albiflorin and peoniflorin

Radix Paeoniae Alba (RPA), a traditional Chinese medicine, has been used frequently in the treatment of asthma. Previous studies demonstrated the dichloromethane fraction of Stir-Frying RPA (FDCM) enhanced the effect of anti-allergic asthma compared with the dichloromethane fraction of RPA (DCM).. The significant increasing of Paeoniflorin (PF), ethyl gallate (EG), 1,2,3,4,6-pentagalloylglucose (PGG) had been observed in FDCM. This study aimed to investigate the effects and mechanisms of these compounds from FDCM in ovalbumin (OVA)-induced allergic asthma mouse model.. The significant difference contents compounds fraction (FB-40) and other fractions in FDCM were enriched by Medium Pressure Liquid Chromatography (MPLC). The pharmacodynamics was verified among all fractions in OVA-induced allergic asthma mice. Moreover, the drug dose dependence of FB-40 (0.42 mg/kg, 0.21 mg/kg, and 0.07 mg/kg), which were the most active fraction from FDCM for anti-allergic asthma, was explored. The expression of IL-6, p-STAT3, and STAT3 was analyzed by Western blot analysis. In addition, the main components of FB-40 were identified by UPLC with standards. Finally, the anti-inflammatory effects of the main components from FB-40 were detected by LPS-stimulated BEAS-2B cells using an Elisa assay.. The results showed that FB-40 was the most active fraction from FDCM, which could significantly improve the lung tissue pathological condition, and decrease the number of inflammatory cells in bronchoalveolar lavage fluid (BALF). It had greater pharmacological activity than its main component PF. FB-40 also showed dose dependence and regulated the IL-6/STAT3 signaling pathway in allergic asthma mice. Besides, PF, Albiflorin (AF), PGG, EG, and 1,2,3,6-Tetra-O-galloyl-β-D-glucose (TGG) from FB-40 were identified by UPLC with the standard. At last, in the LPS-induced BEAS-2B cell experiments, EG, PGG, 1,2,3,6-Tetra-O-galloyl-β-D-glucose (TGG) showed stronger inhibiting activities of cytokine than the monoterpenoid glycosides (PF and AF).. The research proved that FB-40 was an active fraction in FDCM, which regulates IL-6/STAT3 Signaling Pathway to ameliorate allergic asthma. Gallic acids including TGG and PGG, and EG also play a role in the treatment of allergic asthma in FB-40.

Topics: Animals; Anti-Allergic Agents; Asthma; Bronchoalveolar Lavage Fluid; Glucose; Interleukin-6; Lipopolysaccharides; Methylene Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Signal Transduction

Qi-Lin pill (QLP) is an effective traditional Chinese medicine prescription (TCMP) that has been used for the treatment of the oligoasthenozoospermia in China. Recently, some articles described the pharmacological effects of QLP and multiple ingredients in QLP contribute to its effects. However, the pharmacokinetic and target tissue distribution data of QLP are still unknown. In the present study, according to the Bioanalytical Method Validation Guidance of FDA, a sensitive and selective UPLC-MS/MS method was developed and validated for simultaneous determination of multiple constituents in rat plasma and testicular tissue, including morusimic acid A, codonopyrridium B, magnoflorine, emodin, 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (THSG), ecliptasaponin A, paeoniflorin, albiflorin, gallic acid, danshensu, salvianolic acid A, catechin, isosinensetin, nobiletin, formononetin, calycosin, icariside II, icariin and epimedin C. For 19 analytes, the LLOQs reached 0.01-4 ng/mL. And all calibration curves showed favorable linearity (r ≥ 0.9903) in linear ranges. The intra-day and inter-day precision (relative standard deviation) for all analytes was less than 14.92 %, and the accuracies (as relative error) were in the range of - 6.44 % to 6.22 %. Extraction recoveries and matrix effects of analytes and IS were acceptable. All analytes were stable during the assay and storage in plasma samples. The method was successfully applied for the pharmacokinetics and testis distribution of multiple chemical constituents in QLP after a single oral dose. As a result, high exposure of danshensu, gallic acid, paeoniflorin and albiflorin were observed in rat plasma and testicular tissue. Among the flavonoids, isosinensetin and nobiletin had high exposure in testicular tissue. Moreover, alleviation of progesterone reduction was evaluated in H

Topics: Animals; Chromatography, High Pressure Liquid; Chromatography, Liquid; Drugs, Chinese Herbal; Gallic Acid; Hydrogen Peroxide; Male; Rats; Reproducibility of Results; Tandem Mass Spectrometry; Testis; Tissue Distribution

Shen-Wu-Yi-Shen tablets (SWYST), a Chinese patent medicine consisting of 12 herbal medicines, was formulated by a famous TCM nephrologist, Zou Yunxiang. It is clinically used to improve the symptoms of nausea, vomiting, poor appetite, dry mouth and throat, and dry stool in patients with chronic renal failure (CRF) accompanied by qi and yin deficiency, dampness, and turbidity. SWYST can reduce urea nitrogen, blood creatinine, and urinary protein loss, and increase the endogenous creatinine clearance rate. However, little is known about its pharmacokinetics.. To compare the pharmacokinetics of six bioactive components after oral administration of SWYST in normal and adenine-induced CRF rats.. The UPLC-TSQ-MS/MS method was fully validated for its satisfactory linearity (r ≥ 0.9913), good precisions (RSD <11.5%), and accuracy (RE: -13.4∼13.1%), as well as acceptable limits in the extraction recoveries, matrix effects, and stability (RSD <15%). In normal rats, the six analytes were rapidly absorbed (T. A sensitive UPLC-TSQ-MS/MS method was validated and used to investigate the pharmacokinetics of SWYST in normal and CRF rats. This is the first study to investigate the pharmacokinetics of SWYST, and our findings elucidate the causes of their different pharmacokinetic behaviors in CRF rats. Furthermore, the results provide useful information to guide further research on the pharmacokinetic-pharmacodynamic correlation and clinical application of SWYST.

Topics: Administration, Oral; Animals; Chromatography, High Pressure Liquid; Creatinine; Drugs, Chinese Herbal; Emodin; Kidney Failure, Chronic; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Tablets; Tandem Mass Spectrometry

Total glucosides of peony (TGP) is extracted from Paeonia lactiflora Pallas, which has been approved for rheumatoid arthritis (RA) treatment. There were approximately 15 monoterpene glycosides identified in TGP. Pervious researches focused on the effects of TGP and the major ingredient paeoniflorin (PF), but the functions of other monoterpene glycosides and their interactions were not clear. Network pharmacology has been one of the new strategies for multi-target drug discovery. In this study, we investigate the functions of all components of TGP and their interactions in RA treatment based on network pharmacology methods.. The components of TGP were searched out the Web of Science, PubMed, China National Knowledge Infrastructure databases; then we identified the potential targets based of chemical similarity in the Similarity Ensemble Approach. The molecular related with RA were obtained from DrugBank, GeneCards, DisGeNET and Online Mendelian Inheritance in Man (OMIM) databases. The components-targets-disease network was constructed and analyzed with Cytoscape software; Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted with R for function analysis. The hub components-targets interactions were validated with Autodock Vina.. Twenty potential targets of TGP were predicted for RA treatment. The major components of TGP, PF and albiflorin (AF) had more predicted targets. Hub targets of TGP were LGALS3/9, VEGFA, FGF1, FGF2, IL-6, IL-2, SELP, PRKCA and ERAP1. These targets ameliorated RA mainly through inhibiting leukocyte recruitment and angiogenesis. Enriched pathways including VEGFR pathway, signaling by interleukins, PI3K-Akt signaling pathway, platelet activation, extracellular matrix organization, and so on. The combination of PF, AF and lactiflorin (LF) with the hub targets was further validated using docking program.. We investigated the comprehensive mechanism of TGP for RA treatment. We analyzed the different targets of the components in TGP and predicted the new effects of TGP on inhibiting leukocyte recruitment and angiogenesis. This study provides a better understanding of TGP on the RA treatment.

Topics: Aminopeptidases; Arthritis, Rheumatoid; Glucosides; Glycosides; Humans; Leukocytes; Minor Histocompatibility Antigens; Monoterpenes; Network Pharmacology; Paeonia; Phosphatidylinositol 3-Kinases

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Benzopyrans; Bridged-Ring Compounds; Disease Models, Animal; Drugs, Chinese Herbal; Flavonoids; Freund's Adjuvant; Glucosides; Joints; Male; Medicine, Chinese Traditional; Monoterpenes; Rats; Rats, Sprague-Dawley

Topics: Animals; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Coumaric Acids; Drugs, Chinese Herbal; Glucosides; Male; Molecular Structure; Monoterpenes; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

Total glucosides of peony (TGP), extracted from the root and rhizome of Paeonia lactiflora Pall, has well-confirmed immunomodulatory efficacy in the clinic. However, the mechanism and active ingredients remain largely unclear.. Our previous study revealed a low systemic exposure but predominant gut distribution of TGP components. The aim of this study was to investigate involvement of the gut microbiota in the immunoregulatory effects and identify the active component.. Mice received 3% DSS to establish a model of colitis. The treatment group received TGP or single paeoniflorin (PF) or albiflorin (AF). Body weight, colon length, inflammatory and histological changes were assessed. Gut microbiota structure was profiled by 16s rRNA sequencing. Antibiotic treatment and fecal transplantation were used to explore the involvement of gut microbiota. Metabolomic assay of host and microbial metabolites in colon was performed.. TGP improved colonic injury and gut microbial dysbiosis in colitis mice, and PF was responsible for the protective effects. Fecal microbiota transfer from TGP-treated mice conferred resilience to colitis, while antibiotic treatment abrogated the protective effects. Both TGP and PF decreased colonic indole-3-lactate (ILA), a microbial tryptophan metabolite. ILA was further identified as an inhibitor of epithelial autophagy and ILA supplementation compromised the benefits of TGP.. Our findings suggest that TGP acts in part through a gut microbiota-ILA-epithelial autophagy axis to alleviate colitis.

Topics: Animals; Autophagy; Bridged-Ring Compounds; Colitis; Drugs, Chinese Herbal; Dysbiosis; Feces; Gastrointestinal Microbiome; Glucosides; HCT116 Cells; Humans; Immunologic Factors; Indoles; Male; Mice, Inbred BALB C; Monoterpenes; Paeonia; RNA, Ribosomal, 16S

Topics: Animals; Bridged-Ring Compounds; China; Chromatography, High Pressure Liquid; Cinnamates; Drugs, Chinese Herbal; Glucosides; Isoflavones; Male; Medicine, Chinese Traditional; Monoterpenes; Paeonia; Plasma; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

Topics: Administration, Oral; Animals; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Female; Flavanones; Glucosides; Glycyrrhetinic Acid; Glycyrrhizic Acid; Monoterpenes; Polycystic Ovary Syndrome; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

In this study, in-depth observation and investigation of blood-brain barrier permeability and neuroprotective effect of Trichosanthes kirilowii cassia twig particles on rats with cerebral ischemia-reperfusion injury were performed. Focal cerebral ischemia-reperfusion injury model was established by middle cerebral artery occlusion method, reperfusion was implemented 2 hours after ischemia; qualitative analysis and investigation of trichosanthes kirilowii cassia twig particles in plasma, brain tissue and cerebrospinal fluid in normal and middle cerebral artery occlusion (MCAO) rats were done by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS); changes in neurological deficits, cerebral infarction stereometry, blood-brain barrier permeability and histopathological changes of MCAO model rats were observed. Qualitative analysis by HPLC-MS/MS results showed that ingredients, paeoniflorin, albiflorin, liquiritin in Trichosanthes kirilowii cassia twig particles can reach the brain through the blood-brain barrier. In the model group, glycyrrhizin and glycyrrhizic acid can be detected in brain tissue or cerebrospinal fluid. In addition, Trichosanthes kirilowii cassia twig particles can significantly lower neurological deficits of rats in middle cerebral artery occlusion model, reduce the Evans blue penetration, contract infarct size, and reduce pathological tissue injury of cerebral ischemia reperfusion. The ingredients of Trichosanthes kirilowii cassia twig particles can reach the brain tissue through the blood-brain barrier and play a role in neuroprotection of rats with cerebral ischemia-reperfusion injury, which has important research significance and brings scientific experimental, theoretical basis for clinical drug use.

Topics: Animals; Blood-Brain Barrier; Brain; Bridged-Ring Compounds; Flavanones; Glucosides; Glycyrrhizic Acid; Infarction, Middle Cerebral Artery; Male; Monoterpenes; Neuroprotective Agents; Plant Components, Aerial; Plant Extracts; Rats; Recovery of Function; Reperfusion Injury; Trichosanthes

Near-infrared (NIR) spectroscopy with multivariate analysis was used to quantify gallic acid, catechin, albiflorin, and paeoniflorin in Radix Paeoniae Rubra, and the feasibility to classify the samples originating from different areas was investigated. A new high-performance liquid chromatography method was developed and validated to analyze gallic acid, catechin, albiflorin, and paeoniflorin in Radix Paeoniae Rubra as the reference. Partial least squares (PLS), principal component regression (PCR), and stepwise multivariate linear regression (SMLR) were performed to calibrate the regression model. Different data pretreatments such as derivatives (1st and 2nd), multiplicative scatter correction, standard normal variate, Savitzky-Golay filter, and Norris derivative filter were applied to remove the systematic errors. The performance of the model was evaluated according to the root mean square of calibration (RMSEC), root mean square error of prediction (RMSEP), root mean square error of cross-validation (RMSECV), and correlation coefficient (r). The results show that compared to PCR and SMLR, PLS had a lower RMSEC, RMSECV, and RMSEP and higher r for all the four analytes. PLS coupled with proper pretreatments showed good performance in both the fitting and predicting results. Furthermore, the original areas of Radix Paeoniae Rubra samples were partly distinguished by principal component analysis. This study shows that NIR with PLS is a reliable, inexpensive, and rapid tool for the quality assessment of Radix Paeoniae Rubra.

