RESUMEN
In this work, a sensitive and efficient method was established and validated for qualitative and quantitative analysis of major bioactive constituents in Dazhu Hongjingtian capsule by liquid chromatography tandem mass spectrometry. A total of 32 compounds were tentatively identified using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Furthermore, 12 constituents, namely gallic acid, 3,4-dihydroxybenzoic acid, salidroside, p-coumaric acid-4-O-ß-d-glucopyranoside, bergeninum, 4-hydroxybenzoic acid, 4-hydroxyphenylacetic acid, syringate, 6''-O-galloylsalidroside, rhodiosin, rhodionin and kaempferol-7-O-α-l-rhamnoside, were simultaneously quantified by the developed ultra-performance liquid chromatography coupled with a triple quadrupole mass spectrometry method in 9 min. All of them were analyzed on an Agilent ZorBax SB-C18 column (3.0 × 100 mm, 1.8 µm) with linear gradient elution of methanol-0.1% formic acid water. The proposed method was applied to analyze three batches of samples with acceptable linearity (R, 0.9979-0.9997), precision (RSD, 1.3-4.7%), repeatability (RSD, 1.7-4.9%), stability (RSD, 2.2-4.9%) and recovery (RSD, 0.6-4.4%) of the 12 compounds. As a result, the analytical method possessing high throughput and sensitivity is suitable for the quality control of Dazhu Hongjingtian capsule.
Asunto(s)
Cromatografía Liquida/métodos , Medicamentos Herbarios Chinos/química , Espectrometría de Masas en Tándem/métodos , Estándares de Referencia , Reproducibilidad de los ResultadosRESUMEN
Reactive oxygen species are damaging to cardiomyocytes. H9c2 cardiomyocytes are commonly used to study the cellular mechanisms and signal transduction in cardiomyocytes, and to evaluate the cardioprotective effects of drugs following oxidative damage. The present study developed a robust, automated high throughput screening (HTS) assay to identify cardioprotective agents from a traditional Chinese medicine (TCM) library using a H2O2induced oxidative damage model in H9c2 cells. Using this HTS format, several hits were identified as cardioprotective by detecting changes to cell viability using the cell counting kit (CCK)8 assay. Two TCM extracts, KY0520 and KY0538, were further investigated. The results of the present study demonstrated that treatment of oxidatively damaged cells with KY0520 or KY0538 markedly increased the cell viability and superoxide dismutase activity, decreased lactate dehydrogenase activity and malondialdehyde levels, and inhibited early growth response1 (Egr1) protein expression. The present study also demonstrated that KY0520 or KY0538 treatment protected H9c2 cells from H2O2induced apoptosis by altering the Bcl-2/Bax protein expression ratio, and decreasing the levels of cleaved caspase3. In addition, KY0520 and KY0538 reduced the phosphorylation of ERK1/2 and p38MAPK proteins, and inhibited the translocation of Egr1 from the cytoplasm to nucleus in H2O2-treated H9c2 cells. These findings suggested that oxidatively damaged H9c2 cells can be used for the identification of cardioprotective agents that reduce oxidative stress by measuring cell viabilities using CCK8 in an HTS format. The underlying mechanism of the cardioprotective activities of KY0520 and KY0538 may be attributed to their antioxidative activity, regulation of Egr1 and apoptosisassociated proteins, and the inhibition of ERK1/2, p38-MAPK and Egr-1 signaling pathways.
Asunto(s)
Cardiotónicos/farmacología , Descubrimiento de Drogas/métodos , Medicamentos Herbarios Chinos/farmacología , Estrés Oxidativo/efectos de los fármacos , Línea Celular Tumoral , Proteína 1 de la Respuesta de Crecimiento Precoz/genética , Proteína 1 de la Respuesta de Crecimiento Precoz/metabolismo , Ensayos Analíticos de Alto Rendimiento , Humanos , Peróxido de Hidrógeno/farmacología , Lactato Deshidrogenasas/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Malondialdehído/metabolismo , Medicina Tradicional China , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Reproducibilidad de los Resultados , Superóxido Dismutasa/metabolismoRESUMEN
This paper summarized the recent 30 years research progress of the chemical constituents from Notopterygii Rhizoma et Radix. The chemical constituents from Notopterygii Rhizoma et Radix mainly consist of coumarins, polyene-polyacetylenes, sesquiterpenes, phenolic acids, while steroids and flavonoids were less reported. All constituents were confirmed and corrected through SciFinder. We also checked the Chinese name and English name and listed the CAS number of each compound. It can provide some guidelines for the research, development and utilization of Notopterygii Rhizoma et Radix in the future. Whether there is columbianin in the Notopterygii Rhizoma et Radix need to be further researched.
