lignans and pinoresinol-diglucoside

This study is aimed to investigate the effect of pinoresinol diglucoside (PDG) in ameliorating myocardial ischemia-reperfusion injury (MIRI). Hypoxia/reperfusion (H/R)-induced H9c2 cardiomyocytes were used to establish an in-vitro ischemia-reperfusion injury model of cardiomyocytes. Cells were treated with 1 μmol/L of PDG. Reactive oxygen species (ROS) level was detected by a 2',7'-dichlorofluorescein-diacetate assay. The release of lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) was examined by enzyme-linked immunosorbent assay. The viability and apoptosis of H9c2 cells were probed by MTT assay and flow cytometry. Besides this, Western blot and quantitative real-time PCR were used to detect microRNA-142-3p (miR-142-3p) and hypoxia-inducible factor 1 subunit alpha inhibitor (HIF1AN) expression levels. The binding sequence between miR-142-3p and HIF1AN 3'-untranslated region was validated by a dual-luciferase reporter gene assay. PDG treatment significantly reduced the level of ROS, LDH, and CK-MB, promoted viability, and inhibited the apoptosis of H9c2 cells. PDG treatment promoted miR-142-3p expression and inhibited HIF1AN expression in H9c2 cells. MiR-142-3p overexpression enhanced the effects of PDG on ROS, LDH, CK-MB levels, cell viability, and apoptosis in H9c2 cardiomyocytes, while overexpression of HIF1AN reversed the above effects. PDG ameliorates H/R-induced injury of cardiomyocytes by regulating miR-142-3p and HIF1AN.

Topics: Apoptosis; Creatine Kinase; Humans; Hypoxia; Hypoxia-Inducible Factor 1; L-Lactate Dehydrogenase; Lignans; MicroRNAs; Mixed Function Oxygenases; Myocardial Reperfusion Injury; Myocytes, Cardiac; Reactive Oxygen Species; Repressor Proteins; Untranslated Regions

Brain ischemia/reperfusion (I/R) injury is a common pathological process after ischemic stroke. Pinoresinol diglucoside (PDG) has antioxidation and anti-inflammation activities. However, whether PDG ameliorates brain I/R injury is still unclear. In this study, middle cerebral artery occlusion (MCAO) model was established with male C57BL/6 mice, and the mice were treated with 5 and 10 mg/kg PDG via intravenous injection, respectively. The neurological deficit, infarct volume, and brain water content were then evaluated. HE staining and Nissl staining were used to analyze neuron injury. Besides, enzyme-linked immunosorbent assay and colorimetry assay were used to examine the level of inflammatory markers and oxidative stress markers, and Western blot was used to detect the expressions of p-p65, Nrf2, and HO-1. It was revealed that PDG could significantly alleviate the MCAO-induced neurological dysfunction of the mice and reduce the infarct volume, brain water content, and neuron injury. PDG treatment decreased the levels of TNF-α, IL-1β, IL-6, NO, ROS, and MDA, and significantly increased the activities of SOD, GSH, and GSH-Px in the brain tissue of the mice. Additionally, PDG could repress the activation of p65 and promote Nrf2 and HO-1 expressions. In conclusion, PDG exerts anti-inflammatory and antioxidation effects via regulating the NF-κB pathway and Nrf2/HO-1 pathway, thereby reducing the I/R-induced brain injury of mice.

Topics: Animals; Brain; Brain Ischemia; Cytokines; Disease Models, Animal; Heme Oxygenase-1; Lignans; Male; Membrane Proteins; Mice, Inbred C57BL; Middle Cerebral Artery; Neuroinflammatory Diseases; Neurons; NF-E2-Related Factor 2; Oxidative Stress; Reperfusion Injury