Topics: Bridged-Ring Compounds; Catechin; Chromatography, High Pressure Liquid; Gallic Acid; Glucosides; Linear Models; Monoterpenes; Paeonia; Plant Preparations; Reproducibility of Results; Spectroscopy, Near-Infrared

A simple and sensitive HPLC-MS/MS method was developed and fully validated for simultaneous determination of ginsenoside Rb1, ginsenoside Rg1, paeoniflorin, albiflorin and oxypaeoniflorin in rat plasma. Plasma samples were pretreated with protein precipitation using acetonitrile. The chromatographic separation was carried out on a C

Topics: Animals; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Ginsenosides; Glucosides; Linear Models; Male; Monoterpenes; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

The compounds of Radix Paeoniae Rubra (RPR) were isolated and identified by bioassay-guided method, and antithrombotic effects and mechanism were investigated by the acute blood stasis rat model. The RPR extract was evaluated by APTT, TT, PT, and FIB assays in vitro. Results indicated that RPR extract exhibited the anticoagulant activity. In order to find active compounds, six compounds were isolated and identified, and four compounds, paeoniflorin (Pae), pentagalloylglucose (Pen), albiflorin (Ali), and protocatechuic acid (Pro), exhibited the anticoagulant activity in vitro. Therefore, the antithrombosis effects of RPR extract and four active compounds were investigated in vivo by measuring whole blood viscosity (WBV), plasma viscosity (PV), APTT, PT, TT, and FIB. Meanwhile, the levels of TXB

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Anticoagulants; Biological Assay; Bridged-Ring Compounds; Drugs, Chinese Herbal; Endothelin-1; Female; Fibrinolytic Agents; Glucosides; Hydrolyzable Tannins; Hydroxybenzoates; Male; Monoterpenes; Nitric Oxide Synthase Type III; Paeonia; Phytotherapy; Rats; Rats, Sprague-Dawley; Thrombosis; Thromboxane B2; Viscosity

This paper aimed to investigate the antidepressant-like effects and the mechanism of the albiflorin, paeoniflorin on rats with chronic restraint stress model. Fluoxetine and Xiaoyao group served as the positive control, body weight, sucrose preference test and the open field behavioral experiment were measured, the levels of nitric oxide (NO) in hippocampus were detected by Greg colorimetric method. Furthermore, the levels of brain derived neurotrophic factor (BDNF) in hippocampus were detected by ELISA. Finally, the expressions of nNOS mRNA in hippocampus detected by RT-qPCR, the protein levels of nNOS and BDNF in hippocampus were detected by Western blot. Compared with the model group,the pass counts of paeoniflorin group(30 mg•kg⁻¹) and albiflorin group(30 mg•kg⁻¹) were obviously increased(P<0.05,P<0.01). Furthermore, compared with the model group,the levels of NO in paeoniflorin groups(30 mg•kg⁻¹ and 15 mg•kg⁻¹) and albiflorin groups(30 mg•kg⁻¹ and 15 mg•kg⁻¹) were all significantly decreased(P<0.05,P<0.01; P<0.05, P<0.05). BDNF levels of paeoniflorin group(30 mg•kg⁻¹) and albiflorin group(30 mg•kg⁻¹) were obviously increased(P<0.05). Finally, compared with the model group, the expressions of nNOS mRNA of paeoniflorin groups(30 mg•kg⁻¹ and 15 mg•kg⁻¹) (P<0.01, P<0.01) and albiflorin groups(30 mg•kg⁻¹) were significantly decreased(P<0.05). Compared with the model group, the protein exprsssions of BDNF of paeoniflorin group(30 mg•kg⁻¹) and albiflorin group(30 mg•kg⁻¹) were obviously increased(P<0.05). Albiflorin and paeoniflorin have the effects of smooth the liver and dispel stagnation, the mechanism has the relevant with adjusting and controlling the expression of NO and BDNF in hippocampus.

Topics: Animals; Brain-Derived Neurotrophic Factor; Bridged-Ring Compounds; Glucosides; Hippocampus; Monoterpenes; Nitric Oxide; Rats; Rats, Sprague-Dawley; Restraint, Physical; Stress, Physiological

To prepare porosity osmotic pump tablets of total glucosides of paeony( TGP),and to study the behavior on synchronous release of its main components.. Taking the accumulative release of TGP as indexes, through single-factor test and orthogonal design to investigate the optimal formulation porosity osmotic pump tablets of TGP. The main components, paeoniflorin, albiflorin and benzoylpaeoniflorin were employed to study synchronous release of the optimal formulation.. The membrane weight, and the content of PEG 400,and diethyl phthalate( DEP) were the main factors influencing the behavior of TGP release. The accumulated release of the prepared osmotic pump release tablets achieved about 90%. Three main components achieved the desired zero-order release profile and had a synchronized release behavior.. The prepared porosity osmotic pump tablets of TGP can achieve the behavior of synchronized release of multi-components with good reproducibility.

Topics: Bridged-Ring Compounds; Delayed-Action Preparations; Glucosides; Monoterpenes; Osmosis; Paeonia; Polyethylene Glycols; Porosity; Reproducibility of Results; Solubility; Tablets

A simple, rapid and reliable microwave-assisted extraction (MAE) combined with ultra performance liquid chromatography tandem mass spectrometry method was developed for simultaneous determination of the seven bioactive constituents in Guizhi Fuling capsule (GFC), namely gallic acid, amygdalin, albiflorin, paeoniflorin, paeonol, cinnamic acid and pachymic acid, respectively. The operation of MAE optimised through orthogonal array design experiment was performed at 80°C for 10 min with methanol-water (70:30, v/v) as the extracting solvent. The method was validated including intra- and inter-day precision, repeatability and stability, with relative standard deviation less than 3.9%, 3.3%, 4.4% and 3.1%, respectively. All analytes showed the good linearity (r >0.999), and their average recoveries varied between 98.2% and 101.2%. The results indicated that this method was simple, effective and suitable for the quality control of GFC.

Topics: Acetophenones; Amygdalin; Bridged-Ring Compounds; Capsules; Chemical Fractionation; Chromatography, High Pressure Liquid; Cinnamates; Drugs, Chinese Herbal; Gallic Acid; Glucosides; Microwaves; Monoterpenes; Quality Control; Reproducibility of Results; Tandem Mass Spectrometry; Triterpenes

Danggui-Shaoyao San (DSS) is a famous Chinese formula for activating blood circulation and promoting urination. This study was to investigate the difference of material basis between a blood-associated herbs group and a water-associated herbs group. According to the theory of traditional Chinese medicine, the formula can be divided into a blood-associated herbs group (Angelica sinensis, Paeonia lactiflora and Ligusticum chuanxiong) and a water-associated herbs group (Atractylodes macrocephala, Alisma orientale and Poria cocos). The HPLC fingerprint of the formula was established for quality control. Serum samples from rats, orally administrated DSS, and the decomposed recipes of DSS, were analyzed by HPLC-DAD and the transitional blood components of DSS were identified. Twenty-one common peaks were identified in the fingerprint of DSS. Contents of paeoniflorin, albiflorin, ferulic acid and alisol B 23-acetate in co-decoction were significantly higher than those in individual decoction. Eleven peaks belonged to the blood-associated herbs group (four metabolites and seven prototype components; paeoniflorin and ferulic acid appeared in prototype components), whereas six peaks belonged to the water-associated herbs group (three metabolites and three prototype components). It was concluded that the serum pharmacochemistry is a meaningful approach for clarifying the difference between blood-associated and water-associated herbs in chemical composition.

Topics: Animals; Bridged-Ring Compounds; Cholestenones; Chromatography, High Pressure Liquid; Coumaric Acids; Drugs, Chinese Herbal; Glucosides; Male; Monoterpenes; Rats, Sprague-Dawley; Serum; Solubility; Water

A sensitive and reliable method using liquid chromatography tandem mass spectrometry (LC-MS/MS) was established for the simultaneous assay of paeoniflorin and albiflorin in bio-samples of rats after liquid-liquid extraction with ethylacetate. For the first time, the developed method was validated and successfully applied to the pharmacokinetics study of paeoniflorin and albiflorin after oral administration of Total Glucosides Of White Paeony Capsule (TGP). Relative to the intravenous injection, the absolute bio-availabilities of paeoniflorin and albiflorin were 2.8 and 1.7%, while their excretion in feces was 43.06 and 40.87%, respectively. Both paeoniflorin and albiflorin showed dose-dependent exposure in plasma, with a half-life of approximately 1.8h. No significant differences were observed between a single equal dose of paeoniflorin or albiflorin and that of TGP for the pharmacokinetic parameters, including AUC, T1/2 and Cmax. Paeoniflorin and albiflorin were exposed at high levels in immune relevant organ/tissues, such as the spleen, thymus and bone, which could facilitate immuno-regulatory activities.

Topics: Administration, Oral; Animals; Bridged-Ring Compounds; Chromatography, Liquid; Drugs, Chinese Herbal; Female; Glucosides; Isomerism; Limit of Detection; Linear Models; Male; Monoterpenes; Paeonia; Rats; Reproducibility of Results; Tandem Mass Spectrometry

There is evidence suggesting that herbal extracts demonstrate greater bioactivities than their isolated constituents at an equivalent dose. This phenomenon could be attributed to the absence of interacting substances present in the extracts. By measuring the pharmacokinetic parameters of paeoniflorin (PF) and albiflorin (AF) after being orally administered to rats in isolated form, in combination with each other and within total peony glucosides (TPG), respectively, the current study aimed to identify positive pharmacokinetic interactions between components of peony radix extracts. Moreover, the pharmacokinetic profiles of PF and AF under normoxia and hypoxia were also investigated and compared. In order to achieve these goals, a highly sensitive and reproducible ultra-peformance liquid chromatography-mass spectrometry (UPLC-MS) method was developed and validated for simultaneously quantitation of PF and AF in rat plasma. This study found that compared with that of single component (PF/AF), the exposure of PF in rat plasma after combination administration or TPG administration was significantly increased, meanwhile the elimination of PF/AF was remarkably reduced. It was also noticed that AUC and Cmax of PF in hypoxia rats were significantly decreased compared with that of normaxia rats, suggesting that there was a decreased exposure of PF in rats under hypoxia. The current study, for the first time, revealed the pharmacokinetic interactions between PF/AF and other constitutes in TGP and the pharmacokinetic profiles of PF and AF under hypoxia. In view of the current findings, it could be supposed that the clinical performance of total peony glucosides would be better than that of single constitute (PF/AF). The outcomes of this animal study are expected to serve as a basis for development of clinical guidelines on total peony glucosides usage.

Topics: Animals; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drug Stability; Drugs, Chinese Herbal; Glucosides; Hypoxia; Male; Molecular Structure; Monoterpenes; Paeonia; Rats; Reproducibility of Results; Tandem Mass Spectrometry

Monoterpenoids with "cage-like" pinane skeleton are the unique and main bioactive constituents in peony root, the root of Paeonia lactiflora. A liquid chromatography coupled with ion trap and time-of-flight mass spectrometry (LC-IT-TOF-MS) method was developed for characterization and quantification of monoterpenoids in different types of peony root and the roots of related Paeonia species. MS/MS fragmentation patterns of monoterpenoids with paeoniflorin-, albiflorin- and sulfonated paeoniflorin-type of skeletons were elucidated, which provided basic clues enabling subsequent identification of 35 monoterpenoids in LC-MS profiles of Paeonia species. The profiling analysis and further quantification of 15 main monoterpenoids in 56 samples belonged to red peony root (RPR), white peony root (WPR), peony root in Japanese market (PR) and the roots of related Paeonia species revealed that paeoniflorin, benzoylpaeoniflorin, galloylpaeoniflorin, oxypaoniflorin and albiflorin were predominant constituents in all the samples; mudanpioside C was the characteristic component of P. lactiflora, and 4-O-methyl-paeoniflorin was only detected in P. veitchii and P. anomala. Total contents of the 15 monoterpenoids were obviously higher in the roots of P. lactiflora and P. veitchii than in those of P. anomala and P. japonica. Principal component analysis based on the quantitative results showed that the samples derived from P. lactiflora were clearly classified into RPR, WPR/PR, and sulfur-fumigated WPR groups, besides the respective group of P. veitchii and P. anomala. This study clarified the chemical characteristics of the respective type of peony root and the related Paeonia species, as well as the marker constituents for their discrimination.

