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Based on the processing and compatibility, this study explored the effects of components in Corni Fructus(CF) and Astragali Radix(AR) on plasma metabolomics in diabetic nephropathy rats. SD rats were randomly divided into four groups and diabetic nephropathy rat model was induced by high-fat diet combined with 30 mg·kg~(-1) streptozotocin(STZ). Histopathological observations of kidney tissue sections of rats in each group were conducted using HE, PAS, and Masson staining. Ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UHPLC-Q-TOF-MS/MS) metabolomics method was employed to investigate the effects of CF before and after wine-processing combined with AR-related components on plasma metabolites in diabetic nephropathy rats. After drug treatment, kidney tissue damage and interstitial collagen fiber deposition area in diabetic nephropathy rats were improved to varying degrees(P<0.001). The detection results of plasma metabolomics showed that 71 biomarkers related to the pathogenesis of diabetic nephropathy were identified in diseased rats, mainly involving linoleic acid metabolism, caffeine metabolism, glycerophospholipid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, phenylalanine metabolism, retinol metabolism, and ether lipid metabolism. After drug intervention, 26 of them were significantly downregulated, with better efficacy observed in precision processed herb-pair group(P-CG_5). This study elucidated from the perspective of plasma metabolomics that P-CG_5 could improve metabolic disorders in diabetic nephropathy through pathways such as phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, and caffeine metabolism, providing theoretical support and experimental basis for the clinical application of CF and AR compatibility in traditional Chinese medicine.
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Cornus , Nefropatías Diabéticas , Medicamentos Herbarios Chinos , Metabolómica , Ratas Sprague-Dawley , Espectrometría de Masas en Tándem , Animales , Nefropatías Diabéticas/tratamiento farmacológico , Nefropatías Diabéticas/metabolismo , Ratas , Masculino , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/farmacología , Cornus/química , Astragalus propinquus/química , Vino/análisis , Humanos , Riñón/efectos de los fármacos , Riñón/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Cistanche tubulosa (CT) is the dried fleshy stem with scaly leaves of Cistanche tubiflora (Schenk) Wight, which has the effects of tonifying the kidney-yang, benefiting the vital essence and blood, and moisturizing the intestines and laxatives. There are differences in the activity of CT before and after processing, but the mechanism of processing is not clear. AIM OF THE STUDY: The study aimed to compare the strength of action of CT before and after yellow-wine processing in the treatment of constipation and kidney yang deficiency and to identify the active ingredients responsible for the differences in activity before and after yellow-wine processing. MATERIALS AND METHODS: This study established the fingerprints of CT and PCT using HPLC to identify their shared components. Then efficacy of KYDS and FC were carried out to compare the differences between CT and PCT in terms of efficacy. Next, this study established the spectrum-effect relationship between the shared chemical components and the medical effects of CT and PCT using the gray correlation analysis and entropy methods. Ultimately, the activity of the analyzed chemical components was verified using the zebrafish model. RESULTS: CT was more effective than PCT in promoting intestinal peristalsis, regulating gastrointestinal hormone levels, and thus treating FC. PCT was more effective than CT in improving the level of hormone indexes of the hypothalamus-pituitary-target gland axis, replenishing blood, and enhancing immunity. Through the analysis of the spectrum-effect relationship, it was finally found that 5, 6, 12 (tubuloside A), and 13 (isoacteoside) might be more closely related to the activity of tonifying kidney yang, and peaks 9, 10, and 11 (acteoside) are more closely associated with the treatment of constipation, and peaks 3 (salidroside), 4, 1, 2 (geniposidic acid), and 8 (echinacoside) were associated with both kidney yang tonic and treatment of constipation. At the same time, an activity verification experiment showed that echinacoside, geniposidic acid, and salidroside were effective in the treatment of FC and KYDS, while acteoside was very effective in the treatment of FC, and tubuloside A was significant in supplementing the blood, which validated the spectrum-effect relationship analysis. CONCLUSION: This study proved that the raw CT had a better laxative effect, while the yellow-wine processed CT had a better kidney-yang tonic effect; moreover, spectrum-effect relationships were established to analyze the chemical components leading to changes in the activity of CT before and after yellow-wine processing.
