Quality evaluation of Commelina communis from different origins based on multicomponent content determination combined with chemometrics / 中国中药杂志

This research established a high-performance liquid chromatography(HPLC) method for simultaneous determination of isoorientin, orientin, vitexin, and isovitexin in Commelina communis to conduct content difference analysis and quality evaluation of 62 batches of C. communis from different origins. The HPLC content determination was performed on a Dikma Platisil ODS chromatographic column(4.6 mm×250 mm, 5 μm), with acetonitrile-0.1% formic acid(14∶86) as the mobile phase. The detection wavelength was set at 348 nm, the flow rate was 1.0 mL·min~(-1), and the column temperature was 35 ℃. The differences in origins and quality of 62 batches of C. communis were studied by chemometrics. The results showed that the determination of four components mani-fested a good linear relationship in the range of mass concentration(r>0.999 9), and the average recovery rate was 96.17%-103.0%. The relative standard deviations(RSDs) of precision, stability, and repeatability were all less than 2.0%. The content of four components from high to low was isoorientin>isovitexin>orientin>vitexin. Forty-seven batches of C. communis with clear origins were classified into six categories by chemometrics. C. communis from different origins had different qualities. Generally, C. communis from Western China, Central China, and South of China had superior qualities. The HPLC method established in this study is specific, simple, and efficient, which provides references for the comprehensive evaluation of the quality of C. communis. The chemometrics shows that the qualities of C. communis from different origins are largely different. Isoorientin can be used as an index to determine the content of C. communis, and its content limit should be set no less than 0.023%.

HPLC fingerprint of Lomatogonium rotatum / 中草药

Objective To study the fingerprints of 15 batches of Mongolian medicinal materials of Lomatogonium rotatum collected from different places by HPLC, and to evaluate the quality of the medicinal materials of L. rotatum by similarity calculation. Methods A total of 15 batches of ribbed flowers were collected by HPLC. Chromatographic column: YMC C18 column (250 mm × 4.6 mm, 5 μm), mobile phase: acetonitrile-0.4% phosphoric acid-methanol, gradient elution, flow rate was 0.8 mL/min, detection wavelength was 254 nm, column temperature was 30 ℃. The similarity of fingerprints was evaluated by using the “Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine 2004A Edition”. Results The fingerprints of L. rotatum of Mongolian medicine were established, 15 common peaks were identified, five common peaks were identified, and the similarity among 15 batches of L. rotatum and the fingerprints of control was in the range of 0.881—0.997. The fingerprints of erect column flowers have good precision, stability and reproducibility. Conclusion The characteristic fingerprint of L. rotatum was established for the first time, which not only provides a new scientific basis for the identification and quality control of L. rotatum, but also has important significance for the quality evaluation of L. rotatum.

Contents of seven indicative constituents in seven kinds of Gentiana macrophylla by UPLC / 中草药

Objective: An ultra performance liquid chromatographic (UPLC) method was developed for simultaneously determining seven constituents, such as loganic acid, swertiamarin, 6'-O-β-D-glucosyl gentiopicroside, gentiopicroside, sweroside, isoorientin, and isovitexin, from the medicinal plants of Gentiana macrophylla. Methods: The separation was performed on an Acquity UPLC® BEH C18 column (50 mm ×2.1 mm, 1.7 μm) through a gradient elution of methanol-0.04% aqueous phosphorite at a flow rate of 0.3 mL/min. The detection wavelength was 242 nm, and the column temperature was set at 30℃. Results: For the seven analytes, loganic acid, swertiamarin, 6'-O-β-D-glucosyl gentiopicroside, gentiopicroside, sweroside, isoorientin, and sovitexin, a good linearity (r≥0.9995) was obtained in the range of 2.100-537.100, 1.050-270.000, 0.920-236.000, 11.100-2830.000, 0.750-192.000, 0.167-102.000, and 0.216-52.800 μg, respectively. Their average recoveries (n=6) were 97.83%-100.08%, respevtively, with RSD values less than or equal to 3.76%. Conclusion: The UPLC method is simpler and more effective than HPLC, and can be used for the simultaneous determination of seven indicative constituents in medicinal plants of G. macrophylla.