Topics: Bridged Bicyclo Compounds, Heterocyclic; Bridged-Ring Compounds; Chromatography, Liquid; Drugs, Chinese Herbal; Glucosides; Monoterpenes; Paeonia; Plant Roots; Tandem Mass Spectrometry

Paeonia lactiflora root (baishao in Chinese) is a commonly used herb in traditional Chinese medicines (TCM). Two isomers, paeoniflorin (PF) and albiflorin (AF), are isolated from P. lactiflora. The present study aimed to investigate the protective effects of PF and AF on myelosuppression induced by chemotherapy in mice and to explore the underlying mechanisms. The mouse myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide (CP, 200 mg·kg(-1)). The blood cell counts were performed. The thymus index and spleen index were also determined and bone morrow histological examination was performed. The levels of tumor necrosis factor-α (TNF-α) in serum and colony-stimulating factor (G-CSF) in plasma were measured by Enzyme-Linked Immunosorbent Assays (ELISA) and the serum levels of interleukin-3 (IL-3), granulocyte-macrophagecolony-stimulatingfactor (GM-CSF), and interleukin-6 (IL-6) were measured by radioimmunoassay (RIA). The levels of mRNA expression protein of IL-3, GM-CSF and G-CSF in spleen and bone marrow cells were determined respectively. PF and AF significantly increased the white blood cell (WBC) counts and reversed the atrophy of thymus. They also increased the serum levels of GM-CSF and IL-3 and the plasma level of G-CSF and reduced the level of TNF-α in serum. PF enhanced the mRNA level of IL-3 and AF enhanced the mRNA levels of GM-CSF and G-CSF in the spleen. PF and AF both increased the protein levels of GM-CSF and G-CSF in bone marrow cells. In conclusion, our results demonstrated that PF and AF promoted the recovery of bone marrow hemopoietic function in the mouse myelosuppression model.

Topics: Animals; Antineoplastic Agents; Bridged-Ring Compounds; Cyclophosphamide; Drugs, Chinese Herbal; Glucosides; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematologic Diseases; Humans; Interleukin-3; Interleukin-6; Male; Mice; Monoterpenes; Paeonia; Tumor Necrosis Factor-alpha

Zengmian Yiliu (ZMYL), a traditional Chinese formula, is designed to improve clinical efficacy and reduce adverse effects in combination with cisplatin in ovarian cancer chemotherapy. In ZMYL, Radix Paeoniae Alba (RPA, made from root of Paeonia lactiflora Pall.) acts as an adjunctive drug in cancer treatment by ameliorating side effects induced by radio- and chemotherapy. The pharmacokinetics differences between isomer albiflorin and paeoniflorin, the main components of RPA, after oral administration decoction of single-herb RPA and ZMYL were compared using a sensitive and accurate UPLC-MS/MS. The results indicate that there are statistically significant differences between the pharmacokinetic parameters: decreasing area under the plasma concentration-time curve (AUC), maximum concentration (Cmax ), elimination rate constant (Ke ) and increasing apparent volume of distribution (Vd ) and clearance (CL) for albiflorin, increasing distribution half-life (T1/2d ) and decreasing elimination half-life (T1/2e ), distribution rate constant (Kd ) and absorption rate constant (Ka ) for paeoniflorin in the ZMYL group compared with the single-herb RPA group. In comparison with albiflorin, the pharmacokinetic parameters of paeoniflorin included significantly increasing mean residence time (MRT) and Vd , decreasing CL and Ke in the single-herb RPA group and increasing MRT and T1/2d and decreasing CL, Ke and Kd in the ZMYL group. Both paeoniflorin and albiflorin are more likely, as the main active ingredients in RPA and ZMYL, to play a variety of pharmacological effects, and herb-herb interactions occur, resulting in different pharmacokinetics of albiflorin and paeoniflorin in RPA and ZMYL.

Topics: Administration, Oral; Animals; Area Under Curve; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drug Stability; Drugs, Chinese Herbal; Female; Glucosides; Half-Life; Herb-Drug Interactions; Isomerism; Limit of Detection; Male; Monoterpenes; Paeonia; Rats, Sprague-Dawley; Reproducibility of Results; Tandem Mass Spectrometry

Two kinds of peony roots--white peony root (WPR) and red peony root (RPR)--are used for different remedies in traditional Chinese medicine; however, most of them are derived from the same botanical origin, Paeonia lactiflora. The difference between WPR and RPR has been debated for a long time. This study attempted to clarify the genetic and chemical characteristics of WPR and RPR in order to provide a scientific dataset for their identification and effective use. The nucleotide sequence of nrDNA internal transcribed spacer (ITS) and the contents of 8 main bioactive constituents were analyzed from specimens of P. lactiflora, P. veitchii and two related species as well as crude drug samples of WPR, RPR and peony root produced in Japan. Of the samples derived from P. lactiflora, the WPR produced in the southern parts of China and the RPR produced in the northern parts of China were clearly divided into two subgroups within the P. lactiflora group based on similarity of the ITS sequences. The nucleotides at positions 69, 458 and 523 upstream of the ITS sequence served as molecular markers to discriminate between WPR and RPR. Quantitative analysis indicated that the RPR samples obviously contained a higher content of paeoniflorin and paeonol, but a lower content of albiflorin than the WPR produced in the southern parts of China and peony root produced in Japan. The WPR available from Chinese markets was usually processed by sulfur fumigation, which resulted in an extremely low content of paeoniflorin. This study indicated that WPR and RPR were not only geographically isolated, but also genetically and chemically separated. The ITS sequence provided a genetic index for their identification.

Topics: Acetophenones; Bridged-Ring Compounds; DNA, Ribosomal Spacer; Drugs, Chinese Herbal; Glucosides; Monoterpenes; Paeonia; Plant Roots

The metabolic profile of the traditional Chinese medicine, Sinisan, in miniature pig urine via intragastric administration was investigated. In total, 50 compounds, including 10 unchanged parent glycosides, which were not found from Sinisan's metabolic profile in rats' urine, were identified. Among these, 36 compounds were characterized by HPLC-SPE-NMR coupled with HPLC-HRESIMS, five of which are new and nine are endogenous metabolites of miniature pig. Most of phase I and phase II metabolites are hydrolytic products of parent glycosides and glucuronide conjugates, respectively, the latter having been reported as sulfate conjugates while the experimental animal is rat. Benzoic acid, obtained from hydrolysis of albiflorin and paeoniflorin, and phenylpropenoic acids, obtained from oxidative cleavage of flavones, formed phase II glycine conjugates.

Topics: Animals; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Female; Glucosides; Glucuronides; Glycosides; Magnetic Resonance Spectroscopy; Mass Spectrometry; Medicine, Chinese Traditional; Metabolic Detoxication, Phase I; Metabolic Detoxication, Phase II; Monoterpenes; Phenylpropionates; Swine; Swine, Miniature

To compare the chemical change of Paeoniae Radix Alba (PRA) after vinegar-baking processing, as well as the effect of vinegar types exerted on the processing, 1H NMR-based metabolomic approach combined with multivariate statistical analysis was used to investigate the different metabolites between the raw and two vinegar-baked PRA. More than thirty metabolites were identified in the 1H NMR spectrum of PRA, and the multivariate statistical analysis showed that raw and two vinegar-baked PRA could be separated obviously. After vinegar-baking, the contents of isoleucine, lactate, alanine, arginine, albiflorin, and 5-hydroxymethyl furfural (5-HMF) elevated, while those of sucrose, paeoniflorin and its analogues (calculated by benzoate) decreased. The chemical compositions of two vinegar-baked PRA were also different. Shanxi vinegar- baked PRA showed higher levels of leucine, isoleucine, valine, and albiflorin, while rice vinegar-baked PRA contained more sucrose and paeoniflorin's analogues (calculated by benzoate). And the chemical changes in Shanxi vinegar-baked PRA were greater than those of rice vinegar-baked PRA. The results revealed the chemical differences between raw and vinegar-baked PRA, as well as the influence of vinegar type on processing, in a holistic manner, the results obtained suggested that the correlations between the chemical change and the drug action after processing, as well as the vinegar type used in processing, should be further studied.

Topics: Acetic Acid; Benzoates; Bridged-Ring Compounds; Cooking; Drugs, Chinese Herbal; Furaldehyde; Glucosides; Metabolome; Metabolomics; Monoterpenes; Paeonia; Plant Roots

Gan-Sui-Ban-Xia Decoction (GSBXD) was first presented by Zhang Zhongjing in the book Synopsis of Golden Chamber during the Han Dynasty period. The formula was then used for the treatment of persistent fluid retention with floating pulse in Traditional Chinese Medicine (TCM), which in modern medicine is known as malignant ascites. Here, a rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the determination of glycyrrhizinic acid, liquiritin, paeoniflorin, albiflorin after oral administration of GSBXD plus-minus Gansui and Gancao anti-drug combination to investigate the possible pharmacokinetic profile differences of different prescriptions with GSBXD in normal rats. The differences of pharmacokinetic parameters among groups were tested by the Student's t-test with p < 0.05 as the level of significance. Significant differences were found between the Gansui and Gancao anti-drug combination and other herbs in GSBXD on pharmacokinetic profile of glycyrrhizinic acid, liquiritin, paeoniflorin and albiflorin. The obtained knowledge might contribute to the rationality of the clinic use of GSBXD and also reveal the compatibility conditions of the Gansui and Gancao anti-drug combination.

Topics: Administration, Oral; Animals; Ascites; Bridged-Ring Compounds; Chromatography, Liquid; Drug Combinations; Drugs, Chinese Herbal; Euphorbia; Flavanones; Glucosides; Glycyrrhiza uralensis; Glycyrrhizic Acid; Male; Medicine, Chinese Traditional; Monoterpenes; Paeonia; Pinellia; Rats; Rats, Wistar; Tandem Mass Spectrometry

Total glucosides of paeony are the active constituents of Paeoniae Radix Alba. In this study, a novel strategy was proposed to find more metabolites and the differences between paeoniflorin, albiflorin and total glucosides of paeony (TGP). This strategy was characterized as follows: firstly, the animals were divided into three groups (paeoniflorin, albiflorin and TGP) to identify the source of TGP metabolites from paeoniflorin or albiflorin; secondly, a generic information-dependent acquisition scan for the low-level metabolites was triggered by the multiple mass defect filter and dynamic background subtraction; thirdly, the metabolites were identified with a combination of data-processing methods including mass defect filtering, neutral loss filtering and product ion filtering; finally, a comparative study was used in the metabolism of paeoniflorin, albiflorin and TGP. Based on the strategy, 18 metabolites of TGP, 10 metabolites of paeoniflorin and 13 metabolites of albiflorin were identified respectively. The results indicated that the hydrolysis, conjugation reaction and oxidization were the major metabolic pathways, and the metabolic sites were the glycosidic linkage, the ester bond and the benzene ring. This study is first to explore the metabolism of TGP, and these findings enhance our understanding of the metabolism and the interactions of paeoniflrin and albiflorin in TGP.

Topics: Animals; Bridged-Ring Compounds; Chromatography, Liquid; Glucosides; Glycosides; Male; Monoterpenes; Paeonia; Rats; Rats, Wistar; Tandem Mass Spectrometry

To compare the effects and mechanism of blood enriching on mouse model of blood deficiency syndrome induced by cyclophosphamide of albiflorin and paeoniflorin.. Albiflorin and paeoniflorin were determined by using animal models of blood deficiency syndrome induced by cyclophosphamide. The amount of WBC, RBC, HGB, index of thymus gland and spleen, and the changes of GM-CSF, IL-3 and TNF-α in serum were detected after the treatment.. Compared with the model group, the amount of WBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01). The amount of RBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01, P < 0.001), which did not had a significant difference compared with the same dose. The index of thymus gland in the group of 30 mg x kg(-1) albiflorin was superior to the model group (P < 0.01), the difference was significant compared with the same dose of paeoniflorin (P < 0.05). The GM-CSF in serum in all groups of 30 mg x kg(-1) albiflorin, 15 mg x kg(-1) albiflorin, 30 mg x kg(-1) paeoniflorin and 15 mg x kg(-1) paeoniflorin increased obviously (P < 0.01, P < 0.05, P < 0.01, P < 0.05); The IL-3 in serum in both group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin also increased (P < 0.001). The content of TNF-α in group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were reduced (P < 0.01), which showed the obvious difference compared with the same dose group (P < 0.01).. Albiflorin had the effect of blood enriching by regulating the immune function, same with the paeoniflorin. The probable mechanism of nourishing blood and liver of Paeoniae Radix Alba was not only the better effect of adjusting the content of TNF-α, but also might act synergistically with paeoniflorin.

Topics: Animals; Blood Cells; Bridged-Ring Compounds; Cyclophosphamide; Glucosides; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoiesis; Interleukin-3; Male; Mice; Monoterpenes; Tumor Necrosis Factor-alpha

In this work, a simple and efficient protocol for the rapid separation of two pairs of isomeric monoterpenes from Paeoniae Alba Radix was developed by combining macroporous resin and elution-extrusion counter-current chromatography. The crude extract was firstly subjected to a D101 macroporous resin column eluted with water and a series of different concentrations of ethanol. Then, effluents of 30 and 95% ethanol were collected as sample 1 and sample 2 for further counter-current chromatography purification. Finally, a pair of isomers, 96 mg of compound 1 and 48 mg of compound 2 with purities of 91.1 and 96.2%, respectively, was isolated from 200 mg of sample 1. The other pair of isomers, 14 mg of compound 3 and 8 mg of compound 4 with purities of 93.6 and 88.9%, respectively, was isolated from 48 mg of sample 2. Their purities were analyzed by high-performance liquid chromatography, and their chemical structures were identified by mass spectrometry and (1) H NMR spectroscopy. Compared to a normal counter-current chromatography separation, the separation time and solvent consumption of elution-extrusion counter-current chromatography were reduced while the resolutions were still good. The established protocol is promising for the separation of natural products with great disparity of content in herbal medicines.

