RESUMEN
Total steroid saponins isolated from Dioscorea zingiberensis have shown a variety of beneficial bioactivities. However, there are no reports about their neuroprotective effects, until now. Therefore, in the present study, we explored the neuroprotective effects of the total steroid saponins from D. zingiberensis on rats against transient focal cerebral ischemia-reperfusion and their underlying mechanisms. Healthy adult Sprague-Dawley rats were randomly assigned to six groups. After pretreatment with D. zingiberensis total steroid saponins intragastrically for six days, the rats were subjected to an ischemia injury by surgery on the middle cerebral artery occlusion for 90 min. Compared to the ischemia-reperfusion group, the D. zingiberensis total steroid saponin group of rats, especially those given a 30-mg/kg dose, showed not only a marked reduction in neurological deficit scores, cerebral infarct volume, and brain edema, but also an increase in neuron survival (Nissl bodies) in the hippocampal cornuammons 1 and cortex hemisphere of the ipsilateral ischemia. At the same time, the inflammatory cytokines in serum induced by the middle cerebral artery occlusion were significantly decreased by the preadministration of D. zingiberensis total steroid saponins. Furthermore, the increase of caspase-3 was evidently reduced in the hippocampal cornuammons 1 and cortex of the hemisphere injured brain. Finally, the downregulating antiapoptotic Bcl-2 and upregulating proapoptotic Bax proteins were obviously suppressed. Taken together, the current findings suggest that D. zingiberensis total steroid saponins played a potential neuroprotective role against a severe injury induced by transient focal cerebral ischemic reperfusion in a rat experimental model, and this role may be mediated by its anti-inflammatory and antiapoptotic actions.
Asunto(s)
Antiinflamatorios/farmacología , Apoptosis/efectos de los fármacos , Dioscorea/química , Fármacos Neuroprotectores/farmacología , Daño por Reperfusión/prevención & control , Saponinas/farmacología , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Masculino , Fármacos Neuroprotectores/química , Fármacos Neuroprotectores/aislamiento & purificación , Ratas Sprague-Dawley , Saponinas/química , Saponinas/aislamiento & purificaciónRESUMEN
The aim of our present study is to explore the anti-arthritic potential effect of total steroid saponins (TSSNs) extracted from the rhizome of Dioscorea zingiberensis C.H.Wright (DZW) and to investigate the underlying mechanisms. This work was performed using adjuvant-induced arthritis (AIA) rats in vivo and lipopolysaccharide (LPS) simulated 264.7 macrophage cells in vitro. In AIA-induced arthritic rats, TSSN significantly alleviated the arthritic progression through evaluating arthritic score, immune organ indexes, paw swelling, and body weight. This phenomenon was well correlated with significant suppression of the overproduction of inflammation cytokines (IL-1, IL-1ß, IL-6, and TNF-α), oxidant stress makers (MDA and NO), eicosanoids (LTB4 and PGE2), and inflammatory enzymes (5-LOX and COX-2) versus the AIA rats without treatment. On the contrary, the release of SOD and IL-10 was profoundly increased. What's more, TSSN could obviously ameliorate the translocation of NF-κB to the nucleus through phosphorylation of the p65 and IκBα in vivo and in vitro. The current findings demonstrated that TSSN could protect the injured ankle joint from further deterioration and exert its satisfactory anti-arthritis properties through anti-inflammatory and anti-oxidant effects via inactivating the NF-κB signal pathway. This research implies that DZW may be a useful therapeutic agent for the treatment of human arthritis.
