Junbo Xie

Simultaneous determination of jujuboside A, B and betulinic acid in semen Ziziphi spinosae by high performance liquid chromatography-evaporative light scattering detection

A reverse phase high performance liquid chromatographic (HPLC) method with evaporative light scattering detection (ELSD) was developed for simultaneous determination of jujuboside A, B and betulinic acid in semen Ziziphi spinosae. The analysis was performed by gradient elution, using an aqueous mobile phase (containing 0.1% acetic acid) modified by acetonitrile. The evaporator tube temperature of ELSD was set at 45 degrees C, and with the nebulizing gas flow-rate of 1.8l/min. The method was validated for accuracy, reproducibility, precision and limits of detection and quantification. Quantification of the three active compounds in semen Ziziphi spinosae from different locations was performed by this method, which provides a new tool for quality assessment of semen Ziziphi spinosae.

Hplc-ESI-MS/MS analysis of the water-soluble extract from Ziziphi spinosae semen and its ameliorating effect of learning and memory performance in mice

Background: Ziziphi Spinosae Semen (ZSS), the seed of Ziziphus jujuba var. spinosa (Bunge) Hu ex H. F. Chow., is a traditional herb for insomnia and anxiety in eastern Asia. However, few researches have been concerned with its effect on ameliorating memory and learning performance. Objective: To investigate the constituents of ZSS water soluble extract and its ameliorating learning and memory in mice. Materials and Methods: A new high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-ESI-MS/MS) method was developed and validated to determine the main constituents in the extract. The effect of ZSS water soluble extract on memory and learning performance was investigated in mice by Y-maze and passive avoidance test. Results: The extract could significantly decrease the number of errors (NOE), and increase the transfer latency time (TLT) and electrical stimuli time (EST). In addition, spinosin, jujuboside A (JuA) and jujuboside B (JuB) were simultaneously identified and quantified in the extract. Their contents in the extract were as followed: Spinosin (223.51mg/g), JuA (63.76mg/g) and JuB (26.29mg/g). Conclusion: The extract played a promising role in ameliorating memory in mice with alcohol induced memory retrieval disorders, and might help to improve learning capacity to some extent. Spinosin, JuA and JuB were the predominant constituents, which might be mainly responsible for the definite activity.

Composition of fatty oils from Semen Ziziphi Spinosae and its cardiotonic effect on isolated toad hearts

In this study, the composition of fatty oil from Semen Ziziphi Spinosae and its cardiotonic activity on the heart isolated from a toad were studied. Oil-in-water (O/W) emulsions of fatty oil were prepared by the perfusion method. The fatty oil had a positive inotropic effect on isolated rat hearts at a concentration between 5 × 10−3 and 2 × 10−2 mL/10 mL, and the effect was in positive correlation with the concentration of calcium ions. In addition, this effect was inhibited by 2 mg/mL nifedipine, suggesting that the cardiotonic mechanism could be responsible for accelerating the inflow of calcium ions. Gas chromatography–mass spectrometry analysis showed that the main constituents of the fatty oil were 9-octadecenoic acid (43.32%), 9,12-octadecadienoic acid (42.57%), hexadecanoic acid (4.76%), 9-eicosenoic acid (2.95%), stearic acid (2.41%) and arachidic acid (0.81%). This preliminary study revealed that the fatty oil of Semen Ziziphi Spinosae exhibited remarkable cardiotonic activity in the tested models, and it is necessary to further reveal the effective substances of the fatty oil.

Brain Tissue Distribution of Spinosin in Rats Determined by a New High-Performance Liquid Chromatography- Electrospray Ionization -Mass/Mass Spectrometry Method

Spinosin, a flavone-C-glycoside, is a bioactive ingredient isolated from a traditional Chinese herb Zizyphi Spinosi Semen. In this study, a new high-performance liquid chromatography-electrospray ionization-mass/mass spectrometry method was developed and validated to determine spinosin in brain tissues including olfactory region, hippocampus, corpus striatum, cerebrum (cerebral cortex) and cerebellum, after intravenous administration with the dose of 5 mg/kg. The tissue homogenate samples were pretreated and extracted with acetonitrile by a simple protein precipitation method. The separation was performed on a YMC ODS-AQ(TM) column (250 × 2.0 mm, 3.5 μm) with the mobile phase of acetonitrile-aqueous phase (0.1% formic acid) (25 : 75, v/v) at a flow rate of 0.3 mL/min. The retention times of spinosin and naringin (internal standard) were 3.3 and 5.1 min, respectively. Multiple reaction monitoring mode was used to monitor precursor/product ion transitions of m/z 607.2 → 427.0 for spinosin and m/z 579.2 → 271.0 for naringin. The proposed method was successfully applied to the preclinical brain tissue distribution of spinosin in rats. The results showed that there was a wide brain regional tissue distribution of spinosin. The concentrations of spinosin in corpus striatum and hippocampus were higher than that in other areas.