Study on flavonoid glycosides from Gentiana veitchiorum / 中草药

Objective: To study the flavonoid glycosides from the aerial parts of Gentiana veitchiorum. Methods: The compounds were isolated and purified by silica gel and ODS chromatography in addition to p-HPLC. The structures of the isolated compounds were identified on the basis of 1D & 2D NMR spectroscopy methods. Results: Six compounds were isolated from the aerial parts of G. veitchiorum and identified as 5,7,3',4'-tetrahydroxyl flavone-6-C-β-D-glucopyranosyl-4'-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (1), isoorientin (2), isovitexin (3), isoscoparine (4), isoorientin-4'-O-glucoside (5), and isosaponarin (6). Conclusion: Compound 1 is a new compound named isoorientin-4'-diglucoside, and compounds 5 and 6 are isolated from this plant for the first time.

Analysis of main active components in Desmodii Styraciflii Herba by HPLC-Q- TOF-MS and HPLC-DAD / 中草药

Objective: This study is aimed to establish a high performance liquid chromatography coupled with quadrupole-time-of- flight mass spectrometric (HPLC-Q-TOF-MS) method for analyzing the main active components in Desmodii Styraciflii Herba, and to establish an HPLC-DAD method for simultaneously determining its seven flavonoids compounds (schaftoside, isoschaftoside, vicenin-2, isoorientin, isovitexin, luteolin, and apigenin). Methods: The analysis was performed on a Phenomenex Kinetex C18 column with a gradient elution of methanol-0.2% aqueous formic acid at the flow rate of 1.0 mL/min. The column temperature was 40 oC. The Q-TOF-MS discriminant analysis was performed under negative electrospray ion mode and the split ratio was 1∶1. Quantitative analysis of seven compounds in Desmodii Styraciflii Herba was carried by HPLC-DAD. The determination wavelength was at 272 nm. Results: Forty-one main active constituents were separated and identified by HPLC-Q-TOF-MS, including 37 flavonoids compounds and four phenolic acids. A HPLC-DAD method was successfully developed for determining seven flavonoids compounds. The standard curves for all seven compounds showed good linearity with correlation coefficients higher than 0.999 0 within the test ranges. The average recoveries (n = 6) were between 96.7%-102.4%, and RSD ≤ 4.94%. Conclusion: Analyzing the main active components provides a significant guidance for the substance basis research of Desmodii Styraciflii Herba. The HPLC-DAD method for simultaneously determining seven flavonoids compounds is accurate, reproducible, and provides the basis for the quality control of Desmodii Styraciflii Herba.

Chemical constituents from Vitex negundo var. heterophylla / 中草药

Objective: To study the chemical constituents in the twigs of Vitex negundo var. heterophylla. Methods: A variety of silica gel column chromatography, Sephadex LH-20 gel column chromatography, and HPLC methods were used for the separation and purification of chemical composition. Their structures were identified on the basis of physicochemical property and spectral data. Results: Eight compounds were obtained and identified as apigenin-6-C-β-D-glucopyranosyl 8-C-α-L-arabinopyranoside (1), isoorientin (2), luteolin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (3), luteolin-6,8-di-C-α-L-arabinopyranoside (4), apigenin-6-α-L- arabinopyranosyl-8-C-β-D-glucopyranoside (5), luteolin-6-C-α-L-arabinopyranosyl 8-C-β-L-arabinopyranoside (6), luteolin-6-C-β-L- arabinopyranosyl-8-C-β-D-glucopyranoside (7), and luteolin-7-O-β-D-glucopyranoside (8). Conclusion: Compounds 1 and 3-7 are first isolated from the plants of Vitex Linn.

