Fast preparative separation of flavones from capitula of Eriocaulon ligulatum (Vell.) L.B.Smith (Eriocaulaceae) by High-speed countercurrent chromatography (HSCCC)

High-speed countercurrent chromatography (HSCCC) is a leading method for the fast separation of natural products from plants. It was used for the preparative isolation of two flavone monoglucosides present in the capitula of Eriocaulon ligulatum (Veil.) L.B.Smith (Eriocaulaceae). This species, known locally as botão-dourado, is exported to Europe, Japan and North America as an ornamental species, constituting an important source of income for the local population of Minas Gerais State, Brazil. The solvent system, optimized in tests prior to the HSCCC run, consisted of the two phases of the mixture ethyl acetate: n-propanol: water (140:8:80, v/v/v), which led to the successful separation of 6-methoxyluteolin-7-O-β-D-allopyranoside and 6-methoxyapigenin-7-O-β-D-allopyranoside in only 3 hours. The two flavonoids were identified by NMR (1-D and 2-D) and ESI-MS, comparing their spectra with published data.

An efficient combination of supercritical fluid extraction and high-speed counter-current chromatography to extract and purify homoisoflavonoids from Ophiopogon japonicus (Thunb.) Ker-Gawler

Supercritical fluid extraction (SFE) was used to extract homoisoflavonoids from Ophiopogon japonicus (Thunb.) Ker-Gawler. The optimization of parameters was carried out using an orthogonal test L-9 (3)(4) including pressure, temperature, dynamic extraction time and the amount of modifier. The process was then scaled up by 100 times with a preparative SFE system under the optimized conditions of 25 MPa, 55 degrees C, 4.0 h and 25% methanol as a modifier. Then crude extracts were separated and purified by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane/ethyl acetate/methanol/ACN/water (1.8:1.0:1.0:1.2:1.0 v/v). There three homoisoflavonoidal compounds including methylophiopogonanone A 6-aldehydo-isoophiopogonone A, and 6-formyl-isoophiopogonanone A, were successfully isolated and purified in one step. The collected fractions were analyzed by HPLC. In each operation, 140 mg crude extracts was separated and yielded 15.3 mg of methylophiopogonanone A (96.9% purity), 4.1 mg of 6-aldehydo-isoophiopogonone A (98.3% purity) and 13.5 mg of 6-formyl-isoophiopogonanone A (97.3% purity) respectively. The chemical structure of the three homoisoflavonoids are identified by means of ESI-MS and NMR analysis.

One step isolation and purification of liquiritigenin and isoliquiritigenin from Glycyrrhiza uralensis Risch using high-speed counter-current chromatography

High-speed counter-current chromatography (HSCCC) technique in semi-preparative scale has been successfully applied to the separation of bioactive flavonoid compounds, liquiritigenin and isoliquiritigenin in one step from the crude extract of Glycyrrhiza uralensis Risch. The HSCCC was performed using a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-acetonitrile-water (2:2:1:0.6:2, v/v). Yields of liquiritigenin (98.9% purity) and isoliquiritigenin (98.3% purity) obtained were 0.52% and 0.32%. Chemical structures of the purified liquiritigenin and isoliquiritigenin were identified by electrospray ionization-MS (ESI-MS) and NMR analysis. (c) 2005 Published by Elsevier B.V.

Mutagenic activity promoted by amentoflavone and methanolic extract of Byrsonima crassa Niedenzu

Byrsonima crassa is a plant pertaining to the Brazilian central savannah-like belt of vegetation and popularly used for the treatment of gastric dysfunctions and diarrhoea. The methanol extract contains catechin, tannins, terpenes and flavonoids; both mutagenic potential and antioxidant properties have been ascribed to flavonoids. The mutagenicity of some flavonoids is believed to be associated with the formation of reactive oxygen species and seems to depend on the number and position of hydroxyl groups. In the present study the mutagenic activity of the methanol, chloroform and 80% aqueous methanol extracts, as well as acetate and aqueous sub-fractions, of this medicinal plant were evaluated by Salmonella typhimurium assay, using strains 100, TA98, TA102 and TA97a, and in mouse reticulocytes. The results showed mutagenic activity of the methanolic extract in the TA98 strain without S9, but no mutagenicity to mouse cells in any of the extracts. The acetate fraction showed strong signs of mutagenicity without S9, suggesting that in this enriched fraction were concentrated the compounds that induced mutagenic activity. The aqueous fraction showed no mutagenic activity. The TLC and HSCCC analyses of the acetate fraction with some standard compounds permitted the isolation of the quercetin-3-O-beta-D-galactopyranoside, quercetin-3-O-alpha-L-arabinopyranoside, amentoflavone, methyl gallate and (+)-catechin, of which only the amentoflavone exhibited positive mutagenicity to TA98 (+S9, -S9). (c) 2006 Elsevier B.V.. All rights reserved.