Topics: Bridged-Ring Compounds; Chromatography; Chromatography, High Pressure Liquid; Countercurrent Distribution; Ethanol; Glucosides; Glycosides; Magnetic Resonance Spectroscopy; Mass Spectrometry; Monoterpenes; Paeonia; Plant Preparations; Solvents; Ultrasonography

In order to explore the differences of chemical constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra, a qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) was developed for identification of multi-constituents and an HPLC-DAD analytical method was developed for simultaneously determining 14 major compounds (gallic acid, protocatechuic acid, paeoniflorin sulfonate, protocatechuic aldehyde, methyl gallate, oxypaeoniflorin, catechin, albiflorin, and paeoniflorin, ethyl gallate, benzoic acid, pentagaloylglucose, benzoyl-paeoniflorin, and paeonol) in Paeoniae Radix Alba and Paeoniae Radix Rubra. Q-TOF/MS qualitative analysis was performed under negative ion mode and inferred 38 components of Paeoniae Radix Alba and 30 components of Paeoniae Radix Rubra. HPLC-DAD quantitative method result showed the contents of 8 ingredients were different between Paeoniae Radix Alba and Paeoniae Radix Rubra. The results indicated that the new approach was applicable in qualitative and quantitative quality control of Paeoniae Radix Alba and Paeoniae Radix Rubra.

Topics: Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glucosides; Molecular Structure; Monoterpenes; Paeonia; Tandem Mass Spectrometry

The present study aims to predict the action targets of antidepressant active ingredients of Xiaoyaosan to understand the "multi-components, multi-targets and multi-pathways" mechanism. Using network pharmacology, the reported antidepressant active ingredients in Xiaoyaosan (saikosaponin A, saikosaponin C, saikosaponin D, ferulic acid, Z-ligustilide, atractylenolide I, atractylenolide II, atractylenolide III, paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid and pachymic acid), were used to predict the targets of main active ingredients of Xiaoyaosan according to reversed pharmacophore matching method. The prediction was made via screening of the antidepressive drug targets approved by FDA in the DrugBank database and annotating the information of targets with the aid of MAS 3.0 biological molecular function software. The Cytoscape software was used to construct the Xiaoyaosan ingredients-targets-pathways network. The network analysis indicates that the active ingredients in Xiaoyaosan involve 25 targets in the energy metabolism-immune-signal transmutation relevant biological processes. The antidepressant effect of Xiaoyaosan reflects the features of traditional Chinese medicine in multi-components, multi-targets and multi-pathways. This research provides a scientific basis for elucidation of the antidepressant pharmacological mechanism of Xiaoyaosan.

Topics: Antidepressive Agents; Benzoates; Bridged-Ring Compounds; Coumaric Acids; Drug Evaluation, Preclinical; Drugs, Chinese Herbal; Flavanones; Glucosides; Glycyrrhizic Acid; Lactones; Medicine, Chinese Traditional; Monoterpenes; Sesquiterpenes; Software

Palmul-tang, a traditional herbal medicine, is composed of eight herbs (Ginseng radix, Glycyrrhizae radix, Hoelen, Atractylodis rhizoma, Angelicae gigantis radix, Cnidii rhizoma, Paeoniae radix and Rehmanniae radix) and exhibits various bioactivities, including antiallergic and antitumor effects. In this study, an effective, reliable and accurate high-performance liquid chromatography method has been developed for the simultaneous determination of 11 marker components in Palmul-tang: hydroxymethylfurfural, albiflorin, paeoniflorin, ferulic acid, nodakenin, ginsenoside Rg1, decursinol, glycyrrhizin, 6-gingerol, ginsenoside Rg3 and decursin. All calibration curves of the 11 components indicated excellent linearity (correlation coefficient > 0.9997) within the test range. The limits of detection and quantification of each component were in the ranges of 0.08-1.03 and 0.23-3.11 µg/mL, respectively. The intra-day and inter-day relative standard deviation values were within 1.65 and 2.71%, respectively. The mean recovery values were 94.49 to 101.10%. The established method was successfully applied to the simultaneous determination of 11 major components in 12 commercial samples of Palmul-tang. The developed analytical method is simple and suitable for the quality control of Palmul-tang.

Topics: Angelica; Atractylodes; Bridged-Ring Compounds; Catechols; Chromatography, High Pressure Liquid; Coumaric Acids; Coumarins; Drugs, Chinese Herbal; Fatty Alcohols; Furaldehyde; Glucosides; Glycyrrhiza; Glycyrrhizic Acid; Monoterpenes; Panax; Rehmannia; Reproducibility of Results; Tandem Mass Spectrometry

A liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method was developed and validated for simultaneous determination of seven constituents including puerarin, daidzin, daidzein, paeoniflorin, albiflorin, liquiritin and liquiritigenin in rat plasma using schisandrin as the internal standard (IS). The plasma samples were pretreated by a one-step direct protein precipitation with acetonitrile. The chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of acetonitrile and water (containing 0.1% formic acid and 5mM ammonium acetate). All analytes and IS were quantitated through electrospray ionization in positive ion multiple reaction monitoring (MRM) mode. The mass transitions were as follows: m/z 417.5→297.2 for puerarin, m/z 417.1→255.2 for daidzin, m/z 255.2→152.4 for daidzein, m/z 498.1→179.3 for paeoniflorin, m/z 481.1→197.3 for albiflorin, m/z 436.2→257.3 for liquiritin, m/z 257.2→137.3 for liquiritigenin and m/z 415.0→384.2 for IS, respectively. All calibration curves exhibited good linearity (r>0.9979) over a wide concentration range for all components. The intra-day and inter-day precisions (RSD) at three different levels were both less than 14.3% and the accuracies (RE) ranged from -13.2% to 14.8%. The extraction recoveries of the seven compounds ranged from 72.9% to 117.4%. The validated method was successfully applied to pharmacokinetic study of the seven components in female rat plasma after oral administration of Ge-Gen Decoction aqueous extract.

Topics: Animals; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Glucosides; Isoflavones; Monoterpenes; Rats; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Paeonia lactiflora Pall. (Ranunculaceae) has been used for more than 1000 years in traditional Chinese medicine for the treatment of gynecological problems, cramp, pain, giddiness, and congestion. Paeoniflorin, monoterpene glycosides isolated from P. lactiflora, possesses a variety of pharmacological activities. However, the pharmacological activity of the pharmacological activity of albiflorin, another main monoterpene glycoside, has not been well studied.. The present study investigated the anti-inflammatory activities of paeoniflorin and albiflorin using models of lipopolysaccharides (LPS) induced RAW 264.7 cells.. Production of nitric oxide (NO) was measured by the Griess colorimetric method. In addition, prostaglandin E2 (PGE2), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) synthesis were analyzed using an enzyme-linked immunosorbent assay (ELISA). The protein expression of cyclooxygenase-2 (COX-2) was detected by a cell-based ELISA. The gene expression levels of inducible nitric oxide synthase (iNOS), COX-2, TNF-α, and IL-6 were detected by quantitative real-time reverse-transcription polymerase chain reaction (real-time RT-PCR).. Compared with the LPS-induced group, the inhibition rates of NO, PGE2, TNF-α, and IL-6 production were 17.61, 27.56, 20.57, and 29.01% by paeoniflorin and 17.35, 12.94, 15.29, and 10.78% by albiflorin. The IC50 values of paeoniflorin and albiflorin on NO production were 2.2 × 10(-4 )mol/L and 1.3 × 10(-2 )mol/L, respectively. The protein expression of COX-2 was reduced by 50.98% with paeoniflorin and 17.21% with albiflorin. The inhibition rates of gene expression of iNOS, COX-2, IL-6, and TNF-α were 35.65, 38.08, 19.72, and 45.19% by paeoniflorin and 58.36, 47.64, 50.70, and 12.43% by albiflorin, respectively.. These results show that albiflorin has similar anti-inflammatory effects to paeoniflorin, which provides new evidence that albiflorin can serve as a new chemical marker for the quality control of Paeoniae Radix and the Chinese Pharmacopoeia can be updated.

Topics: Animals; Anti-Inflammatory Agents; Bridged-Ring Compounds; Cell Line; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation; Glucosides; Lipopolysaccharides; Macrophages; Medicine, Chinese Traditional; Mice; Monoterpenes; Nitric Oxide; Paeonia; Reverse Transcriptase Polymerase Chain Reaction

In this present study, a sensitive and rapid UPLC-MS/MS method was developed for simultaneous quantification of paeoniflorin, albiflorin, ferulic acid, tetrahydropalmatine, protopine, typhaneoside and senkyunolide I in Beagle dog plasma after oral administration of the Shao-Fu-Zhu-Yu Decoction. Chloramphenicol and clarithromycin were used as internal standards. Plasma samples were processed by protein precipitation with methanol. The separation was performed on an Acquity BEH C18 column (100mm×2.1mm, 1.7μm) at a flow-rate of 0.4mL/min, using 0.1% formic acid-acetonitrile as mobile phase. Method validation was performed as per Food and Drug Administration guidelines and the results met the acceptance criteria. After validation, this method was successfully applied to a pharmacokinetic study. The results showed that the apparent plasma clearance of paeoniflorin, albiflorin, typhaneoside and senkyunolide I were significantly higher than others. Double peak was observed in plasma concentration curves of tetrahydropalmatine, the ferulic acid had a good absorption in Beagle dog plasma, and senkyunolide I was detected in plasma from the first blood sampling time (15min) and rapidly reached Tmax. The compound of typhaneoside has a low bioavailability according to the results.

Topics: Administration, Oral; Animals; Benzofurans; Benzophenanthridines; Berberine Alkaloids; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Coumaric Acids; Dogs; Drugs, Chinese Herbal; Female; Glucosides; Glycosides; Monoterpenes; Tandem Mass Spectrometry

One hundred and forty-four samples of Chishao and Baishao, which represented six species of Paeonia L. were evaluated for their genetic variation, genetic differentiation and phylogenetic relationship, based on the nuclear ribosomal DNA internal transcribed spacer (ITS) region. Samples from representative of the population were then used to do a cultivation comparison experiment, and then to identify the contents of the active ingredients. The results showed there were differences in the haplotype distribution and frequency between populations of Chishao and Baishao. An analysis of molecular variance (AMOVA) indicated statistically significant (p<0.001) genetic differentiation between the populations of wild and cultivated Paeonia lactiflora PALL. The albiflorin content between Chishao and Baishao was also significantly different (p<0.05). All the results clearly illustrate that currently cultivated P. lactiflora cannot be used as a substitute for Chishao.

Topics: Base Sequence; Benzoic Acid; Bridged-Ring Compounds; Catechin; China; DNA, Plant; DNA, Ribosomal; Gallic Acid; Genetic Variation; Glucosides; Haplotypes; Hydrolyzable Tannins; Molecular Sequence Data; Monoterpenes; Paeonia; Phylogeny; Sequence Alignment; Sequence Analysis, DNA

To study the chemical constituents of Paeonia lactiflora flowers.. [corrected] The chemical constituents were isolated and purified by various chromatography methods,and the structures were identified by physicochemical and modem spectroscopic.. 11 compounds were identified as gallic acid(1),methyl gallate(2),ethyl gallate(3),1,2,3,6-tetragalloyl-beta-D-glucopyranoside(4), 1,2,3,4,6-pentagalloyl-beta-D-glucopyranoside(5), quercetin-3-O-glucoside-6"-gallate(6), kaempferol-3-O-glucoside-6"-gallate(7), 1-O-galloyl-beta-D-glucose (8), kaempferol-3, 7-di-O-beta-D-glucoside(9), paeoniflorin(10) and albiflorin(11).. Compounds 1-8, 10 and 11 are obtained from the flowers of Paeonia lactiflora for the first time,compounds 6 and 7 are obtained from Paeonia genus for the first time.

