RESUMEN
Herba Cistanche (Rou Cong Rong in Chinese), dried succulent stems of Cistanche deserticola or C. tubulosa, is a famous Chinese herbal medicine and has been recorded in the Chinese Pharmacopoeia. In recent years, another two non-official species, C. salsa and C. sinensis have also been used as Herba Cistanche in some regions of China. To investigate the possibility of using these two non-official species as alternatives to the official species, a high-performance liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS) fingerprint method was developed to comparatively analyze the crude herbs of these four species. The fingerprint of C. deserticola, a historically certified species of Herba Cistanche, serves as 'standard pattern' for comparing the similarities with the other species by means of similarity and Principle Component Analysis. Additionally, 18 characteristic peaks in the fingerprints were identified by comparing their retention times, UV spectra and ESI-MS data with those of the reference substances and/or the data in the literatures. The comparative results demonstrate that the fingerprints of C. tubulosa and C. salsa possess high similarity to the standard pattern, suggesting that these two species may be used as alternative species; while that of C. sinensis has low similarity (0.053 correlation coefficient) to the standard pattern, indicating that it cannot be used as the substitute of the official herb. However, the varying fingerprint patterns among the samples of C. deserticola collected from various habitats illustrate that the quality consistency of crude herbs is still a problem worthy of serious concern.
Asunto(s)
Cromatografía Líquida de Alta Presión , Cistanche/química , Medicamentos Herbarios Chinos/química , Espectrometría de Masas , Medicamentos Herbarios Chinos/normas , Análisis de Componente Principal , Control de Calidad , Reproducibilidad de los Resultados , Especificidad de la EspecieRESUMEN
Herba Epimedii (family Berberidaceae), Yinyanghuo in Chinese, is one of commonly used Chinese medicines. Flavonoids are considered as its active components. In this study, a rapid ultra-performance liquid chromatography (UPLC) method was developed for simultaneous determination of 15 flavonoids, including hexandraside E, kaempferol-3-O-rhamnoside, hexandraside F, epimedin A, epimedin B, epimedin C, icariin, epimedoside C, baohuoside II, caohuoside C, baohuoside VII, sagittatoside A, sagittatoside B, 2''-O-rhamnosyl icariside II and baohuoside I in different species of Epimedium. The analysis was performed on Waters Acquity UPLC system with an Acquity UPLC BEH C18 column (50 mm x 2.1mm I.D., 1.7 microm) and gradient elution of 50mM acetic acid aqueous solution and acetonitrile within 12 min. All calibration curves showed good linearity (R2>0.9997) within test ranges. The LOD and LOQ were lower than 0.13 and 0.52 ng on column, respectively. The R.S.D.s for intra- and inter-day of 15 analytes were less than 5.0% at three levels, and the recoveries were 95.0-103.7%. The validated method was successfully applied to quantitatively analyze 15 flavonoids in different species of Epimedium. The results showed there were great variations among the contents of investigated flavonoids. Hierarchical clustering analysis based on characteristics of 15 investigated compounds peaks in UPLC profiles showed that 37 samples were divided into 3 main clusters, which were in accordance with their flavonoids contents. The simulative mean chromatogram of the high content cluster was generated to compare the samples from different species and/or locations of Epimedium. Four flavonoids including epimedin A, B, C and icariin were selected as markers for quality control of the species of Epimedium used as Yinyanghuo.
Asunto(s)
Cromatografía Liquida/métodos , Epimedium/química , Flavonoides/análisis , Calibración , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Presión , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrofotometría UltravioletaRESUMEN
Herba Epimedii (family Berberidaceae), Yinyanghuo in Chinese, is one of the commonly used Chinese medicines. Flavonoids are considered as its active components. In this study, a reliable pressurized liquid extraction (PLE) and HPLC method was developed for simultaneous determination of 15 flavonoids, namely hexandraside E, kaempferol-3-O-rhamnoside, hexandraside F, epimedin A, epimedin B, epimedin C, icariin, epimedoside C, baohuoside II, caohuoside C, baohuoside VII, sagittatoside A, sagittatoside B, 2''-O-rhamnosyl icariside II and baohuoside I in different species of Epimedium. The analysis was performed by using a Zorbax SB-C18 analytical column (250 mm x 4.6 mm I.D., 5 microm) at gradient elution of water and acetonitrile with diode-array detection (270 nm). All calibration curves showed good linearity (r(2)>0.9997) within test ranges. The LOD and LOQ were lower than 1.31 ng and 2.62 ng on column, respectively. The RSD for intra- and inter-day of 15 analytes was less than 3.8% at three levels, and the recoveries were 90.5-106.8%. The validated method was successfully applied for the analysis of 15 flavonoids in different species of Epimedium which had great variation on the contents of investigated flavonoids. Hierarchical clustering analysis based on the characteristics of 15 investigated compound peaks in HPLC profiles showed that 26 samples were divided into three main clusters, which were in accordance with their flavonoid contents. Four flavonoids including epimedin A, B, C and icariin were optimized as markers for quality control of the species of Epimedium used as Yinyanghuo.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Epimedium/química , Flavonoides/análisis , Flavonoides/química , Reproducibilidad de los ResultadosRESUMEN
One new phenylethanoid glycoside, cistansinenside A and one new oligosaccharide, cistansinensose A1/A2, were isolated from the stems of Cistanche sinensis, together with six known compounds. The structures of the new compounds were elucidated on the basis of spectral data.