For Alzheimer's disease (AD), there is still no effective treatment strategy. Pinoresinol diglucoside (PDG) is one of the major lignans isolated from Eucommia ulmoides. It is endowed with multiple pharmacological activities, including anti-inflammatory, antioxidant and anticancer activities. In this study, we investigated the potential neuroprotective functions of PDG in AD. Mice model with AD was established adopting stereotactic hippocampal injection of Aβ1-42 (410 pmol/mouse), and 3 days later, mice were administrated with 5 and 10 mg/kg PDG by intragastric administration every day for 3 weeks. Morris water maze and Y-maze tests demonstrated that PDG treatment could markedly reverse Aβ1-42-induced memory impairment in mice. It is found that PDG restrained the release of proinflammatory cytokines (tumor necrosis factor α and interleukin 1β), reactive oxygen species and malondialdehyde, and promoted the activity of the antioxidant enzyme (superoxide dismutase and catalase) by quantitative real-time-PCR, colorimetric method and ELISA assay. Western blot assay results have shown that PDG could also upregulate the ratio of Bcl-2/Bax and downregulate cytochrome c and cleaved caspase-3 expressions, thereby inhibiting neuronal apoptosis. Furthermore, PDG also significantly reduced the expression of Toll-like receptor 4 (TLR4) and the activation of nuclear factor-κB (NF-κB) p65, and promoted nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) expressions. In conclusion, PDG can attenuate neuroinflammation, neuronal apoptosis and oxidative stress through the TLR4/NF-κB and Nrf2/HO-1 pathways, and ameliorate memory dysfunction induced by Aβ1-42 in mice.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Apoptosis; Catalase; Disease Models, Animal; Hippocampus; Inflammation; Injections; Interleukin-1beta; Lignans; Malondialdehyde; Mice; Morris Water Maze Test; Oxidative Stress; Peptide Fragments; Reactive Oxygen Species; Stereotaxic Techniques; Superoxide Dismutase; Toll-Like Receptor 4; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Pinoresinol diglucoside (PDG), the active compound extracted from Eucommia ulmoides, Styrax sp. and Forsythia suspensa, plays the roles in regulating hypertension, inflammation and oxidative stress.. Considering that hypertension and inflammation has been proved to contribute to cardiac remodeling, we tested the effects of PDG on cardiac hypertrophy (CM).. Male Sprague Dawley (SD) rats were used to construct hypertrophic rats by partial abdominal aortic constriction (AAC)-surgery. PDG solution (2 mg/ml) was used to treat AAC-induced rats by intraperitoneal injection at low dose (L-PDG, 2.5 mg/kg per day), medium dose (M-PDG, 5 mg/kg per day), and high dose (H-PDG, 7.5 mg/kg per day) for 3 weeks post AAC-surgery. CM was evaluated by the ratio of left ventricular weight to body weight ratio (LVW/BW), left ventricular wall thickness by H&E staining, and collagen content deposit by Masson's staining. Further, isoproterenol (ISO) and phenylephrine (PE) were used to produce cellular models of CM in neonatal rat ventricular cardiomyocytes (NRVMs). PDG pre-treated NRVMs 2 h at low dose (L-PDG, 2.5 μg/ml), medium dose (M-PDG, 5 μg/ml), and high dose (H-PDG, 7.5 μg/ml) for 24 h with or without PE- and ISO-stimulation. CM was evaluated by the expressions of hypertrophic biomarkers. Next, the hypertrophic biomarkers and pro-inflammatory cytokines were measured using quantitative real-time PCR (qRT-PCR), the expressions of protein kinase B (AKT)/mammalian target of rapamycin (mTOR)/transcription factor nuclear factor-kappa B (NF-kB) signaling pathway were determined by Western blotting.. PDG treatment prevented cardiac histomorphology damages, decreased upregulations of hypertrophic biomarkers, and prevented fibrosis and inflammation after pressure overload resulting from AAC-surgery. Consistently, PDG remarkably inhibited the changes of cardiomyocyte hypertrophic biomarkers and inflammatory responses in cellular models of CM. Interestingly, PDG administration inhibited the activation of AKT/mTOR/NF-kB signaling pathway both in vivo and in vitro.. PDG prevents AAC-induced CM in vivo, PE- and ISO-induced CM in vitro. The AKT/mTOR/NF-kB signaling pathway could be the potential therapeutic target involved in the protection of PDG. These findings provide novel evidence that PDG might be a promising therapeutic strategy for CM.