Topics: Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flowers; Glucosides; Kaempferols; Molecular Structure; Monosaccharides; Monoterpenes; Paeonia; Plants, Medicinal

To study the blood enriching effects of Paeoniae Radix Rubra and Paeoniae Radix Alba, paeoniflorin and albiflorin on mouse model of blood deficiency caused by γ-ray radiation.. Build mouse model of blood deficiency induced by γ-ray radiation. Paeoniae Radix Rubra and Paeoniae Radix Alba were given during modeling. The amount of WBC was detected af- ter the treatment. Based on the result of WBC and paeoniflorin content, albiflorin content in Paeoniae Radix Rubra and Paeoniae Radix Alba, the same model and the same method were used to comparatively study the effect of blood enriching of paeoniflorin and albiflorin.. On the 7th day, the amount of WBC in model mice treated with 2 g x kg(-1) Paeoniae Radix Alba and 2 g x kg(-1) Paeoniae Radix Rubra significantly increased compared with that of model group (P < 0.05). In another experiment with the same model, the amount of WBC in model mice treated with 120 mg x kg(-1) paeoflorin and 120 mg x kg(-1) albiflorin significantly increased (P < 0.05) compared with that of model group on the 7th day. On the 10th day, the amount of WBC in rats treated with 120 mg x kg(-1) paeoflorin increased significantly (P < 0.05) compared with that of model group. Compared with the same dose of paeoniflorin, the amount of WBC in mice treated with albiflorin had no significant difference.. All Paeoniae Radix Alba, Paeoniae Radix Rubra, paeoniflorin and al- biflorin can raise the amount of WBC and have the effect of enriching blood induced by radiation, while paeoniflorin and albiflorin have a similar result in this model. The result indicated that both paeoniflorin and albiflorin are effective constituents in Paeoniae Radix Alba, and paeoniflorin work as the common effective constituent in both Paeoniae Radix Rubra and Paeoniae Radix Alba.

Topics: Animals; Bridged-Ring Compounds; Gamma Rays; Glucosides; Leukocyte Count; Leukocytes; Male; Mice; Monoterpenes; Rats

The root of Paeonia lactiflora Pall (family Ranunculaceae) or peony root, a herbal medicine, possesses therapeutic potential for neurodegenerative diseases. The isomers paeoniflorin (PF) and albiflorin (AF) are major constituents contained in peony root. Our previous study has shown notable neuroprotective effects of PF. In the present study, we further compared the effects of AF and PF against glutamate (Glu)-induced cell damage and the underlying mechanisms in differentiated PC12 cells. Both AF and PF significantly ameliorated Glu-induced reduction of cell viability, nuclear and mitochondrial apoptotic alteration, reactive oxygen species accumulation, and B-cell lymphoma 2 (Bcl-2)/Bax ratio. The two isomers also enhanced phosphorylation of AKT and its downstream element glycogen synthase kinase-3β, and this effect was abrogated by the AKT inhibitor LY294002. PF, but not AF, however, suppressed intracellular Ca(2+) overload and the expression of calcium/calmodulin protein kinase II (CaMKII). The improvement of cell damage by the CaMKII inhibitor KN93 further confirms the role of CaMKII in PF-mediated neuroprotection. These results suggest that both AF and PF possess robust effects in protecting neuronal cells against Glu toxicity. PF further displayed remarkable effects in preventing intracellular Ca(2+) overload and suppressing overexpression of CaMKII. Differential mechanisms may be involved in neuroprotective action of the two isomers.

Topics: Animals; Apoptosis; Benzoates; Bridged-Ring Compounds; Calcium; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Cell Survival; Cytoplasm; Glucosides; Glutamic Acid; Monoterpenes; Neuroprotective Agents; PC12 Cells; Protein Kinase Inhibitors; Rats; Reactive Oxygen Species

Sixty peony root training samples of the same age were collected from various regions in Korea and China, and their genetic diversity was investigated for 23 chloroplast intergenic space regions. All samples were genetically indistinguishable, indicating that the DNA-based techniques employed were not appropriate for determining the samples' regions of origin. In contrast, (1)H-nuclear magnetic resonance ((1)H-NMR) spectroscopy-based metabolomics coupled with multivariate statistical analysis revealed a clear difference between the metabolic profiles of the Korean and Chinese samples. Orthogonal projections on the latent structure-discrimination analysis allowed the identification of potential metabolite markers, including γ-aminobutyric acid, arginine, alanine, paeoniflorin, and albiflorin, that could be useful for classifying the samples' regions of origin. The validity of the discrimination model was tested using the response permutation test and blind prediction test for internal and external validations, respectively. Metabolomic data of 21 blended samples consisting of Korean and Chinese samples mixed at various proportions were also acquired by (1)H-NMR analysis. After data preprocessing which was designed to eliminate uncontrolled deviations in the spectral data between the testing and training sets, a new statistical procedure for estimating the mixing proportions of blended samples was established using the constrained least squares method for the first time. The predictive procedure exhibited relatively good predictability (adjusted R (2) = 0.7669), and thus has the potential to be used in the quality control of peony root by providing correct indications for a sample's geographical origins.

Topics: Alanine; Arginine; Biomarkers; Bridged-Ring Compounds; China; Chloroplasts; DNA, Intergenic; DNA, Plant; gamma-Aminobutyric Acid; Glucosides; Magnetic Resonance Spectroscopy; Metabolomics; Monoterpenes; Multivariate Analysis; Paeonia; Phylogeny; Phylogeography; Plant Roots; Quality Control; Republic of Korea

To control the quality of Qizhiweitong granules with the all-time multi-wavelength fusion fingerprint quantification as the major technique.. Agilent TC-C18 (4.6 mm x 250 mm, 5 microm) chromatographic column was adopted, with 0.02% formic acid water-acetonitrile as the mobile phase for linear gradient elution. The flow rate was 1 mL x min(-1), column temperature was 30 degrees C, and detector wavelength was 230, 254, 283 nm. Matlab was adopted for all-time multiple-wavelength fusion for data in dif format.. A good relationship was shown for albiflorin in 56.5-452 mg x L(-1) (r = 0.999 8), paeoniflorin in 107-856 mg x L(-1) (r = 0.999 8), licorice glycoside in 73.4-687 mg x L(-1) (r = 0.999 8), naringin in 109-872 mg x L(-1) (r = 0.999 8), neohesperidin in 48.0-384 mg L(-1) (r = 0.999 8), and glycyrrhizic acid in 38.6-308 mg x L(-1) (r = 0.999 8), with recoveries of 0.999 8.. The method is simple, accurate and highly reproducible, and can provide basis for quality control of Qizhiweitong granules.

Topics: Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glucosides; Glycosides; Monoterpenes; Quality Control

The objective of the present study was to establish a method based on principal component analysis (PCA) for the study of transdermal delivery of multiple components in Chinese medicine, and to choose the best penetration enhancers for the active fraction of Xiangfusiwu decoction (BW) with this method. Improved Franz diffusion cells with isolated rat abdomen skins were carried out to experiment on the transdermal delivery of six active components, including ferulic acid, paeoniflorin, albiflorin, protopine, tetrahydropalmatine and tetrahydrocolumbamine. The concentrations of these components were determined by LC-MS/MS, then the total factor scores of the concentrations at different times were calculated using PCA and were employed instead of the concentrations to compute the cumulative amounts and steady fluxes, the latter of which were considered as the indexes for optimizing penetration enhancers. The results showed that compared to the control group, the steady fluxes of the other groups increased significantly and furthermore, 4% azone with 1% propylene glycol manifested the best effect. The six components could penetrate through skin well under the action of penetration enhancers. The method established in this study has been proved to be suitable for the study of transdermal delivery of multiple components, and it provided a scientific basis for preparation research of Xiangfusiwu decoction and moreover, it could be a reference for Chinese medicine research.

Topics: Administration, Cutaneous; Alkenes; Animals; Azepines; Benzophenanthridines; Berberine Alkaloids; Bridged-Ring Compounds; Coumaric Acids; Drug Combinations; Drug Synergism; Drugs, Chinese Herbal; Glucosides; In Vitro Techniques; Male; Monoterpenes; Permeability; Plants, Medicinal; Principal Component Analysis; Rats; Rats, Sprague-Dawley; Skin Absorption

To study the effects of blood enriching on mouse model of blood deficiency syndrome of Paeoniae Radix Alba and Paeoniae Radix Rubra, paeoniflorin and albiflorin.. Building the mouse model of blood deficiency syndrome induced by compound method of bleeding, starved feeding and exhausting of swimming, extract from Paeoniae Radix Alba and Paeoniae Radix Rubra were given during modeling. The amount of RBC, HGB were detected after the treatment. Based on the amount results of RBC, HGB and the paeoniflorin content, albiflorin content in Paeoniae Radix Alba and Paeoniae Radix Rubra, the same model and the same method were used to comparatively study the effect of blood enriching of paeoniflorin and albiflorin.. At the 7th day, the amount of RBC and HGB in model mice was significantly increased (P <0. 01) by 2 g kg-1 Paeoniae Radix Alba and 2 g kg-1 Paeoniae Radix Rubra. At the 14th day, the amount of RBC and HGB in model mice was significantly increased (P <0. 01) by 2 g kg-1 Paeoniae Radix Alba. The amount of RBC and HGB in mice treated with Paeoniae Radix Rubra had an increasing trend compared with the same dose of Paeoniae Radix Rubra, but the difference was not significant. In another experiment with the same model, the amount of RBC and HGB in model mice was significantly increased (P<0.01) by 120 mg kg-1 paeoflorin and 120 mg kg-1 albiflorin at the 7th day, meanwhile, 60 mg kg-1 and 30 mg kg-1 albiflorin also increased the amount of RBC and HGB. At the 14th day, 120 mg kg-1 paeoflorin and all doses of albiflorin increased the amount of RBC and HGB. Comepared with that of the same dose of paeoniflorin, the amount of RBC in mice was significantly increased (P <0. 05) by 30 mg kg-1 albiflorin and 120 mg kg-1 albiflorin; the amount of HGB was significantly increased (P <0. 05) by 30 mg kg -1 albiflorin.. Paeoniae Radix Alba has a better effect of blood enriching than Paeoniae Radix Rubra. Albiflorin is more effective in blood enriching than paeoniflorin. Combining these, it infers that albiflorin involves in the better blood enriching effect of Paeoniae Radix Alba.

Topics: Animals; Benzoates; Bridged-Ring Compounds; Disease Models, Animal; Drugs, Chinese Herbal; Erythrocytes; Glucosides; Hemoglobins; Male; Mice; Monoterpenes; Paeonia

Paeoniae radix is one of the most important crude drugs used in Kampo medicines (KMs). A part of its pharmaceutical properties is due to the presence of paeoniflorin (PF) and albiflorin (AF).. For the specific and easy identification of PF and AF, an immunostaining technique was developed using anti-PF monoclonal antibody (MAb).. PF and AF were treated with a NaIO₄ solution and reacted with bovine serum albumin (BSA) preparing PF- and AF-BSA conjugates on the polyethersulphone (PES) membrane. Anti-PF MAb was bound and then antibody labelled with peroxidase directed against anti-PF MAb. Finally, a substrate was added and then PF and AF were detected.. Anti-PF MAb recognised not only PF but also AF when 10 µg was present on the PES membrane. As little as 0.5 µg of PF and AF were still detected under immunostaining. Various Paeoniae radix samples and KMs were qualitatively analysed, and total amounts of PF and AF were visually detected by immunostaining technique. This method was applied to investigate the distribution of PF and AF in fresh peony root using immunoblotting by transferred from peony root to the PES membrane.. The technique permitted the visualisation of PF and AF on PES membrane using immunostaining. The immunostaining technique established would be a powerful tool for probing the sources of PF and AF in plant extracts.

Topics: Animals; Antibodies, Monoclonal; Benzoates; Bridged-Ring Compounds; Cattle; Glucosides; Immunoblotting; Membranes, Artificial; Molecular Structure; Monoterpenes; Paeonia; Perchlorates; Plant Extracts; Polymers; Reproducibility of Results; Serum Albumin, Bovine; Sodium Compounds; Staining and Labeling; Sulfones

A rapid, highly sensitive, and selective method was applied in a non-invasive way to investigate the antidepressant action of Xiaoyaosan (XYS) using ultra performance liquid chromatography-mass spectrometry (UPLC-MS) and chemometrics. Many significantly altered metabolites were used to explain the mechanism. Venlafaxine HCl and fluoxetine HCl were used as chemical positive control drugs with a relatively clear mechanism of action to evaluate the efficiency and to predict the mechanism of action of XYS. Urine obtained from rats subjected to chronic unpredictable mild stress (CUMS) was analyzed by UPLC-MS. Distinct changes in the pattern of metabolites in the rat urine after CUMS production and drug intervention were observed using partial least squares-discriminant analysis. The results of behavioral tests and multivariate analysis showed that CUMS was successfully reproduced, and a moderate-dose XYS produced significant therapeutic effects in the rodent model, equivalent to those of the positive control drugs, venlafaxine HCl and fluoxetine HCl. Metabolites with significant changes induced by CUMS were identified, and 17 biomarker candidates for stress and drug intervention were identified. The therapeutic effect of XYS on depression may involve regulation of the dysfunctions of energy metabolism, amino acid metabolism, and gut microflora changes. Metabonomic methods are valuable tools for measuring efficacy and mechanisms of action in the study of traditional Chinese medicines.

