OBJECTIVETo identify the endophyte strain E8 with high activity from Curcuma wenyujin and study its secondary metabolites.

METHODThe strain E8 was identified by morphological observation and ITS sequence analysis. Manifold chromatographic methods were used to separate and purify the chemical constituents of fermentation broth from strain E8, and their structures were identified by physiochemical properties and spectral data.

RESULTThe strain E8 belongs to P. oxalicum. Four compounds were isolated from the fermentation broth of this strain and elucidated as chrysophanol, emodin, secalonic acid A and beta-sitosterol.

CONCLUSIONThe endophyte P. oxalicum was isolated from medicinal plant Curcuma wenyujin for the first time. Four compounds were first isolated from endophytic fungus in C. wenyujin. Thus, microbial fermentation is a new access for these compounds production.

Anthraquinones ; analysis ; Curcuma ; microbiology ; Emodin ; analysis ; Fermentation ; Penicillium ; genetics ; isolation & purification ; metabolism ; Sitosterols ; analysis ; Xanthones ; analysis

AIM: To develop and validate a high performance liquid chromatography (HPLC) coupled with diode array and evaporative light scattering detectors (DAD-ELSD) method for the quantitative determination and fingerprint analysis of ten active constituents in three chemical classes (namely, xanthone glycosides, steroidal saponins, and alkaloids) in Zhimu-Huangbai herb pair (ZB). METHOD: Chromatographic separation was performed on a Diamonsil C18 column (4.6 mm × 250 mm, 5 μm, Dikma) by gradient elution using acetic acid in acetonitrile solution at a flow rate of 1.0 mL·min(-1) at 260 nm. The drift tube temperature of ELSD was set to 60 °C and nebulizer gas pressure was 4.0 Bar. Method validation was performed to assure its linearity, limits of detection and quantification, precision, repeatability, stability, and accuracy. RESULTS: The HPLC-DAD-ELSD method allowed the quantification of ten compounds (phellodendrine, jatrorrhizine, palmatine, berberine, neomangiferin, mangiferin, timosaponin E-I, timosaponin B-II, timosaponin B, and timosaponin A-III), and was successfully applied to fingerprint analysis for ten batches of ZB samples. CONCLUSION This was the first time to apply the combination of DAD and ELSD for the simultaneous determination of ten active ingredients in ZB. The results showed that the combination of quantitative analysis for marker ingredients and chemical fingerprint for the TCM herb pair provides a potentially powerful, widely introduced, and internationally accepted strategy for assessment of complex TCM formulas.
Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Glycosides ; analysis ; Xanthones ; analysis

An HPLC method for simultaneous determination of chlorogenic acid and mangiferin in original medicinal materials and decoction pieces of Pyrrosiae Folium was developed. The assay was performed on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column eluted with a mobile phase consisted of acetonitrile and 0.5% phosphoric acid solution in gradient elution at a flow rate of 1.0 mL x min(-1). The column temperature was set at 25 degrees C. The detection wavelength was 320 nm. The results showed that The linear ranges of chlorogenic acid and mangiferin were 5.2-130 mg x L(-1) (r = 0.9999) and 1.2-18 microg x mL(-1) (r = 0.9999), and the average recoveries (n=6) were 97.9% (RSD 1.9%) and 99.6% (RSD 2.9%), respectively. The method was simple, reproducible and valid. It can be used for quality evaluation and control of original medicinal materials and decoction pieces of Pyrrosiae Folium.
Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Xanthones ; analysis

The xanthones in the ethyl acetate extract of Comastoma pedunlulatum were investigated. The chromatographic and spectroscopic techniques were used to isolate and identify the constituents. Nine xanthones were isolated from the active parts of the ethyl acetate portion of the 70% ethanolic extract of C. Pedunlulatum, which possess the protective activity against hepatocyte damage caused by DL-GalN, and identified as 1,8-dihydroxy-2,6-dimethoxyxanthone (1), 8-hydroxy-1,2,6-trimethoxyxanthone (2), 1,6,8-trihydroxy-2-methoxyxanthone (3), 1,8-dihydroxy-3,5-dimethoxyxanthone (4), 1-hydroxy-3,5,8-trimethoxyxanthone (5), 1 -hydroxy-3,7-dimethoxyxanthone (6), 1,2,6,8-tetrahydroxyxanthone (7), 1,3,7-trihydroxy4- methoxyxanthone (8), 6,8-dihydroxy-1, 2-dimethoxyxanthone (9). Among them, compounds 6-9 were isolated from the genus Comastoma for the fist time.
Gentianaceae ; chemistry ; Plant Extracts ; analysis ; isolation & purification ; Xanthones ; analysis ; isolation & purification

OBJECTIVETo establish a method for determing the content of two isomers containd in Garcinia hanburyi by HPLC.