Topics: Animals; Animals, Newborn; Aorta, Abdominal; Cardiomegaly; Constriction, Pathologic; Disease Models, Animal; Fibrosis; Inflammation; Isoproterenol; Lignans; Male; Myocytes, Cardiac; NF-kappa B; Phenylephrine; Pressure; Primary Cell Culture; Proto-Oncogene Proteins c-akt; Rats, Sprague-Dawley; Signal Transduction; TOR Serine-Threonine Kinases; Ventricular Remodeling

The effects of roasting and in vitro digestion on total phenolic content (TPC), total flavonoid content (TFC), phenolic profiles, and antioxidant activity of water-soluble extracts from six varieties of sesame were investigated in this study. Our results showed that the major phenolic compounds in raw, roasted and digested sesame were gallic acid (GA), protocatechuic acid (PA), 4-hydroxybenzoic acid (4 HBA), ferulic acid (FA) and quercetin (Quer). Roasting significantly increased the TPC, pinoresinol diglucoside (PD), sesamol, as well as the content of phenolic compounds (especially GA, PA, 4 HBA and Quer) in sesame, but kept or reduced the TFC, sesamin and sesamolin. After roasting, the antioxidant potency composite index (ACI) of six varieties of sesame was significantly increased by 29.8%-216.6%. Additionally, the ACI of gastric digestion was significantly higher than that of oral and intestinal digestion during the in vitro digestion of the roasted-sesame, except for the varieties of Ganzhi 9 and Ganzhi 17. This study showed that five phenolic compounds (GA, PA, 4 HBA, p-coumaric acid, Quer) and sesamol of the water-soluble extracts contributed to the antioxidant activities of the digestive products of sesame.

Topics: Anticarcinogenic Agents; Antioxidants; Benzodioxoles; Coumaric Acids; Digestion; Dioxoles; Flavonoids; Gallic Acid; Hydroxybenzoates; Lignans; Parabens; Phenol; Phenols; Plant Extracts; Principal Component Analysis; Propionates; Quercetin; Seeds; Sesamum

Phomopsis sp. XP-8, an endophytic fungus from the bark of Tu-Chung (Eucommia ulmoides Oliv) showed capability to biosynthesize pinoresinol (Pin) and pinoresinol diglucoside (PDG) from glucose (glu) and phenylalanine (Phe). To verify the mass flow in the biosynthesis pathway, [

Topics: Ascomycota; Endophytes; Eucommiaceae; Furans; Glucose; Glycosides; Lignans; Mass Spectrometry; Phenylalanine

Defatted-sesame meal (DSM), a byproduct of sesame oil, has attracted considerable interest in the food industry because of its strong antioxidant activity. The aim of this study was to measure the content and distribution of lignans in DSM and evaluate their antioxidant activity after thermal processing and in vitro digestion. The results showed that the sesame lignans (SL) content and antioxidant activity were significantly influenced by the temperature and time during thermal preparation, and the maximum antioxidant potency composite index (ACI) was obtained after roasting the samples at 240 °C for 20 min. As sesame seed was processed with longer time and higher temperature, more pinoresinol diglucoside (PD) and sesamol were measured in DSM. According to the correlation matrix under thermal preparation, a significant contribution to the antioxidant potency of DSM was discovered. After in vitro digestion, the release amount of lignans increased by 19.6%, and the values of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and ACI gradually declined after digestion, with a 40% decrease in both the DPPH radical scavenging activity and the ACI from oral to intestinal phase. These results could be used to help improve the bioavailability of SL and obtaining high quality sesame byproducts.

Topics: Antioxidants; Biological Availability; Biphenyl Compounds; Hot Temperature; Hydrolysis; In Vitro Techniques; Lignans; Models, Biological; Picrates; Seeds; Sesamum

Topics: Anti-Bacterial Agents; Antioxidants; Bacteria; China; Chlorogenic Acid; Chromatography, High Pressure Liquid; Eucommiaceae; Flavonoids; Fungi; Gutta-Percha; Iridoid Glucosides; Iridoids; Kaempferols; Lignans; Metabolome; Phenols; Plant Bark; Plant Extracts; Plant Leaves; Plants, Medicinal; Quercetin; Rutin

Topics: Antihypertensive Agents; Ascomycota; Biotransformation; Freeze Drying; Glucose; Hydrogen-Ion Concentration; Lignans; Temperature

To develop a quantitative models for simultaneous determination of pinoresinol diglucoside (PDG) and geniposidic acid (GPA) in Eucommia ulmoides with near-infrared (NIR) spectroscopy combined with chemometrics. The NIR spectra were collected in diffuse reflection mode and pretreated with various spectra preprocessing methods including first-order differentiator, multiplicative scatter correction and so on. The optimal wavelength variables were screened out by competition adaptive weighted sampling method. The quantitative models for the simultaneous determination of PDG and GPA in Eucommia ulmoides were established with partial least squares (PLS) algorithm and cross validation methods. The quantitative prediction models for simultaneous determination of PDG and GPA in Eucommia ulmoides showed good predictive ability. The correlation coefficients (R(2)) of the two calibration models were 0.961 5, 0.958 3 while the roots mean square of cross-validation (RMSECV) were 0.001 5, 0.006 4, respectively. The quantitative prediction models proved that near infrared spectra method used for the quantitative analysis of PDG and GPA in Eucommia ulmoides owned high prediction accuracy and can meet the precision need of rapid determinations of PDG and GPA in Eucommia ulmoides in reality so t it provides a new method to realize the real time on line of quality control of Eucommia ulmoides.