Preparative separation of flavonoids from the medicinal plant Davilla elliptica St. Hill. by high-speed counter-current chromatography

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Preparative isolation of a novel flavonoid from an infusion of Byrsonima basiloba leaves by high-speed counter-current chromatography

A method for the isolation of three compounds from the infusion of leaves of Byrsonima basiloba A. Juss. by high-speed counter-current chromatography (HSCCC) was developed. This technique led to the separation of a novel compound, quercetin 3-O-alpha-L-rhamnopyranosyl-(1 -> 3)-O-[alpha-L-rhamnopyranosyl-(1 -> 6)]-beta-D-allopyranoside, and two known compounds quercetin3-O-(X-L-rhamnopyranosyl-(1 -> 6)-beta-D-galactopyranoside and (+)-catechin in 4 h with purities of over 92%. The structures of the compounds were determined by one- and two-dimensional NMR spectroscopy and HPLC.

Quick preparative separation of natural naphthopyranones with antioxidant activity by high-speed counter-current chromatography

The natural naphthopyranones paepalantine (1), paepalantine-9O-β-D-glucopyranoside (2) and paepalantine-9-O-β-D-allopyranosyl-(1→6)-O-β-D-glucopyranoside (3) were separated in a preparative scale from the ethanolic extract of the capitula of Paepalanthus bromelioides by high-speed counter-current chromatography (HSCCC). The solvent system used was composed of water-ethanol-ethyl acetate-hexane (10:4:10:4, v/v/v/v). This technique led to the separation of the three different naphthopyranone glycosides in pure form in approximately 7 hours. Paepalantine showed a good antioxidant activity when assayed by the DPPH radical spectrophotometric assay.

Separation of the toxic zierin from Zollernia ilicifolia by high speed countercurrent chromatography

Preliminary pharmacological assays of the 70% methanol extract from the leaves of the Brazilian medicinal plant Zollernia ilicifolia Vog. (Fabaceae) showed analgesic and antiulcerogenic effects. Previous analyses have shown that this extract contains, besides flavonoid glycosides and saponins, a toxic cyanogenic glycoside. Flavonoids and saponins are compounds reported in literature with antiulcerogenic activity. In this work, we developed a methodology to separate the cyanogenic glycoside from these compounds in order to obtain enough amount of material to perform pharmacological assays. The cyanogenic glycoside zierin (2S)-β-D-glucopyranosyloxy-(3-hydroxy-phenyl)- acetonitrile was separated from the other components by high speed countercurrent chromatography (HSCCC). The solvent system used was composed of chloroform-methanol-n-propanol-water (5:6:1:4, v/v/v/v). This technique led to the separation of zierin from the possible active compounds of Zollernia ilicifolia.

Compostos fenólicos e atividade antioxidante de Leiothrix flavescens (Bong.) Ruhland (Eriocaulaceae)

This paper describes a chemical investigation (by high-speed counter-current chromatography) of an extract in methanol of the capitula (flower-heads) of the endemic Brazilian herb Leiothrix flavescens (Bong.) Ruhland (Eriocaulaceae). Fractionation of this extract by preparative chromatography and identification of the isolated compounds by spectrometric methods (IR, UV, ESI-MS, NMR) led to the identification of flavones (apigenin, luteolin and 6-methoxyluteolin) and 1,3-di-O-feruloyl glycerol. The antioxidant activity of the extract was determined by DPPH reduction and the total phenolic content by the Folin-Ciocalteau assay. It was found that the methanolic extract of L. flavescens possesses strong antioxidant activity. Additionally, the chemical profile provided useful data for a discussion of the taxonomy of the Eriocaulaceae.

Bioprospecção em Eugenia jambolana (Myrtaceae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo químico e atividades mutagênica e antiulcerogênica de Syngonanthus nitens (Bong.) Ruhland

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Investigação fitoquímica e avaliação da atividade antioxidante das folhas e frutos de Eugenia jambolana Lam. (Myrtaceae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Uso sustentável da biodiversidade brasileira: prospecção químico-farmacológica em plantas superiores: Miconia spp

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Uso sustentável da biodiversidade brasileira: prospecção químico-farmacológica em plantas superiores: metodologia para estabelecimento de perfis quali e quantitativos para extratos vegetais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo químico e biológico de plantas da família Eriocaulaceae

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)