Asunto(s)
Cistanche/química , Glicósidos/aislamiento & purificación , Oligosacáridos/aislamiento & purificación , Fenoles/aislamiento & purificación , Procesamiento Automatizado de Datos , Glicósidos/química , Espectrometría de Masas , Resonancia Magnética Nuclear Biomolecular , Oligosacáridos/química , Fenoles/química , Tallos de la Planta/química , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
As an alternative of traditional extraction method, pressurized liquid extraction (PLE) was applied for five flavanoids extraction from Lysimachia clethroide. The operational parameters of PLE, such as extraction solvent, temperature, pressure, static extraction time, flush volume and cycles were optimized by univariate approach coupled with central composite design (CCD) in order to obtain the highest extraction efficiency. The optimized result employed 50% acetonitrile aqueous as extraction solvent, 100 degrees C of extraction temperature, 1500 psi of extraction pressure, 25 min of static time, 70% flush volume, and only one cycle to extract the target compounds completely. Finally, the contents of five major flavanoids in L. clethroides from different sources were determined simultaneously by the combination of the presented PLE and HPLC method.
Asunto(s)
Flavonoides/aislamiento & purificación , Primulaceae/química , Calibración , Cromatografía Líquida de Alta Presión , Modelos Estadísticos , Extractos Vegetales/análisis , Solventes , UltrasonidoRESUMEN
Two new acylated presenegenin glycosides E-onjisaponin H (5) and Z-onjisaponin (6) together with seven known saponins were isolated from the roots of Polygala tenuifolia Willd. Compounds 5 and 6 were obtained as a pair of isomers due to trans and cis-p-methoxycinnamoyl. Their structures were elucidated mainly by 2D-NMR techniques including 1H-1HCOSY, TOCSY, HSQC, HMBC as 3-O-(beta-D-glucopyranosyl) presenegenin 28-[O-beta-D-apiofuranosyl-(1 --> 3)-O-[beta-D-xylopyranosyl-(1 --> 4)]-O-alpha-L-rhamnopyranosyl-(1 --> 2)-O-[alpha-L-rhamnopyranosyl-(1 --> 3)]-4-O-[(E)-p-methoxycinnamoyl]-beta-D-fucopyranosyl] ester (5) and its (Z)-isomer (6).
Asunto(s)
Polygala/química , Saponinas/química , Triterpenos/química , Acilación , Hidrólisis , Espectroscopía de Resonancia Magnética , Raíces de Plantas/química , Saponinas/aislamiento & purificación , Espectrometría de Masa por Ionización de Electrospray , Espectrofotometría Ultravioleta , Triterpenos/aislamiento & purificaciónRESUMEN
A new method based on pressurized liquid extraction (PLE) followed by a sensitive and specific HPLC-DAD analysis is developed for determination of seven compounds in Cortex Dictamni. The operational parameters of PLE, such as extraction solvent, extraction temperature, extraction pressure, static extraction time, flush volume and extraction cycles were optimized, using the extraction efficiencies of dictamnine, obacunone and fraxinellone as targets. The optimized procedure employed MeOH as extraction solvent, 150 degrees C of extraction temperature, 1,500 psi extraction pressure, 5 min of static extraction time, 60% flush volume and the extraction recoveries of the three compounds were nearly to 100% for only one cycle. The following HPLC analysis was performed on a reversed-phase C(18) column with methanol-water as mobile phase in gradient manner, detected at 236 and 218 nm. The limits of detection (LOD) and limits of quantification (LOQ) of the seven compounds were in the range of 0.4-15.6 ng and 1.2-38.8 ng. This assay can be readily utilized as a quality control method for Cortex Dictamni and other related medicinal plants.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Dictamnus/química , Raíces de Plantas/química , Espectroscopía de Resonancia Magnética , Presión , Espectrofotometría UltravioletaRESUMEN
Three unique polysaccharides (1-3) have been obtained from the 0.5 M NaOH extract of the stem of Cistanche deserticola Y. C. Ma. The results of methylation analysis, partial acid hydrolysis, 13C, 1H NMR, 1H-1H COSY, HMQC and HMBC spectroscopic analyses indicate that they are all composed of glucose, having a backbone of alpha-(1 --> 6)-glucan, and have different molecular weights. Their structures differ from that of linear starch.
