β2-Adrenoceptor affinity chromatography and its application in the screening of the active compounds from Semen Armeniacae Amarum

β2-Adrenoceptor （β2-AR） was purified from the rabbit lung tissue by sepherose-salbutamol affinity chromatographic column. To prepare the β2-AR stationery phase, β2-AR was evenly immobilized on the surface of macro-pore silica with e mild chemical coupling method through covalent bond. The retention properties of β2-AR stationery phase were characterized by four ligends, selbutamol sulfate, noredreneline bitartrete, adrenaline hydrochloride end proprenolol hydrochloride, to establish the β2-AR affinity chromatography. Then, the method was used to screen the active compounds from the total extracts of Semen Armeniacae Amarum. The results showed that β2-AR on the surface of the stationary phase could keep its original bioectivity end selectivity. Amygdalin retained in the chromatographic column was proved to be the active compound of the total extracts of Semen Armeniacae Amarum. Compared with the existing chromatographic screening approaches, this method showed e good stability end high selectivity. The active compounds which could interact with β2-AR in traditional Chinese medicine （TCM） could be screened efficiently by this method, providing e new way to screen the active compounds in complicated samples such as TCM.     

Establishment of the model of vascular endothelial cell membrane chromatography and its preliminary application

A model of vascular endothelial cell membrane chromatography was established by using an ECV304 cell membrane stationary phase (ECV304 CMSP) prepared by immobilizing the ECV304 cell membrane onto the surface of silica carrier. The surface and chromatographic characteristics of ECV304 CMSP were studied. The active component from Caulophyllum robustum was screened by using the model of vascular endothelial cell membrane chromatography. The interaction between the active component and membrane receptor was determined by using a replace experiments. The effect of the active component was tested by using tube formation of ECV304 cell. The results indicated that the model of ECV304 cell membrane chromatograph (ECV304 CMC) can stimulate the interaction between drug and receptor in vitro and the retention characteristics of taspine as active component was similar to that of model molecule in the model of ECV304 CMC. And therefore, taspine acted on VEGFR2 and inhibited the tube formation of ECV304 cell induced by VEGF. This model can be used to screen definite active component as a screening model.     

Development of the model of pancreatic β-cell membrane chromatography and its chromatographic characteristics

A new model of pancreatic β-cell membrane chromatography has been established by using a β-cell membrane stationary phase （β-CMSP） prepared by immobilizing the β-cell membrane onto the surface of silica carrier. The protein level and K^＋, Na^＋-ATP enzymatic activity of the β-CMSP were detected respectively. The surface characteristics of the β-CMSP were tested by using the scanning electron microscope and surface energy spectrometer. In this model, the column （10 mm × 2 mm, I.D.） packed with β-CMSP, 25 mmol/ammonium sulfate buffer solution （pH 7.4） as mobile phase with the flow rate of 0.2 mL/min at 37 ± 0.5℃ were used in the following studies. The retention characteristics of the sulfonylureas （gliquidone, glibenclamide, gliclazide and glipizide） were investigated under the chromatographic conditions above. The affinities of the sulfonylureas on β-cell membrane and receptor will be expressed by using the logk＇ values （the logarithm of capacity factor of a solute） in the model. The correlation of the affinity with the pharmacological effect of the sulfonylureas was analyzed also.     

Affinity ultrafiltration of DNA topoisomerases-targeted compounds determined with HPLC／ESI-MS for drug candidate screening

A method of screening assay is demonstrated. The approach is based on the affinity of antitumor candidates for topoisomerases. In this method, antitumor candidates are fished out using topoisomerases as targets. Traditional analysis of complex compounds typically encounters signal suppression due to the relatively low concentrations, but enzyme-affinity screening for the active compounds can effectively concentrate the desired analysts into a small volume of high concen-tration. Active compounds are separated from non-affinity compounds by ultrafiltration. The molecules-enzymes complexes that are retained on the filter are subsequently separated by acidification to obtain the topoisomerases-affinity compounds for analysis on High Performance Liquid Chromatography coupled with electrospray ionization mass spectrometric detec-tion (ESI-MS). This enzyme-affinity based screening assay provides a highly specific and efficient method that can directly screen, identify, and acquire drug candidates thus improving the accuracy and speed of high-throughput screening activities.     