Asunto(s)
Artritis/inducido químicamente , Dioscorea/química , Extractos Vegetales/farmacología , Rizoma/química , Saponinas/farmacología , Esteroides/farmacología , Animales , Antirreumáticos/administración & dosificación , Antirreumáticos/farmacología , Artritis/tratamiento farmacológico , Relación Dosis-Respuesta a Droga , Adyuvante de Freund/toxicidad , Metotrexato/farmacología , Estructura Molecular , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Saponinas/administración & dosificación , Saponinas/química , Esteroides/administración & dosificación , Esteroides/químicaRESUMEN
To establish a HPLC-ELSD fingerprint for total steroid saponins in herbs of Dioscorea zingiberensis. Welchrom C,8 (4. 6 mm x 250 mm,5 microm) chromatographic column was adopted and eluted with the mobile phase of acetonitrile (A)-water (B) at the flow rate of 1.0 mL min-1. The column temperature was room temperature. The ELSD conditions were as follows: the temperature of drift tube was 90.0 degreeC, the flow rate of carrier gas (N2) was 2. 8 L min-1, and the injection volume was 10 microL. After the detection of 10 batches of samples,the common mode of HPLC-ELSD fingerprint for total steroid saponins in herbs of D. zingiberensis was established for the first time,and 25 common peaks were determined. Among them, 10 peaks were identified by comparing with reference substances. The similarities of 10 batches of herbs were evaluated in the common mode. All of them were higher than 0. 80. This method is so accurate, reliable and highly repeatable that it can provide scientific basis for evaluating and controlling the quality of total steroid saponins in herbs of D. zingiberensis.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Dioscorea/química , Dioscorea/clasificación , Dioscorea/citología , Saponinas/análisisRESUMEN
AIMS: Gentiopicroside (Gent) is one of the secoiridoid compound isolated from Gentiana lutea. This compound exhibits analgesic activities and inhibits the expression of GluN2B-containing N-methyl-D-aspartate (NMDA) receptors in the anterior cingulate cortex in mice. Nucleus accumbens (NAc) is a forebrain structure known for its role in drug addiction. However, little is known about the role of Gent on morphine dependence and synaptic transmission changes in the NAc. METHODS: Conditioned place preference (CPP) test and behavioral sensitization of locomotor activity were used to investigate drug-seeking related behaviors. Brain slices containing NAc were prepared, and whole-cell patch-clamp recordings were performed to record the excitatory postsynaptic currents (EPSCs). Expression of proteins was detected by Western blot analysis. RESULTS: Systemic administration of Gent attenuated the CPP effect induced by morphine, but had no effect on morphine-induced behavioral sensitization. Gent significantly reversed overexpression of GluN2B-containing NMDA receptors and dopamine D2 receptors in NAc during the first week of morphine withdrawal. However, the compound did not affect the overexpression of GluN2A-containing NMDA receptors, GluA1, and dopamine D1 receptors. Lastly, Gent significantly reduced NMDA receptors-mediated EPSCs in the NAc. CONCLUSION: Our study provides strong evidence that Gent inhibits morphine dependence through downregulation of GluN2B-containing NMDA receptors in the NAc.
Asunto(s)
Analgésicos Opioides/antagonistas & inhibidores , Analgésicos Opioides/farmacología , Glucósidos Iridoides/farmacología , Morfina/antagonistas & inhibidores , Morfina/farmacología , Núcleo Accumbens/metabolismo , Receptores de N-Metil-D-Aspartato/biosíntesis , Animales , Conducta Adictiva/tratamiento farmacológico , Conducta Adictiva/psicología , Western Blotting , Condicionamiento Operante/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Actividad Motora/efectos de los fármacos , Núcleo Accumbens/efectos de los fármacos , Técnicas de Placa-Clamp , Receptores de Dopamina D2/metabolismo , Receptores de N-Metil-D-Aspartato/efectos de los fármacos , RecompensaRESUMEN
An expeditious and effective HPLC-UV method has been developed for the simultaneous determination of seven major flavonoids in Apocyni Veneti Folium (AVF) extract. The chemical profile of seven flavonoids, including quercetin-3-O-ß-D-glc(2 â 1)-ß-D-glucoside, rutin, isoquercetin, kaempferol-3-O-ß-D-glucoside, quercetin-3-O-(6â³-O-malonyl)-ß-D-glucoside, quercetin and kaempferol was acquired by HPLC-UV. The analysis was performed on a Diamosil C18 analytical column with a gradient solvent system of acetonitrile-0.1% aqueous acetic acid. Full validation of the method was carried out (linearity, reproducibility, repeatability, accuracy and limit of detection). The results indicated that the contents of investigated flavonoids in Apocyni Veneti Folium varied significantly from habitat to habitat, with contents ranging from 0.01 to 5.57 mg g⻹. The antioxidant activity results demonstrate that the seven flavonoids showed great efficiency in scavenging DPPH radicals. The high content of flavonoid components of AVF could be responsible for its high antioxidant activity. This study provides powerful evidence for the relationship between the chemical ingredients of and bioactivity in AVF.
