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

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

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

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

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

A HPLC method was developed for the determination of mangiferin in rat plasma and aqueous humor. 4-Nitrophenol was used as internal standard. Analysis was performed on a Cosmosil ODS C18 analytical column (4.6 mm x 250 mm, 5 microm) with mobile phase consisting of methanol-water (40: 60) with 2% glacial acetic acid at a flow rate of 1.0 mL x min(-1). The calibration curve of mangiferin in rat plasma and aqueous humor showed excellent linear behaviors over the investigated concentration of 0. 50-250.00 mg x L(-1) in plasma and 0.10-10.00 mg x L(-1) in aqueous humor, respectively, and the correlation coefficients were all above 0.995 4. The intra-day and inter-day precisions for all samples were measured to be below 12%. The limit of quantitation was 0.10 mg x L(-1) and low enough for the determination of mangiferin in all samples. The validated method has been successfully applied to preliminary pharmacokinetics study of mangiferin in rat plasma and aqueous humor.
Animals ; Aqueous Humor ; chemistry ; Chromatography, High Pressure Liquid ; Female ; Male ; Nitrophenols ; analysis ; Rats ; Rats, Wistar ; Reproducibility of Results ; Sensitivity and Specificity ; Xanthones ; blood ; pharmacokinetics

OBJECTIVETo study the chemical constituents of Halenia elliptica.

METHODThe air-dried whole plants of Halenia elliptica were extracted with 90% EtOH. The EtOH extract was condensed to a small amount of volume and extracted with petroleum ether, EtOAc and n-BuOH, successively. The compounds were isolated and purified by column chromatography from the EtOAc fraction, and identified based on spectral analyses (MS, 1H-NMR, 13C-NMR).

RESULT12 compounds were isolated from H. elliptica, and characterized as 8-hydroxy-2-methylchromone (1), 5-methoxy-2-methylchromone (2), 7-epi-vogeloside (3), coniferl aldehyde (4), sinapaldehyde (5), norbellidifolin (6), 1-hydroxyl-2,3,4,6-tetramethoxyxanthone (7), 1-hydroxyl-2,3,4,7-tetramethoxyxanthone (8), 1-hydroxyl-2,3,5-trimethoxyxanthone (9), together with azelaic acid, beta-sitosterol, and oleanolic acid.

CONCLUSIONCompounds 1, 2 were new natural compounds and compounds 3-6, 10 were obtained from H. elliptica for the first time and compound 6 showed inhibitory activities against HBsAg and HBeAg secretion with IC50 value of 0.77 and < 0.62 mmol x L(-1), respectively.

Acrolein ; analogs & derivatives ; analysis ; isolation & purification ; Chromatography ; Dicarboxylic Acids ; analysis ; isolation & purification ; Gentianaceae ; chemistry ; Iridoid Glycosides ; isolation & purification ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Oleanolic Acid ; analysis ; isolation & purification ; Plant Extracts ; isolation & purification ; Sitosterols ; analysis ; isolation & purification ; Xanthones ; analysis ; isolation & purification

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

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

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