Topics: Calibration; Eucommiaceae; Iridoid Glucosides; Least-Squares Analysis; Lignans; Spectroscopy, Near-Infrared

Phomopsis sp. XP-8 is an endophytic fungus with the ability to produce pinoresinol diglucoside (PDG) in vitro and thus has potential application in biosynthesis of PDG independent of plants. In order to enhance the production of PDG, 18 different natural materials were tested in solid-state cultivation of Phomopsis sp. XP-8.. Most of the tested natural materials promoted the production of PDG. A supplement derived from mung beans produced the highest PDG yield and better fungal growth than the other materials. Also, pinoresinol monoglucoside, pinoresinol and other substrates (phenylalanine, p-coumaric acid, cinnamic acid, caffeic acid, and ferulic acid) were obtained after fermentation on mung beans. Furthermore, PDG production was much higher when mung beans were incorporated into solid state agar versus a liquid medium. The highest pinoresinol diglucoside production (72.1 mg kg(-1) in fresh culture) was obtained in 9 days using a solid state culture of Phomopsis sp. XP-8 on a mung bean grain medium containing 100 g kg(-1) glucose. Mung bean water-soluble polysaccharide was identified as a major promoter of PDG production by Phomopsis sp. XP-8.. Mung bean, especially its water-soluble polysaccharide fraction, was an efficient natural material to promote PDG production by Phomopsis sp. XP-8. © 2015 Society of Chemical Industry.

Topics: Ascomycota; Culture Media; Food Microbiology; Humans; Lignans; Vigna

Phomopsis sp. XP-8 is an endophytic fungus that has the ability to produce pinoresinol diglucoside (PDG) in vitro and thus has potential application for the biosynthesis of PDG independent of plants. When cultivated in mung bean medium, PDG production was significantly improved and pinoresinol monoglucoside (PMG) and pinoresinol (Pin) were also found in the culture medium. In this experiment, starch, protein, and polysaccharides were isolated from mung beans and separately used as the sole substrate in order to explore the mechanism of fermentation and identify the major substrates that attributed to the biotransformation of PDG, PMG, and Pin. The production of PDG, PMG, and Pin was monitored using high-performance liquid chromatography (HPLC) and confirmed using HPLC-MS. Activities of related enzymes, including phenylalanine ammonia-lyase (PAL), trans-cinnamate 4-hydroxylase (C4H), and 4-coumarate-CoA ligase (4CL) were analyzed and tracked during the cultivation. The reaction system contained the compounds isolated from mung bean in the designed amount. Accumulation of phenylalanine, cinnamic acid, p-coumaric acid, PDG, PMG, and Pin and the activities of PAL, C4H, and 4CL were measured during the bioconversion. PMG was found only when mung bean polysaccharide was analyzed, while production of PDG and Pin were found when both polysaccharide and starch were analyzed. After examining the monosaccharide composition of the mung bean polysaccharide and the effect of the different monosaccharides had on the production of PMG, PDG, and Pin, galactose in mung bean polysaccharide proved to be the major factor that stimulates the production of PMG.

Topics: Ascomycota; Biotransformation; Chromatography, High Pressure Liquid; Fabaceae; Furans; Glycosides; Lignans; Mass Spectrometry; Plant Extracts; Plant Proteins; Starch