Asunto(s)
Antioxidantes/química , Apocynaceae/química , Antioxidantes/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Flavonoides/análisis , Flavonoides/aislamiento & purificación , Extractos Vegetales/químicaRESUMEN
An ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC Q-TOF MS) metabonomics approach was employed to study the serum metabolic profiling of adenine-induced chronic renal failure (CRF) rats. Acquired data were subjected to principal component analysis (PCA) for differentiating the CRF and the normal control groups. Potential biomarkers were screened by using S-plot and were identified by the accurate mass, isotopic pattern and MS/MS fragments information obtained from UPLC Q-TOF MS analysis. Significant differences in the serum level of creatinine, amino acids and LysoPCs were observed, indicating the perturbations of amino acid metabolism and phospholipid metabolism in adenine-induced CRF rats. This research proved that metabonomics is a promising tool for disease research.
Asunto(s)
Biomarcadores/sangre , Creatinina/sangre , Fallo Renal Crónico/sangre , Lisofosfolípidos/sangre , Metabolómica/métodos , Triptófano/sangre , Adenina/farmacología , Animales , Biomarcadores/metabolismo , Cromatografía Liquida/métodos , Creatinina/metabolismo , Fallo Renal Crónico/inducido químicamente , Lisofosfolípidos/metabolismo , Masculino , Metaboloma , Análisis de Componente Principal , Ratas , Ratas Sprague-Dawley , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Triptófano/metabolismoRESUMEN
OBJECTIVES: Aristolochic acid (AA) nephropathy, first reported as Chinese herbs nephropathy, is a rapidly progressive tubulointerstitial nephropathy that results in severe anemia, interstitial fibrosis and end-stage renal disease. Tubulointerstitial injury was studied in a rat model of AA nephropathy to determine whether ergosta-4,6,8(14),22-tetraen-3-one (ergone) treatment prevents early renal injury in rats with aristolochic acid I-induced nephropathy. METHODS: Early renal injury via renal interstitial fibrosis was induced in rats by administration of aristolochic acid I (AAI) solution intragastrically for 8 weeks. Ninety-six rats were randomly divided into four groups (n = 24/group): (1) control (2) AAI (3) AAI + ergone (10 mg/kg) and (4) AAI + ergone (20 mg/kg). Blood and urine samples were collected and rat were sacrificed for histological assessment of the kidneys on at the end of weeks 2, 4, 6 and 8. KEY FINDINGS: AAI caused progressive elevation of blood urea nitrogen, creatinine, potassium, sodium, chlorine, proteinuria and urinary N-acetyl-ß-D-glucosaminidase (NAG). Ergone suppressed elevation of blood urea, nitrogen, creatinine, proteinuria and urinary NAG to some degree, but the AAI-ergone-treated group did not differ from AAI-treated group for body weight, serum potassium, sodium and chlorine. The progress of the lesions in the kidney after AAI administration was also observed by histopathological examinations, but kidneys from rats of AAI-ergone-treated group displayed fewer lesions. CONCLUSIONS: Ergone treatment conferred protection against early renal injury in a rat model of AA nephropathy. Early administration of ergone may prevent the progression of renal injury and the subsequent renal fibrosis in AA nephropathy.
Asunto(s)
Lesión Renal Aguda/prevención & control , Ácidos Aristolóquicos , Colestenonas/aislamiento & purificación , Colestenonas/farmacología , Polyporus/química , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/patología , Animales , Peso Corporal/efectos de los fármacos , Fibrosis , Riñón/patología , Pruebas de Función Renal , Masculino , Nefritis Intersticial/inducido químicamente , Nefritis Intersticial/patología , Proteinuria/inducido químicamente , Proteinuria/patología , Proteinuria/orina , Ratas , Ratas Sprague-DawleyRESUMEN
A new triterpenoid saponin, named esculentoside T (1), together with five known compounds (2-5), was isolated from the roots of Phytolacca acinosa. The structures of these compounds were elucidated on the basis of spectral data.