Topics: Animals; Antidepressive Agents; Benzoates; Biomarkers; Bridged-Ring Compounds; Catechin; Chalcone; Chromatography, Liquid; Citric Acid; Citric Acid Cycle; Coumaric Acids; Creatine Kinase; Creatinine; Cyclohexanols; Drugs, Chinese Herbal; Flavanones; Fluoxetine; Gallic Acid; Gastrointestinal Tract; Glucosides; Glycine; Hippurates; Ketoglutaric Acids; Kynurenic Acid; Male; Mass Spectrometry; Metabolic Networks and Pathways; Metabolomics; Microbiota; Monoterpenes; Phytotherapy; Plant Extracts; Rats; Rats, Sprague-Dawley; Stress, Psychological; Tryptophan; Tyrosine; Venlafaxine Hydrochloride

A simple and accurate high-performance liquid chromatographic method was applied to the quantitative analysis of seven components of the traditional herbal prescription Jaeumganghwa-tang (JGT), including 5-hydroxymethyl-2-furaldehyde, albiflorin, paeoniflorin, liquiritin, ferulic acid, nodakenin, and glycyrrhizin. All seven compounds were separated in less than 40 min on a Gemini C18 column at 40°C by gradient elution using 1.0% (v/v) aqueous acetic acid and acetonitrile containing 1.0% (v/v) acetic acid as mobile phase. The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 230 nm, 254 nm, 280 nm, and 330 nm. The calibration curves showed good linearity (r (2) > 0.9998) in different concentration ranges. The recovery of each component was in the range of 91.47-102.62%, with relative standard deviations (RSDs, %) less than 4.5%. The RSDs (%) for intra- and interday precision were 0.06-2.85% and 0.06-2.83%, respectively. The concentrations of the seven components in JGT were in the range 0.74-5.48 mg/g.

Topics: Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Coumaric Acids; Coumarins; Drugs, Chinese Herbal; Flavanones; Furaldehyde; Glucosides; Glycyrrhizic Acid; Magnoliopsida; Monoterpenes

Shaoyao-Gancao-Tang (SGT) is a traditional Chinese prescription containing Radix Paeoniae alba and Radix Glycyrrhizae and is commonly used to relieve pains. Albiflorin and paeoniflorin are the main effective compounds of Radix Paeoniae alba, and the pharmacokinetic differences of the two compounds in rats after oral administration of SGT and single herb Paeony decoction were studied. At different time points (5, 10, 15, 20, 30, 45, 60, 90, 120, 180, 240, 360, and 540 min) after administration, plasma concentrations of albiflorin and paeoniflorin were determined using a simple and reliable UPLC method, and main pharmacokinetic parameters were evaluated. It was found that there were significant differences (p < 0.05 or p < 0.01) between the two groups. The results indicated that some components in the other ingredient herb of SGT (Radix Glycyrrhizae) had a pharmacokinetic interaction with albiflorin and paeoniflorin and hence reduced their systematic exposure level.

Topics: Animals; Benzoates; Bridged-Ring Compounds; Drugs, Chinese Herbal; Glucosides; Glycyrrhiza; Male; Monoterpenes; Paeonia; Phytotherapy; Plant Extracts; Plants, Medicinal; Rats; Rats, Sprague-Dawley

A comparative study was designed and conducted to compare the pharmacokinetic difference of paeoniflorin and albiflorin after oral administration of Radix Paeoniae Rubra to normal rats and the acute cholestasis hepatitis rats induced by alpha-naphthylisothiocyanate (ANIT). UPLC-ESI-MS/MS method was employed to determine the level of paeoniflorin and albiflorin in rat plasma using geniposide as the internal standard (IS). Unpaired Student's t-test was used for the statistical comparison. The investigation showed that there were significant differences between the normal rats and the acute cholestasis hepatitis rat groups in calculated parameters, such as AUC(0-t), AUC(0-∞), T(max) and CLz/F. The results indicated that acute liver injury in rats could alter the pharmacokinetics of drug. Since patients are the final users of the drug, it is essential to investigate the pharmacokinetics of the drug in disease status. Therefore, we used normal rats and the acute cholestasis hepatitis rats to study pharmacokinetics of Radix Paeoniae Rubra with the purpose of examining the influence of disease on the metabolic course.

Topics: Acute Disease; Administration, Oral; Animals; Anti-Inflammatory Agents, Non-Steroidal; Benzoates; Bridged-Ring Compounds; Cholestasis; Drugs, Chinese Herbal; Glucosides; Hepatitis; Humans; Male; Monoterpenes; Paeonia; Random Allocation; Rats; Rats, Sprague-Dawley; Sensitivity and Specificity; Time Factors

A high-performance liquid chromatographic (HPLC) method was developed for quantitative analysis of seven components, 5-hydroxymethyl-2-furaldehyde (1), albiflorin (2), paeoniflorin (3), liquiritin (4), ferulic acid (5), nodakenin (6), and glycyrrhizin (7) of Palmul-tang (PMT), a traditional Korean medicine. HPLC analysis was performed using a Gemini C18 column at 40°C, and photodiode array (PDA) detection at 230 nm, 254 nm, 280 nm, 320 nm, and 330 nm was used for quantification of the seven components in PMT. The mobile phase was a gradient flow composed of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile containing 1.0% (v/v) acetic acid. Calibration curves were acquired with r (2) values > 0.9998, and the relative standard deviations (RSDs, %) for intra- and interday precision were both less than 6.0%. The recovery of each component was in the range of 90.66-103.79%, with a RSD less than 5.0%. The contents of the seven components in PMT range form 0.61-6.21 mg/g. Additionally, we investigated the cytotoxicity of the extract against the RBL-1 and BEAS-2B cell lines, as well as splenocytes.

Topics: Animals; Benzoates; Bridged-Ring Compounds; Cell Line, Tumor; Chemistry, Pharmaceutical; Chromatography, High Pressure Liquid; Coumaric Acids; Coumarins; Cytotoxins; Drug Evaluation, Preclinical; Flavanones; Furaldehyde; Glucosides; Glycyrrhizic Acid; Medicine, Korean Traditional; Mice; Mice, Inbred BALB C; Monoterpenes; Plant Preparations; Rats

Peoniflorin (PF) and albiflorin (AF) are two principal components of Paeonia species, which exhibit various biological activities such as improvement of blood circulation and immunoregulating function. To further utilization of waste parts of peony plants, an efficient method for preparative purification of these two ingredients from white peony rhizome was developed based on macroporous resin (MAR) and medium-pressure liquid chromatography (MPLC). The separation characteristics of nine typical MARs were investigated by static adsorption/desorption experiments, and LX38 was revealed as optimal one. Further static experiments with LX38 resin indicated that the adsorbents fitted well to the pseudo-second-order kinetics model and both Langmuir and Freundlich isotherm models. Based on the optimal process parameters, a large-scale preparation was successfully applied. After one run treatment with LX38, the contents of PF and AF were increased 15-fold to 24.5 and 16.8% in the refined extract, respectively. Both purified compounds were obtained from refined extract by reversed-phase MPLC at second-stage separation. The process developed is better because of its low cost, high efficiency, and procedural simplicity making it a potential approach for large-scale production of PF and AF for their further applications in functional foods and pharmaceuticals.

Topics: Adsorption; Benzoates; Bridged-Ring Compounds; Chromatography, Liquid; Glucosides; Monoterpenes; Paeonia; Plant Extracts; Porosity; Resins, Synthetic; Rhizome

To study the dissolution rate of active components of different extracting solvents of Danggui Chishao drug pair.. The dissolution rates of phenolic acids (ferulic acid, vanillic acid and gallic acid), monoterpenes (gallic acid, peoniflorin, albiflorin, hydroxypeoniflorin and galloylpaeoniflorin) and phthalates (senkyunolide and ligustilide) contained in Danggui Chishao drug pair were determined by quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) and ultra high performance liquid chromatography in combination with triple-quadrupole mass spectrometry (UPLC-TQ-MS).. The dissolution rates of phenolic acids and monoterpenes decreased with the increase in alcohol concentration, on the contrary the rates of phthalates increased. The relative dissolution rates of most active components were relatively high in water and low concentration alcohol than other solvents.. Liquid chromatography-mass spectrometry is practical for comprehensive multi-component assessment on traditional Chinese medicine preparation processes and can provide reference for optimization of processing parameters.

Topics: 4-Butyrolactone; Benzoates; Benzofurans; Bridged-Ring Compounds; Chromatography, Liquid; Drugs, Chinese Herbal; Gallic Acid; Glucosides; Hydroxybenzoates; Mass Spectrometry; Monoterpenes; Phthalic Acids; Solvents; Vanillic Acid

The paper is to report the development of a method of quantitative analysis of multi-components by high performance liquid chromatography (HPLC) for simultaneously determining paeoniflorin sulfonate (PS), paeoniflorin (PF) and albiflorin (AF) in sulfated Paeoniae Radix Alba. Moreover, the cytotoxicity of paeoniflorin sulfonate by MTT-assay and the acute toxicity of mice by administration of paeoniflorin sulfonate were evaluated. Chromatographic separation of paeoniflorin sulfonate, PF and AF were performed on a SHISEIDO CAPCELL PAK C18 column (250 mm x 4.6 mm, 5 microm) for HPLC and a mixture of acetonitrile and 0.02% phosphoric acid solution (15 : 85) as the mobile phase. As detector a spectrophotometer set at 230 nm; column temperature 30 degrees C; flow rate 1.0 mL x min(-1). The toxicity of paeoniflorin sulfonate was evaluated by in vitro cytotoxicity carried out on mouse and human primary hepatocytes, and by acute oral toxicity test carried out on mice. The calibration curve of paeoniflorin sulfonate, PF and AF revealed linearity in the range of 0.041 8 - 1.045 0, 0.023 5 - 0.587 5, and 0.039 8 - 0.995 0 mg x mL(-1), respectively (r > 0.999 8). The average recovery was ranged from 99.11% to 101.71%, RSD < 2%. Paeoniflorin sulfonate does not have any cytotoxicity to cells at all the tested concentrations (< or = 300 micromol x L(-1)) in the in vitro cytotoxicity assay. The maximum tolerance dose of paeoniflorin sulfonate solution and extraction of Paeoniae Radix Alba to mouse is 5 g x kg(-1) and 80 g x kg(-1) respectively. The contents of these three components in the samples were determined with the developed method. It is a rapid, convenient and accurate method to determine multi-components. The content of PF in sulfated Paeoniae Radix Alba is significantly lower, and there is negative correlationship between the content of paeoniflorin sulfonate and PF. The in vitro cytotoxicity assay and in vivo mouse acute toxicity test showed that there is no obvious toxicity of paeoniflorin sulfonate and water-soluble extract of sulfated Paeoniae Radix Alba.

Topics: Animals; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Glucosides; Hepatocytes; Humans; Mice; Mice, Inbred ICR; Monoterpenes; Paeonia; Plant Roots; Plants, Medicinal; Sulfur; Volatilization

This study compared the pharmacokinetics of albiflorin (ALB) and paeoniflorin (PAE), respectively, after oral administration of ALB, PAE, Radix Paeoniae alba (RPA) extract, and Danggui-Shaoyao-San (DSS) extract to rats on separate occasions. Analytes were detected simultaneously with liquid chromatography-tandem mass spectrometry. Noncompartmental pharmacokinetic parameters were calculated. After oral administration of RPA and DSS extract to rats, ALB reached maximum concentrations of 4637 ± 2774 ng/ml (0.40 ± 0.14 h) and 226 ± 122 ng/ml (0.35 ± 0.14 h) and PAE reached maximum concentrations of 2132 ± 560 ng/ml (0.40 ± 0.14 h) and 143 ± 65 ng/ml (0.45 ± 0.11 h), respectively. Compared to the AUC(0 - t) value (1122 ± 351 and 722 ± 158 ng h/ml for ALB and PAE, respectively) after administration of monomers, larger AUC(0 - t) value of ALB (4755 ± 2560 ng h/ml) and PAE (2259 ± 910 ng h/ml) after administration of RPA extract and smaller AUC(0 - t) value of ALB (411 ± 118 ng h/ml) and PAE (242 ± 126 ng h/ml) after administration of DSS extract were obtained. The C(max), AUC, and K(el) of ALB and PAE were remarkably increased (P < 0.05, 0.01 or 0.005) during oral administration of RPA extract in comparison to that of DSS extract.

Topics: Administration, Oral; Animals; Benzoates; Bridged-Ring Compounds; Drugs, Chinese Herbal; Glucosides; Male; Molecular Structure; Monoterpenes; Paeonia; Rats; Rats, Sprague-Dawley

Central composite design (CCD), together with multiple linear regression, was successfully used to optimize the electrophoretic buffer system of micellar electrokinetic capillary chromatography (MEKC) for the determination of albiflorin, paeoniflorin, liquiritin, and glycyrrhizic acid in the traditional Chinese medicine (TCM) prescription, Yangwei granule. Concentrations of sodium deoxycholate (SDC) and borate, and proportions of ammonia, acetonitrile, and methanol were optimized. The total resolutions of peaks between the analytes and their adjacent peaks in real samples were integrated into the evaluation index of separation efficiency. The optimum electrophoretic buffer contained 80 mmol/L SDC, 20 mmol/L borate, 5% (v/v) methanol, 0.5% (v/v) ammonia, and 5% (v/v) acetonitrile. The correlation coefficients (R(2)) between the peak areas and the corresponding concentrations of analytes were greater than 0.9956. The limits of detection (LODs) (S/N=3) of the analytes were 0.97-4.00 μg/ml. The results indicate the superiority of CCD in optimizing the separation conditions of complex samples such as TCM prescriptions.

