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Aim: To evaluate the association between genetic polymorphisms and plasma concentration-to-dose ratio of valproic acid (CDRV) in Chinese epileptic patients. Methods: A total of 46 epileptic patients treated with valproic acid therapy were enrolled. 18 SNPs in nine genes related to valproic acid were directly sequenced with Sanger methods. Results: Patients carrying UGT1A6 heterozygous genotypes had significantly lower CDRV than those carrying the wild-type genotypes. In contrast, patients with the homozygote genotypes of CYP2C9 and ABAT had higher CDRV than those with the wild-type genotypes and patients with the heterozygous genotypes of CYP2C19 had higher CDRV. Conclusion: Detection of genetic polymorphism in these genes might facilitate an appropriate dose of valproic acid for epileptic patients. Further studies with larger cohorts are necessary to underpin these observations.

We aim to establish a simple and easy high-performance liquid chromatography system coupled with an ultraviolet detector suitable for simultaneous determination of 24 antiepileptic drugs in human plasma. Optimized chromatographic separation was performed on a ZORBAX Eclipse Plus-C18 (4.6 × 150 mm

Gut microbiota plays a key role in the pathophysiology of type 2 diabetes mellitus (T2D). Mulberry leaf has a hypoglycemic effect, but the potential mechanism is not fully understood. This study aimed to explore the influences and potential mechanisms of mulberry leaf water extract (MLWE) intervention on mice with T2D induced through a high-fat and high-sucrose diet combined with streptozotocin by the combination of fecal metabolomics and gut microbiota analysis. Results showed that MLWE could decrease fasting blood glucose and body weight while ameliorating lipid profiles, insulin resistance, liver inflammation, and the accumulation of lipid droplets in T2D mice. MLWE could reverse the abundances of the phyla Actinobacteria and Bacteroidetes and the ratio of Firmicutes/Bacteroidetes, and increase the abundances of the phyla Cyanobacteria and Epsilonbacteraeota in the feces of T2D mice. The abundances of genera Alloprevotella, Parabacteroides, Muribaculaceae, and Romboutsia in the feces of T2D mice could be reversed, while Oscillatoriales_cyanobacterium and Gastranaerophilales could be reinforced by MLWE supplementation. The levels of nine metabolites in the feces of T2D mice were improved, among which glycine, Phe-Pro, urocanic acid, phylloquinone, and lactate were correlated with Romboutsia and Gastranaerophilales. Taken together, we conclude that MLWE can effectively alleviate T2D by mediating the host-microbial metabolic axis.

A dual-template magnetic molecularly imprinted polymer (Dt-MMIP) with a specific recognition capability for carbamazepine (CBZ) and lamotrigine (LTG) was synthesized using methacrylic acid as a functional monomer, and ethylene glycol dimethylmethacrylate as a cross-linking agent. A magnetic non-molecularly imprinted polymer without templates (MNIP) was also prepared using the same procedure. The prepared polymers were characterized using scanning electron microscopy, Fourier-transform infrared spectroscopy and adsorption experiments. Results indicated that both Dt-MMIPs and MNIPs were microspherical nanoparticles, and the surface of the Dt-MMIP was rougher than that of the MNIP. In addition, the prepared Dt-MMIPs possessed a higher adsorption capacity and better selectivity for CBZ and LTG than the MNIPs. The maximum static adsorption capacities of Dt-MMIP for CBZ and LTG were 249.5 and 647.9 μg g

Lichens are the result of a symbiotic interaction between fungi (mycobionts) and algae (phycobionts). Aside from mycobionts, lichen thalli can also contain non-lichenised fungal species, such as lichenicolous and endolichenic fungi. For this study, three surveys were conducted in China’s Yunnan Province and Inner Mongolia Autonomous Region between 2017 and 2020. Several samples of four lichen species were collected during these surveys: Candelaria fibrosa, Flavoparmelia caperata, Flavopunctelia flaventior and Ramalina sinensis. Six isolates of Coniochaeta were recovered from these four lichen species. The phylogenetic and morphological analyses revealed that two of these isolates were previously identified species, Coniochaeta velutinosa and C. acaciae. Those remaining were from potentially unknown species. We used molecular and morphological data to describe these previously-unknown species as Coniochaeta fibrosaesp. nov., C. mongoliaesp. nov. and C. sinensissp. nov. The findings of this study significantly improve our understanding of the variety and habitat preferences of Coniochaeta in China and globally.

The “shuttle effect” of long-chain polysulfides and the low conductivity of elemental sulfur lead to the inferior cycling stability of lithium–sulfur batteries and imped their practical applications. Herein, Co3O4 nanoflakes with uniform macro pores distribution were synthesized via facile oil bath and calcination methods. Coupled with super P and coated on common polypropylene separators, they were expected to hinder the migration of lithium polysulfides (LiPSs) and accelerate the redox kinetics of polysulfides. Coin cells assembled with the Co3O4–super P interlayer exhibited a capacity of 760 mA h g−1 at 1 C, maintained 598 mA h g−1 after 350 cycles, and the decay rate of discharge capacity was only about 0.062% per cycle. Such high performance can be attributed to the synergistic effects between polar Co3O4 and conductive super P. The facile fabrication method and high performance make the Co3O4-super P interlayer a feasible material to apply in lithium-sulfur batteries.