METHODChromatographic column of SunFire (Waters) C8 (2.1 mm x 150 mm, 3.5 microm) was adopted, with acetonitrile-methanol-0.3% trifluoroacetic acid (36: 37:27) as the mobile phase. The detection wavelength was 360 nm,the flow rate was 0.3 mL x min(-1), and the column temperature was 28 degrees C.

RESULTThe linear regression equation of r-gambogic acid was Y = 2.87 x 10(6) X - 2.24 x 10(5), r = 0.999 9. The linear regression equation of S-gambogic acid was Y = 3.31 x 10(6) X - 1.44 x 10(5), r = 0.999 9. The average recoveries were 100.0% and 100.9%, with RSD being 2.1% and 2.5% (n = 6), respectivley. The average contents of two gambogic acid in G. hanburyi were 30.06% and 21.45%, respectively.

CONCLUSIONThe method was so convenient and stable that it can be used for identification and content determination of two isomers containd in G. hanburyi.

Chromatography, High Pressure Liquid ; methods ; Garcinia ; chemistry ; Isomerism ; Linear Models ; Xanthones ; analysis

The chemical constituents from the stems and leaves of Cratoxylum cochinchinense were isolated and purified using silica gel, ODS gel, and Sephadex LH-20 gel column chromatography, as well as preparative HPLC. The chemical structures of all isolated compounds were identified on the basis of their physicochemical properties, spectroscopic analyses, and the comparison of their physicochemical and spectroscopic data with the reported data in literature. As a result, 21 compounds were isolated from the 90% ethanol extract of the stems and leaves of C. cochinchinense, which were identified as cratocochine(1), 1-hydroxy-3,7-dimethoxyxanthone(2), 1-hydroxy-5,6,7-trimethoxyxanthone(3), ferrxanthone(4), 3,6-dihydroxy-1,5-dimethoxyxanthone(5), 3,6-dihydroxy-1,7-dimethoxyxanthone(6), 1,2,5-trihydroxy-6,8-dimethoxyxanthone(7), securixanthone G(8), gentisein(9), 3,7-dihydroxy-1-methoxyxanthone(10), pancixanthone B(11), garcimangosxanthone A(12), pruniflorone L(13), 9-hydroxy alabaxanthone(14), cochinchinone A(15), luteolin(16), 3,5'-dimethoxy-4',7-epoxy-8,3'-neolignane-5,9,9'-triol(17), N-benzyl-9-oxo-10E,12E-octadecadienamide(18), 15-hydroxy-7,13E-labdadiene(19), stigmasta-4,22-dien-3-one(20), and stigmast-5-en-3β-ol(21). Among these isolates, compound 1 was a new xanthone, compounds 2-5, 7, 8, 12, and 16-21 were isolated from the Cratoxylum plant for the first time, and compounds 11 and 13 were obtained from C. cochinchinense for the first time. Furthermore, all isolated compounds 1-21 were appraised for their anti-rheumatoid arthritis activities by MTS method through measuring their anti-proliferative effect on synoviocytes in vitro. As a result, xanthones 1-15 displayed notable anti-rheumatoid arthritis activities, which showed inhibitory effects on the proliferation of MH7A synoviocytes with the IC_(50) values ranging from(8.98±0.12) to(228.68±0.32) μmol·L~(-1).
Synoviocytes ; Clusiaceae/chemistry* ; Xanthones/analysis* ; Plant Leaves/chemistry* ; Cell Proliferation ; Arthritis

A HPLC method was developed for simultaneous determination of 1-hydroxy-2, 3, 4, 7-trimethoxyxanthone (1), 1-hydroxy-2,3, 7- trimethoxyxanthone (2), 1-hydroxy-2, 3, 4, 5-trimethoxyxanthone (3), and 1-hydroxy-2, 3, 5- trimethoxyxanthone (4) in Halenia elliptica. The analytical column was Welchrom C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase was acetonitrile- water (43:57). The detection wavelength was 265 nm. The flow rate was 1 mL x min(-1) and the column temperature was set at 40 degrees C. There was good linearity between the peak areas and concentration at the ranges of 0.414-16.6, 1.73-69.6, 5.89-117, 3.01-120.5 mg x L(-1) for 1, 2, 3 and 4 respectively. The average recoveries (n = 6) of 1, 2, 3 and 4 were 102.5%, 100.5%, 97.9% and 101.2%. Those four xanthones in thirty samples of H. elliptica. were determined by this method. The method is simple, accurate, repeatable, which could be used for the quality evaluation of H. elliptica. The total content of those four xanthones in H. elliptica should not less than 1.80% by comprehensive analysis.
Chromatography, High Pressure Liquid ; methods ; Gentianaceae ; chemistry ; Plant Extracts ; analysis ; isolation & purification ; Tibet ; Xanthones ; analysis ; isolation & purification

OBJECTIVETo develop an HPLC method for the quantification of six active components in three species (Swertia davidi, S. nervosa and S. mussotii) .

