Laser microdissection hyphenated with high performance gel permeation chromatography-charged aerosol detector and ultra performance liquid chromatography-triple quadrupole mass spectrometry for histochemical analysis of polysaccharides in herbal medicine: Ginseng, a case study.

This study establishes a new combinatorial approach for histochemical analysis of polysaccharides in herbal medicines using laser microdissection followed by high performance gel permeation chromatography coupled with charged aerosol detector and ultra-performance liquid chromatography hyphenated with triple quadrupole mass spectrometry. Ginseng was employed as a study model. Tissue-specific qualitative and quantitative characterization of ginseng polysaccharides was performed by determining their molar masses and monosaccharide compositions in three macro-dissected parts (rhizome, main and branched roots) and five micro-dissected tissues (cork, cortex, xylem, phloem and resin canal). The results showed that ginseng "flesh" (xylem, phloem and resin canal) contained more polysaccharides with larger molecular weights and higher ratios of glucose residue, whereas ginseng "skin" (cork and cortex) had fewer polysaccharides with smaller molecular weights and higher ratios of non-glucose constituents (e.g. galacturonic acid, galactose, arabinose and rhamnose). These findings suggested that the polysaccharides of the "flesh" were predominantly starch-like glucans, while those of the "skin" were of a higher proportion of acidic pectins. The revealed histologic distribution and accumulation pattern of ginseng polysaccharides contributes to the scientific understanding of ginseng regarding the biosynthesis and transportation of polysaccharides, medicinal quality evaluation as well as empirical clinical application.

Chemical Constituents of Pyrrosia calvata.

A novel flavanone glycoside, 3',5',5,7-tetrahydroxy-6-C-ß-D-glucopyranosyl-flavanone (1), along with 16 known compounds, (R/S)-eriodictyol-8-C-ß-D-glucopyranoside (2), quercetin-3-O-α-L-rhamnosyl (1''' --> 3''')-ß-D-glucopyranoside (3), hemipholin (4), 4ß-carboxymethyl-(-)-epicatechin methyl ester (5), kaempferol (6), quercetin (7), mangiferin (8), chlorogenic acid (9), 1,5-O-dicaffeoylquinic acid (10), 3,5-O-dicaffeoylquinic acid (11), 3-O-caffeoylquinic acid methyl ester (12), 1-O-caffeoyl glycoside (13), 4-O-ß-D-glucopyranosyl-caffeic acid (14), 3'-O-methyleplcatechin-7-O-ß-D-glucopyranoside (15), hop-22(29)-en-30-ol (16) and diploptene (17), were isolated from the whole plant of Pyrrosia calvata (Backer) Ching. Among them, compounds 2, 3, 4, 10, 11, 13 and 14 were isolated from the family Polypodiaceae for the first time, and compound 5 has not been recorded previously from the genus Pyrrosia.

[Chemical constituents from rhizomes of Iris germanica].

Twenty-one compounds were isolated from the rhizomes of Iris germanica by various chromatographic techniques such as silica gel, ODS and Sephadex LH-20 chromatography. Their structures were established on basis of physical properties, MS and NMR spectroscopic data Their structures were identified as ombuin (1), 5, 3, 3'-trihydroxy-7, 4'-dimethoxyflavanone (2), naringenin (3), cirsiliol-4'-glucoside (4), 3beta, 4'-dihydroxy-7,3'-dimethoxyflavonone-5-O-beta-D-glucopyranoside (5), genistein (6), irilin D (7), muningin (8), 5, 7, 4'-trihydroxy-6, 3', 5'-trimethoxyisoflavone (9), tectorigenin (10), irigenin (11), tectoridin (12), iridin (13), mangiferin (14), irisxanthone (15), pyroglutamic acid (16), 2, 4', 6-trihydroxy-4-methoxybenzophenone-2-O-beta-D-glucoside (17), apocynin (18), androsin (19), beta-sitosterol (20), and daucosterol (21). Among them, compounds 1-9, 16, 17 were obtained from this plant for the first time, compounds 8 and 9 were separated from Iris species for the first time, compounds 1, 4, and 17 were obtained from the family for the first time.

Anticancer activity of total flavonoids isolated from Xianhe Yanling Recipe (仙鹤延龄方).

OBJECTIVE: To investigate the anticancer activity of the total flavonoids isolated from a herbal formula, Xianhe Yanling Recipe (仙鹤延龄方), a recipe commonly used in cancer patients in China. METHODS: The in vitro anticancer activity of the total flavonoids was determined using the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on three cancer cell lines: MCF-7 (a human breast adenocarcinoma cell line), HepG-2 (a human hepatocellular carcinoma cell line) and ES-2 (a human ovarian cancer cell line). The in vivo anticancer effect of the total flavonoids was assessed in a mouse tumor model bearing H22-induced hepatocellular carcinoma, and cisplatin was used as a positive control. RESULTS: The total flavonoids exerted a powerful inhibitory effect on the three cell lines, with 50% inhibiting concentrations (IC(50)) of 24.948, 31.569 and 6.923 µg/mL, respectively. In vivo studies showed that the total flavonoids had dose-dependent inhibitory effects on hepatocellular carcinoma in mice. CONCLUSION: The total flavonoids from Xianhe Yanling Recipe have potential anticancer activity, and further researches and development are warranted.