The research of high-performance flexible supercapacitors is urgent due to the rapid development of wearable and portable electronics. The key challenge is the preparation of flexible electrodes with high areal capacitance since electrodes are the most important part of supercapacitors. Compared to those conventional electrodes loading with typical flexible substrates such as textile, PET, paper et al, free-standing electrodes have many advantages such as more efficient capacity contribution, solidly embedded active materials and thinner thickness. Herein, we have successfully fabricated a novel sandwich-like structure free-standing MoO3-rGO (reduced graphene oxide) composite film electrode for flexible supercapacitors using simple vacuum filtration method followed by HI reduction process. The obtained MoO3-rGO composite film electrode shows excellent electrochemical performance, whose areal specific capacitance reaches 8972 mF·cm-2 (1.5 mA·cm-2). Here, MoO3 provides pseudocapacitance and rGO provides double-layer capacitance. After cycling for 2000 cycles, the capacity retention is 86.7%, showing good cycle stability. Besides, the as-prepared composite film has good flexibility and will not break easily during following bending, rolling, folding or twisting steps. This study has been approved to be an important step for the high-performance electrode design for free-standing flexible supercapacitors.

Oxygen vacancy-enriched MoO3−x nanobelts with oxygen vacancies up to 20% (equal to MoO2.4) were synthesized and demonstrated fast reaction kinetics. An asymmetric supercapacitor was assembled with active carbon (AC) as the anode and the as-prepared MoO3−x nanobelts as the cathode. A new aqueous electrolyte of H2SO4/ethylene glycol (EG) was investigated and found to work appropriately at low temperatures, even at −25 °C. The MoO3−x nanobelts electrode possesses excellent specific capacitance, rate capacity (1230 F g−1 and 1220 F g−1 at 5 A g−1 and 50 A g−1, respectively) and cycle performance (100% after 38 000 cycles). Its energy and power densities reached 111 W h kg−1 and 803 W kg−1 or 50 W h kg−1 and 27 321 W kg−1 and maintained 80 W h kg−1 and 794 W kg−1 or 17 W h kg−1 and 23 565 W kg−1 at −25 °C. It is hypothesized that the coexistence of oxygen vacancies and low valence Mo ions enhanced both mass and charge transport kinetics and catalyzed the redox reactions.

MoO3−x nanobelts with different concentrations of oxygen vacancies were synthesized by a one-step hydrothermal process. XPS test results show that oxygen vacancies are distributed from the exterior to the interior of the MoO3−x nanobelts. As an anode material for lithium-ion batteries, MoO3−x-10 releases excellent rate capacitance. It can maintain a high specific capacitance of about 500 mA h·g−1 at a high current density of 1000 mA·g−1. In the aspect of cycling stability, MoO3−x-10 can retain a high specific capacity of 641 mA h·g−1 after cycling for 50 times at 100 mA·g−1 and 420 mA h·g−1 after cycling for 100 times at 500 mA·g−1. The coexistence of oxygen vacancies and low-valence Mo ions is conducive to the intercalation/de-intercalation of Li ions and to promoting redox reactions. It has been proved to be a significantly effective way in which oxygen vacancies can improve the integrated performance of MoO3−x nanobelts as anode materials.

3D-networked CNT–Ni3S2–CNT-A supported by Ni foam was rationally designed and synthesized via a multistep transformation approach. The intriguing and individual nanostructures endow the hybrid composites with outstanding electrochemical performance with an areal specific capacitance of 13 400 mF cm−2 at the current density of 10 mA cm−2 and an excellent rate capacity of 96.04% at 50 mA cm−2. The corresponding asymmetric supercapacitor assembled with CNT–Ni3S2–CNT-A (positive electrode) and active carbon (negative electrode) delivered a high energy density of 75.2 W h kg−1 at the power density of 2416 W kg−1, an excellent areal capacitance (425 mF cm−2 at 3 mA cm−2) and electrochemical cycling stability (90.6% retention after 10 000 cycles). The dual function of CNTs is responsible for the impressive electrochemical performance; the inner CNT provides more electrodeposition sites for higher capacitance while the outer high electrical conductivity CNT acts as highways for faster electron transportation. The rational design concept of this work is versatile and scalable for combining transition-metal sulfides with carbon materials for high performance supercapacitors.

Three classical Fe-MOFs, viz., MIL-100(Fe), MIL-101(Fe), and MIL-53(Fe), were synthesized to serve as platforms for the investigation of structure-activity relationship and catalytic mechanism in the selective conversion of H

Obesity is a worldwide problem that resulted from the excessive fat accumulation in adipose tissue, leading to the impairment of individual health. Mulberry leaf is an important traditional Chinese medicine and has been used to alleviate obesity for a long term. However, its underlying molecular mechanisms have not been fully elucidated yet.In this study, we aimed to investigate the inhibition effects of mulberry leaf water extract (MLWE) on lipid accumulation during the process of differentiation of 3T3-L1 preadipocytes and development of mature adipocytes through the combination of molecular biology assays and metabolomic analysis.The quality consistency and main chemical ingredients of MLWE were analyzed by high performance liquid chromatography and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), respectively. Oil red O staining was used to mirror lipid accumulation. Lipogenesis-, lipolysis- and inflammation-related genes were evaluated by real-time PCR and western blot, respectively. Untargeted metabolomics were performed by LC-MS/MS.Prepared method and quality of MLWE were stable and reliable. A total of 34 compounds were identified and 14 of them were undoubtedly confirmed. MLWE supplementation could dose-dependently inhibit the aggregation of lipid droplets, and the expressions of sterol regulatory element-binding protein (SREBP)-1c, peroxisome proliferator-activated receptor (PPAR) γ, acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), tumor necrosis factor (TNF)-α and interleukin (IL)-6, and increase the expressions of adenosine monophosphate-activated protein kinase (AMPK), hormone-sensitive lipase (HSL) and IL-10 in the differentiation of preadipocytes. Furthermore, MLWE treatment could dose-dependently decrease the level of triglycerides and the expressions of ACC, FAS, TNF-α, and IL-6, and up-regulate the level of glycerol and the expressions of PPARα, adiponectin (ADPN), adiponectin receptor (AdipoR) 1, AdipoR2, AMPK, HSL, and IL-10 in the development of mature adipocytes. Untargeted metabolomics showed that a total of 5 and 18 differential metabolites were reversed by MLWE intervention in the differentiation of preadipocytes and the development of mature adipocytes, respectively, which involved in the biosynthesis of unsaturated fatty acids, arachidonic acid metabolism and glycerophospholipids metabolism.Taken together, this study firstly verified that MLWE could effectively alleviate lipid accumulation and inflammation by regulating ADPN/AMPK-mediated signaling pathways and relevant metabolic disturbances including biosynthesis of unsaturated fatty acids, arachidonic acid metabolism and glycerophospholipids metabolism.