Topics: Acetonitriles; Ammonia; Benzoates; Borates; Bridged-Ring Compounds; Buffers; Chemistry, Pharmaceutical; Chromatography, Micellar Electrokinetic Capillary; Deoxycholic Acid; Drugs, Chinese Herbal; Flavanones; Glucosides; Glycyrrhizic Acid; Methanol; Micelles; Monoterpenes; Reproducibility of Results

To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneously determining paeoniflorin and albiflorin in Paeoniae Radix Alba.. Using paeoniflorin as the internal reference substance, the relative correction factor (RCF) of albiflorin was determined by HPLC and UPLC with good reproducibility. The contents of paeoniflorin in 16 samples of Paeoniae Radix Alba were authentically determined by the external standard method, and the content of albiflorin was calculated according to the RCF. The contents of these two components in the samples were determined with the external standard method.. No siginificant differences between the quantitative results of QAMS method and external standard method were observe.. It is a convenient and accurate method to determine multi-components when some authentic standard substances were unavailable. It can be used to control the quality of Paeoniae Radix Alba

Topics: Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Glucosides; Monoterpenes; Paeonia

To investigate pharmacokinetic parameters of peoniflorin, albiflorin and amygdaloside after administration of Guizhi Fuling capsule in beagle dogs.. Plasma was collected from forelimb vein of Beagle dogs after oral administration of Guizhi Fuling capsule. HPLC-MS/MS method was used to determine the concentrations of constituents in plasma. The pharmacokinetic parameters were analyzed by program DAS 2.0.. The limit of quantitation of peoniflorin, albiflorin and amygdaloside were 0.25, 2.64, 0.04 microg x L(-1), respectively. After administrated with different doses, half-life of peoniflorin in dogs were 4.33, 3.62 h, albiflorin were 6.16, 5.91 h, amygdaloside were 2.43, 1.32 h. The AUC(0-t) of all components were related to dose.. The pharmacokinetic course of peoniflorin, albiflorin and amygdaloside can be described by two-compartment model, and these components have high expose.

Topics: Administration, Oral; Amygdalin; Animals; Area Under Curve; Benzoates; Bridged-Ring Compounds; Capsules; Chromatography, High Pressure Liquid; Dogs; Drugs, Chinese Herbal; Glucosides; Half-Life; Male; Monoterpenes; Tandem Mass Spectrometry

The aim of this study was to investigate the therapeutic effects of the combination of paeoniflorin and albiflorin (CPA) extracted from Paeonia radix on radiation and chemotherapy induced myelosuppression in two animal models: mice and rabbits. Mice were exposed to X-ray radiation (400 Roentgen), and both mice and rabbits were intraperitoneally injected with cyclophosphamide (100.0 mg/kg) and cytarabine chloride (92.7 mg/kg), respectively, for 3 days to induce myelosuppression. CPA was subsequently administrated intravenously at low (15.0 mg/kg for mice, 6.00 mg/kg for rabbits), intermediate (30.0 mg/kg for mice, 12.0 mg/kg for rabbits) and high (60.0 mg/kg for mice, 24.0 mg/kg for rabbits) doses, as well as orally (60.0 mg/kg for mice, 24.0 mg/kg for rabbits) for 7 days. Shenqi tablets were used as positive controls (oral administration of 936.0 mg/kg for mice, 336.0 mg/kg for rabbits). The administration of CPA significantly ameliorated myelosuppression in all cases. For the X-ray irradiated mice and the chemotherapy treated mice and rabbits, high dosages of CPA resulted in the recovery of, respectively, 94.4%, 95.3% and 97.7% of hemoglobin content; 67.7%, 92.0% and 94.3% of platelet numbers; 26.8%, 137.1% and 107.3% of white blood cell counts; as well as a reversal in the reduction of peripheral differential white blood cell counts. There was also a recovery of 50.9%, 146.1% and 92.3%, respectively, in the animals' relative spleen weight. Additionally, a recovery of 35.7% and 87.2% in the number of bone marrow nucleated cells was observed in the radio- and chemotherapy treated mice, respectively. Bone marrow white blood cell counts also resumed to normal levels. These results substantiate the marked therapeutic effects of CPA to ameliorate myelosuppression induced by radio and chemotherapy.

Topics: Animals; Benzoates; Blood Platelets; Bone Marrow Cells; Bridged-Ring Compounds; Chemotherapy, Adjuvant; Cyclophosphamide; Cytarabine; Drug Synergism; Drugs, Chinese Herbal; Female; Glucosides; Hemoglobins; Herb-Drug Interactions; Leukocytes; Male; Mice; Mice, Inbred BALB C; Monoterpenes; Paeonia; Plant Extracts; Plant Roots; Rabbits; X-Rays

To investigate the microbiological transformation of paeoniflorin and albiflorin.. The bacteria strains able to transform paeoniflorin and albiflorin were screened from 18 strains of microorganisms. The products were isolated by chromatography method and their structures were elucidated by spectral technology.. It was found that Cunninghamella blakesleana (AS 3.970) and Syncephalastrum racemosum (AS 3.264) could convert paeoniflorin and albiflorin efficiently, respectively. C. blakesleana could convert paeoniflorin to produce albiflorin, while S. racemosum could convert albiflorin to produce paeoniflorin.. Paeoniflorin and albiflorin could be converted each other in definited condition.

Topics: Benzoates; Biotransformation; Bridged-Ring Compounds; Cunninghamella; Glucosides; Monoterpenes; Mucorales

To establish a HPLC-MS method and investigate the pharmacokinetic properties of paeoniflorin, albiflorin and oxypaeoniflorin and the pharmacokinetics difference of Radix Paeoniae Rubra and Radix Paeoniae Alba.. The extracts of Radix Paeoniae Rubra and Radix Paeoniae Alba were separately administrated to rats. The concentrations of paeoniflorin, albiflorin and oxypaeoniflorin in rat plasma were determined by HPLC-ESI-MS method. The plasma samples were pretreated by protein precipitation with methanol and chromatographic separation was performed on a C(18) column with a mobile phase consisted of 0.1% formic acid and methanol (67:33, v/v). The detection was accomplished by multiple-reaction monitoring (MRM) scanning via electrospray ionization (ESI) source operating in the negative ionization mode. Main pharmacokinetic parameters were estimated and the total AUC of the three components were compared.. The pharmacokinetic parameters of paeoniflorin, albiflorin and oxypaeoniflorin were significantly different. There was significant difference between the pharmacokinetic characteristics of Radix Paeoniae Rubra and Radix Paeoniae Alba.. A specific and sensitive HPLC-ESI-MS method was developed for simultaneous determination of paeoniflorin, albiflorin and oxypaeoniflorin in rat plasma and was successfully applied to pharmacokinetic study. The results might be helpful for the investigation of different effects of Radix Paeoniae Rubra and Radix Paeoniae Alba.

Topics: Administration, Oral; Animals; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glucosides; Male; Monoterpenes; Paeonia; Plant Extracts; Plant Roots; Rats; Rats, Sprague-Dawley; Reference Standards; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization

We developed a method to simultaneously determine paeoniflorin and albiflorin levels using high-performance anion-exchange liquid chromatography with pulsed amperometric detection (HPAEC-PAD). The main principle of our method includes solid-phase extraction step using Amberlite XAD-2 sorbent to remove sugars and to selectively determine glycosides by PAD. Under these conditions, the linear dynamic range was 0.01-100 microg/mL, and the albiflorin and paeoniflorin detection limits (S/N=3) were 5 and 10 pg, respectively. The intra- and inter-day precisions (RSDs) were <5.07%, and the average recoveries from Paeoniae Radix and Si-ni-san ranged from 97.12 to 101.15%. Our method showed high selectivity, high sensitivity, and good repeatability for analyzing albiflorin and paeoniflorin in oriental medicinal preparation.

Topics: Adsorption; Anions; Benzoates; Bridged-Ring Compounds; Carbohydrates; Chromatography, Ion Exchange; Electrochemical Techniques; Glucosides; Linear Models; Monoterpenes; Paeonia; Plant Extracts; Plant Roots; Reproducibility of Results; Sensitivity and Specificity; Solid Phase Extraction

A rapid, sensitive and selective liquid chromatography/tandem mass spectrometry method (LC-MS/MS) was developed and validated for simultaneous determination of albiflorin and paeoniflorin in rat plasma using geniposide as an internal standard. Plasma samples were extracted by solid-phase extraction. Chromatographic separation was carried out on a Zorbax SB-C(18) analytical column (150 × 2.1 mm × 5 µm) with 0.1% formic acid-acetonitrile (70:30, v/v) as the mobile phase. Detection was performed by multiple reaction monitoring mode using electrospray ionization in the positive ion mode. The total run time was 3.0 min between injections. The calibration curves were linear over a range of 1-1000 ng/mL for albiflorin and 2-2000 ng/mL for paeoniflorin. The overall precision and accuracy for all concentrations of quality controls and standards were better than 15%. Mean recovery was determined to be 87.7% for albiflorin and 88.8% for paeoniflorin. The validated method was successfully applied to the pharmacokinetic study of albiflorin and paeoniflorin in rat plasma after oral administration of Radix Paeoniae Alba extract and Tang-Min-Ling-Wan. The pharmacokinetic parameters showed that albiflorin and paeoniflorin from Tang-Min-Ling-Wan were absorbed more rapidly with higher concentrations in plasma than that from Radix Paeoniae Alba extract. The results provided a meaningful basis for evaluating the clinical applications of traditional Chinese medicine.

Topics: Animals; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glucosides; Male; Monoterpenes; Paeonia; Rats; Rats, Wistar; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

The contents of gallic acid, catechin, albiflorin, paeoniflorin, benzoic acid and paeonol extracted in different growth years, collecting season and of different parts of Paeonia lactiflora were determined. The results showed that the contents of catechin and paeoniflorin in Paeonia lactiflora collected in autumn are the highest, and the contents of benzoic acid was lower than that of those collected at other time. The longer is the age of Paeonia lactiflora, the higher is the contents of catechin and paeoniflorin. The contents of catechin and paeoniflorin in the root of Paeonia lactiflora were higher than those in other parts of the plant. There is a certain content of paeoniflorin in the leaves of Paeonia lactiflora. Judging from the result, paeoniflorin is synthesized in the leaf and then transported to the root. Catechin is not synthesized in the leaf, but mainly in the root. Paeonia lactiflora should be collected in autumn, and immature plant should not be collected.

Topics: Acetophenones; Benzoates; Benzoic Acid; Bridged-Ring Compounds; Catechin; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Gallic Acid; Glucosides; Monoterpenes; Paeonia; Plant Leaves; Plant Roots; Plants, Medicinal; Seasons

We have developed a reversed-phase high-performance liquid chromatography-pulsed amperometric detection (RP-HPLC-PAD) method for the detection of albiflorin and paeoniflorin in Paeoniae Radix and Wu-ji-san. Albiflorin and paeoniflorin were completely separated using 10% acetonitrile in 5mM sodium phosphate buffer (pH 3.0) as an eluent and detected by PAD under alkaline conditions after using a post-column delivery system. The limit of detection (S/N=3) and the limit of quantification (S/N=10) were 0.10 and 0.35 ng for albiflorin, and 0.20 and 0.50 ng for paeoniflorin, respectively. The coefficients of linear regression were 0.9995 and 0.9999 for concentrations between 0.035 and 100 microg/mL. The intra- and inter-day precision (RSDs) was less than 3.56% in Paeoniae Radix and Wu-ji-san. The average recoveries from Paeoniae Radix and Wu-ji-san were 99.01-100.94% and 99.46-100.64%. This method shows higher selectivity than HPLC-UV method for analyzing albiflorin and paeoniflorin in Chinese medicinal preparation.

Topics: Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Electrochemical Techniques; Glucosides; Isoflavones; Monoterpenes; Paeonia; Reference Standards; Reproducibility of Results; Solvents; Spectrophotometry, Ultraviolet

Peony, the processed root of Paeonia lactiflora Pall. (Ranunculaceae), is a component herb of many traditional formulae for the treatment of depression-like disorders.. The present study aimed to investigate whether the total glycosides of peony (TGP) could prevent depression induced by chronic stress.. Mice were subjected to an experimental setting of chronic unpredictable stress (CUS). The effect of TGP treatment on CUS-induced depression was examined by measuring behavioral and neurochemical parameters of depression and the antioxidant status of brain tissue.. CUS-induced depression, as indicated by a significant increase in immobility time in the tail suspension test, was associated with increases in the activities of monoamine oxidases, depletion of reduced glutathione, and an increase in malondialdehyde level, in mice brains. TGP treatment alleviated the extent of CUS-induced depression and the associated impairment of antioxidant status in the mouse brain.. The results suggest that TGP alleviates depression induced by chronic unpredictable stress. The antidepressant-like activity of TGP is probably mediated by inhibition of monoamine oxidases and the attenuation of oxidative stress in mouse brain.

Topics: Animals; Antidepressive Agents; Antioxidants; Benzoates; Bridged-Ring Compounds; Depressive Disorder; Glucosides; Glutathione; Glycosides; Male; Malondialdehyde; Mice; Mice, Inbred ICR; Monoamine Oxidase; Monoterpenes; Paeonia; Phytotherapy; Plant Extracts; Stress, Psychological

To investigate the purification process of total paeony glycoside (TPG) from Radix Paeoniae Rubra by macroporous resin with double indices of albiflorin and paeoniflorin.. HPLC was used for simultaneous determination of albiflorin and paeoniflorin and the results were used as main indices to investigate the technological parameters for purifying TPG by D101 macroporous resin. Then the result was verified by ampliate test.. The applicable technological conditions of purification for TPG by D101 macroporous resin were as follows: the concentrations of albiflorin and paeoniflorin in original solution were 0.12 mg/mL, 2.58 mg/mL with a flow rate of 3.6 BV/h, and the adsorption capacity were 0.71 mg/mL, 15.43 mg/mL, respectively. The eluant was 20% alcohol with 8 times of resin volume, and the elution ratio were 72.18%, 90.84%, respectively. The result of ampliate test accorded with that of small test.. D101 macroporous adsorption resin can be used to purify TPG from crude extracts of Radix Paeoniae Rubra.