Pinoresinol diglucoside (PDG) and pinoresinol (Pin) are normally produced by plant cells via the phenylpropanoid pathway. This study reveals the existence of a related pathway in Phomopsis sp. XP-8, a PDG-producing fungal strain isolated from the bark of the Tu-chung tree (Eucommiaulmoides Oliv.). After addition of 0.15 g/L glucose to Phomopsis sp. XP-8, PDG and Pin formed when phenylalanine, tyrosine, leucine, cinnamic acid, and p-coumaric acid were used as the substrates respectively. No PDG formed in the absence of glucose, but Pin was detected after addition of all these substrates except leucine. In all systems in the presence of glucose, production of PDG and/or Pin and the accumulation of phenylalanine, cinnamic acid, or p-coumaric acid correlated directly with added substrate in a time- and substrate concentration- dependent manner. After analysis of products produced after addition of each substrate, the mass flow sequence for PDG and Pin biosynthesis was defined as: glucose to phenylalanine, phenylalanine to cinnamic acid, then to p-coumaric acid, and finally to Pin or PDG. During the bioconversion, the activities of four key enzymes in the phenylpropanoid pathway were also determined and correlated with accumulation of their corresponding products. PDG production by Phomopsis sp. exhibits greater efficiency and cost effectiveness than the currently-used plant-based system and will pave the way for large scale production of PDG and/or Pin for medical applications.

Topics: Amino Acids; Ascomycota; Furans; Glucose; Lignans; Propionates

Pinoresinol diglucoside (PD), a typical marker compound in Ecommia ulmoides Oliv., is an important and natural antihypertensive drug. A selective, sensitive, and rapid liquid chromatography tandem mass spectrometric (LC-MS/MS) analytical method was developed for the determination of PD in rats. After simple protein precipitation with acetonitrile, chromatographic separation of PD was conducted using a reversed-phase ZORBAX SB C18 analytical column (4.6mm × 150 mm, 5 μm particles) with a mobile phase of 10mM ammonium acetate-methanol-acetic acid (50:50:0.15, v/v/v) and quantified by selected reaction monitoring mode under positive electrospray ionization condition. The chromatographic run time was 3.4 min for each sample, in which the retention times of PD and the internal standard were 2.87 and 2.65 min, respectively. The calibration curves were linear over the range of 1.00~3000 ng/mL and the lower limit of quantification was 1.00 ng/mL in rat plasma. The precision expressed by relative standard deviations were <8.9% for intra-batch precision and <2.0% for inter-batch precision, and the intra- and inter-batch accuracy by relative error was within the range of -3.9% ~7.3%, which met acceptable criteria. The LC-MS/MS method was successfully applied to investigate the pharmacokinetics and oral bioavailability of PD in rats, with the bioavailability being only 2.5%.

Topics: Administration, Oral; Animals; Antihypertensive Agents; Biological Availability; Calibration; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Drug Stability; Injections, Intravenous; Lignans; Linear Models; Male; Rats, Wistar; Reference Standards; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

To establish and apply a new LC/MS/MS method for the simultaneous, quantitative determination of six ingredients, aucubin (AU), geniposide (GP), geniposidic acid (GPA), pinoresinol diglucoside (PDG), secologanin (SLG), and loganin (LG) in single and combined extracts of Eucommia ulmoides and Dipsacus asperoides.. Using the LC/MS/MS-ESI(-)-MRM mode to detect the six compounds, chromatographic separation was achieved on an Agilent Eclipse plus C18 column, and the mobile phase consisted of solvent A (CH3CN) and solvent B (H2O containing 0.01% CH3COOH V/V).. This method was successfully applied to quantify the six compounds in rat plasma after oral administration, and showed good precision, accuracy, reproducibility, and linear regression (r(2)>0.99).. The results showed that following the use of the two medicinal plants, for AU and GP, the values of Cmax markedly increased, and the values of cmax markedly decreased. It was found that the compatibility of the medicinal plants might affect their pharmacokinetic properties of their constituents.

Topics: Animals; Chromatography, High Pressure Liquid; Dipsacaceae; Drug Combinations; Eucommiaceae; Female; Iridoid Glucosides; Iridoids; Lignans; Plant Extracts; Rats, Sprague-Dawley; Tandem Mass Spectrometry

To study the ecological suitability regionalization of Eucommia ulmoides, for selecting artificial planting base and high-quality industrial raw material purchase area of the herb in Guizhou.. Based on the investigation of 14 Eucommia ulmoides producing areas, pinoresinol diglucoside content and ecological factors were obtained. Using spatial analysis method to carry on ecological suitability regionalization. Meanwhile, combining pinoresinol diglucoside content, the correlation of major active components and environmental factors were analyzed by statistical analysis.. The most suitability planting area of Eucommia ulmoides was the northwest of Guizhou. The distribution of Eucommia ulmoides was mainly affected by the type and pH value of soil, and monthly precipitation. The spatial structure of major active components in Eucommia ulmoides were randomly distributed in global space, but had only one aggregation point which had a high positive correlation in local space. The major active components of Eucommia ulmoides had no correlation with altitude, longitude or latitude.. Using the spatial analysis method and statistical analysis method, based on environmental factor and pinoresinol diglucoside content, the ecological suitability regionalization of Eucommia ulmoides can provide reference for the selection of suitable planting area, artificial planting base and directing production layout.