CeO2: an active and selective catalyst for the oxidative dehydrogenation of ethane with CO2

It was assumed that the homogeneous dehydrogenation of CiHg occurred in the gas phase was independent of the presence of catalysts and this is applicable to the oxidative dehydrogenation of ethane with CO2 over CeO2 . On the basis of this assumption, a method was developed to analyze the contribution of C2rtj formation from heterogeneous catalysis. In this study, ceria was found to be active and selective for the oxidative dehydrogenation of ethane with CO2 and the selectivity to C, H4 was above 60 % and the actual contribution for C, H4 formation from heterogeneous catalysis was about 55%— 75% in the range of 953—993 K.     

β-Adrenergic activation enhances NMDA-induced current in pyramidal cells of the basolateral nucleus of amygdala

NMDA receptor （NMDA-R） in the amygdala complex is critical for both long-term potentiation （LTP） and formation of conditioned fear memory. It is reported that activation of β-adrenoceptors （β-AR） in the amygdala facilitates LTP and enhances memory consolidation. The present study examined the regulatory effect of β-AR activation on NMDA-R mediated current in pyramidal cells of the basolateral nucleus of amygdala （BLA）, using whole-cell recording technique. Bath application of the β-AR agonist isoproterenol enhanced NMDA-induced current, and this facilitatory effect was blocked by co-administered propranolol, a β-AR antagonist. The facilitatory effect of isoproterenol on NMDA-induced current could not be induced when the protein kinase A （PKA） inhibitor Rp-cAMPs was added in electrode internal solution.The present results suggest that β-AR activation in the BLA could modulate NMDA-R activity directly and positively, probably via PKA.     

New approach for screening anti—tumor compounds

A new screening approach for anticancer active compounds is presented.In this method,a target enzyme is incubated with a mixture of compounds,and then the com-plex formed by the target and small molecules is separated from the rest of the mixture by ultra-filtration.The complex that is retained on the membrane is subsequently washed with acid and small molecules that are specifically bound to the target are released and collected,then analyzed by high-performance liquid chromatograph combined with electrospray ionization mass spectrometry (HPLC/ESI-MS) analysis.We have successfully applied this method to screen anti-cancer compounds.DNA topoisomerase Ⅱ is used as a target to capture anti-tumor candidates from a mixture of combinatorial compounds,such as doxorubicin,daunorubi-cin and pravastain.     

A new chromatographic method for fast separation of active proteins

A new method with only a single chromatographic column to accomplish the separation of intact protein molecules by the combination of ion exchange chromatography and hydrophobic interaction chromatography is firstly presented. By selecting twice a suitable stationary phase, mobile phase, buffer exchange condition, and sample injection, some proteins usually required to separate with two-dimensional chromatography can now be accomplished only using the single column in one hour. In addition, the separated proteins can maintain their original three-or four-dimensional molecular structure. It would be expected that this method can also provide a new thought for sample pre-fractionation in proteomic investigation, especially, for the limitation of sample source.     

Chiral separation of racemic drugs using molecular imprinting

Molecularly imprinted polymers (MIPs) of (S)-ketoprofen and (S)-naproxen are prepared using non-covalent imprinting in the presence of template molecules. The prepared MIPs are used as the chiral stationary phase to separate ramemic naproxen and ketoprofen. The results show that racemic naproxen and ketoprofen are efficiently resolved on MIPs. The effect of concentration of acetic acid in the mobile phase is studied, and the data are analyzed using the affinity chromatography model, and the close agreement is achieved between the simulated and experimental curves. The results suggest that the affinity chromatography mechanism controls the retention in this system. Moreover, the affinity chromatography equilibrium constants on (S)-naproxen and (S)-ketoprofen are estimated.     

Determination of Polysaccharide in Radix pseudostellariae Extract by Size-Exclusion High-Performance Liquid Chromatography

A simple size-exclusion high-performance liquid chromatographic method was developed for rapid molecular mass screening to determine the average molecular mass, polydispersity, and quantity of Taizishen polysaccharides extract. The screening used a TOSOH TSK-GEL G3000SW high-performance liquid chromatographic column with 0.1 mol/L NaNO3 buffer as the mobile phase and refractive index detec- tor. The molecular mass calibration curve was linear for polysaccharide standards from 10 to 100 kDa with a correlation coefficient of 0.9991. The method can be used to analyze the quantity, average molecular mass, and polydispersity of polysaccharides. In addition, the screening method is suitable for quality control of polysaccharide preparations in Chinese medicines and pharmaceutical products.     