Topics: Absorption; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Ethanol; Glucosides; Glycosides; Humans; Monoterpenes; Paeonia; Plant Roots; Plants, Medicinal; Resins, Synthetic; Technology, Pharmaceutical

Paeony root has long been used for its anti-inflammatory effects. In this study, the effects of albiflorin, paeoniflorin, and paeonol, compounds from paeony root, on gene expression profiles were examined in macrophages challenged with the inflammation inducer lipopolysaccharide (LPS).. The RAW264.7 macrophages were treated with LPS in the presence or absence of albiflorin, paeoniflorin, or paeonol. Global mRNA expression levels were detected by using an oligonucleotide microarray platform covering the mouse whole genome.. Treatment with LPS caused expression level changes in 1,270 genes by 2 folds or more. Paeonol attenuated the induction level of 355 LPS-responsive genes. Classification of the genes targeted by paeonol according to the Panther group analysis revealed 20 biological processes, 24 molecular functions, and 22 signaling pathways. The Panther signaling pathways highly affected by paeonol included the 'inflammation mediated by chemokine and cytokine signaling', 'interleukin signaling', and 'Toll receptor signaling'.. Our results demonstrate that paeonol has extensive inhibitory effects on the regulation of inflammation associated gene expression by LPS in macrophages. In addition, the predominant effect of paeonol among the tested compounds suggests that paeonol may be a major ingredient for the anti-inflammatory effect of paeony root.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Benzoates; Bridged-Ring Compounds; Cells, Cultured; Chemokines; Drugs, Chinese Herbal; Gene Expression Profiling; Gene Expression Regulation; Glucosides; Inflammation; Interleukins; Lipopolysaccharides; Macrophages; Mice; Monoterpenes; Oligonucleotide Array Sequence Analysis; Toll-Like Receptors

In order to analyze paeoniflorin, albiflorin and their derivatives (PADs) in Paeonia Lactiflora rapidly and effectively, (+)ESI-MS(n) experiments were conducted, from which two diagnostic fragment patterns were acquired. Meanwhile, the dehydration ability of aglycones of PADs was obtained by calculating their activation energy using density functional theory, through which the unique dehydration phenomenon of benzoylalbiflorin, compared with benzoylpaeoniflorin, was interpreted. In addition, a computer-aided mass spectrometry analysis program was developed to facilitate the analysis of the unknown compound by suggesting the possible structure of the analyte.

Topics: Algorithms; Benzoates; Bridged-Ring Compounds; Computer Simulation; Glucosides; Models, Chemical; Monoterpenes; Paeonia; Sodium; Software; Spectrometry, Mass, Electrospray Ionization

A complex formula composed of Paeonia lactiflora PALL. and Glycyrrhiza uralensis Fisch., which is called as Jakyak-Gamcho Decoction (JGD), has been used for a pain-relieving function and muscle spasms due to blood deficiency in the traditional medicine. In this study, the anti-inflammatory activity of JGD was evaluated based on the quantitative determinations and the relative proportions of six major constituents in the decoction mixture extracted by orthogonal array methods. Our results suggest that the three parameters are all crucial factors. The optimized conditions for extraction were therefore established [solvent (water); pH value (4); extraction number (4)]. We also optimized the extraction conditions related to anti-inflammatory activity [solvent (70% EtOH); pH value (6); extraction number (4)]. So, we found that the bioactivity was responsible for mixed components but not individual one. It was proportionally associated with the amounts of some components in the extracts of herbal medicines. When the proportion of the active components was similar to each other, they had the similar functions. Furthermore, the results could establish a model system for the quality assurance of herbal preparations, and provided a new paradigm of active components-pharmacodynamics, which is used for illustrating the connections between the bioactivities and the proportion of active constituents in the extracts of herbal medicines.

Topics: Alkaline Phosphatase; Animals; Benzoates; Bridged-Ring Compounds; Cell Line; Cell Survival; Chalcone; Chemical Fractionation; Chromatography, High Pressure Liquid; Ethanol; Flavanones; Glucosides; Glycyrrhiza; Glycyrrhizic Acid; Herbal Medicine; Lipopolysaccharides; Macrophages; Monoterpenes; NF-kappa B; Nitric Oxide; Paeonia; Plant Extracts; Research Design; Solubility; Technology, Pharmaceutical; Water

A new monoterpene glycoside, 6'-O-beta-D-glucopyranosylalbiflorin (1), and four known compounds; albiflorin (2), 6'-O-benzoylalbiflorin (3), paeoniflorin (4) and benzoyl paeoniflorin (5), were isolated from the methanolic extract of the roots of Paeonia lactiflora Pall.. Their chemical structures were completely elucidated using a combination of 2D NMR techniques (COSY, HMQC and HMBC) and HRESI-MS analyses. To investigate the bioactivities of these compounds, their effects on the differentiation of osteoblastic MC3T3-E1 cells were tested. Compound 1 (0.01-10 microM) significantly increased the alkaline phosphatase activity and nodules mineralization of MC3T3-E1 cells compared to those of the control (P<0.05). These results suggest that newly isolated compound 1 has a direct stimulatory effect on bone formation in vitro and may contribute to the prevention for osteoporosis.

Topics: Alkaline Phosphatase; Animals; Benzoates; Bone Density Conservation Agents; Bridged-Ring Compounds; Calcification, Physiologic; Cell Differentiation; Cell Line; Dose-Response Relationship, Drug; Glucosides; Glycosides; Magnetic Resonance Spectroscopy; Mice; Molecular Structure; Monoterpenes; Osteoblasts; Paeonia; Plant Extracts; Plant Roots; Spectrometry, Mass, Electrospray Ionization; Vietnam

A recombinant single chain variable-fragment (scFv) antibody against paeoniflorin (PF) was produced using the hybridoma cell line C31B9. Variable regions of heavy (V (H)) and light (V (L)) chain antibody genes were directly cloned from cDNA resources of hybridoma C31B9 and assembled using splicing by overlap extension (SOE)-PCR using a (Gly (4)Ser) (3) linker DNA. The constructed scFv genes were cloned into pET28a vectors for the generation of recombinant proteins in Escherichia coli. Most of the recombinant proteins were expressed in inclusion bodies. The yield of refolded and purified scFv was 1.89 mg per 100 mL of cell culture. The recombinant scFv displayed cross-reactivity as its mother monoclonal antibody (MAb) C31B9. Therefore, the newly expressed scFv protein was applied to quantitative ELISA to determine the total paeoniflorin (PF) and albiflorin (Alb) concentrations in peony root samples. Using PF as a standard compound, the full linear range of the assay was extended from 0.78 to 25 microg/mL. The results obtained by ELISA employing both the recombinant scFv and the original MAbC31B9 showed a reasonably good agreement with each other.

Topics: Antibodies, Monoclonal; Antibody Affinity; Benzoates; Bridged-Ring Compounds; Cell Line; Cross Reactions; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Glucosides; Immunoglobulin Fragments; Immunoglobulin Variable Region; Monoterpenes; Paeonia; Recombinant Proteins

A sensitive and rapid high-performance liquid chromatography (HPLC) method with solid-phase extraction (SPE) to simultaneously determine albiflorin and paeoniflorin in rat serum was described. Serum samples were pretreated with solid-phase extraction using Extract-Clean cartridges, and the extracts were analyzed by HPLC on a reversed-phase C(18) column and a mobile phase of acetonitrile-0.03% formic acid (17:83 (v/v)) with ultraviolet detection at 230 nm. Pentoxifylline was used as the internal standard (IS). The linear ranges of the calibration curves were 29-1450 ng/ml for albiflorin and 10-2000 ng/ml for paeoniflorin. The intra- and inter-day precisions (R.S.D.) were

Topics: Administration, Oral; Animals; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glucosides; Monoterpenes; Rats; Reproducibility of Results

A high performance liquid chromatographic method (HPLC), together with solid phase extraction (SPE), was developed for simultaneous determination of albiflorin and paeoniflorin in rat urine after oral administration of Si-Wu decoction. The samples were pretreated with solid phase extraction using Extract-Cleantrade mark cartridges. Analysis of the extract was performed on a reversed-phase C18 column and a mobile phase made up of acetonitrile and 0.03% formic acid (17:83, v/v). UV detection was set at 230 nm. The assay was linear over the range 2.625-52.50 mg/mL for albiflorin and 3.875-77.50 microg/mL for paeoniflorin. The average percentage recoveries of three spiked urines were 97.01 +/- 3.32 and 102.32 +/- 6.97 for albiflorin and paeoniflorin, respectively. The intra-day precision (RSD) ranged from 0.21 to 1.79% at concentrations of 4.20, 10.50, 26.25 and 39.375 microg/mL of albiflorin and 0.12 to 2.92% at concentrations of 3.875, 10.85, 23.25 and 58.125 microg/mL of paeoniflorin, and inter-day precision (RSD) was from 1.02 to 1.86% for albiflorin and 0.94 to 3.30% for paeoniflorin, at the same four concentrations. This method was applied in order to analyze albiflorin and paeoniflorin in rat urine following oral administration of traditional Chinese medicinal preparation of Si-Wu decoction.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glucosides; Male; Monoterpenes; Rats; Rats, Sprague-Dawley; Sensitivity and Specificity

To study the chemical constituents of the root of Paeonia sinjiangensis.. The constituents were isolated by silica column chromatography, and their structures were identified on the basis of spectral analysis and their physical-chemical constants.. Five compounds, paeoniflorin( I ), albiflorin (II), lactiflorin(III), daucosterol(IV), sucrose (V), were obtained.. All of the compounds were obtained from this plant for the first time.

Topics: Benzoates; Bridged-Ring Compounds; Glucosides; Glycosides; Molecular Conformation; Molecular Structure; Monoterpenes; Paeonia; Plant Roots; Plants, Medicinal

This work describes an immunochemical approach for the quality control of Paeoniae Radix by an enzyme-linked immunosorbent assay (ELISA) based on determination of the total concentration of paeoniflorin (PF) and albiflorin (Alb), which are major bioactive constituents in Paeoniae Radix. Four hybridromas secreting monoclonal antibodies against PF and Alb were produced by fusing splenocytes from a mouse immunized by a PF-bovine serum albumin (BSA) conjugate with the hypoxanthine-aminopterin-thymidine (HAT)-sensitive mouse myeloma cell line, P3-X63-Ag8-653. A relatively higher reactivity of monoclonal antibodies (MAbs) with PF and Alb than oxypaeoniflorin (OP) and benzoylpaeoniflorin (BP) was observed, while other monoterpenes and benzoic acid did not cross-react. When PF was used as a standard, the assay can cover a measuring range of 20-600 ng/ml for PF and Alb. A series of Paeoniae Radix samples have been determined, and the results showed good agreement with that determined by traditional high-performance liquid chromatography (HPLC). The developed competitive ELISA was 100 times more sensitive than the HPLC method. Meanwhile, fifteen Chinese traditional prescriptions were determined by the competitive ELISA.

Topics: Animals; Antibodies, Monoclonal; Benzoates; Bridged-Ring Compounds; Cell Fusion; Chromatography, High Pressure Liquid; Cross Reactions; Enzyme-Linked Immunosorbent Assay; Glucosides; Hybridomas; Medicine, Chinese Traditional; Mice; Monoterpenes; Tumor Cells, Cultured

We previously reported that paeoniflorin but not albiflorin, components of peony root, produced ameliorative effects on scopolamine-induced spatial cognitive impairment in rats. In this study, we examined the effects of paeoniflorin and muscarinic receptor antagonists on long-term potentiation (LTP) of population spike recorded from the CA1 region of rat hippocampal slices. Bath applications of an M1- and M2-receptor antagonist scopolamine and a selective M1-receptor antagonist pirenzepine, at a concentration of 10 microM, significantly suppressed LTP, whereas AF-DX116, a selective M2-receptor antagonist, failed to affect it. Paeoniflorin (0.1-1 microM), which alone was ineffective on LTP induction, significantly reversed the suppressive effects of scopolamine and pirenzepine (10 microM). In contrast, albiflorin (0.1- 1 microM) had no effect on the scopolamine-induced LTP suppression. These results suggest that paeoniflorin reversal of the muscarinic M1-receptor-mediated inhibition of LTP may be implicated in the ameliorative effect of paeoniflorin on spatial cognitive impairment caused by cholinergic dysfunction.

Topics: (4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethylammonium Chloride; Animals; Benzoates; Bridged-Ring Compounds; Cognition; Glucosides; Hippocampus; Long-Term Potentiation; Male; Monoterpenes; Muscarinic Antagonists; Rats; Rats, Wistar; Receptor, Muscarinic M1; Receptors, Muscarinic