Topics: Adaptation, Biological; China; Climate; Ecology; Eucommiaceae; Geographic Information Systems; Geography, Medical; Lignans; Models, Theoretical; Plant Bark; Plants, Medicinal; Soil

Pinoresinol diglucoside (PDG) is the important antihypertensive compound in Eucommia ulmoides Oliv., a traditional Chinese herb medicine. The research objective was to certify the possibility of producing PDG through fermentation. PDG-producing endophytic fungi were isolated from E. ulmoides Oliv., and the highest PDG-yielding (11.65 mg/L) isolate, XP-8, was identified as Phomopsis sp. according to the morphological characteristics and the phylogenetic tree constructed on the basis of the gene sequence in the internal transcribed spacers district. The microbial PDG was isolated by using S-8 resin and purified to a purity of 98.7% using preparative high-performance liquid chromatography (HPLC). Information obtained from the UV spectrum (277 and 227 nm, in water solution), infra-red spectrum (3,428; 2,930; 2,877; 1,637; 1,600; and 1,513; 1,460; 1,421; 1,269; 1,223; 1,075; 658 cm(-1), in powder), molecular weight (682 Da, measured using HPLC-electrospray ionization mass spectrometry (ESI/MS) and tandem mass spectrometry), and nuclear magnetic resonance analysis show the microbial PDG is (+)-1-pinoresinol 4,4'-di-O-β-D-glucopyranoside, same as the plant-derived PDG. The microbial PDG is stable in pH range from 3 to 11 but less stable at temperature higher than 90 °C and in light exposure. During the fermentation, PDG production outside cells starts at the later stage of cell growth when the residual sugar in the medium was low. The study reveals the possibility for production of PDG by fermentation.

Topics: Ascomycota; Chromatography, High Pressure Liquid; DNA, Fungal; DNA, Ribosomal Spacer; Endophytes; Eucommiaceae; Fermentation; Lignans; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Molecular Weight; Phylogeny; Sequence Analysis, DNA; Spectrometry, Mass, Electrospray Ionization; Spectrophotometry, Infrared; Spectrophotometry, Ultraviolet

To establish the quality criteria of decoction pieces of salt Eucommia.. Decoction pieces of salt Eucommia were measured with moisture, total ash and alcohol-extract, and were analyzed by TLC and HPLC for research.. We obtained 13 batches of decoction pieces of salt Eucommia moisture, total ash and alcohol-extract measurements; Meanwhile we set the TLC and HPLC method.. By studying the development of the decoction pieces of salt Eucommia, we established the quality control standards.

Topics: Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Drugs, Chinese Herbal; Eucommiaceae; Lignans; Methanol; Plant Bark; Plants, Medicinal; Quality Control; Reproducibility of Results; Sodium Chloride; Solvents; Technology, Pharmaceutical; Water

(+)-Pinoresinol 4,4'-di-O-beta-D-glucopyranoside ((+)-PDG) is one of the major lignans with various pharmacological activities which could be isolated from Duzhong and other plant species. In this study, a diastereomeric impurity, (-)-pinoresinol 4,4'-di-O-beta-D-glucopyranoside ((-)-PDG), the main impurity was identified in (+)-PDG chemical reference substance (CRS) and a reliable chromatographic method for rapid purity determination of (+)-PDG CRS was firstly developed. The optimal chromatographic condition was found to be using ACN/1,4-dioxane-water (2.5:6:91.5, v/v/v) as mobile phase on a Waters Acquity UPLC HSS T3 column (2.1 mm x 100 mm, 1.8 microm) with column temperature of 37 degrees C. The method was validated and applied to determine the chromatographic purity of five (+)-PDG CRS samples. The content of (-)-PDG in four commercial (+)-PDG CRS was 8.47-20.30%, whereas no (-)-PDG was detected in our in-house prepared (+)-PDG CRS in which purity was confirmed to be 99.80%. The above results confirmed that this method is fast and highly efficient for purity determination of the (+)-PDG CRS.