Functional Polymers for Chromatographic Enantiomer Separation

Today, methods to obtain pure enantiomers of organic compounds are of extreme importance, not the least due to the demands imposed on the pharmaceutical industry. Separation of the two enantiomers in a racemate is necessary in order to study possible differences in biological activity. Consequently, research on methods for direct chromatographic resolution by means of columns containing a chiral stationary phase （CSP） has been intense during the last decades and a wide variety of CSPs,many of which commercially available,exist today. Polymeric CSPs have been made from naturally occurring polymers （cellulose, amylose） via suitable derivatization, as well as either by polymerization of chiral monomers or via asymmetric catalysis to yield linear, isotactic polymers of a predominantly single-handed helicity. An excellent review of the field, with special emphasis on oreoarative applications, was published in 1994.     

Fabrication and characterization of novel tentacle-type adsorbent for resolution of chiral drugs

Novel tentacle-type chiral adsorbent based on silica gels modified with β-cyclodextrin (β-CD) and polyvinyl alcohol(PVA) is synthesized by surface grafting technique. The adsorption behavior of chiral pharmaceuticals using the resulting adsorbents is investigated. In the study, FT-IR and thermogravimetric analysis (TGA) are employed to characterize the chemical and physical properties of the adsorbent; mandelic acid (MA) serves as model solute to evaluate the sorption capacity and selectivity of the adsorbent. The experimental results show that the novel tentacle-type adsorbent is possessed of larger equilibrium adsorption capacity and better selectivity over the conventional one and could be as potential chiral stationary phase (CSPs).     

Numerical simulation for volatile organic compound removal in rotating drum biofilter

Rotating drum biofilters (RDBs) could effectively remove volatile organic compounds (VOCs) from waste gas streams. A mathematical model was developed on the basis of mass transport and mass balance equations in an RDB,the two-film theory,and the Monod kinetics. This model took account of mass transfer and biodegradation of VOC in the gas-water-biofilm three-phase system in the biofilter,and could simulate variations of VOC removal efficiency with a changing specific surface area and porosity of the media due to the increasing of biofilm thickness in the biofilter. Toluene was used as a model VOC. This model was further simplified by introducing a coefficient of the gas velocity and ne-glecting the water phase due to the complexity of operating conditions. The equations for the biofilm phase,gas phase,and biofilm accumulation in this model were solved using collocation method,ana-lytic method,and the Runge-Kutta method separately. A computer program was written down as MATLAB to solve this model. Results of numerical solutions showed that toluene removal efficiency in the RDB increased and reached the maximum values of 97% on day 4 after the startup,and then de-creased and remained at 90% after 5 more days of operation. Toluene concentration was high at the outermost layer where more than 70% toluene was removed,and was low at the innermost layer where less than 10% toluene was removed. The dynamic removal efficiencies from this model correlated reasonably well with experimental results for toluene removal in a multi-layered RDB.     

Separation of C6-Olefin Isomers in Reactive Extractants

Chemical complexation, in which certain metal ions （especially Ag^＋ and Cu^＋） reversibly and selectively complex olefin isomers, was used to separate 1-hexene from a mixture of internal hexenes as an attractive alternative to traditional distillation. Several potential reactive extractants were investigated for their selectivity of both 1-hexene to 2-hexene and 1-hexene to 3-hexene. With 3 mol/L AgNO3-N-methyl-pyrrolidone （NMP） solution as extractant, the selectivity of 1-hexene to 3-hexane was increased to about 2.0 and the selectivity of 1-hexene to 2-hexene reached 1.5. Both the raffinate phase and the extracted phase were analyzed using a gas chromatograph equipped with a flame ionization detector and a 50-m capillary column. The experimental results show that the silver nitrate NMP solution has the better selectivity than other reactive extractions.     

Study on metabolites of bioactive compounds assisted by LC-SPE-NMR

Aim To study the metabolites of active compound dicentrine in miniature pig.Method Solvent extraction and column chromatographic methods were used to give the metabolites containing fractions of pig urine.UV absorption,mass spectrometry and HPLC-SPE-NMR were used to identify the metabolites of dicentrine.Results Twenty-four in total,including nine phase I metabolites(MI-1～9) and 15 phase II metabolites(MII-1～15),were characterized.Conclusion Dicentrine easily metabolizes into all kinds of compounds structurally related to pig body.     