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Cistanche , Glucósidos , Glucósidos Iridoides , Fenoles , Polifenoles , Animales , Quimiometría , Pez Cebra , Glicósidos/farmacología , Glicósidos/uso terapéutico , EstreñimientoRESUMEN
This study aimed to explore the differences in raw Angelica Sinensis (RAS), wine washing AS (WAS), and wine stir-frying AS (WSAS). The results showed there were differences among the three AS in color and aroma, and 34 aroma compounds were identified. The content determination results revealed the ferulic acid and Z-ligustilide levels of RAS decreased after processing, and those in WAS were higher than in WSAS. Furthermore, 85 representative common components and 37 unique components were tentatively identified in three AS. Finally, the free radical scavenging assay results indicated the antioxidant capacity of RAS was reduced after processing, and the antioxidant capacity of WAS was better than WSAS. Collectively, the RAS undergoes significant changes in color, aromas, components, and antioxidant ability after processing, and the different processing methods also result in significant differences between WAS and WSAS.
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ETHNOPHARMACOLOGICAL RELEVANCE: Taohong Siwu Decoction (THSWD) is based on the "First Recipe of Gynecology." It is widely used in various blood stasis and deficiency syndromes, mainly in gynecological blood stasis, irregular menstruation, and dysmenorrhea. THSWD has great demand in traditional Chinese medicine (TCM), gynecology, orthopedics, and internal medicine. According to classical records, three medicinal materials, namely Rehmanniae radix, Angelica sinensis, and Carthamus tinctorius, used in THSWD need to be "washed with yellow rice wine." In the study of TCM prescriptions, the processing methods of medicinal materials not only needed to follow traditional records but also should consider modern technical conditions. Many medicinal materials in the repertoire of classical prescriptions involve yellow rice wine processing. Determining the processing method for medicinal materials is a key and difficult problem in the research and development of classical prescriptions. AIM OF THE STUDY: With THSWD as the representative, this study analyzed differences between no processing method, the modern processing method of "stir-frying the materials with yellow rice wine," and the traditional processing method of "washing with yellow rice wine." We focused on three aspects: composition, efficacy, and endogenous metabolism. This study aimed to provide a reference for research on the processing methods of medicinal materials used in classical prescriptions. MATERIALS AND METHODS: UPLC-Q-Orbitrap HRMS was used to quickly identify and classify the main chemical compounds of THSWD. A model of primary dysmenorrhea (PD) was established using estradiol benzoate combined with oxytocin. The latent period and writhing time; the levels of serum PGF2α, PGE2, ET-1, and ß-EP; and the pathological sections of the uterus were observed to determine their pharmacodynamic differences. GC-TOF/MS was used to analyze the differences in serum metabolites in rats. RESULTS: A total of 54 active compounds were identified, and the results showed that catalpol and rehmapicroside disappeared following yellow rice wine processing. Compared with materials processed by the traditional method, the relative contents of 15 components, such as 5-hydroxymethylfurfural and digitalis C, increased in materials processed by the modern method. However, the relative contents of 16 components, such as hydroxysafflor yellow A, verbascoside, and ferulic acid, decreased in the modern processing method. The modern and classic processing methods acted on PD through different metabolic pathways. THSWD obtained by classical processing methods mainly treated PD through anti-inflammatory and estrogen metabolism pathways, whereas THSWD obtained by modern processing methods mainly treated PD through anti-inflammatory metabolic pathways. CONCLUSION: The study revealed the differences in different yellow rice wine processing methods in terms of chemical composition of the THSWD obtained, as well as the mechanisms of action for the treatment of PD. This study provides a reference for the clinical application of THSWD and development of classical prescription preparations.
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Química Farmacéutica/métodos , Medicamentos Herbarios Chinos/química , Oryza/química , Vino , Angelica sinensis , Carthamus tinctorius , Chalcona/análogos & derivados , Chalcona/química , Humanos , Quinonas/química , RehmanniaRESUMEN
Wine-processed Angelica Sinensis is a widely used Chinese medicinal decoction piece in China. However, there are hardly any robust markers indicating the processing procedure of wine-processed Angelica Sinensis, including the amount of rice wine and processing degree. A strategy integrating untargeted metabolomics and chemometric analysis for deducing robust markers was provided and applied to the discrimination of processing procedure. First, 86 compounds were tentatively identified in wine-processed Angelica Sinensis by ultra-high-performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry. Second, 93 potential chemical markers were selected using multivariate analysis, among which nine robust chemical markers were selected by verification with commercial samples. Finally, the effects of processing temperature, time, and amount of rice wine on the three selected chemical markers were investigated through a rapid analytical method. It was demonstrated that both m/z 258.1097 and 238.1189 were positively correlated with the amount of rice wine and processing degree. In summary, this study introduced two candidate processing markers as robust markers for discriminating the processing procedures of wine-processed Angelica sinensis. It also proposed a strategy to provide the reference for the research of other decoction pieces.