Classical amino-functionalized Fe-terephthalate metal–organic framework NH2-MIL-53(Fe) and its parent framework MIL-53(Fe) were prepared via simple hydrothermal methods. The catalytic performaces o...

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Ginkgo biloba leaves extract (GBE), one of the most widely used traditional Chinese medicines worldwide, can be used for the treatment of diabetes mellitus (DM). However, its biotransformation in liver is not fully known under the state of DM. In this study, an off-line hydrophilic interaction × reversed-phase two-dimensional liquid chromatography (HILIC × RP 2D-LC) system coupled with diode array detection (DAD) and quadrupole time-of-flight mass spectrometry (q/TOF-MS) was established for the qualification and quantification of the biotransformation of GBE in normal and diabetic rat liver microsomes (RLMs). 6 metabolites were tentatively identified according to the exact molecular weights and the characteristic fragment ions provided by q/TOF-MS data. The results of metabolic stability showed that the metabolic ratio of four target compounds including quercetin, genistein, kaempferol and isorhamnetin in diabetic RLMs were significantly enhanced when comparing with normal RLMs. The results of enzyme kinetics showed that compared with normal RLMs, the Michaelis-Menten constant (Km) value of genistein was obvious increased while its maximal velocity (Vmax) and intrinsic clearance (CLint) values were significantly decreased by diabetic RLMs, and the Vmax and CLint values of kaempferol and isorhamnetin were notably enhanced while their Km values were markedly reduced. For the half-time (t1/2) values of four target compounds and the Km, Vmax and CLint values of quercetin, there were not statistically significant changes between normal and diabetic RLMs. The results suggest that the developed off-line 2D LC-DAD-q/TOF-MS method is an easy and accurate approach for the study of GBE biotransformation in RLMs and may provide the essential data for further pharmacological and clinical studies of GBE.

Classical amino-functionalized Fe-terephthalate metal-organic framework NH

Rat renal tubular epithelial cell (RTEC) cultured with high glucose has been used to observe the protective effect of Ginkgo biloba extract (GBE) against diabetic nephropathy (DN). The compounds in GBE binding with cell membrane or entering into cell are still unknown, which may be potential bioactive components. In this paper, a powerful method for screening and analyzing the potential bioactive components from GBE was developed using cell extraction coupled with high performance liquid chromatography tandem mass spectrometry (LC-MS/MS). 8 prototype compounds and 5 metabolites were obtained, among which 6 prototype compounds and 1 metabolite were identified or tentatively characterized as rutin, bilobalide, ginkgolide B, ginkgolide C, genkwanin, apigenin and diosmetin by comparing their retention times and MS spectra with those of authentic standards or literature data. The 6 prototype compounds were further quantitatively analyzed using electrospray ionization in negative mode multiple reaction monitoring (MRM). The results showed that high glucose changed the Tmax, MRT(0-t), Cmax and AUC(0-t) of all observed compounds and decreased the t1/2 of genkwanin and apigenin, significantly. The overall findings indicate that 8 prototype compounds may be the potential bioactive components of GBE with preventive effect against DN and the method of RTEC extraction coupled with LC-MS/MS technology screening method we developed is a feasible, rapid, and useful tool for screening and analyzing potential bioactive components.

A novel UPLC-DAD method was developed and validated for the simultaneous determination of baicalin (baicalein-7-glucuronide, BG), oroxylin A-7-O-glucuronide (OAG) and wogonoside (WG) in rat plasma using rutin as the internal standard. Plasma samples were precipitated using acetonitrile containing 0.1% formic acid. Separation was performed on an Agilent Eclipse Plus C18 column (2.1 × 50 mm, 1.8 µm) using gradient acetonitrile and 0.2% formic acid water solution as mobile phase. The flow-rate was set at 0.4 mL/min and the eluate was detected at 275 nm. The method was linear over the ranges of 0.075-17.50, 0.050-12.60 and 0.056-14.10 µg/mL for BG, OAG and WG, respectively. The intra- and inter-day precisions were respectively

The chromatographic separation of traditional Chinese medicines is still a highly challenging task in analytical science with respect to its hundreds and thousands of chemical compounds, while increase of separation efficiency can greatly improve the separation power of chromatographic column for traditional Chinese medicine. In this study, 13 bioactive components in HuanglianShangqing pill were selected as an index to optimize the separation conditions and evaluate the system suitability of three commercially available columns packed with 1.8, 3.5, and 5.0 m particles. The chromatographic separations were obtained by the most appropriate Eclipse Plus C18 column (100 × 2.1 mm, 3.5 m) within 45 min using gradient elution with aqueous-ammonium acetate (10 mmol/L, pH 5.0) and acetonitrile, at a flow rate of 0.3 mL/min and an operating temperature of 30C. The quality of HuanglianShangqing pill was assessed through combining simultaneous quantification of 13 compounds with fingerprint analysis. For the qualitative analysis, mass spectrometry was used to confirm the 13 compounds. All the validation data conformed to the acceptable requirements. For the fingerprint analysis, 32 peaks were selected as the common peaks at 254 nm to evaluate the similarities among HuanglianShangqing pills obtained from ten manufacturers.