Tissue Distribution of Jujuboside A in Sprague-Dawley Rats Determined by an Efficient HPLC–ESI-MS/MS Method

Jujuboside A (JuA), a dammarane-type triterpene glycoside, is a main bioactive saponin in Zizyphi Spinosi Semen (a traditional Chinese herbal food). In this research, the distribution of JuA in Sprague-Dawley rats was investigated with a new and efficient HPLC–ESI-MS/MS method. The results showed that JuA distributed rapidly and widely in various rat tissues (including heart, liver, spleen, kidney, and lung) after intravenous administration. In addition, it was initially found that JuA could pass through the blood-brain barrier and reach various areas of the brain quickly. At 0.25 hr, the concentration of JuA in the hippocampus (204.10 ng · g−1) was significantly higher than in the cerebrum (144.27 ng · g−1), olfactory bulb (98.42 ng · g−1), and corpus striatum (76.04 ng · g−1) (p < 0.05), indicating that the hippocampus was the main accumulation area of JuA in the brain.

Degradation Kinetics of Jujuboside a by Rat Intestinal Flora and Identification of the Metabolites by HPLC-Ms/Ms

Jujuboside A (JuA) is a representative saponin with significant sedative and hypnotic effects. Up to now, there were many arguments on its metabolic mechanism. In this study, a high performance liquid chromatography-tandem mass spectrometry (MS/MS) method was developed for investigating the degradation characteristics of JuA incubated with rat feces. With the method, the degradation kinetics of JuA was studied. The results showed JuA decomposed rapidly. It could decompose nearly 100% in only 4–6 h. The degradation regularity was consistent with the first dynamic process. In addition, seven metabolites were determined and identified to be the serial hydrolysis products of JuA. The results revealed that gastrointestinal microbiota played an indispensable role in the metabolic process of JuA. JuA may be a supplier of sugars under the hydrolysis effect induced by the bacterial enzymes. At the same time, its aglycone was produced as a by-product, which could be easier to be absorbed into the body to perform its specific bioactivities.

Degradation Kinetics of Jujuboside B by Rat Intestinal Flora In vitro with an RRLC-MS-MS Method

Jujuboside B (JuB) is a main bioactive saponin constituent of Ziziphi Spinosae Semen, which is a traditional herb for the treatment of insomnia and anxiety. However, the detailed metabolic mechanism of JuB is poorly understood. In this study, a novel method of rapid resolution liquid chromatography-triple quadrupole mass spectrometry was developed and validated for the analysis of JuB. With the method, the degradation kinetics of JuB by rat intestinal flora in vitro was investigated. The analysis was performed with an Agilent Eclipse Plus C18 (2.1 mm × 50 mm, 1.8 μm) column and an aqueous mobile phase (containing 0.1% formic acid) modified by methanol. The analyte was measured by multiple reaction-monitoring (MRM) modes with m/z 1043.3 → m/z 749.2. This method was validated with perfect accuracy, precision and limit of quantitation. It showed that jujuboside B (JuB) degradation started slowly as incubation with rat feces. The rate constant was correlated greatly with the concentration of sample solutions. Furthermore, some metabolites were elucidated with their chromatographic behavior and typical fragment ions. The results might help better interpret the metabolic and pharmacological mechanism of JuB.