Effects of calcium compounds on the carbothermic reduction of vanadium titanomagnetite concentrate

Effects of calcium compounds on the carbothermic reduction of vanadium titanomagnetite concentrate(VTC) were investigated. It was found that calcium compounds had great effects on the metallization rate of the reduction product, the order of the metallization rate of reduction product being CaCO_3 no additive CaSO_4 CaCl_2, which indicated that the addition of CaCO_3 was more conducive to promoting the reduction of iron than other calcium compounds. Gas analysis showed that there were mainly two processes in the carbothermic reduction of VTC, a solid–solid and a solid–gas reaction. The concentrations of CO and CO_2 were highest when CaCO_3 was added, while that in a roasting system decreased the most when CaCl_2 was added. X-ray diffraction(XRD) analysis showed that calcium compounds could change the reduction process of ilmenite in VTC. The phase compositions of the reduction products were changed from metallic iron(Fe) and anosovite(FeTi_2O_5) to metallic iron(Fe) and perovekite(CaTiO_3) when calcium compounds were added. Additionally, CaSO_4 and CaCl_2 could significantly promote the growth of metallic iron particles, though the existence of Fe-bearing Mg_2TiO_4 in reduction products was not conducive to the reduction of iron. The formation of FeS would further hinder the reduction of iron after adding CaSO_4.     

Enzyme-Mediated Hydrolysis of Poly（ethylene glycol）-Supported Carbonates

Enzymatic kinetic resolution of racemic alcohols or esters is known as a useful method for the preparation of optically active secondary alcohols. However, the work-up including the separation of the mixture of the remaining substrate and the resulting compound spend a lot of time and waste much amount of solvents. On the other hand, organic synthesis based on polymer supports has made rapid progress. Although the methodology is potentially useful for the easy separation of compounds obtained by the enzymatic reaction, there have been relatively few reports on enzymatic resolutions of using a polymer so far. We have noticed that using a watersoluble polymer could be suitable for enzymatic transformation. Here, we report the first example of an enzyme-mediated enantioseleetive hydrolysis of poly（ethylene glycol）（PEG）-supported substrates with a carbonate moiety to afford optically active compounds, and the method enables us to achieve the easy separation of the products. See Scheme 1.     

Enzyme-Mediated Hydrolysis of Poly(ethylene glycol)-Supported Carbonates

1Introduction Enzymatic kinetic resolution of racemic alcohols or esters is known as a useful method for the preparation of optically active secondary alcohols. However, the work-up including the separation of the mixture of the remaining substrate and the resulting compound spend a lot of time and waste much amount of solvents. On the other hand, organic synthesis based on polymer supports has made rapid progress. Although the methodology is potentially useful for the easy separation of compounds obtained by the enzymatic reaction, there have been relatively few reports on enzymatic resolutions of using a polymer so far. We have noticed that using a watersoluble polymer could be suitable for enzymatic transformation. Here, we report the first example of an enzyme-mediated enantioselective hydrolysis of poly(ethylene glycol)(PEG)-supported substrates with a carbonate moiety to afford optically active compounds, and the method enables us to achieve the easy separation of the products[1]. See Scheme 1.     

钙化合物对钒钛磁铁矿精矿碳热还原的影响

Effects of calcium compounds on the carbothermic reduction of vanadium titanomagnetite concentrate (VTC) were investigated. It was found that calcium compounds had great effects on the metallization rate of the reduction product, the order of the metallization rate of reduction product being CaCO₃ > no additive > CaSO₄ > CaCl₂, which indicated that the addition of CaCO₃ was more conducive to promoting the reduction of iron than other calcium compounds. Gas analysis showed that there were mainly two processes in the carbothermic reduction of VTC, a solid–solid and a solid–gas reaction. The concentrations of CO and CO₂ were highest when CaCO₃ was added, while that in a roasting system decreased the most when CaCl₂ was added. X-ray diffraction (XRD) analysis showed that calcium compounds could change the reduction process of ilmenite in VTC. The phase compositions of the reduction products were changed from metallic iron (Fe) and anosovite (FeTi₂O₅) to metallic iron (Fe) and perovekite (CaTiO₃) when calcium compounds were added. Additionally, CaSO₄ and CaCl₂ could significantly promote the growth of metallic iron particles, though the existence of Fe-bearing Mg₂TiO₄ in reduction products was not conducive to the reduction of iron. The formation of FeS would further hinder the reduction of iron after adding CaSO₄.