In this work, two high-performance liquid chromatography (HPLC) assays were developed and validated for the independent determination of edaravone and taurine using 3-methyl-1-p-tolyl-5-pyrazolone and L-glutamine as internal standards. In in vitro experiments, human plasma was separately spiked with a mixture of edaravone and taurine, edaravone or taurine alone. Plasma was precipitated with acetonitrile containing 0.1% formic acid. Ultrafiltration was employed to obtain the unbound ingredients of the two drugs. The factors that might influence the ultrafiltration effiency were elaborately optimized. Plasma supernatant and ultrafiltrate containing taurine were derivated with o-phthalaldehyde and ethanethiol in the presence of 40 mmol/L sodium borate buffer (pH 10.2) at room temperature within 1 min. Chromatographic separations were achieved on an InertSustain C18 column (250 × 4.6 mm, 5 µm). Isocratic 50 mmol/L ammonium acetate-acetonitrile and gradient 50 mmol/L sodium acetate (pH 5.3)-methanol were respectively selected as the mobile phase for the determination of edaravone and taurine. All of the validation data including linearity, extraction recovery, precision, accuracy and stability conformed to the requirements. Results showed that there were no significant alterations in the plasma protein binding rate of taurine and edaravone, implying that the proposed combination therapy was pharmacologically feasible.

Most herbal medicines will be metabolized by intestinal bacteria in the gastrointestinal tract before absorbed by the small intestine. Ginkgo biloba extract (GBE) possesses protective effects on the glomerulosclerosis of diabetic nephropathy (DN), but its biotransformation in diabetes and DN intestinal bacteria has not yet been recognized. In this work, a validated liquid chromatography-tandem mass spectrometry (LC-MS) method was established for the simultaneous quantification of 14 components in GBE in rat intestinal bacteria matrix, namely ginkgolides A, ginkgolides B, ginkgolides C, bilobalide, rutin, myricetin, quercitrin, quercetin, luteolin, genistein, kaempferol, apigenin, isorhamnetin and genkwanin. Chromatographic separation was performed on a Kromasil-C18 (4.6mm×250mm i.d., 5.0μm) analytical column maintained at 35°C. The mobile phase was a mixture of methanol (A) and 0.1% formic acid in water (B) with a step linear gradient at a flow rate of 1.0mlmin(-1). The calibration curves of these 14 analytes demonstrated good linearity within the test range (R>0.99). This validated method has successfully been applied into the pharmacokinetic study of the 14 components. More importantly, in the pharmacokinetic study, by comparing the time course of the biotransformation by normal, diabetes and DN rat intestinal bacteria, we found that the biotransformation speed and residence time of the 14 compounds in diabetes and DN rats differed obviously from that obtained in normal group, which provided valuable chemical information for further pharmacology and active mechanism research on GBE.

A rapid and useful approach for screening potential bioactive components in Ginkgo biloba extract (GBE) with preventive effect against diabetic nephropathy (DN) was developed using mesangial cells extraction coupled with high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. Mesangial cells were first divided into two groups according to their treatments with high glucose or high glucose plus GBE. After incubation for 4, 8, 12, 16, 24 and 48 h, the cells were harvested and extracted with 40% acetic acid in water before LC-MS/MS analysis. Then, 19 compounds and five metabolites were found to selectively combine with mesangial cells. Notably, compounds including quercetin and rutin were identified or tentatively characterized according to the results of retention time and MS spectra, which is highly consistent with our previous reports that quercetin and rutin are potent protective agents against glomerulosclerosis in DN. Therefore, all these results indicate that target cell extraction coupled with LC-MS/MS analysis can be successfully applied for predicting the bioactive components in GBE with preventive effect against DN. Copyright © 2014 John Wiley & Sons, Ltd.

Three liquid chromatography–tandem mass spectrometry (LC-MS/MS) methods were respectively developed and validated for the simultaneous or independent determination of taurine and edaravone in rat plasma using 3-methyl-1-p-tolyl-5-pyrazolone and sulfanilic acid as the internal standards (IS). Chromatographic separations were achieved on an Agilent Zorbax SB-Aq (100 × 2.1 mm, 3.5 µm) column. Gradient 0.03% formic acid–methanol, isocratic 0.1% formic acid–methanol (90:10) and 0.02% formic acid–methanol (40:60) were respectively selected as the mobile phase for the simultaneous determination of two analytes, taurine or edaravone alone. The MS acquisition was performed in multiple reaction monitoring mode with a positive and negative electrospray ionization source. The mass transitions monitored were m/z [M + H]+ 175.1 133.0 and [M + H]+ 189.2 147.0 for edaravone and its IS, m/z [M − H]− 124.1 80.0 and [M − H]− 172.0 80.0 for taurine and its IS, respectively. The validated methods were successfully applied to study the pharmacokinetic interaction of taurine and edaravone in rats after independent intravenous administration and co-administration with a single dose. Our collective results showed that there were no significant alterations on the main pharmacokinetic parameters (area under concentration–time curve, mean residence time, half-life and clearance) of taurine and edaravone, implying that the proposed combination therapy was pharmacologically feasible. Copyright © 2014 John Wiley & Sons, Ltd.