Asunto(s)
Apiaceae/química , Medicamentos Herbarios Chinos/análisis , Rizoma/químicaRESUMEN
1,2,3,4,6-penta-O-galloyl-D-glucose (PGG) is one of the main active compounds of Guizhi Fuling capsule. Molecularly imprinted polymers (MIP) have high affinity toward template molecules synthesized by molecularly imprinted technology for its specific combined sites, which can overcome the shortcoming of traditional separation methods, such as complex operation, low efficiency, using large quantity of solvent and environmental pollution. In this paper, surface molecularly imprinted polymer (SMIP) was prepared by surface imprinting with PGG as the template molecule. Its adsorption capacity was measured by the scatchard equation. The separation of PGG from Guizhi Fuling capsule at preparatived scale was achieved with molecularly imprinted polymer as stationary phase and the purity was 90.2% by HPLC. This method can be used to prepare PGG from Guizhi Fuling capsule with large capacity and is easy to operate. It provides a new method for efficient separation and purification for other natural products.
Asunto(s)
Medicamentos Herbarios Chinos/química , Taninos Hidrolizables/aislamiento & purificación , Polímeros/química , Adsorción , Cápsulas/química , Cromatografía Líquida de Alta Presión , Taninos Hidrolizables/química , Impresión Molecular , Polímeros/síntesis químicaRESUMEN
Tyrosol, crenulatin and salidroside are the main active constituents of Rhodiola crenulata, with extensive pharmacological activities. In the study, grams of high purity tyrosol, crenulatin and salidroside were simultaneously separated from R. crenulata by the first time. Firstly, R. crenulata was extracted by 70% alcohol. Then, with the yields of three compounds as the index, the macroporous resin was optimized. At last, grams of high purity tyrosol, crenulatin and salidroside were isolated by D-101 macroporousresin, purified by column chromatography. Detected by HPLC, the purity of three compounds were higher than 98%. This method has the advantages of simple process and operation, less dosage of organic solvent, highly yield and reproducibility, suitable for the simultaneously preparation of tyrosol, crenulatin and salidroside.
Asunto(s)
Fraccionamiento Químico/métodos , Cumarinas/aislamiento & purificación , Medicamentos Herbarios Chinos/aislamiento & purificación , Glucósidos/aislamiento & purificación , Fenoles/aislamiento & purificación , Alcohol Feniletílico/análogos & derivados , Rhodiola/química , Química Farmacéutica , Cromatografía Líquida de Alta Presión , Cumarinas/análisis , Medicamentos Herbarios Chinos/análisis , Glucósidos/análisis , Fenoles/análisis , Alcohol Feniletílico/análisis , Alcohol Feniletílico/aislamiento & purificaciónRESUMEN
OBJECTIVE: To study the anti-complementary phenolic acids from Lonicera japonica. METHOD: The anti-complementary activity-directed isolation was carried out with the hemolysis test as guide. All isolation was evaluated for their in vitro anti-complementary activities. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR, 13C-NMR data. RESULT: Fourteen compounds were isolated from the EtOAc fraction of L. japonica extracts, including 8 phenolic acids: 5-O-caffeoylquinic acid (1), chlorogenic (2), 4-O-caffeoylquinic acid (3), 3,5-di-O-caffeoylquinic acid (4), 4,5-di-O-caffeoylquinic acid (5), 3,4-di-O-caffeoylquinic acid (6), caffeic acid (7) and methyl caffeate acid (8); 3 iridoids: secologanoside (9), sweroside (10) and secoxyloganin (11); and 3 flavonoids: luteolin (12), quercetin (13) and kaempferol (14). Compounds 1-9 and 11-14 showed anti-complementary activity in different extents and 3,5-di-O-caffeoylquinic acid (4) exhibited the most significant activity against the classical pathway. CONCLUSION: Compound 14 is obtained from this plant for the first time, phenolic acids are the main anti-complementary constituents of L. japonica and 3,5-di-O-caffeoylquinic acid(4) is a potential complement inhibitor with strong activity, which worthy to be studied further in the future.
Asunto(s)
Inactivadores del Complemento/aislamiento & purificación , Hidroxibenzoatos/aislamiento & purificación , Lonicera/química , Inactivadores del Complemento/química , Inactivadores del Complemento/farmacología , Hidroxibenzoatos/química , Hidroxibenzoatos/farmacologíaRESUMEN
The extracting technology of salidroside, tyrosol, crenulatin and gallic acid from Rhodiolae Crenulatae Radix et Rhizoma was optimized. With extraction rate of salidroside, tyrosol, crenulatin and gallic acid as indexes, orthogonal test was used to evaluate effect of 4 factors on extracting technology, including concentration of solvent, the dosage of solvent, duration of extraction, and frequency of extraction. The results showed that, the best extracting technology was to extract in 70% alcohol with 8 times the weight of herbal medicine for 2 times, with 3 hours once. High extraction rate of salidroside, tyrosol, crenulatin and gallic acid were obtained with the present technology. The extracting technology was stable and feasible with high extraction rate of four compounds from Rhodiolae Crenulatae Radix et Rhizoma, it was suitable for industrial production.