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Angelica sinensis/química , Fermentación , Metabolómica , Biomarcadores/análisis , Quimiometría , Medicamentos Herbarios Chinos/química , Espectrometría de Masas , Medicina Tradicional China , Vino/análisisRESUMEN
An accurate and sensitive ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry method was established and validated for the determination of nine bioactive compounds of Ligustri Lucidi Fructus in rat plasma. Separation was performed on Halo® C18 column with a mobile phase of acetonitrile and 0.1% formic acid in water. The eluate was detected by multiple reaction monitoring scanning operating in the negative ionization mode. This assay method was validated for selectivity, linearity, intra- and interday precision, accuracy, recovery, matrix effect, and stability, and all methodological parameters fulfilled the Food and Drug Administration criteria for bioanalytical validation. The established method was successfully applied to a comparative pharmacokinetic study of raw and wine-processed Ligustri Lucidi Fructus in rats for the first time. It was found that the AUC0-24 and Cmax value of salidroside, hydroxytyrosol, and nuezhenidic acid were increased significantly after processing, while the AUC0-24 and Cmax value of oleoside 11-methyl ester, 1'''-O-ß-d-glucosylformoside, specnuezhenide, G13, oleonuezhenide, and oleanolic acid were decreased, which suggested that processing affects the absorption and bioavailability of Ligustri Lucidi Fructus. The results might be valuable for the clinical reasonable application and understanding the processing mechanism of Ligustri Lucidi Fructus.
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Medicamentos Herbarios Chinos/análisis , Frutas/química , Ligustrum/química , Vino/análisis , Administración Oral , Animales , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/administración & dosificación , Masculino , Espectrometría de Masas , Estructura Molecular , Ratas , Ratas Sprague-DawleyRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Wine-processed Radix scutellariae (RS) is the processed product of RS, which is the dried root of Scutellaria baicalensis Georgi. It is recorded in Chinese traditional formula that wine-processed RS has the effect of anti-migraine, while the effect has not been confirmed and the possible mechanism remains unclear. AIM OF THE STUDY: To verify the anti-migraine effect of wine-processed RS in nitroglycerin (NTG)-induced rats and explore the correlation between compounds dissolution and the pore structure based on fractal theory. MATERIALS AND METHODS: In the validation of pharmacodynamics, the effects of wine-processed RS on migraines were firstly evaluated by observing the number of head-scratching of rats, then investigated by determining the levels of nitric oxide (NO), calcitonin gene-related peptide (CGRP) and the expression of c-Fos in the brain of NTG-induced rat models using ELISA and immunohistochemical assessments. In the correlation study, the stir-frying time of RS was set to 5 min, 10 min and 15 min. The scanning electron microscope (SEM) and mercury intrusion method were used to explore the pore structure and main parameters of the pore structure including pore size distribution, pore volume, porosity, surface area and fractal dimension. The compounds dissolution of total flavonoids and five major components containing baicalein, baicalin, scutellarin, wogonin and wogonoside was determined by UV-Vis spectrophotometry and HPLC separately. RESULTS: The animal experiments had shown that wine-processed RS could significantly reduce the head-scratching times of NTG-induced rat models (p < 0.01) and markedly decrease the levels of NO (p < 0.01), CGRP (p < 0.05) and the expression of c-Fos (p < 0.01) compared with model group. The data indicated that wine-processing would affect the dissolution of compounds by changing the pore structure of RS. The order of positive correlation between pore structure parameters and compounds' dissolution was total surface area > fractal dimension (r > 0) and the order of negative correlation was average pore size > total porosity > total volume (r < 0). Compared with the other sample groups (p < 0.05), the wine-processed RS stir-fried for 10 min had a pore structure which was more favorable for compounds dissolution. CONCLUSIONS: Wine-processing could strengthen the anti-migraine effect of RS by changing the pore structure of RS, which is linked to the dissolution of compounds. The RS stir-fried for 10 min may be more effective in treating migraine.