Asunto(s)
Medicamentos Herbarios Chinos/aislamiento & purificación , Phytolacca/química , Raíces de Plantas/química , Saponinas/aislamiento & purificación , Triterpenos/aislamiento & purificación , China , Medicamentos Herbarios Chinos/análisis , Medicamentos Herbarios Chinos/química , Etanol , Estructura Molecular , Saponinas/análisis , Saponinas/química , Triterpenos/análisis , Triterpenos/químicaRESUMEN
Rapid resolution liquid chromatography/tandem multi-stage mass spectrometry (RRLC-MS(n)) and rapid resolution liquid chromatography/tandem mass spectrometry (RRLC/MS/MS) methods were developed for the identification and quantification of ergosterol and its metabolites from rat plasma, urine and faeces. Two metabolites (ERG1 and ERG2) were identified by RRLC/MS(n). The concentrations of the ergosterol were determined by RRLC/MS/MS. The separation was performed on an Agilent Zorbax SB-C18 with the mobile phase consisting of methanol and water (containing 0.1% formic acid). The detection was carried out by means of atmospheric pressure chemical ionization mass spectrometry in positive ion mode with multiple reaction monitoring (MRM). Linear calibration curves were obtained in the concentration range of 7-2000, 6-2000 and 8-7500 ng/mL for plasma, urine and faecal homogenate, respectively. The intra- and inter-day precision values (RSD) were below 10%. The method was applied to the pharmacokinetic properties and elimination pathway of ergosterol in rats.
Asunto(s)
Cromatografía Liquida/métodos , Ergosterol/farmacocinética , Espectrometría de Masas en Tándem/métodos , Animales , Estabilidad de Medicamentos , Ergosterol/análisis , Ergosterol/orina , Heces/química , Análisis de los Mínimos Cuadrados , Masculino , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los ResultadosRESUMEN
Du-Zhong is one of the most important tonic herbs in traditional Chinese medicine for the treatment of bone fractures and other bone diseases. The present study examined whether Du-Zhong seed extract named total glycosides from Eucommia ulmoides seed (TGEUS) could display an increased effect on bone density and bone strength of the femur in rats. Sprague-Dawley rats were used and randomly assigned to the normal group and the TGEUS group (400 mg/kg body weight/day). Daily oral administration of TGEUS was found to increase significantly the biomechanical quality of the femur. The mechanical changes were associated with a bone mineral density (BMD) increase or even with some improvements in microarchitecture. Micro-CT analysis of the distal femur showed that TGEUS significantly increased the bone volume/tissue volume (BV/TV), connectivity density (Conn.D), trabecular number (Tb.N) and trabecular thickness (Tb.Th), and decreased trabecular separation (Tb.Sp) and the structure model index (SMI) in normal rats. To conclude, TGEUS taken orally increased bone density and altered bone histomorphology, suggesting that TGEUS might be a potential alternative medicine for the treatment of postmenopausal osteoporosis.
Asunto(s)
Densidad Ósea/efectos de los fármacos , Eucommiaceae/química , Fémur/efectos de los fármacos , Glicósidos/farmacología , Extractos Vegetales/farmacología , Animales , Femenino , Ratas , Ratas Sprague-Dawley , Semillas/químicaRESUMEN
BACKGROUND: Mushrooms have been used in Asia as traditional foods and medicines for a long time. Ergosta-4,6,8(14),22-tetraen-3-one (ergone) is one of the well-known bioactive steroids, which exists widely in various medicinal fungi such as Polyporus umbellatus, Russula cyanoxantha, and Cordyceps sinensis. Ergone has been demonstrated to possess cytotoxic activity. However, the molecular mechanisms by which ergone exerts its cytotoxic activity are currently unknown. METHODS: In the present study, ergone possessed a remarkable anti-proliferative activity toward human hepatocellular carcinoma HepG2 cells. We assayed the cell cycle by flow cytometry using PI staining; investigated the exposure of phosphatidylserine at the outer layer of the cytoplasmic membrane by the FITC-annexin V/PI staining; observed the nuclear fragmentation by Hoechst 33258 staining and studied the protein expression of Bax, Bcl-2, p-53, procaspase-3, -8, -9, PARP and cleaved PARP by Western blotting analysis. RESULTS: Cells treated with ergone showed typical markers of apoptosis: G2/M cell cycle arrest, chromatin condensation, nuclear fragmentation, and phosphatidylserine exposure. Furthermore, PARP-cleavage; activation of caspase-3, -8, -9; up-regulation of Bax and down-regulation of Bcl-2 were observed in HepG2 cells treated with ergone, which show that both the intrinsic and extrinsic apoptotic pathways are involved in ergone-induced apoptosis in HepG2 cells. Ergosta-4,6,8(14),22-tetraen-3-one induces G2/M cell cycle arrest and apoptosis in HepG2 cells in a caspase-dependent manner. GENERAL SIGNIFICANCE: In this study, we reported for the first time that ergone-induced apoptosis through activating the caspase. These results would be useful for the further utilization of many medicinal fungi in cancer treatment.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Carcinoma Hepatocelular/tratamiento farmacológico , Ciclo Celular/efectos de los fármacos , Colestenonas/farmacología , Polyporus/química , Antineoplásicos/síntesis química , Antineoplásicos/aislamiento & purificación , Colestenonas/síntesis química , Colestenonas/aislamiento & purificación , Células Hep G2 , HumanosRESUMEN
The crystal structure of natural diterpenoid alkaloid ranaconitine isolated from Aconitum sinomontanum Nakai has been determined by single crystal X-ray diffraction analysis. The crystal presents a monoclinic system, space group C2 with Z = 4, unit cell dimensions a = 30.972(19) angstrom, b = 7.688(5) angstrom, and c = 19.632(12) angstrom. Moreover, the intermolecular O-H...O hydrogen bonds and weak pi-pi interactions play a critical role in expanding the dimensionality.