An LC-MS/MS method was developed and validated for the simultaneous quantification of edaravone and taurine in beagle plasma. The plasma sample was deproteinized using acetonitrile containing formic acid. Chromatographic separations were achieved on an Agilent Zorbax SB-Aq (100 × 2.1 mm, 3.5 μm) column, with a gradient of water (containing 0.03% formic acid) and methanol as the mobile phase at a flow rate of 0.3 mL/min. The analyte detection was carried out in multiple reaction monitoring mode and the optimized precursor-to-product transitions of m/z [M+H](+) 175.1 → 133.0 (edaravone), m/z [M+H](+) 189.1 → 147.0 (3-methyl-1-p-tolyl-5-pyrazolone, internal standard, IS), m/z [M-H](-) 124.1→80.0 (taurine), and m/z [M-H](-) 172.0 → 80.0 (sulfanilic acid, IS) were employed to quantify edaravone, taurine, and their corresponding ISs, respectively. The LOD and the lower LOQ were 0.01 and 0.05 μg/mL for edaravone and 0.66 and 2 μg/mL for taurine, respectively. The calibration curves of these two analytes demonstrated good linearity (r ＞ 0.99). All the validation data including the specificity, precision, recovery, and stability conformed to the acceptable requirements. This validated method has successfully been applied in the pharmacokinetic study of edaravone and taurine mixture in beagle dogs.

Steam reforming of acetic acid, one model compounds of bio-oil, was studied on the Ni/ZrO2–CeO2 catalysts which were prepared by the impregnation method. The results showed that high acetic acid conversion and hydrogen yield were obtained in the temperature range of 650–750 °C when H2O/HAC ratio was 3. Nevertheless, the catalyst deactivation was caused by carbon deposition eventually with time-on-stream. In order to discuss the behavior of the carbon deposition on the Ni/ZrO2–CeO2 catalyst during steam reforming of bio-oil, the structure and morphology of carbon deposition were investigated by BET, XRD, TG/DTA, TPR, SEM and EDX techniques. All the experimental results showed acetone and CO were the important carbon precursors of acetic acid reforming and the graphitic-like carbon was the main type of carbon deposition on the surface of the deactivated 12%Ni/CeO2–ZrO2 catalyst.

Hydrogen production from catalytic steam reforming of model compound acetic acid of bio-oil was investigated under atmospheric pressure in a bench-scale fixed-bed reactor and a series of Ni catalysts supported on ceria–zirconia were prepared via co-precipitation with varying contents of Ni and Ce loading. Simultaneously, effects of reaction temperature, and molar ratio of steam to carbon fed, liquid hourly space velocity (LHSV) were also explored. The results indicated that the 12Ni/7.5Ce-Zr-O catalyst was superior to other catalysts with different Ni and Ce loading. At T = 650 °C, S / C = 3, LHSV = 2.8 h −1 , the reaction over Ni catalyst supported on ceria–zirconia turns out that the highest activity and the maximum hydrogen selectivity of 83.4% and the minimum methane selectivity of 0.39% are attained. Besides, Ni catalyst supported on ceria–zirconia shows good stability during 25 h continuous reaction without any deactivation.

The quantitative analysis of taurine and edaravone in biological sample is critical in pharmaceutical studies. Although each of them can be individually analyzed by different approaches, concurrent quantification is still a highly challenging task with respect to their great polarity variation and the complex composition of tissue sample. In the present study, to simultaneously determine taurine and edaravone in rat tissue, the sample preparation and chromatographic separation conditions were evaluated and discussed in detail. As for the sample preparation, four kinds of solvent and the volume ratio of the optimal solvent to biological sample were both tested and evaluated based on the chromatographic profile, extraction recovery, and matrix effect (ME). The chromatographic separation was performed in a reverse phase (RP) and two hydrophilic interaction liquid chromatography (HILIC) modes, and the corresponding separation efficiencies were assessed using chromatographic parameters like half-width (W 1/2 ), tailing factor (f t), theoretical plates number (N), and ME. Furthermore, adopted composition of two mobile phase systems and the concentrations of the additives in the optimum buffer system were also investigated on an Atlantis HILIC silica column according to the resultant chromatographic profiles and peak areas of the analytes. The optimal results were obtained when the biological samples were deproteined by 4-fold volume of methanol/acetonitrile (1:3, v/v) and separated on a HILIC column with a gradient elution of acetonitrile/water containing 0.2 % formic acid and 10 mM ammonium formate. The proposed approach was validated and successfully applied to the parallel determination of the tissue distribution of edaravone and taurine in rat tissues.

A novel UPLC-DAD method was developed and validated for the simultaneous determination of baicalin (baicalein-7-glucuronide, BG), oroxylin A-7-O-glucuronide (OAG) and wogonoside (WG) in rat plasma using rutin as the internal standard. Plasma samples were precipitated using acetonitrile containing 0.1% formic acid. Separation was performed on an Agilent Eclipse Plus C18 column (2.1 × 50 mm, 1.8 µm) using gradient acetonitrile and 0.2% formic acid water solution as mobile phase. The flow-rate was set at 0.4 mL/min and the eluate was detected at 275 nm. The method was linear over the ranges of 0.075-17.50, 0.050-12.60 and 0.056-14.10 µg/mL for BG, OAG and WG, respectively. The intra- and inter-day precisions were respectively4.8% and 6.4%. All of the limits of detection of three analytes in rat plasma were 0.01 µg/mL, whereas the limits of quantification were, respectively, 0.035, 0.025 and, 0.025 µg/mL. This assay has been successfully applied to pharmacokinetics of BG, OAG and WG in rats after oral administration of Yinhuang granule (YHG) and comparative pharmacokinetics of BG in rats following oral administration of the pure BG, Radix Scutellariae (RS) or YHG. We speculate that some co-existing ingredients in RS or YHG may increase the absorption and elimination of BG in rat. This work may be helpful for the quality control of Yinhuang granule.