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Trastornos Migrañosos/inducido químicamente , Trastornos Migrañosos/prevención & control , Nitroglicerina/toxicidad , Extractos Vegetales/uso terapéutico , Scutellaria baicalensis , Vino , Animales , Fractales , Masculino , Extractos Vegetales/aislamiento & purificación , Ratas , Ratas Sprague-Dawley , Solubilidad , Vasodilatadores/toxicidad , Vino/análisisRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Scutellariae radix (SR) is one of the most popular traditional Chinese medicines (TCM). Crude SR (CSR) and wine-processed SR (WSR) are the two most common commercial specifications. According to the theories of TCM, wine-processing increases the inclination and direction of SR's actions, thereby strengthening its efficacy in clearing the upper-energizer lung damp heat. The pharmacological mechanism-related research on WSR for the treatment of lung disease is limited and needs to be expanded. AIM OF THE STUDY: The aim of this report was to identify the relevant biological pathways by assessing changes in plasma metabolites between CSR and WSR in a lipopolysaccharide (LPS)-induced acute lung injury (ALI) model, and thus, revealed the potential mechanism of wine processing in SR. MATERIALS AND METHODS: Rats with LPS-induced ALI were treated with CSR and WSR. The contents of inflammatory cytokines and histopathological examination were determined to explore the effects of CSR and WSR. Next, the metabolic profiling of rat plasma samples was performed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Then, principal component analysis (PCA) were used to provide an overview for all of the groups and orthogonal partial least squares-discriminant analysis (OPLS-DA) was utilized to maximize the discrimination and present the differences in the metabolite between all of the groups. RESULTS: WSR exhibited a more remarkable effect on improving ALI than CSR by reducing the levels of inflammatory factors, including nitric oxide (NO), tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and interleukin-8 (IL-8). On the basis of UPLC-QTOF-MS technology, an unequal curative effect was revealed by nontargeting metabolomics. Sixteen biomarkers were discovered in the plasma of LPS-induced rats. Pathway analysis indicated that CSR acted on ALI by regulating the abnormal sphingolipid metabolism pathways; however, an WSR-mediated cure of ALI was linked primarily to reversing the abnormality of retinol metabolism pathways and tryptophan metabolism pathways. CONCLUSIONS: This report examined the underlying wine-processing mechanism of SR from the perspective of plasma metabolites. In addition, this work provided a novel and valuable insight into interpretation of the processing mechanisms of TCM in a holistic way.
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Lesión Pulmonar Aguda/tratamiento farmacológico , Scutellaria baicalensis , Vino , Lesión Pulmonar Aguda/sangre , Lesión Pulmonar Aguda/inmunología , Lesión Pulmonar Aguda/patología , Animales , Citocinas/inmunología , Lipopolisacáridos , Pulmón/efectos de los fármacos , Pulmón/inmunología , Pulmón/patología , Masculino , Redes y Vías Metabólicas/efectos de los fármacos , Metabolómica , Óxido Nítrico/sangre , Ratas Sprague-DawleyRESUMEN
Polygonatum sibiricum (PS) is a traditional Chinese herb used in both food and medicine with great bioactivity. The wine-processed pieces of PS are the main form for clinical application, while research has focused on the polysaccharides of their crude form. This study evaluated the physicochemical properties and immunological activities of water-soluble polysaccharides from both crude (PSPC) and wine-processed PS (PSPW). PSPC and PSPW had significant differences in their physicochemical properties. PSPC was mainly composed of galactose, mannose, glucose, and galacturonic acid, in molar ratios of 29.63:36.10:15.09:10.20, while PSPW was mainly composed of galactose, mannose, and galacturonic acid, in molar ratios of 78.77:5.50:13.84. Both kinds of polysaccharides can enhance the cells viability, phagocytic capacity, acid phosphatase activity, and NO production of RAW264.7 cells. We found that PSPC and PSPW enhanced the immune functions of the immunosuppressive model for spleen deficient mice and reversed the decline of the secretions of IL-2, IL-6, TNF-α, and IFN-γ to a normal range. The PSPW showed more potent immunological activities than PSPC. The results of the study identify the importance of wine-processing for PS and provide application foundations for the further development of PSPW as a functional food.