Asunto(s)
Aconitina/análogos & derivados , Aconitum/química , Aconitina/química , Aconitina/aislamiento & purificación , Cristalografía por Rayos X , Estructura MolecularRESUMEN
A simple and specific HPLC method with dual wavelength UV detection for the determination of ergosta-4,6,8(14),22-tetraen-3-one (ergone) in rat plasma was developed and proved to be efficient. The method used ergosterol as internal standard (IS). Following a single-step protein precipitation, the analyte and IS were separated on an Inertsil ODS-3 column with a mobile phase containing methanol-water (99:1, v/v) at a flow rate of 1 mL/min. The analytes were detected by using UV detection at wavelength of 350 (ergone) and 283 (IS) nm, respectively. The calibration curve was linear over the range of 0.1-2.0 µg/mL and the lower limit of quantification was 0.1 µg/mL. The intra-day and inter-day precision studies showed good reproducibility with RSD less than 8.5%. The intra-day and inter-day accuracy ranged from 95.6 to 104%. Mean extraction recovery was above 95% at the low, medium and high concentrations. The present HPLC-UV method was simple and reliable. The method described herein had been successfully applied for the pharmacokinetic studies in male SD rats after administration of 20 mg/kg dose of solution of ergone.
Asunto(s)
Colestenonas/sangre , Cromatografía Líquida de Alta Presión/métodos , Polyporus/química , Animales , Colestenonas/química , Colestenonas/farmacocinética , Estabilidad de Medicamentos , Medicamentos Herbarios Chinos , Ergosterol/análisis , Ergosterol/química , Análisis de los Mínimos Cuadrados , Masculino , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Ergosta-4,6,8(14),22-tetraen-3-one (ergone) from many medicinal plants has been demonstrated to possess a variety of pharmacological activities in vivo and in vitro, including cytotoxic, diuretic and immunosuppressive activity. Metabolism and pharmacokinetic studies on rat were conducted for ergone. Rapid resolution liquid chromatography with atmospheric pressure chemical ionization tandem multi-stage mass spectrometry (RRLC-APCI-MS(n)) and high-performance liquid chromatography with fluorescence detection (HPLC-FLD) methods were applied for the identification and quantification of ergone and its metabolite from rat plasma, faeces and urine. A metabolite was identified by RRLC-DAD-APCI-MS(n): 22,23-epoxy-ergosta-4,6,8(14)-triaen-3-one (epoxyergone). The concentrations of the analyte with its metabolites were determined by HPLC-FLD at excitation wavelength of 370 nm and emission wavelength of 485 nm. The samples were deproteinized with methanol after addition of camptothecin as internal standard (IS). The analysis was performed on a Diamonsil C18 column (150 mm x 4.6 mm x 5 microm) with a mobile phase gradient consisting of methanol and water at a flow rate of 1 mL min(-1). The assay was linear over the concentration range of 42-1500, 36-7500 and 42-1500 ng mL(-1) for plasma, faecal homogenate and urine respectively. The absolute recoveries were found to be 97.0+/-1.2%, 98.1+/-0.7% and 96.6+/-1.8% for plasma, faecal homogenate and urine respectively. The intra-day and inter-day relative standard deviations (RSD) were less than 10%. The previous HPLC-MS/MS method is not affordable for most laboratories because of the specialty requirement and high equipment cost. However, the HPLC-FLD method is economic and operating simply for quantitative determination of ergone and its metabolite in rat plasma, faeces and urine. In addition, liquid chromatography coupled with ion trap multi-stage mass spectrometry is becoming a useful technique for ergone metabolite identification.