Effect of Gypsophila elegans isoorientin on proliferation and apoptosis of human HepG2 cells / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect and underlying mechanism of Gypsophila elegans isoorientin on the proliferation and apoptosis of human HepG2 cells. METHODS HepG2 cells were treated with isoorientin 5,10,20,40,80 and 160μmol?L-1 for 24,48 and 72 h,respectively. Cell survival was analyzed by MTT assay. HepG2 cells were treated with isoorientin 5,10 and 20μmol?L-1 for 48 h before the lactate dehydrogenase(LDH)level was detected. After treatment with isoorientin for 24 h, the variation of reactive oxygen species (ROS) was monitored by a fluorescence probe H2DCF-DA. HepG2 apoptosis and mitochondria membrane potential(MMP)were evaluated by flow cytometry. The activi?ties of caspase 3,8 and 9 were determined by colorimetry. The mRNA expression of Bcl-2 and Bax was determined by RT-PCR,and the protein expression of Bcl-2, Bax and cytochrome c was detected by Western blotting. RESULTS Isoorientin(5-160μmol?L-1)inhibited HepG2 cell survial in a concen?tration-dependent manner,the 50%inhibitory concentration(IC50)was 62.7±9.1,47.2±11.4 and(18.2± 7.5)μmol?L-1 after treatment with isoorientin for 24,48 and 72 h,respectively. Compared with the cell control group,treatment with isoorientin 5,10 and 20μmol?L-1 significantly increased the LDH level and cell apoptosis rate(P<0.05). Moreover,isoorientin 10 and 20μmol?L-1 notably increased the production of ROS,decreased the MMP(P<0.05),and increased the activities of caspase 3 and 9. RT-PCR analysis and Western blotting showed that isoorientin significantly decreased the mRNA and protein expressions of Bcl-2,decreased the mRNA and protein expressions of Bax(P<0.05),and inhibited the protein expression of cytochrome c(P<0.05). CONCLUSION Isoorientin inhibits HepG2 cell prolif?eration,but promotes cell apoptosis,which is closely related to the regulation of the mitochondrial apoptosis pathway.

Determination of seven components in flowers of Gentiana tibetica and Gentiana dendrologi by HPLC / 中草药

Objective: To establish an HPLC method for simultaneously determining seven components, such as loganic acid, 6′-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, swertiamarin, isoorientin, and isovitexin, from the flowers of Gentiana tibetica and G. dendrology. Methods: Chromatographic analysis was achieved on an Agilent Zorbax ODS column (150 mm × 4.6 mm, 5 μm) by gradient elution of acetonitrile-0.4% acetic acid in water at 30℃. The flow rate was 0.4 mL/min and the detection wavelength was 254 nm. Results: The calibration curves of all the seven constituents showed good linearity in a relatively wide concentration range. The linear ranges of loganic acid, 6′-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, gentiopicroside, isoorientin, and isovitexin were 0.228-2.280, 0.680-6.800, 0.220-2.20, 1.476-14.760, 0.200-2.000, 0.436-4.360, and 0.276-2.760 μg (R2 ≥ 0.999 2), respectively. The recoveries (n = 9) of loganic acid, 6′-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, gentiopicroside, isoorientin, and isovitexin were 100.9%, 99.4%, 101.0%, 105.7%, 103.1%, 98.4%, 104.2%. Conclusion: This method is simple, accurate, and specific, and can be used for the determination of seven constituents in the flowers of G. tibetica and G. dendrologi.