Topics: Chromatography, High Pressure Liquid; Drug Contamination; Lignans; Molecular Structure; Stereoisomerism

A new ultra performance liquid chromatographic (UPLC) method was established for the determination of pinoresinol diglucoside (PDG) in Qing' e Pills. After extracted by the Soxhlet's method, the methanol extracts of the samples were passed through a Waters Oasis HLB SPE column to achieve good chromatographic performance. The separation was performed on a Waters Acquity C18 BEH column (100 mm x 1.0 mm, 1.7 microm) with acetonitrile-water (the pH adjusted to 4.0 with phosphoric acid) (9: 91, v/v) as the mobile phase at a flow rate of 0.1 mL/min. The detection wavelength was set at 227 nm, the column temperature was 25 degrees C and the injection volume was 0.5 microL. Under the optimized conditions, there was good linear relationship between the mass concentration and the peak area of PDG in the range of 1.40 - 506.00 mg/L with the correlation coefficient of 1. The average recoveries of PDG at three levels ranged from 100.10% to 102.37%. The method is accurate, sensitive, highly reproducible and suitable for the quality control of Qing' e Pills.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Lignans; Sensitivity and Specificity; Tablets

To establish a rapid determination method of pinoresinol diglucoside in Eucommiae unloads by near-infrared reflectance spectroscopy (NIRS).. Forty-one samples of E. unloads were collected from three different producing areas and their main component, namely pinoresinol diglucoside, was determined by HPLC. Corresponding data of samples were collected from 12 000 to 4 000 cm(-1) by near-infrared reflectance spectroscopy. The spectral pretreatment was optimized by OPUS software and the calibration equations between the content of pinoresinol diglucoside and spectrum data were constructed by partial least squares regression.. Available information could be extracted from spectra in the range from 7 502 to 4 597.6 cm(-1) after corrected by applying second derivative transformation and subtract a linear correction. Cross validation was used to prevent over-fitting. Good correlation existed between pinoresinol diglucoside content and NIR spectra ( R2 = 0.926 4, SEC = 0.029 and SEP = 0.066 2).. NIRS calibration equations developed in this study could be applied to the rapid analysis of the pinoresinol diglucoside content.

Topics: Eucommiaceae; Lignans; Spectrophotometry, Infrared; Time Factors

To study the quality changes in pre- and post-processed pieces of Eucommia ulmoides Oliv.. The changes of the content of pinoresinol diglucoside, extract and fingerprint were studied.. Pinoresinol diglucoside contents in post-processed pieces were lower than those in pre-processed and alcohol extract had different changes because of its different habitats. Eucommia ulmoides consists of 11 common peaks, the one processed by salt-water consists of 8 Peaks.. Processing can reduce the content of pinoresinol diglucoside. Alcohol extract has different changes. Eucommia ulmoides common peaks of its fingerprint reduce and mostly components descend after processed by salt-water.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Ethanol; Eucommiaceae; Lignans; Plant Bark; Plants, Medicinal; Sodium Chloride; Technology, Pharmaceutical; Time Factors; Water

To evaluate the reborn barks quality, antihypertensive effective component of pinoresnol diglucoside (PDG) in three times reborn barks were determined.. A YWG C18 column (10 microm, 250 x 4.6 mm) was used with a mobile phase of methanol-water (30:75) and flow rate of 1.0 (ml/min). The detective wave-length was set at 277nm and the column temperature at room temperature.. PDG in the first reborn barks are slightly higher than the original ones, and in the second reborn barks are similar with the barks before girdling (the fist reborn barks), but in the third reborn barks, PDG are much lower than the barks before girdling (the second reborn barks).. In order to ensure reborn barks quality, we suggest that the girdling bark regeneration can be made two times, the time between the first and the second girdling is not less than five years. PDG in the third reborn barks should be enhanced.