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Polygonatum/química , Polisacáridos/química , Polisacáridos/farmacología , Animales , Supervivencia Celular/efectos de los fármacos , Galactosa/química , Glucosa/química , Manosa/química , Ratones , Polygonatum/inmunología , Polisacáridos/inmunología , Células RAW 264.7 , Vino/microbiologíaRESUMEN
1. The aim of this study was to develop a selective, rapid, accurate and sensitive ultrahigh performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for pharmacokinetic (PK) studies of phytoecdysones and triterpenoid saponins after oral administration of five monomers, crude, wine-processed and salt-processed Radix Achyranthis bidentatae (RAB).2. A Thermo Hypersil GOLD C18 column (100 mm × 2.1 mm, 1.9 µm) coupled with a mobile phase of (A) acetonitrile and (B) water (both containing 0.3% acetic acid) was used for sample separation. The mass analysis was performed in a triple quadruple mass spectrometer using selected reaction monitoring (SRM) with negative scan mode.3. The results showed that this method exhibited desirable sensitivity, precision, stability and repeatability. The extraction recoveries of the compounds ranged from 94.2 to 99.8% and the matrix effects ranged from 93.3 to 100.5%. Comparing the Cmax and AUC of five analytes in those groups showed this tendency: salt-processed RAB > wine-processed RAB > crude RAB > monomer group. The results confirmed the feasibility of TCM theory to enhance the efficacy of processed RAB.
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Ecdisona/farmacocinética , Fitosteroles/farmacocinética , Saponinas/farmacocinética , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Medicamentos Herbarios Chinos/farmacocinética , Espectrometría de Masa por Ionización de Electrospray , Espectrometría de Masas en Tándem , TriterpenosRESUMEN
ABSTRACT This study was designed to explore the pharmacokinetic regularity of the plasma concentration, tissue distribution and excretion of orcinol glucoside from aqueous extracts of raw and processed Curculigo orchioides Gaertn., Hypoxidaceae. The experiment first used an ultrahigh-performance liquid chromatography-tandem mass spectrometry approach with multiple reaction monitoring and a positive mode to separate orcinol glucoside from naringin to obtain the plasma concentration curves, bar graph of tissue distribution and excretion curves. These results might be beneficial for reasonable clinical application of C. orchioides and for further development of its wine and salt-processing mechanism.
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Objective:To establish the quality control method of the wine-processing Salviae Miltiorrhizae Radix et Rhizoma standard decoction,in order to provide reference for the quality evaluation of the wine-processing Salviae Miltiorrhizae Radix et Rhizoma formula granules and other related products. Method:Totally 15 batches of representative wine-processing Salviae Miltiorrhizae Radix et Rhizoma pieces were collected to prepare the standard decoction, establish the HPLC fingerprint and determine the content of five components(sodium danshensu,caffeic acid,rosmarinic acid,lithospermic acid,salvianolic acid B). The main common peaks in fingerprint were identified to define the main chemical constituents in the standard decoction, the parameters,such as dry extract rate,transfer rate of index components and pH of the standard decoction were calculated, and the comprehensive evaluation index was established to evaluate the stability of the preparation process. Result:The main component standard decoction was phenolic acids. The concentrations of five components(sodium danshensu,caffeic acid,rosmarinic acid,lithospermic acid,salvianolic acid B)in the standard decoction were 0.21%-0.37%,0.03%-0.10%,0.08%-0.18%,0.07%-0.13%,2.68%-4.34%, the dry extract rates of standard decoction were 71.8%-85.4%,50.0%-71.4%,68.2%-81.0%,66.7%-84.6%,67.5%-79.6%,the transfer rates were between 45.1%-55.3%,and pH value was between 5.91-6.05. The fingerprint similarities of the 15 batches of standard decoction with reference fingerprints were>0.98,the fingerprint showed 12 common peaks,7 of which were considered to be sodium danshensu,protocatechuic aldehyde,caffeic acid,ferulic acid,rosmarinic acid,lithosperic acid and salvianolic acid B. Conclusion:The established systematic evaluation for the quality of wine-processing Salviae Miltiorrhizae Radix et Rhizoma standard decoction is stable and feasible,and provides a reference for the quality control of relevant preparations of wine-processing Salviae Miltiorrhizae Radix et Rhizoma decoction.
