Study On Chemical Constituents And HPLC Stimultaneous Determination Of The Effective Constituents In Lysimachia Clethroides

This thesis is composed of four chapters. The first chapter reported the chemical constituents from the 70% actone extract of Lysimachia clethroides. The second chapter described the extraction method and HPLC stimultaneous determination of four flavonoids in Lysimachia Clethroides. The third chapter described HPLC stimultaneous determination of quercetin and kaempferol in Lysimachia Clethroides. The last chapter summarized Lysimachia L's chemical constituents, pharmacological actions and the progress of application study of the HPLC in pharmaceutical analysis.As a result,30 compounds were isolated from this plant. Among them,26 compounds were identified on the basis of spectral methods, These compounds covered flavonoids,lignans,triterpenoids,phenolic acids, and so on.30 compounds were isolated from the 70% actone extract of Lysimachia clethroides, the isolated compounds were deduced as follows:kaempferol (1), kaempferol 3-β-D-glucopyra noside (2), kaempferol 3-O-β-D-galactoside (3), kaempferol 3-O-β-D-(6"-p-coumaroyl)-gluco pyranoside (4), quercetin (5), Isorhamnetin (6), quercetin 3-O-β-D-glucopyranoside (7), quercetin 3'-methoxy-3-O-β-D-galactoside (8), quercetin 3-O-β-D-(6"-p-coumaroyl)-galactos ide (9),4'-methoxy-5,6-dihydroxyisoflavone-7-O-β-D-glucopyranoside (10),7-O-glucosyl liquiritigenin (11), (+)-IsolaricIresinol-9'-β-D-glucopyranoside (12), Urolignoside (13),23-hy droxyursolic acid (14), Isotachioside (15),p-Hydroxybenzoic acid (16),3-Methoxy-4-hydrox ybenzoic acid (17),3,4-Dihydroxybenzoic acid (18),3,5-Dihydroxybenzoic acid (19), Methylα-D-fructofuranoside (20), Uridine (21), Stearic acid glyceride (22), linolenic acid glyceride (23), Oleanolic acid (24),β-Sitosterol (25), Daucosterol (26). Compounds 3,6,16,18,19,22, 23 were isolated from this plant for the first time, Compounds 8～11,14,15,17,20,21 were isolated from this genus for the first time.Using total content of rutin,isoquercitrin,kaempferol 3-O-β-D-galactoside and astragalin as a marker, the optimum extraction technology of raw material was selected with the single factor experiment and orthogonal experiment. As a result, the optimum parameters were 70% ethanol,45minitues,45℃and 1:100 materials (g/mL).To establish RP-HPLC method for the determination of ruti,isoquercitrin,kaempferol 3-O-β-D-galactoside and astragalin, this analysis was performed on a LiChrospher RP-C18 column (4.6mm×250mm,5μm) with a gradient mobile phase of acetonitrile-0.4% phosphoric acid solution (0～15min,15:85-17:83,15～25min,17:83-30:70) at a flow rate of 1mL·min-1, the cloumn was maintained at 35℃, and the detecting wavelength was set at 360nm.To establish RP-HPLC method for the determination of quercetin and kaempferol, this analysis was performed on a ZORBAX SB-C18 column (4.6mm×150mm,5μm) with a gradient mobile phase of acetonitrile-0.4% phosphoric acid solution (0～15min,17:83-40:60) at a flow rate of 1 mL·min-1, the cloumn was maintained at 40℃, and the detecting wavelength was set at 360nm.The review summarized Lysimachia L's chemical constituents, pharmacological actions and the progress of application study of the HPLC in pharmaceutical analysis.