Topics: Chromatography, High Pressure Liquid; Conservation of Natural Resources; Drugs, Chinese Herbal; Eucommiaceae; Lignans; Plant Bark; Plants, Medicinal; Regeneration; Time Factors; Ultrasonics

By anaerobic incubation of pinoresinol diglucoside (1) from the bark of Eucommia ulmoides with a fecal suspension of humans, eleven metabolites were formed, and their structures were identified as (+)-pinoresinol (2), (+)-lariciresinol (3), 3'-demethyl-(+)-lariciresinol (4), (-)-secoisolariciresinol (5), (-)-3-(3", 4"-dihydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butane-1, 4-diol (6), 2-(3', 4'-dihydroxybenzyl)-3-(3", 4"-dihydroxybenzyl)butane-1, 4-diol (7), 3-(3"-hydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butane-1, 4-diol (8), 2-(3', 4'-dihydroxybenzyl)-3-(3"-hydroxybenzyl)butane-1, 4-diol (9), (-)-enterodiol (10), (-)-(2R, 3R)-3-(3", 4"-dihydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butyrolactone (11), (-)-(2R, 3R)-2-(3', 4'-dihydroxybenzyl)-3-(3", 4"-dihydroxybenzyl)butyrolactone (12), (-)-(2R, 3R)-3-(3"-hydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butyrolactone (13), 2-(3', 4'-dihydroxybenzyl)-3-(3"-hydroxybenzyl)butyrolactone (14), 2-(3'-hydroxybenzyl)-3-(3", 4"-dihydroxybenzyl)butyrolactone (15) and (-)-(2R, 3R)-enterolactone (16) by various spectroscopic means, including two dimensional (2D)-NMR, mass spectrometry and circular dichroism. A possible metabolic pathway was proposed on the basis of their structures and time course experiments monitored by thin-layer chromatography. Furthermore, a bacterial strain responsible for the transformation of (+)-pinoresinol to (+)-lariciresinol was isolated from a human fecal suspension and identified as Enterococcus faecalis strain PDG-1.

Topics: Anticarcinogenic Agents; Biotransformation; Chromatography, High Pressure Liquid; Circular Dichroism; Dealkylation; Enterococcus faecalis; Feces; Furans; Humans; Intestines; Lignans; Magnetic Resonance Spectroscopy; Mass Spectrometry; Plant Bark; Plants, Medicinal; Spectrophotometry, Ultraviolet

A reversed-phase high performance liquid chromatographic method for the determination of (+)-pinoresinol-di-O-beta-D-glucopyranoside in the bark of Eucommia Ulmoides was established. The air-dried bark of E. Ulmoides was ground into powder and extracted continuously with methanol. The extract was evaporated under reduced pressure, then suspended in water. This suspension being subjected to column chromatography was eluted successively with water, 30%, 50% and 90% aqueous solution of ethanol. The 30% EtOH eluate was concentrated and then injected into HPLC system. The conditions of HPLC were: YWG-C18 column (250 mm x 4.6 mm i.d. 10 microns); 28%(V/V) methanol mobile phase and UV detector at 232 nm. The calibration curve was linear in the range of 0.068 microgram-0.68 microgram of(+)-pinoresinol-di-O-beta-D-glucopyranoside with correlation coefficient of 0.9999. The coefficient of variation and average recovery were 0.50%-0.74% (n = 5) and 99.22% (n = 3) respectively.

Topics: Antihypertensive Agents; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Furans; Lignans

The contents of pinoresinol diglucoside in different processed products of Cortex Eucommiae were determined by HPLC. The result has shown that the contents of pinoresinol diglucoside in post-processed drugs are higher than those in pre-processed ones.

Topics: Antihypertensive Agents; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Furans; Hot Temperature; Lignans; Plants, Medicinal; Technology, Pharmaceutical; Trees

Three new phenolic compounds were isolated from methanol extracts of the leaves of Daphne pseudo-mezereum (Thymelaeaceae) and characterized as (-)-pinoresinol di-O-glucoside (1), 5-hydroxy-7-methoxy coumarin 8-O-beta-D-glucoside (2), diosmetin 7-O-beta-D-xylopyranosyl (1-->6) beta-D-glucopyranoside (3), respectively, together with six known compounds, using 1H- and 13C-NMR spectra.

Topics: Coumarins; Flavonoids; Furans; Lignans; Magnetic Resonance Spectroscopy; Phenols; Plant Extracts

Topics: Antihypertensive Agents; Chemical Phenomena; Chemistry; Chromatography, High Pressure Liquid; Furans; Lignans; Medicine, Chinese Traditional; Plants, Medicinal

Topics: Antihypertensive Agents; China; Furans; Glucosides; Glycosides; Lignans; Medicine, Chinese Traditional; Plants, Medicinal