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Objective:To explore the difference of effect of raw and wine-processed products of Rhei Radix et Rhizoma extract on brain protection of rats with intracerebral hemorrhage(ICH) and the connotation of the theory of "wine processing could promote efficacy" of Rhei Radix et Rhizoma. Method:Wistar male rats were used to establish the ICH model,extracts of raw and wine-processed products of Rhei Radix et Rhizoma were used for intervention,and these rats were randomly divided into 6 groups(blank group,sham-operated group,model group,raw Rhei Radix et Rhizoma group,wine-processed Rhei Radix et Rhizoma group and Angong Niuhuangwan group).Then the apoptosis of hippocampal neurons was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay.The expression of zonula occludens-1(ZO-1) and vascular endothelial cadherin-2(VE-cadherin-2) in the hemorrhagic brain tissue was detected by immunohistochemical assay.The expression of reduced form of nicotinamide-adenine dinucleotide phosphate(NADPH) oxidase 2(NOX2) and tumor necrosis factor-α stimulated gene-6(TSG-6) in the hemorrhagic brain tissue was detected by Western blot.The content of glutathione(GSH) in the serum of ICH rats was detected by automatic enzyme-linked immunosorbent assay systems.Through these above methods,we investigated the differences between raw products and wine-processed products on brain protection of ICH rats. Result:Compared with the sham-operated group,the apoptosis index of the hippocampal neurons in the model group were increased significantly(PPPPPPPPPPPPConclusion:The raw and wine-processed products of Rhei Radix et Rhizoma all have brain protective effect on ICH rat model,but the protective effect of wine-processed products is slightly better than that of raw products.The result of this study provides experimental basis for exploring the theory of "wine processing could promote efficacy" of this herb.
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Gentiana radix is used in traditional Chinese medicine and has functions of clearing heat and drying dampness, as well as purging liver and gallbladder fire. A highly sensitive and effective strategy for rapid screening and identification of target constituents has been developed by using ultra high-performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap) in crude and wine-processed Gentiana radix. Based on the accurate mass measurement (<5 ppm), retention times, and MS fragmentation ions, 52 constituents were unambiguously or tentatively characterized from Gentiana radix, including 21 iridoids, 11 flavonoids, 19 xanthones, and a triterpenoid. This study demonstrated that the established method could be a rapid, effective analytical tool for screening and characterization of compounds in the complex systems of Gentiana radix. By comparing the structure and peak areas of chemical constituents in crude and wine-processed Gentiana radix, we found that some compounds in crude and wine-processed Gentiana radix were significantly different.
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Gentiana/química , Vino , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/química , Flavonoides/análisis , Raíces de Plantas/química , Espectrometría de Masa por Ionización de Electrospray , Terpenos/análisis , Xantonas/análisisRESUMEN
Radix Scutellariae (RS) is a herbal medicine with various pharmacological activities to treat inflammation, respiratory and gastrointestinal infections, etc. In this study, a rapid, sensitive and selective UPLC-ESI-MS/MS method was developed for simultaneous determination of 10 flavonoids - scutellarin, scutellarein, chrysin, wogonin, baicalein, apigenin, wogonoside, oroxylin A-7-O-glucuronide, oroxylin A and baicalin - from RS aqueous extracts in rat plasma with propyl paraben as internal standard (IS). Chromatographic separation was achieved on a C18 column using gradient elution with the mobile phase consisting of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. The detection was performed in multiple reaction monitoring mode using electrospray ionization in negative mode. The validated method showed good linearity over a wide concentration range (r >0.9935). The intra- and interday assay variabilities were <9.5% and <12.4% for all analytes, respectively. The extraction recovery ranged from 71.2 to 89.7% for each analyte and IS. This method was successfully applied to pharmacokinetic comparision after oral administration of crude and wine-processed RS aqueous extracts. There were significant differences in some pharmacokinetic parameters of most analytes between crude and wine-processed RS. This suggested that wine-processing exerted effects absorption of most flavonoids.