In this work, two high-performance liquid chromatography (HPLC) assays were developed and validated for the independent determination of edaravone and taurine using 3-methyl-1-p-tolyl-5-pyrazolone and L-glutamine as internal standards. In in vitro experiments, human plasma was separately spiked with a mixture of edaravone and taurine, edaravone or taurine alone. Plasma was precipitated with acetonitrile containing 0.1% formic acid. Ultrafiltration was employed to obtain the unbound ingredients of the two drugs. The factors that might influence the ultrafiltration effiency were elaborately optimized. Plasma supernatant and ultrafiltrate containing taurine were derivated with o-phthalaldehyde and ethanethiol in the presence of 40 mmol/L sodium borate buffer (pH 10.2) at room temperature within 1 min. Chromatographic separations were achieved on an InertSustain C18 column (250 × 4.6 mm, 5 µm). Isocratic 50 mmol/L ammonium acetate-acetonitrile and gradient 50 mmol/L sodium acetate (pH 5.3)-methanol were respectively selected as the mobile phase for the determination of edaravone and taurine. All of the validation data including linearity, extraction recovery, precision, accuracy and stability conformed to the requirements. Results showed that there were no significant alterations in the plasma protein binding rate of taurine and edaravone, implying that the proposed combination therapy was pharmacologically feasible.

In this study, two independent and complementary liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were respectively developed and validated for the determination of edaravone or taurine in rat urine, feces and bile after intravenous administration, using 3-methyl-l-p-tolyl-5-pyrazolone and sulfanilic acid as the internal standards (IS). Edaravone was separated on an Agilent Eclipse Plus C18 column (100×2.1 mm, 3.5 μm) using methanol and water (containing 5 mM ammonium formate and 0.02% formic acid) as mobile phase, while taurine was performed on a Waters Atlantis HILIC Silica column (150×2.1 mm, 3 μm) using acetonitrile and water (containing 5mM ammonium formate and 0.2% formic acid) as mobile phase. The mass analysis was performed in a Triple Quadrupole mass spectrometer via multiple reaction monitoring (MRM) with negative ionization mode. The optimized mass transition ion pairs (m/z) for quantification were 173.1→92.2 and 187.2→106.0 for edaravone and its IS, 124.1→80.0 and 172.0→80.0 for taurine and its IS, respectively. The validated methods have been successfully applied to the excretion and metabolism interaction study of edaravone and taurine in rats after independent intravenous administration and co-administration with a single dose. The results demonstrated that there were no significant alternations on the metabolism and cumulative excretion rate of edaravone and taurine, implying that the proposed combination therapy was pharmacologically viable.

A rapid and useful approach for screening potential bioactive components in Ginkgo biloba extract (GBE) with preventive effect against diabetic nephropathy (DN) was developed using mesangial cells extraction coupled with high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. Mesangial cells were first divided into two groups according to their treatments with high glucose or high glucose plus GBE. After incubation for 4, 8, 12, 16, 24 and 48 h, the cells were harvested and extracted with 40% acetic acid in water before LC-MS/MS analysis. Then, 19 compounds and five metabolites were found to selectively combine with mesangial cells. Notably, compounds including quercetin and rutin were identified or tentatively characterized according to the results of retention time and MS spectra, which is highly consistent with our previous reports that quercetin and rutin are potent protective agents against glomerulosclerosis in DN. Therefore, all these results indicate that target cell extraction coupled with LC-MS/MS analysis can be successfully applied for predicting the bioactive components in GBE with preventive effect against DN.

Background: Traditional Chinese Medicine Constitution (TCMC) refers to an integrated, metastable and natural specialty of individual in morphosis, physiological functions and psychological conditions. It is formed on the basis of innate and acquired endowments in the human life process, which can be divided into normal constitution and unbalanced ones. The aim of this study was to investigate the distribution of TCMCs of Chinese women in Hong Kong and its acquired influencing factors. Methods: Local Chinese women between 30 to 65years old, were recruited from 18 districts of Hong Kong (n = 944), and were assessed using the Traditional Chinese Medicine Physical Constitution Scale for their TCMC types. Social-demographic, reproductive, lifestyle, systemic health and emotional status information were collected through structured questionnaire. The associations between different independent factors and each TCMC type, as well as the complex unbalanced TCMC types were tested individually. Significant factors related to unbalanced TCMC types were identified in final models using multiple factor analysis. Results: A total of 764 (80.9%) participants were diagnosed with unbalanced TCMCs. The most common TCMC type was Qi-deficiency constitution (53.9%), followed by Phlegm-wetness (38.9%), Yang-deficiency (38.2%), Yin-deficiency (35.5), Blood-stasis (35.4) and Qi-depressed (31%) constitution. Six hundred and eleven participants (64.7%) had at least two types of combined and unbalanced constitutions. Stepwise logistic analysis indicated that poor systemic health condition (OR, 1.76-2.89), negative emotions (OR = 1.39), overweight (OR = 1.58), high educational level (OR = 1.18) and mental work (OR = 1.44) were significantly positively correlated with certain unbalanced TCMCs. Meanwhile, aging (OR, 0.59-0.73), exercise habit (OR, 0.61-0.79) and reproductive history (OR = 0.72) showed inverse associations with unbalanced constitutions. In addition, systemic health condition and emotional status, exercise habit and age were significantly associated with the combined unbalanced TCMC types. Conclusion: The majority of middle-aged Chinese women in Hong Kong had unbalanced and complex TCMCs. Qi-deficiency, Phlegm-wetness and Yang-deficiency constitutions are the most common constitutions. Poor systemic health condition, less-than-satisfactory emotional life, overweight and mental work are associated with and may be contributors for the formation of unbalanced TCMCs, while regular physical exercise was found to be a potential protective factor for unbalanced TCMCs.