METHODThe determination was performed on a Hypersil BDS colunm (4. 6 mm x 200 mm, 5 microm). Acetonitrile and 0.5% phosphoric acid solution were used as the mobile phases with a gradient elution. The flow rate was 1.0 mL x min(-1). The UV detection wavelength was at 240, 274, 325 and 334 nm. The column oven temperature was at 25 degrees C.

RESULTSix components were separated commendably in 60 minutes. The calibration curves of swertiamarin, gentiopicroside, norswertianolin, swertianolin, demethylbellidifolin and bellidifolin were in good linearity over the range of 0.520-20.8, 0.202-8.06, 0.107-4.28, 0.097-3.86, 0.094-3.77, 0.101-4.02 microg, respectively (r = 0.999 9). The average recoveries were 98.7%, 98.1%, 98.3%, 98.8%, 98.1% and 98.6%, respectively, and the RSD were less than 3.0% (n = 6).

CONCLUSIONThe method is accurate,simple and reproducible, and can be used to control the quality of Swertia.

Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Iridoid Glucosides ; Iridoids ; analysis ; Pyrones ; analysis ; Swertia ; chemistry ; Xanthones ; analysis

OBJECTIVETo develop a new method for determination of Mangiferin in the leafs of Folium mangiferae. By this new method, Mangiferin in F. mangiferae sampled in different months and in different regions was determinated.

METHODA RP-HPLC method was set up, using Shim pack CLC-ODS column, methanol-0.05 mol.L-1 H3PO4(65:134, pH 3.5) as mobile phase, with 258 nm of detection wave, at room temperature, 1 mL.min-1. F. mangiferae sampled in Nanning, Qinzhou and Tianyang, Guangxi province and sampled respectively in January to December were determinated.

RESULTThe average recovery of the RP-HPLC was 99.2%, RSD = 1.05% (n = 5). The content of Mangiferin in F. mangiferae was statistically different due to different sample-regions or sample-time.

CONCLUSIONThis RP-HPLC method is simple, specific and exact. The contents of Mangiferin in the leafs of F. mangiferae sample in Nanning and Tinayang were statistically similar, but higher than that in Qinzhou. The contents of Mangiferin in the leafs of F. mangiferae sampled in July to October were higher than that in the other months. The content in September was the highest, the content in February was the lowest.

China ; Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Mangifera ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Seasons ; Xanthones ; analysis

This study is to develop a sensitive method by using reversed-phase high performance liquid chromatography coupled with UV detector (HPLC-UV) to simultaneously determine four bioactive compounds, iriflophenone 3-C-beta-D-glucoside, iriflophenone 3,5-C-beta-D-diglucoside, mangiferin, and iriflophenone 2-O-alpha-L-rhamnoside in the leaves of Aquilaria sinensis. An Agilent Zorbax SB-C, column (4, 6 mm x 250 mm, 5 microm) was used, and the gradient elution was performed with mobile phase of 0.1% aqueous phosphoric acid and acetonitrile at a flow rate of 1 mL x min(-1). The detection wavelength was 280 nm, and the column temperature was 25 degrees C. The four marker compounds were well separated with good linearity (R2 > 0.9990), precision, stability and repeatabili y. The-recovery rates were in the range of 98.80%-101.39%. For 15 branch of the leaves, the contents of iriflophenone 3-C-beta-D-gluoside, iriflophenone 3,5-C-beta-D-diglucoside, mangiferin, and iriflophenone 2-O-alpha-L-rhamnoside were between 0.41-14.48, 0.72-3.85, 4.30-29.07, 0.24-5.06 mg, respectivley. This method is precise, accurate and reliable, which provides an efficient way for the quality control of the leaves of A. sinensis. This will promote the comprehensive usage of this plant.
Benzophenones ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry ; Spectrophotometry, Ultraviolet ; methods ; Thymelaeaceae ; chemistry ; Xanthones ; analysis