Asunto(s)
Cistanche/química , Glucanos/aislamiento & purificación , Secuencia de Carbohidratos , Glucanos/química , Datos de Secuencia Molecular , Peso Molecular , Resonancia Magnética Nuclear Biomolecular , Tallos de la Planta/química , Plantas Medicinales/química , Espectrofotometría InfrarrojaRESUMEN
Radix Adenophorae (Shashen), a traditional Chinese medicine commonly used as an antitussive and expectorant, is derived from roots of Adenophora stricta Miq. and Adenophora tetraphylla (Thunb.) Fisch. Twelve species and varieties of Adenophora and Glehnia, however, could act as substitutes or adulterants of Radix Adenophorae on the commercial markets in South East Asia, and roots of Adenophora hunanensis Nannf. and Glihnia littoralis F. Schmidt ex Miq. are the most common examples. The authentic identification of dried roots of A. stricta and A. tetraphylla, however, is difficult on the basis of appearance and morphology. A molecular genetic approach was developed here to identify the species of Radix Adenophorae. The 5S-rRNA spacer domains (approximately 250 bp) were amplified by the polymerase chain reaction (PCR) from genomic DNAs isolated from A. stricta, A. tetraphylla, A. hunanensis and G. littoralis, and subsequently, the nucleotide sequences were determined. Diversity in DNA sequence and restriction enzyme mapping among various species were found in their 5S-rRNA spacer domains, which could serve as markers for authentic identification of Radix Adenophorae.
Asunto(s)
Campanulaceae/genética , Medicamentos Herbarios Chinos/normas , Raíces de Plantas/genética , ARN Ribosómico 5S/genética , Antitusígenos/normas , Secuencia de Bases , Campanulaceae/clasificación , Industria Farmacéutica/normas , Expectorantes/normas , Marcadores Genéticos , Datos de Secuencia Molecular , ARN de Planta/análisisRESUMEN
AIM: To investigate the inhibition capacities of carnosol, rosmanol, and epirosmanol, which are phenolic diterpenes from Rosmarinus officinalis, to oxidized low-density lipoprotein (LDL) formation in human blood and detect their scavenging activities to lipid free radical and superoxide anion in vitro. METHODS: The antioxidant activities which were expressed with the inhibilities to lipid free radicals in the membrane lipid of cell and oxidized LDL formation were evaluated by TBARS assay and ESR method. The inhibition on the Cu2+-mediated oxidization of apo B formation in LDL was investigated by fluorescence spectroscopy. RESULTS: Carnosol, rosmanol, and epirosmanol had an inhibitory activity to lipid peroxidation and oxidized apo B formation in human bloods LDL. The IC50 were 7-10 micromol/L. The antioxidant mechanism was related to the scavenging activities to lipid free radical. CONCLUSION: carnosol, rosmanol, and epirosmanol showed the activity in inhibiting LDL oxidation.
Asunto(s)
Antioxidantes/farmacología , Diterpenos/farmacología , Lipoproteínas LDL/metabolismo , Fenantrenos/farmacología , Rosmarinus/química , Abietanos , Membrana Celular/efectos de los fármacos , Radicales Libres/metabolismo , Humanos , Hidroxibenzoatos/farmacología , Peroxidación de Lípido/efectos de los fármacos , Superóxidos/metabolismoRESUMEN
A new sesquiterpene lactone (1) was obtained from the cytotoxic fraction of 95% ethanol extract of root barks of Tsoongiodendron odorum Chun together with two known sesquiterpene lactones, costunolide (2) and parthenolide (3). The structure of 1 was elucidated as 5alpha, 6alpha, 7beta, 10beta- 11alpha, 13-dihydro-4(15)-eudesmene-12, 6-olide on the basis of chemical and spectral evidence including X-ray diffraction analysis. Costunolide showed cytotoxic activity against human leukemia (HL-60) cell line. Parthenolide showed promising cytotoxic activities in vitro against HCT-8, Bel-7402, SKOV3, KB, HELA and EJ cell lines. Also, the cytotoxic ethyl acetate fraction of ethanol extract of the root barks from which three chemical components were isolated showed promising cytotoxic activities in vitro against KB, BGC-823, Bel-7402, HCT-8, HL-60 cell lines.