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Medicamentos Herbarios Chinos/administración & dosificación , Flavonoides/sangre , Scutellaria baicalensis/química , Vino/análisis , Animales , Cromatografía Líquida de Alta Presión/métodos , Estabilidad de Medicamentos , Medicamentos Herbarios Chinos/farmacocinética , Flavonoides/química , Flavonoides/farmacocinética , Modelos Lineales , Masculino , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masas en Tándem/métodosRESUMEN
Rhizoma coptidis (R.C.), a widely used traditional Chinese medicine, has been used for centuries in the treatment of hypertension, inflammation, dysentery and liver diseases, etc. Wine-processing is a specialized technology by sautéing crude herbal medicine using Chinese rice wine. This paper was designed to establish a simultaneous quantitative method of ten alkaloids (berberine, coptisine, palmatine, jatrorrhizine, epiberberine, magnoflorine, columbamine, noroxyhydrastinine, oxyberberine and 8-oxocoptisine) in rat plasma. Furthermore, the pharmacokinetics of those alkaloids after administration of crude and wine-processed R.C. aqueous extracts was compared. As a result, a ultra high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) method was developed and validated for the first time. Chromatographic separation was achieved on a C18 column using gradient elution with the mobile phase consisting of acetonitrile and water (containing 0.2% formic acid) at a flow rate of 0.2 ml/min. The validated method showed good linearity over a wide concentration range (r>0.99), and lower limits of quantification less than 5.46 ng/ml for the each analyte. The intra- and inter-day assay variability was below 9.9% and 10.5% for all analytes, respectively. The extraction recovery of those alkaloids and I.S. ranged from 65.3% to 90.7%. The validated method has been successfully applied to pharmacokinetic comparison after administration of crude and wine-processed R.C. aqueous extracts. Pharmacokinetic comparative study showed that Cmax of coptisine and 8-oxocoptisine and AUC0-t of coptisine, palmatine and 8-oxocoptisine were increased significantly (p<0.05) after wine-processing, while other compounds didn't show significant difference, which suggested that wine-processing exerted limited effects on the absorption of alkaloids. These results might be helpful for R.C.' clinical reasonable application and further studies on its wine-processing mechanism.
Asunto(s)
Alcaloides/sangre , Cromatografía Líquida de Alta Presión/métodos , Coptis/química , Medicamentos Herbarios Chinos/química , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masas en Tándem/métodos , Vino , Alcaloides/farmacocinética , Animales , Calibración , Masculino , Ratas Sprague-Dawley , Estándares de Referencia , Rizoma/química , Sensibilidad y Especificidad , Agua/químicaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Ascending and descending theory is a core principle of traditional Chinese medicine (TCM) theories. It plays an essential role in TCM clinical applications. Some TCM medicine has specific properties, which could alter the inclination and direction of their actions. The properties of the ascending and floating process of one herbal medicine are affected by means of herb processing. Wine-processing, which is sautéing with rice wine, is one of the most popular technologies of herb processing. Wine-processing increases the inclination and direction of its actions, thereby producing or strengthening their efficacy in cleaning the upper-energizer heat. Radix scutellariae, the dried roots of Scutellaria baicalensis Georgi, is a well-known TCM used for the treatment of inflammation, pyrexia, jaundice, etc. Recently, wine-processed Radix scutellariae was normally applied in clinical studies for the treatment of upper-energizer syndrome. In order to investigate the effects of wine-processing on ascending and descending of Radix scutellariae, the comparative study of distribution of flavonoids in rat tissues of triple energizers (SanJiao-upper, middle, lower jiao) after oral administration of crude and wine-processed Radix scutellariae aqueous extracts was carried out. MATERIALS AND METHODS: The rats were randomly assigned to two groups and orally administered with crude and wine-processed Radix scutellariae aqueous extracts, respectively. At different pre-determined time points after administration, the concentrations of compounds in rat tissue homogenate were determined, and the main tissue pharmacokinetic parameters were investigated. Tissue pharmacokinetic parameters including AUC0-t, t1/2, Tmax and Cmax were calculated using DAS 2.0. An unpaired Student t-test was used to compare the differences in tissue pharmacokinetic parameters between the two groups. All the results were expressed as arithmetic mean±S.D. RESULTS: The parameters of Cmax and AUC0-t of some flavonoids in wine-processed Radix scutellariae were remarkably increased (p<0.05, p<0.01, p<0.001) in the rat upper-energizer tissues (lung and heart) compared with those of the crude group. However, in the rat middle- and lower-energizer tissues (spleen, liver and kidney), the Cmax and AUC0-t of some flavonoids were significantly decreased (p<0.05, p<0.01) compared with the crude group. The main explanation for these differences seems to the effects of wine-processing on ascending and descending theory. CONCLUSIONS: All of these differences in the distribution of triple energizers after oral administration of crude and wine-processed Radix scutellariae aqueous extracts may lead to the increase of efficacy on the upper-energizer tissues and were in compliance with the ascending and descending theory. Therefore, wine-processing was recommended when Radix scutellariae was used for cleaning the upper-energizer heat and humidity. The obtained knowledge can be used to evaluate the impact of these differences on the efficacy of both the drugs in clinical applications and might be helpful in explaining the effects of wine-processing on ascending and descending theory.