An LC-MS/MS method was developed and validated for the simultaneous quantification of edaravone and taurine in beagle plasma. The plasma sample was deproteinized using acetonitrile containing formic acid. Chromatographic separations were achieved on an Agilent Zorbax SB-Aq (100 × 2.1 mm, 3.5 μm) column, with a gradient of water (containing 0.03% formic acid) and methanol as the mobile phase at a flow rate of 0.3 mL/min. The analyte detection was carried out in multiple reaction monitoring mode and the optimized precursor-to-product transitions of m/z [M+H](+) 175.1 → 133.0 (edaravone), m/z [M+H](+) 189.1 → 147.0 (3-methyl-1-p-tolyl-5-pyrazolone, internal standard, IS), m/z [M-H](-) 124.1→80.0 (taurine), and m/z [M-H](-) 172.0 → 80.0 (sulfanilic acid, IS) were employed to quantify edaravone, taurine, and their corresponding ISs, respectively. The LOD and the lower LOQ were 0.01 and 0.05 μg/mL for edaravone and 0.66 and 2 μg/mL for taurine, respectively. The calibration curves of these two analytes demonstrated good linearity (r ＞ 0.99). All the validation data including the specificity, precision, recovery, and stability conformed to the acceptable requirements. This validated method has successfully been applied in the pharmacokinetic study of edaravone and taurine mixture in beagle dogs.

Three liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were respectively developed and validated for the simultaneous or independent determination of taurine and edaravone in rat plasma using 3-methyl-1-p-tolyl-5-pyrazolone and sulfanilic acid as the internal standards (IS). Chromatographic separations were achieved on an Agilent Zorbax SB-Aq (100 × 2.1 mm, 3.5 µm) column. Gradient 0.03% formic acid-methanol, isocratic 0.1% formic acid-methanol (90:10) and 0.02% formic acid-methanol (40:60) were respectively selected as the mobile phase for the simultaneous determination of two analytes, taurine or edaravone alone. The MS acquisition was performed in multiple reaction monitoring mode with a positive and negative electrospray ionization source. The mass transitions monitored were m/z [M + H](+) 175.1 → 133.0 and [M + H](+) 189.2 → 147.0 for edaravone and its IS, m/z [M - H](-) 124.1 → 80.0 and [M - H](-) 172.0 → 80.0 for taurine and its IS, respectively. The validated methods were successfully applied to study the pharmacokinetic interaction of taurine and edaravone in rats after independent intravenous administration and co-administration with a single dose. Our collective results showed that there were no significant alterations on the main pharmacokinetic parameters (area under concentration-time curve, mean residence time, half-life and clearance) of taurine and edaravone, implying that the proposed combination therapy was pharmacologically feasible.

The dog-faced water snake (Cerberus rynchops) is a viviparous colubrid snake. This species occurs in coastal South Asia, peninsular India, Sri Lanka, Bangladesh, Myanmar, the Andaman Islands, eastward to the coastal areas of the Indochinese and Malaysian Peninsulas, southwards into Indonesia and eastwards to Palau and Micronesia. It has been listed on Appendix III of CITES. In India, this species is protected under the Indian Wildlife Act, due to its historic trade as skin. Accordingly, we aimed to characterize new microsatellite markers, which could be helpful for us to get a better understanding of population structure and evolutionary history of the snake. Here, eight polymorphic microsatellite loci were isolated from an enriched genomic library of C. rynchops. One hundred and fifteen individuals were collected from Singapore. These markers revealed a high degree of genetic diversity (5–14 alleles per locus) and heterozygosity (H O ranged from 0.565 to 0.965, and H E ranged from 0.701 to 0.882). No locus exhibited significant deviations from Hardy–Weinberg equilibrium. There was no evidence of linkage disequilibrium between any pair of loci. These loci are first identified for C. rynchops and will be useful for future study of population structure, genetic diversity and conservation strategy design of this species.

The rice paddy snake (also known as Boie’s mud snake) Enhydris plumbea is a viviparous colubrid snake. This is a widely distributed species found from the Andaman Islands (India) and Myanmar eastwards across the Southeast Asian Peninsula and southern China southwards into Indonesia; and it is present on most of the Indonesian islands and eastwards to Sulawesi and the Moluccas. The snake has been overexploited by local people for traditional medicine to treat skin diseases, and is listed on the “Lists of terrestrial wildlife under state protection, which are beneficial or of important economic or scientific value” promulgated by China’s State Forestry Administration in 2000. Knowledge of population structure and genetic diversity is crucial for effective wildlife preservation. Specific molecular markers such as mitochondrial DNA and microsatellites are useful tools for achieving this goal. Here, eight polymorphic microsatellite loci were isolated from an enriched genomic library of E. plumbea. One hundred and twenty individuals were collected from Wenzhou population in Zhejiang, China. These markers revealed a high degree of genetic diversity (4–8 alleles per locus) and heterozygosity (H O ranged from 0.148 to 0.789, and H E ranged from 0.347 to 0.766). No locus exhibited significant deviations from Hardy-Weinberg equilibrium. There was no evidence of linkage disequilibrium among pairs of loci. These loci will be useful for future study of population structure, genetic diversity and conservation strategy design of E. plumbea.