Asunto(s)
Antineoplásicos Fitogénicos/aislamiento & purificación , Lactonas/aislamiento & purificación , Magnoliaceae/química , Sesquiterpenos de Eudesmano , Sesquiterpenos/aislamiento & purificación , Terpenos/aislamiento & purificación , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/farmacología , Cromatografía en Gel , Cristalografía por Rayos X , Formazáns , Cromatografía de Gases y Espectrometría de Masas , Células HL-60 , Células HeLa , Humanos , Células KB , Lactonas/química , Lactonas/farmacología , Espectroscopía de Resonancia Magnética , Modelos Moleculares , Raíces de Plantas/química , Sesquiterpenos/química , Sesquiterpenos/farmacología , Terpenos/química , Terpenos/farmacología , Sales de TetrazolioRESUMEN
Echinacoside, one kind of phenylethanoid glycosides (PhGs), was isolated from the stems of Cistanche tubulosa (Schenk) R. Wight with a series of steps, including solvent extraction, D101 polymer adsorption column separation, Sephadex LH-20 separation, C18 column reversed-phase preparative high performance liquid chromatographic (RP-prep-HPLC) preparation and polyamide thin-layer chromatographic detection. The purity of the product was over 98%. The chemical structure of echinacoside was identified by 1H NMR and 13C NMR. To find out the optimum condition of mobile phase of RP-prep-HPLC, several systems were used in this work. Finally acetonitrile-1% HCOOH water (18:82, V/V) system was found to be the best. On the other hand D101 polymer adsorption column and Sephadex LH-20 were also effective for PhGs separation.
Asunto(s)
Cistanche/química , Glicósidos/aislamiento & purificación , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/química , Glicósidos/química , Tallos de la Planta/químicaRESUMEN
AIM: To study the metabolic process of phenylethanoid glycosides (PhGs) in the gastrointestine of beagle dogs that were administered intragastrially process, and develop some new methods of biopharmacology on the effective position of traditional Chinese medicine. METHODS: High-performance liquid chromatography was used to purify constituents from faeces and analyze relative contents of the three main compounds in the gastrointestinal tract at different times and in the faeces of dogs. Every sample was collected, extracted with methanol and analyzed with integration. RESULTS: Four compounds, based on reference substances, were identified as echinacoside, acteoside, isoacteoside, and 2'-acetylacteoside from extraction of faces of dogs. Quantitative "with HPLC" analysis reveals that variation of ratios of the three main compounds is not distinct when moving in the gastrointestinal tract 7 h, that is quite different from those in faeces, in which the content of echinacoside fell from 48.0% to 16.0%, and acteoside increased from 11.0% to 34.7%. CONCLUSION: PhGs are mainly metabolized in large intestine. Among them, a portion of echinacoside is transformed into aceteoside.
Asunto(s)
Glicósidos/metabolismo , Intestino Grueso/metabolismo , Alcohol Feniletílico/metabolismo , Animales , Cistanche/química , Perros , Heces/química , Mucosa Gástrica/metabolismo , Glucósidos/química , Glucósidos/aislamiento & purificación , Glicósidos/química , Glicósidos/aislamiento & purificación , Fenoles/química , Fenoles/aislamiento & purificación , Alcohol Feniletílico/aislamiento & purificación , Plantas Medicinales/químicaRESUMEN
OBJECTIVE: To investigate the chemical constituents of Cistanche tubulosa. METHOD: The chemical constituents were isolated by solvent extraction together with various chromatographic techniques including preparative HPLC. The structures were elucidated on the basis of chemical evidence and spectral data. RESULTS: Four iridoid glycosides, one lignan glycoside and one monoterpenoid were isolated from the 95% ethanol extract from the stem of C. tubulosa and identified as adoxosidic acid(I), 8-epiloganic acid(II), geniposidic acid (III), mussaenosidic acid(IV), (+)-syringaresinol-O-beta-D-gluco pyranoside(V) and 8-hydroxygeraniol(VI). CONCLUSION: Compounds I and VI were isolated from the genus of Cistanche for the first time. Compounds III, IV and V were isolated from this plant for the first time.