Asunto(s)
Colestenonas/farmacocinética , Cromatografía Líquida de Alta Presión/métodos , Citotoxinas/farmacocinética , Inmunosupresores/farmacocinética , Espectrometría de Masas/métodos , Polyporus/química , Animales , Colestenonas/química , Colestenonas/aislamiento & purificación , Colestenonas/metabolismo , Cromatografía Líquida de Alta Presión/economía , Citotoxinas/química , Citotoxinas/aislamiento & purificación , Citotoxinas/metabolismo , Fluorescencia , Inmunosupresores/química , Inmunosupresores/aislamiento & purificación , Inmunosupresores/metabolismo , Modelos Lineales , Espectrometría de Masas/economía , Ratas , Sensibilidad y Especificidad , Factores de TiempoRESUMEN
The steroids ergone (1), (22E, 24R)-ergosta-7,22-dien-3ß-ol (2), 5α,8α-epidioxy-(22E,24R) -ergosta-6,22-dien-3ß-ol (3), ergosta-6,22-dien-3ß,5α,6ß-triol (4), and polyporusterone B (5) were isolated from Polyporus umbellatus by bioassay-guided approach. They showed potent anticancer activity against HepG2 cells. Ergone displayed remarkable anticancer activity against HepG2, Hep-2, and Hela cancer cells, of which HepG2 cells were the most sensitive. Furthermore, the cytotoxic effects of ergone on normal human cells (HUVEC) were smaller than on cancer cells. The results showed that ergone had more selective cytotoxic activity against cancer cells than against normal cells.
Asunto(s)
Antineoplásicos/farmacología , Citotoxinas/farmacología , Polyporus/química , Esteroides/farmacología , Antineoplásicos/química , Antineoplásicos/aislamiento & purificación , Citotoxinas/química , Citotoxinas/aislamiento & purificación , Células HeLa , Células Hep G2 , Humanos , Esteroides/química , Esteroides/aislamiento & purificaciónRESUMEN
Polyporus umbellatus is a widely used diuretic herbal medicine. In this study, a high-performance liquid chromatography coupled with atmospheric pressure chemical ionization-mass spectrometric detection (HPLC-APCI-MS) method was developed for qualitative and quantitative analysis of steroids, as well as for the quality control of Polyporus umbellatus. The selectivity, reproducibility and sensitivity were compared with HPLC with photodiode array detection and evaporative light scattering detection (ELSD). Selective ion monitoring in positive mode was used for qualitative and quantitative analysis of eight major components and beta-ecdysterone was used as the internal standard. Limits of detection and quantification fell in the ranges 7-21 and 18-63 ng/mL for the eight analytes with an injection of 10 microL samples, and all calibration curves showed good linear regression (r(2) > 0.9919) within the test range. The quantitative results demonstrated that samples from different localities showed different qualities. Advantages, in comparison with conventional HPLC-diode array detection and HPLC-ELSD, are that reliable identification of target compounds could be achieved by accurate mass measurements along with characteristic retention time, and the great enhancement in selectivity and sensitivity allows identification and quantification of low levels of constituents in complex Polyporus umbellatus matrixes.
