liquidliquid partition method 液-液

4、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 spee

5、d: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),

6、sample solution,dissolving the crude extract in the lower phase,Experimental,Experimental,Separation procedure,Prep-LC,reversed phase C18 column (19300 mm, 7 m, Symmetry Prep TM),eluent A:methanoleluent B:water(45:55, v/v),flow rate:5 ml/min,30min,inject 2ml sample solution,Experimental,HPLC analyse

7、s and identification of HSCCC peak fractions,YMCPack ODS-A, 1504.6 mm I.D. ODS column;,eluent A: MeOHeluent B: 0.08% H3PO4 in water,gradient elution,flow rate:0.8 ml/min,sample quantity:10l,Flavonoids黄酮类 UV detection wavelength：254nm,Results and discussion,Fig.1,Peak 1: epigallocatechin. Peak 2 (a,

8、b, c): hyperoside, isoquercitrin and miquelianin. Peak 3: quercitrin. Peak 4: avicularin. Peak 5: quercetin.,Fig.2,ethyl acetate-methanol-water 50:1:50 (v/v) (a)； 10:1:10 (v/v) (b)； 5:1:5 (v/v) (c),Peak 2 (in Fig.2a, 2b, 2c): mixture of hyperoside,isoquercitrin and miquelianin. Peak 3: mixture of epigallocatechin and avicularin. Peak 4:quercitrin,Fig.3,Fig.4,Conclusions,900 mg crude extract,7.8mg quercetrin槲皮素(a purity of over 97%),12.6mg quercetin槲皮苷(a purity of over 93%),38.9mg hyperoside, isoquercitrin and miquelianin mixture (over 97% of the fractio