Asunto(s)
Cistanche/química , Iridoides/aislamiento & purificación , Magnoliopsida/química , Ácidos Carboxílicos/química , Ácidos Carboxílicos/aislamiento & purificación , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/aislamiento & purificación , Glucósidos/química , Glucósidos/aislamiento & purificación , Glucósidos Iridoides , Iridoides/química , Tallos de la Planta/química , Terpenos/química , Terpenos/aislamiento & purificaciónRESUMEN
AIM: To investigate the antifungal effect of three natural products on the genetic substance of Saccharomyces cerevisiae GL7 and Prototheca wickerhamii. METHODS: The normal and treated cells were observed by confocal laser scanning microscope (CLSM) and image analysis to quantitatively described the cell morphology, area, DNA and RNA content. RESULTS: The morphology, area, DNA and RNA contents were changed greatly in the treated cells. CONCLUSION: Solasodine, 4'-hydroxy-3, 5-dimethoxystilbene and dictamnine directly or indirectly interfered the synthesis and function of genetic substance in S. cerevisiae and P. wickerhamii.
Asunto(s)
Antifúngicos/farmacología , ADN de Hongos/efectos de los fármacos , Prototheca/efectos de los fármacos , Quinolinas , ARN de Hongos/efectos de los fármacos , Saccharomyces cerevisiae/efectos de los fármacos , Alcaloides/farmacología , Prototheca/genética , Saccharomyces cerevisiae/genética , Alcaloides Solanáceos/farmacologíaRESUMEN
The Chinese drug "Rou Cong-rong" (Herba Cistanchis) is one of the commonly used drugs in Chinese traditional medicine. It is used to reinforce the vital function of kidney, especially that of the sexual organs and induce laxation, for the treatment of impotence, premature ejaculation in men, infertility, morbid leukorrhea, profuse metrorrhagia in women, and chronic constipation in the aged. This paper deals with the qualitative and quantitative analysis of phenylethanoid glycosides of four species and one variety of Genus Cistanche and 23 lots of commercial crude drugs of Herba Cistanchis by RP-HPLC. The results were as follows: the chemical constituents of Cistanche deserticola Ma, C. salsa (C. A. Mey) G. Beck, C. salsa var. albiflora P. F. Tu et Z. C. Lou and C. tubulosa were similar while those of C. sinensis were different from the others; the contents of echinacoside and acteoside of C. salsa, which were 2.13% and 1.51%, were the highest of the genus Cistanche. An ODS column (Alltima C18, 5 microns, 250 x 4.6 mm) was employed. Linear gradient elution of acetonitrile--1.5% acetic acid was used as mobile phase, and concentration of acetontrile was from 8% to 20% (0-60 min) in the qualitative analysis, and from 11.5 to 20% (0-35 min) in the quantitative analysis. The flow rate was 1.2 ml.min-1. The detection wavelength was set at 335 nm.
Asunto(s)
Medicamentos Herbarios Chinos/química , Glucósidos/análisis , Glicósidos/análisis , Fenoles , Plantas Medicinales/química , Cromatografía Líquida de Alta Presión/métodos , Contaminación de Medicamentos , Inmunosupresores/análisis , Magnoliopsida/química , Plantas Medicinales/clasificación , Control de CalidadRESUMEN
The commercial crude drug of Radix Adenophorae bought from 15 provinces and autonomous regions, and Radix Glehniae bought from 17 provinces and autonomous regions were surveyed. The original plants of 60 samples of Radix Adenophorae were identified as 9 species and 4 subspecies of genus Adenophora. The main species are Adenophora stricta, A. stricta subsp. sessilifolia, A. potaninii and A. hunanensis. Sixty two samples of Radix Glehniae were all identified as the roots of Glehnia littoralis.
Asunto(s)
Medicamentos Herbarios Chinos , China , Conservación de los Recursos NaturalesRESUMEN
This paper deals with the investigation of the original plants of Rou Congrong and Cao Congrong recorded in the herbalogical works of the past dynasties. The results have shown that Rou Congrong is the dried fleshy stem of Cistanche deserticola and C. salsa, and Cao Congrong is the dried fleshy stem of Orobanche coerulescens. The substitutes and false drugs of Rou Congrong were also investigated.