Chemical constituents from flower of Gentiana tibetica / 中草药

Objective: To investigate the chemical constituents in flowers of Gentiana tibetica. Methods: The chemical constituents were isolated from the 95% alcohol extract of G. tibetica flowers by silica column chromatography, Sephadex LH-20, C18 and RP-HPLC. Their chemical structures were elucidated on the basis of IR, ESI-MS, 1H-MNR, and 13C-MNR spectroscopic data. Results: Twelve compounds, including chromene, flavones C-glycosides, secoiridoid glycosides, iridoid glycosides, aliphatic alcohol, and disaccharide, were obtained from the 95% alcohol extract of G. tibetica flowers, Their structures were identified as macrophylloside D (1), orientin 7-caffeate (2), 7-O-feruloylorientin (3), isovitexin (4), saponarin (5), isoorientin (6), 6'-O-β-D-glucopyranosyl gentiopicroside (7), sweroside (8), swertiamarine (9), loganic acid (10), 1-heneicosanol (11), and sucrose (12). Conclusion: Compounds 1-12 are isolated from G. tibetica for the first time.

Effects of isoorientin on autophagy of gastric cancer cell / 中国生化药物杂志

Objective To explore effect of isoorientin on gastric cancer cell autophagy.Methods Isoorientin and autophagy activator and effect of inhibitors on proliferation of human gastric cancer cell line SGC-7901 by MTT assay.Cell apoptosis was detected by flow cytometry.Production of autophagy lysosomal in gastric cancer SGC-7901 cells was obseroved by AO and MDC staining.Characteristic expression of autophagy protein was analysed by Western blot.Results MTT assay indicated that isoorientin could inhibit gastric cancer cell growth, RAP has the same effects, but 3-MA inhibition of apoptosis.Flow cytometry was used to detect the apoptosis rate of isoorientin and RAP could promote cell apoptosis , while 3-MA had the opposite effect.In AO staining, the red light appeared in the cells, and the green fluorescence appeared in the cells of MDC staining, which showed that there was an autophagy in the cells.Western blot test found the isoorientin was cell autophagy specific protein LC-3II, Beclin-1 expression increased, 3-MA suppressed the expression of the two proteins, and RAP had the opposite effect.Conclusion Isoorientin could induce apoptosis in gastric cancer SGC-7901 cells, autophagy is involved in the process of death.

Seasonal variations in the amount of isoorientin and isovitexin in Cecropia glaziovii Sneth. leaves over a two-year period / Variación estacionaria en el contenido de isoorientina e isovitexina en hojas de Cecropia glaziovii Sneth. durante un período de dos años

Cecropia glaziovii Sneth. (Urticaceae) is a common tree from Southeast and South of Brazil, being widely used in traditional medicine to treat heart and respiratory conditions. C-glycosylflavonoids have being described as the major compounds for this genus, however, no seasonality studies of individual flavonoids was conducted for any Cecropia specie. In this work, the content of isoorientin and isovitexin in aqueous extract from the leaves of C glaziovii during a two-year period was determined by high-performance liquid chromatography with diode array detector (HPLC-DAD). Seasonal alterations in the content of these two majority C-glycosylflavonoids as well its possible correlation with the pluviosity in the period of January/2008 to January/2010 were determined. Isoorientin presented higher content in November/09 (6.04 mg/g of extract) and lower content in May/08 (1.01 mg/g of extract). The higher content of isovitexin was observed in March/09 and the lower in September/08 (11.42 and 4.47 mg/g of extract, respectively). Although they have distinct behaviors, it was not observed correlation between the values of pluviosity and the production of these C-glycosylflavonoids.

Cecropia glaziovii Sneth. (Urticaceae) es un árbol comúnmente encontrada en el Suroriente y Sur de Brasil, siendo ampliamente utilizada en la medicina tradicional para el tratamiento de problemas cardíacos y respiratorios. Flavonoides C-glucósidos vienen siendo descritos como los compuestos mayoritarios del género, sin embargo, ningún estudio de estacionalidad de flavonoides analizados de modo individual fue realizado con ninguna especie de Cecropia. En el presente trabajo, el contenido de isoorientina e isovitexina en el extracto acuoso de las hojas de C. glaziovii durante un período de dos años fue determinado por HPLC-DAD. Variaciones estacionarias en el contenido de esos dos flavonoides C-glucósidos así como la posible correlación con la pluviosidad en el periodo de Enero/2008 hasta Enero/2010 fueron determinados. Isoorientina presentó mayor contenido en Noviembre/09 (6,04 mg/g de extracto) y menos contenido en Mayo/08 (1,01 mg/g de extracto). El mayor contenido de isovitexina fue observado in Marzo/09 y el menor contenido in Septiembre/08 (11,42 y 4,47 mg/g de extracto, respectivamente). Aunque los flavonoides poseen distintos comportamientos, no fue observada correlación entre los valores de pluviosidad y la producción de esos compuestos.

