RESUMEN
As a kind of commonly used Traditional Chinese Medicine in clinical, Spatholobi Caulis (SPC) contains a wide variety of bioactive compounds, including protocatechuate (1), nicotinic acid (2), p-hydroxybenzoic acid (3), salicylic acid (4), 6,9-dihydroxy megastigma-4,7-dien-3-one (5), 8,9-dihydroxy megastigma-4,6-dien-3-one (6), daidzin (7), genistin (8), isolariciresinol (9), ononin (10), 4',8-dimethoxy-7-O-ß-d-glucopyranosyl isoflavone (11), 3'-methoxydaidzein (12), odoratin (13), spasuberol A (14), (+)-pinoresinol (15), 4-hydroxy-3-methoxy cinnamic acid methyl ester (16), (+)-epipinoresinol (17), calycosin (18), 8-O-methylretusin (19), formononetin sodium (20), formononetin (21), biochanin A (22), butesuperin A (23), homovanillyl-4-oxo-nonanoate (24) and (6aR,11aR)-maackiain (25). The pharmacokinetic characteristics of these twenty-five compounds in rat plasma were quantitatively and simultaneously studied using a fast, sensitive and precise ultra fast liquid chromatography combined with electrospray ionization triple quadrupole tandem mass spectrometry (UFLC-MS/MS) method after oral administration of aqueous extract of SPC to rats. The mobile phase consists of acetonitrile and 0.5 mM ammonium acetate in water, and these compounds were well separated at a gradient elution program with flow rate of 0.35 mL/min. Carbamazepine was employed as the internal standard (IS) and all samples were precipitated with MeOH-ACN (2:1, v/v). The analytical method has been proved to be good linearity (R2 ≥ 0.9957), precise, accurate, stable, recovery and matrix effect, which applicated becomingly to study the pharmacokinetic processes of these compounds in rat plasma. In addition, these twenty-five compounds exhibited anti-inflammatory activity on the inflammatory model of NO over production in RAW264.7 cells stimulated by lipopolysaccharide (LPS). Isoflavones, especially compounds 20-22 (The IC50 of which were 22.75 µM, 21.11 µM and 48.29 µM, respectively.) might be the important constituents for anti-inflammatory activity of SPC. This study provides reference values for the clinical application, in-depth study on new dosage forms and pharmacological activities of SPC.
Asunto(s)
Medicamentos Herbarios Chinos , Espectrometría de Masas en Tándem , Animales , Antiinflamatorios/farmacología , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Medicamentos Herbarios Chinos/farmacología , Ratas , Reproducibilidad de los ResultadosRESUMEN
A method of ultra-fast liquid chromatography in series with tandem mass spectrometry for the rapid and sensitive detection of 57 compounds in Spatholobi Caulis (the vine stem of Spatholobus suberectus Dunn) within 35 min was established. This assay can simultaneously determine a variety of compounds without matrix interference in multiple reaction monitoring mode including evaluating the quality of different batches of Spatholobi Caulis from several areas and further identifying the characteristic compounds efficiently. After comprehensive validation, this method can be used to determinate samples rapidly, precisely, accurately, repeatably, and sensitivity. There were significant content differences in 12 batches of Spatholobi Caulis, which were further classified and systematically differentiated applying multivariate statistical analysis. Furthermore, orthogonal partial least squares discrimination analysis results indicated that (-)-gallocatechin (10), (-)-epiafzelechin (20), 4,7,2'-trihydroxy-4'-methoxyisoflavanol (51), and biochanin A (53) characterize compounds to discernment internal quality of Spatholobi Caulis, and recommended as quality control indicators. Hence, presented work provides a method for further study on pharmaceutic preparation, metabolism, as well as for the design, production optimization process, and clinical application.