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Fraccionamiento Químico/métodos , Química Farmacéutica/métodos , Cumarinas/aislamiento & purificación , Medicamentos Herbarios Chinos/aislamiento & purificación , Ácido Gálico/aislamiento & purificación , Glucósidos/aislamiento & purificación , Fenoles/aislamiento & purificación , Alcohol Feniletílico/análogos & derivados , Rizoma/química , Rhodiola/química , Alcohol Feniletílico/aislamiento & purificaciónRESUMEN
The reaction conditions of baicalin hydrolyzed into baicalein by a kind of thermophilic and sugar-tolerant beta-glucosidase were studied in this paper. The beta-glucosidase could catalyze baicalin into baicalein well in the acetic acid-sodium acetate buffer. The optimal enzyme activity was at 85 degrees C and pH 5.5. The enzyme was stable at the temperature less than 85 degrees C and pH range of 5-7.5. The maximum reaction rate V. and michaelis constant K. were 0.41 mmol x L(-1) x min(-1) and 3.31 mmol x L(-1) respectively. Different metal ions had different effects on the activity of enzyme. Na+ existing in acetic acid-sodium acetate buffer had an activation effect on enzyme. The enzyme activity was enhanced by the concentrations of glucose below 0.6 mol x L(-1), and was gradually inhibited when monosaccharide concentration was over 0.6 mol x L(-1). When the monosaccharide concentration reached 1.2 mol x L(-1), the inhibition rate of enzyme activity was about 50%, which showed good glucose tolerance. The good reaction conditions through the experiment have been determined as follows, the substrate: enzyme dose was 1 g: 0.2 mL, acetic acid-sodium acetate buffer pH 5.5, reaction temperature 85 degrees C, reaction time 10 h, and the enzymatic hydrolyzation ratio could reach 97%.
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Flavanonas/química , beta-Glucosidasa/química , Biocatálisis , Estabilidad de Enzimas , Flavonoides/química , Glucosa/química , Calor , Hidrólisis , CinéticaRESUMEN
Four new triterpenoids, 2-O-acetyl-3-O-(4'-O-acetyl)-α-l-arabinopyranosylmaslinic acid (1), 2-O-acetyl-3-O-(3'-O-acetyl)-α-l-arabinopyranosylmaslinic acid (2), 2-O-acetyl-3-O-(3',4'-O-diacetyl)-α-l-arabinopyranosylmaslinic acid (3), and 3-O-(3'-O-acetyl)-α-l-arabinopyranosyloleanolic acid (4), together with six known triterpenoids, 3-O-(4'-O-acetyl)-α-l-arabinopyranosyloleanolic acid (5), maslinic acid (6), 2-O-acetylmaslinic acid (7), 3-O-acetylmaslinic acid (8), betulinic acid (9), and 2α-hydroxy-3ß-O-acetylbetulinic acid (10), were isolated from the EtOAc extract of Garcinia hanburyi resin. Their structures were elucidated by analysis of the spectroscopic data and chemical methods.
Asunto(s)
Garcinia/química , Triterpenos/aislamiento & purificación , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Triterpenos Pentacíclicos , Resinas de Plantas/química , Triterpenos/química , Vietnam , Ácido BetulínicoRESUMEN
OBJECTIVE: To investigate the chemical constituents of dried whole plants of Artemisia annua. METHOD: The chemical constituents were isolated by repeated silica gel chromatography, medium pressure column chromatography, and semi-preparative HPLC, and their structures were elucidated by spectroscopic analyses and comparison of NMR data with those reported in literature. RESULT: 15 compounds were isolated and identified to be 5-O-[(E)-Caffeoyl] quinic acid(l), 1,3-di-O-caffeoylquinic acid(2), 4 5-di-O-caffeoylquinic acid(3), 3, 5-di-O-caffeoylquinic acid (4), 3, 4-di-O-caffeoylquinic acid (5), methyl-3,4-di-O-caffeoylquinic acid(6), methyl-3,5-di-O-caffeoylquinic acid(7), 3,6'-O-diferuloylsucrose(8), 5'-ß-D-glucopyranosyloxyjasmonic acid(9), Scopoletin(10), scoparone (11), 4-O-ß-D-glucopyranosyl-2-hydroxyl-6-methoxyacetophenone (12), chrysosplenol D (13), casticin (14), chrysosplenetin(15). CONCLUSION: Compounds 2, 6, 8 and 9 are obtained from the Artemisia genus for the first time. Compounds 7 and 15 are obtained from this plant for the first time.