Isolation of isoorientin and orientin from bamboo leaf flavones using preparative high-performance liquid chromatography / 中草药

Objective: To establish a method for preparation of orientin and isoorientin from bamboo leaf flavones (BLF) using preparative high-performance liquid chromatography (PHPLC). Methods: After BLF was purified by AB-8 resin-based column chromatography, PHPLC was employed to isolate orientin and isoorientin, using a mobile phase consisting of methyl alcohol-water with 0.3% acetic acid (32:68). The flow rate was 5 mL/min while the column temperature was maintained at 20℃. The injection volume was 400 μL and the concentration of sample was 15.3 mg/mL. Signal was monitored at 330 nm with the diode array detector. Results: HPLC analysis showed that the purities of isoorientin and orientin were >99% and >93%, respectively, and the recovery rates were (92.7±2.92)% and (85.1±3.45)%, respectively. The confirmation of chemical structures was performed by 1H-NMR and 13C-NMR. Conclusion: The method is simple, with low toxicity but high recovery rate, and can be applied to the separation and preparation of isoorientin and orientin.

Chemical constituents in leaves of Vaccinium bracteatum and their anti-complementary activity / 中草药

Objective: To study the chemical constituents in the leaves of Vaccinium bracteatum and their anti-complementary activity. Methods: The isolation was guided by anti-complementary activity test, compounds were isolated and purified by silica gel and Sephadex LH-20 column chromatographies as well as preparative HPLC. The structures were identified by the various spectroscopic data. The compounds were evaluated for the anti-complementary activity in vitro. Results: Twenty-eight compounds were isolated and identified as pentatriacontane (1), hentriantane (2), glutinone (3), friedelin (4), epifriedelanol (5), lupeol (6), oleanolic acid (7), ursolic acid (8), scopoletin (9), trans-p-hydroxycinnamic acid (10), chrysoeriol (11), apigenin (12), kaempferol (13), maslinic acid (14), corosolic acid (15), euscaphic acid (16), 2α, 3α-dihydroxyurs-12-en-oic acid (17), 19α-hydroxyoleanolic acid (18), caffeic acid (19), isoorientin (20), orientin (21), vitexin (22), isovitexin (23), quercetin-3-O-α-L-rhamnoside (24), isoquerecitrin (25), chrysoeriol-7-O-β-D-glucopyranoside (26), quceritin (27), and luteolin (28). Among them 25 compounds were tested for their anti-complementary activity and 5, 7, 8, 11-13, 15, 18-20, 23-25, 27, and 28 show the various inhibitions on the complement system towards the classical pathway. Conclusion: Compounds 1, 3, 6, 14-18, 23, 25 are obtained from this plant for the first time. Ursolic acid shows the strongest activity in vitro with the CH50 of 0.014 mg/mL.

Determination of orientin,isoorientin,isovitexin in Bamboo Leaf from different sources by HPLC / 中成药

AIM: To determine the contents of orientin,isoorientin,isovitexin in Bamboo Leaf from different sources. METHODS : Orientin,isoorientin,isovitexin were determined by HPLC. The mobile phase was MeOH-H_2O-HAc (35∶65∶1). The detective wavelength was at 340nm. RESULTS : The highest content of three favonoids was 0.89% and the lowest content was 0.07% from different sources. CONCLUSION : This method is simple,quick and accurate for determination of the three favonoids in Bamboo Leaf from different sources.