Asunto(s)
Medicamentos Herbarios Chinos/análisis , Fabaceae/química , Extractos Vegetales/análisis , Cromatografía Líquida de Alta Presión , Estructura Molecular , Espectrometría de Masas en TándemRESUMEN
The stems of Mahonia fortunei (MF) are commonly used in Chinese Traditional Medicine and contain multiple bioactive compounds, including 3,4,5-trimethoxyphenol-1-O-ß-d-glucopyranoside (1), 5-hydroxypicolinic acid methyl ester (2), acortatarin A (3), syringic acid (4), 9-epi-acortatarin A (5), vomifoliol (6), corydaldine (7), noroxyhydrastinine (8), columbamine (9), jatrorrhizine (10), palmatine (11), berberine (12) and schisandrin (13). The pharmacokinetics of these 13 compounds in the rat plasma were assessed using a novel sensitive, rapid, and specific UPLC-ESI-MS/MS method after oral administration of an aqueous extract of MF stems. Carbamazepine was employed as the internal standard (IS) and all samples were precipitated with acetonitrile. Chromatographic separation was performed on a C18 column using a gradient elution at 0.3â¯mL/min, with the mobile phase consisting of acetonitrile and 0.06 % formic acid and 5â¯mM ammonium acetate aqueous solution. The calibration curves showed satisfactory linearity in the examination area (r2â¯≥â¯0.99). The accuracy, precision, extraction recovery, matrix effect, and stability were within acceptable ranges. The method successfully assessed the pharmacokinetics of these 13 compounds. In vitro, compound 12 exhibited potent inhibitory activity against production of nitric oxide (NO) in the RAW264.7 cell line when stimulated by lipopolysaccharide (LPS), while compounds 7, 12, and 13 were the most potent inhibitors of NO production in the BV2 cell line when stimulated by LPS. The IC50 values of compounds 7, 12 and 13 were 42.81, 20.55 and 22.74⯵M. We conclude that these compounds have promise for clinical application, although their synergistic action may be more effective than that by any single compound alone.
Asunto(s)
Antiinflamatorios/análisis , Mahonia/química , Extractos Vegetales/análisis , Administración Oral , Animales , Antiinflamatorios/farmacocinética , Antiinflamatorios/farmacología , Cromatografía Líquida de Alta Presión/métodos , Concentración 50 Inhibidora , Masculino , Ratones , Óxido Nítrico/metabolismo , Extractos Vegetales/farmacocinética , Extractos Vegetales/farmacología , Células RAW 264.7 , Ratas , Ratas Sprague-Dawley , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masas en Tándem/métodosRESUMEN
The dried vine stems of Spatholobus suberectus are commonly used in traditional Chinese medicine for treating gynecological and cardiovascular diseases. In this study, five new compounds named spasuberol A (2), homovanillyl-4-oxo-nonanoate (5), spasuberol C (6), spasuberoside A (14), and spasuberoside B (15), together with ten known compounds (1, 3, 4, 7-13), were isolated from the dried vine stems of S. suberectus. Their chemical structures were analyzed using spectroscopic assays. This is the first study interpreting the detailed structural information of 4. The anti-inflammatory activity of these compounds was evaluated by reducing nitric oxide overproduction in RAW264.7 macrophages stimulated by lipopolysaccharide. Compounds 1 and 8-10 showed strong inhibitory activity with half maximal inhibitory concentration (IC50) values of 5.69, 16.34, 16.87, and 6.78 µM, respectively, exhibiting higher activity than the positive drug l-N6-(1-iminoethyl)-lysine (l-NIL) with an IC50 value of 19.08 µM. The IC50 values of inhibitory activity of compounds 2 and 4-6 were 46.26, 40.05, 45.87, and 28.29 µM respectively, which were lower than l-NIL, but better than that of positive drug indomethacin with an IC50 value of 55.44 µM. Quantitative real-time polymerase chain reaction analysis revealed that assayed compounds with good anti-inflammatory activity, such as 1, 6, 9, and 10 at different concentrations, can reduce the messenger RNA (mRNA) expression of some pro-inflammatory cytokines such as tumor necrosis factor α (TNF-α), nitric oxide synthase (iNOS), and cyclooxygenase 2 (COX-2). The anti-inflammatory activity and the possible mechanism of the compounds mentioned in this paper were studied preliminarily.