The Chinese water snake Enhydris chinensis is an ovoviviparous colubrid snake distributed in China and Vietnam. It is commonly used in the production of Chinese snake oil, which is known for treating ailments such as fever, joint pain, and headache. It is listed as a vulnerable species according to the China red data book of endangered animals. Specific molecular markers such as mitochondrial DNA and microsatellite loci are helpful for us to get a better understanding of population structure and evolutionary history of the snake and take effective conservation action for this species. Here, we characterize eight polymorphic microsatellite loci isolated from E. chinensis genomic libraries. Forty-seven individuals were collected from Ji’an population in Jiangxi, China. These markers revealed a high degree of genetic diversity (3–10 alleles per locus) and heterozygosity (H O ranged from 0.532 to 0.957, and H E ranged from 0.565 to 0.806). No locus exhibited significant deviations from Hardy–Weinberg equilibrium. There was no evidence of linkage disequilibrium among pairs of loci. The results of this paper will be useful for future study of population structure and genetic diversity of E. chinensis.

Eight polymorphic microsatellite loci are described for the endangered Amur ratsnake, Elaphe schrenckii (Colubridae). Loci were screened in 31 unrelated individuals of E. schrenckii collected from Liaoning Province, China. These markers revealed a high degree of genetic diversity (6–20 alleles per locus) and heterozygosity (H O ranged from 0.296 to 1.000, and H E ranged from 0.469 to 0.850). No significant linkage disequilibrium between pairs of loci was found, except one of them significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction. These loci are the first time to be characterized and will be useful for population genetic structure investigation, conservation management and paternity identification study for E. schrenckii.

Ptyas korros is an oviparous colubrid snake that has a distribution covering the southeastern part of China (including Taiwan, Hainan and Hongkong), Indo-China Peninsula, India and Indonesia. The snake is listed as a vulnerable species according to the China Red Data Book of Endangered Animals. To develop effective conservation strategies for the snake, we need a better understanding of gene flow, population structure and evolutionary history of this species by using kinds of molecular markers such as mitochondrial DNA and microsatellite loci. Here, we characterize ten polymorphic microsatellite loci isolated from P. korros genomic libraries. Fifty-seven individuals were collected from Quanzhou population in Guangxi, China. These markers revealed a high degree of genetic diversity (4–14 alleles per locus) and heterozygosity (H O ranged from 0.296 to 1.000, and H E ranged from 0.419 to 0.868). No locus exhibited significant deviations from Hardy–Weinberg equilibrium. There was no evidence of linkage disequilibrium among pairs of loci. These microsatellite markers will be useful for the study of gene flow, population structure and evolutionary history of P. korros.

The beauty rat snake Elaphe taeniura is an oviparous colubrid snake, the distributional range of which covers most provinces of China (including Taiwan and Hainan), India, Indochina, and the northern half of Malay Peninsula. The snake is listed on the “Lists of terrestrial wildlife under state protection, which are beneficial or of important economic or scientific value” promulgated by China’s State Forestry Administration in 2000. To develop some more effective conservation strategies for the snake, we need a better understanding of population structure and evolutionary history of this species through kinds of specific molecular markers such as microsatellites or short, tandemly repeated base-pair sequences. Here, we characterize 11 polymorphic microsatellite loci isolated from E. taeniura genomic libraries. Forty-eight individuals were collected from Quanzhou population in Guangxi, China. These markers revealed a high degree of genetic diversity (5–19 alleles per locus) and heterozygosity (H O ranged from 0.121 to 0.875, and H E ranged from 0.276 to 0.922). No locus exhibited significant deviations from Hardy–Weinberg equilibrium. There was no evidence of linkage disequilibrium among pairs of loci. These microsatellite markers will be useful for future study of population structure and evolutionary history of E. taeniura.

A quality control (QC) strategy for quantitative and qualitative analysis of âcommon peaksâ in chemical fingerprint was proposed to analyze Yuanhu Zhitong tablet (YZT), using high performance liquid chromatography with diode array detector and tandem mass spectrometry (HPLC-DADâMS/MS). The chromatographic separation was achieved on an Agilent Eclipse plus C18 column with a gradient elution using a mixture of 0.4â° ammonium acetate aqueous (pH 6.0 adjusted with glacial acetic acid) and acetonitrile. In chemical fingerprint, 40 peaks were assigned as the âcommon peaksâ. For quantification of âcommon peaksâ, the detection wavelength was set at 254Â nm, 270Â nm, 280Â nm and 345Â nm, respectively. The method was validated and good results were obtained to simultaneously determine 10 analytes (protopine, jatrorrhizine, coptisine, palmatine, berberine, xanthotoxin, bergapten, tetrahydropalmatine, imperatorin and isoimperatorin). For qualification of âcommon peaksâ, 33 compounds including 10 quantitative analytes were identified or tentatively characterized using LCâMS/MS. These results demonstrated that the present approach may be a powerful and useful tool to tackle the complex quality issue of YZT. Keywords: Yuanhu Zhitong tablet, Alkaloids, Coumarins, Quality control, HPLC-DADâMS/MS