Asunto(s)
Polyporaceae/química , Esteroides/análisis , Calibración , Cromatografía Líquida de Alta Presión , Indicadores y Reactivos , Espectrometría de Masas , Estándares de Referencia , Reproducibilidad de los Resultados , Soluciones , UltrasonidoRESUMEN
A fast and sensitive HPLC-APCI-MS/MS method was developed for the determination of ergosta-4,6,8(14),22-tetraen-3-one (ergone) in rat plasma. The plasma sample containing ergone and ergosterol (internal standard) were simply treated with acetone to precipitate and remove proteins and the isolated supernatants were directly injected into the HPLC-APCI-MS/MS system. Chromatographic separation was performed on a 1.8microm Zorbax SB-C18 column (100mm x 3.0mm) with a 97:3 (v/v) mixed solution of methanol and 0.1% aqueous formic acid being used as mobile phase. Quantification was performed by multiple selected reactions monitoring (MRM) of the transitions with (m/z)(+) 393-268 for ergone and (m/z)(+) 379-69 for the IS. The method was validated in the concentration range of 5-1600ng/mL for ergone. The precision of the assay (RSD%) was less than 10.5% at all concentrations levels within the tested range and adequate accuracy, and the limit of detection was 1.5ng/mL. The absolute recoveries of both ergone and ergosterol from the plasma were more than 95%. The developed method has been successfully applied to the pharmacokinetic study of the drug in SD rats.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Ergosterol/análogos & derivados , Espectrometría de Masas en Tándem/métodos , Animales , Colestenonas , Estabilidad de Medicamentos , Ergosterol/sangre , Ergosterol/química , Ergosterol/farmacocinética , Análisis de los Mínimos Cuadrados , Masculino , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: To determine the content of rosmarinic acid in seeds and its processed products of Perilla frutescens var. typica (PFT), Perilla frutescens var. acuta (PFA) and Perilla frutescens var. acuta form. discolor (PFAD). METHODS: The content of rosmarinic acid was determined by RP-HPLC with Welch C18 (250 mm x 4.6 mm, 5 microm) column, using methanol-phosphonic acid solution (39:61) as mobile phase at flow rate of 1.0 mL/min. UV detection wavelength was set at 330 nm and column temperature was set at 28 degrees C. RESULTS: The content of rosmarinic acid in roasted seeds of PFT is higher than in its seeds, but that of PFA and PFAD are lower; The content of rosmarinic acid in seeds of PFT, PFA and PFAD decline after the seeds are honey-pocessed and made into frost-like powder, and that of honey-processed one decline by a larger margin. CONCLUSION: The content of rosmarinic acid in seeds of PFT, PFA and PFAD are prone to decline by heat-processing.
Asunto(s)
Cinamatos/análisis , Depsidos/análisis , Medicamentos Herbarios Chinos/química , Perilla frutescens/química , Semillas/química , Tecnología Farmacéutica/métodos , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/aislamiento & purificación , Calor , Farmacognosia , Control de Calidad , Reproducibilidad de los Resultados , Ácido RosmarínicoRESUMEN
X-ray diffraction analyses of iridoid glycoside aucubin (1) and its aglycone aucubigenin (2) are reported. It was found that crystals of 1 are orthorhombic, with P2(1)2(1)2(1) space group, both cyclopentane ring and pyran ring adopt envelope conformations, and the Glc moiety is in the (4)C(1) conformation. Crystals of 2 are monoclinic, with space group P2(1), the cyclopentane and pyran rings also adopt the envelope conformation. The absolute configurations of 1 and 2 were also determined. Intensive O-H . . . O hydrogen bonds in both crystal lattices were observed.
Asunto(s)
Glucósidos/química , Iridoides/química , Piranos/química , Cristalografía por Rayos X , Enlace de Hidrógeno , Glucósidos IridoidesRESUMEN
AIM OF THE STUDY: Polyporus umbellatus is a fungus used as a diuretic medicine. The objective of this study was to isolate and elucidate the diuretic constituents of n-hexane, ethyl acetate, n-butanol and water extracts of Polyporus umbellatus and to evaluate their diuretic activity. MATERIALS AND METHODS: The n-hexane, ethyl acetate, n-butanol and water extracts of Polyporus umbellatus were tested by diuretic experiment of normal rats in metabolic cage. The n-hexane extract and n-butanol extract were prepared separately by the bioassay-guided approach. Three isolated compounds doses (5, 10 and 20 mg/kg BW) were orally administered to normal rats. Water excretion rate, pH and content of Na(+), K(+) and Cl(-) were measured in the urine of saline-loaded rats. RESULTS: n-Hexane extract (P<0.05), n-butanol extract (P<0.05) and three isolated compounds (ergosta-4,6,8(14),22-tetraen-3-one, ergosterol and d-mannitol) displayed diuretic activity. CONCLUSIONS: The ergosta-4,6,8(14),22-tetraen-3-one was the strongest diuretic constituent in the three compounds. Ergosterol and D-mannitol were found to be also responsible for duiretic effects in Polyporus umbellatus for the first time. Data show that 20 mg/kg dose of the ergosterol for urine out put became significantly higher than in the control rats, but the ratio of Na(+)/K(+) almost unaltered in the three doses. The highest dose of the D-mannitol was significant and increased the cumulative urine output. Regarding the electrolyte excretion, data show that the doses 10 and 20 mg/kg produce significant increase for excretion of Na(+) and Cl(-). The present results provide a quantitative basis explaining application of Polyporus umbellatus as a diuretic medicine. The result proved that its diuretic effects were also due to the contribution of multi-components in clinical application.