Asunto(s)
Antiinflamatorios/farmacología , Fabaceae/química , Expresión Génica/efectos de los fármacos , Glicósidos/farmacología , Lipopolisacáridos/antagonistas & inhibidores , Óxido Nítrico/antagonistas & inhibidores , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Glicósidos/química , Glicósidos/aislamiento & purificación , Humanos , Indometacina/farmacología , Concentración 50 Inhibidora , Lipopolisacáridos/farmacología , Lisina/análogos & derivados , Lisina/farmacología , Medicina Tradicional China , Ratones , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Extractos Vegetales/química , Tallos de la Planta/química , Células RAW 264.7 , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
To establish an HPLC characteristic fingerprint method of Fuke Qianjin Capsules,and determine the contents of its main components. The analysis was carried out on a Kromasil 100-5-C18 analytical column(4. 6 mm ×250 mm,5 µm) with gradient elution by acetonitrile(A)-0. 1% phosphoric acid aqueous solution(B),a flow rate at 1. 0 m L·min-1 and the detection wavelength of 254 nm.The column temperature was 30 â,and the injection volume was 10 µL. The determination method of genistin,jatrorrhizine,andrographolide and 14-deoxy-11,12-didehydroandrographolide index components were studied methodologically. The common mode of the characteristic fingerprint of Fuke Qianjin Capsules was set up with 8 common peaks,which were identified as genistin,jatrorrhizine,palmatine,berberine,andrographolide,14-deoxy-11,12-didehydroandrographolide,Z-ligustilide,and Z-3-butylidenephthalide,respectively,in comparison with the references. The similarities of 20 batches of Fuke Qianjin Capsules samples were above 0. 95. All of the above-mentioned 4 analytes could be well separated under the optimized chromatographic conditions. RSD of precision and repeatability experiment were both less than 1. 5%,and the sample solution was stable during 72 h. All of the compounds had a good linearity and linear range. The contents of genistin,jatrorrhizine,andrographolide,and 14-deoxy-11,12-didehydroandrographolide in 20 batches of Fuke Qianjin Capsules samples were 28. 66-56. 04,94. 77-197. 92,1 705. 33-4 148. 93 and 462. 16-1 225. 96 µg in each capsule,respectively. The developed HPLC characteristic fingerprint and quantitative analysis methods were reliable,accurate and sensitive,and could be used effectively evaluate the quality of Fuke Qianjin Capsules samples.
Asunto(s)
Medicamentos Herbarios Chinos/química , Fitoquímicos/análisis , Cápsulas , Cromatografía Líquida de Alta PresiónRESUMEN
Four new sesquiterpenes including the novel sesquiterpene lactone (6R)-dehydroxysipanolinolide (1), two new eudesmane-type sesquiterpenes, curcolide (2) and curcodione (3), and a new xanthane-type sesquiterpene, curcumadionol (4), were isolated from Curcuma wenyujin Y. H. Chen et C. Ling, together with five known ones, 5-9. The structures of the new compounds were elucidated by spectroscopic methods. The inhibitory effects of compounds 1-9 on nitric-oxide production in lipopolysaccaride-activated macrophages were evaluated.
Asunto(s)
Curcuma/química , Óxido Nítrico/metabolismo , Sesquiterpenos/química , Animales , Línea Celular Tumoral , Macrófagos/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Ratones , Conformación Molecular , Rizoma/química , Sesquiterpenos/aislamiento & purificación , Sesquiterpenos/farmacologíaRESUMEN
Four new guaiane-type sesquiterpenes (1S,4S,5S,10R)-zedoarondiol, zedoalactones D, E, and F (1-4), along with 10 known ones (5-14), were isolated from Curcuma wenyujin Y.H. Chen et C. Ling. The structures of these new compounds were elucidated by spectroscopic methods. The inhibitory effects of compounds 1-14 on nitric oxide production in lipopolysaccharide-activated macrophages were evaluated.