A galactomannoglucan derived from Agaricus brasiliensis: Purification, characterization and macrophage activation via MAPK and IκB/NFκB pathways

In this study, a novel galactomannoglucan named as TJ2 was isolated from Agaricus brasiliensis with microwave extraction, macroporous resin, ion exchange resin and high resolution gel chromatography. TJ2 is composed of glucose, mannose and galactose in the ratio 99.2:0.2:0.6. Infrared spectra (IR), methylation analysis and nuclear magnetic resonance spectra indicated that TJ2 mainly contained a β-(1→3) - linked glucopyranosyl backbone. Interestingly, TJ2 significantly promoted RAW264.7 cell proliferation, and was able to activate the cells to engulf E. coli. In addition, TJ2 induced the expression of Interleukin 1β (IL-1β), Interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) and cyclooxygenase-2 (Cox-2) in the cells. TJ2 also promoted the production of nitric oxide (NO) by inducing the expression of inducible nitric oxide synthase (iNOS). Moreover, TJ2 is a potent inducer in activating the mitogen-activated protein kinase (MAPK) and inhibitor of nuclear factor-kappa B (IκB)/nuclear factor-kappa B (NFκB) pathways.

Royal Sun Medicinal Mushroom, Agaricus brasiliensis (Agaricomycetidae), Derived Polysaccharides Exert Immunomodulatory Activities In Vitro and In Vivo

The royal sun mushroom, Agaricus brasiliensis is a widely consumed mushroom around the world. In this study, the immunoregulatory potential of A. brasiliensis polysaccharides was investigated in vitro and in vivo. In vivo, the polysaccharides remarkably increased the spleen and thymus indexes in mice, and this effect was influenced significantly by age (the adult and the juvenile). The spleen index increased by 27.28% in adult mice treated with the polysaccharides, whereas the increase in juvenile mice was just 12.59% at the dose of 150 mg·kg-1·d-1. Moreover, the effect of the polysaccharides on the thymus and spleen indexes in adult mice was obvious both in males and females. The carbon clearance ability (phagocytic index) was improved with increasing doses, (32.81% at 120 mg·kg-1·d-1, and 38.34% at 150 mg·kg-1·d-1) in mice treated with the polysaccharides. In vitro, the polysaccharides increased the RAW264.7 cell proliferation with 34.78% at 25 µg/mL and 26.78% at 50 µg/mL. Furthermore, the polysaccharides also promoted mRNA expressions of interleukin (IL)-6, IL-1β, cyclooxygenase-2, and Toll-like receptor 4 (TLR4), myeloid differentiation 88 (MYD88), and TIR-domain-containing adapter-inducing interferon-β (TRIF) in the cells, indicating that the polysaccharides induce the secretion of inflammatory cytokines by stimulating TLR4/MyD88 and TLR4/TRIF pathways. In conclusion, these results suggest that A. brasiliensis polysaccharides induce a very promising immunostimulation effect in vivo and in vitro. Therefore, it should be explored as a novel natural functional food additive.

The pharmacokinetics and tissue distribution of coumaroylspinosin in rat: A novel flavone C-glycoside derived from Zizyphi Spinosi Semen

Zizyphi Spinosi Semen (ZSS) has a long history of sedative-hypnotic use in China. As a novel flavone C-glycoside, coumaroylspinosin is a main flavonoid only found in ZSS. Up to now, its pharmacokinetic information and tissue distribution in vivo are not available yet. With a simple, rapid and sensitive HPLC-MS/MS method, the pharmacokinetics and tissue distribution of coumaroylspinosin were investigated in Sprague-Dawley (SD) rats after its intravenous administration. Puerarin was used as the internal standard (IS). The samples were extracted by a simple protein precipitation method with methanol. The MS analysis was performed with multiple reaction monitoring (MRM), and the transitions were set at m/z 753.3→427.0 for coumaroylspinosin and m/z 415.3→295.3 for IS, respectively. The method was successfully applied for investigating the pharmacokinetics and tissue distribution of coumaroylspinosin in Sprague Dawley (SD) rats after tail vein injection with 4.0mg/kg of the flavonoid. The calibration curves covered over the range of 0.02-10μg/mL in plasma and various tissues samples with good linearity(r≥0.9956). The lower limit of quantification (LLOQ) in all samples was less than 20ng/mL. The intra- and inter-day precisions were below 15% and accuracy was from -3.78% to 4.68%. No significant matrix effect was observed, and the average extraction recovery was acceptable. Coumaroylspinosin could be cleared quickly from the rat plasma with the half-life (t1/2) of 1.86±0.15h. It was distribute widely in vivo, and the main tissue depots of coumaroylspinosin in rats were found to be intestine, muscle and lung. With the method, the pharmacokinetic parameters and tissue distribution of coumaroylspinosin in SD rats were investigated for the first time. The results demonstrated that coumaroylspinosin was distributed widely and rapidly in various rat tissues after intravenous administration.