Exopolysaccharide La0.1-1 was extracted from the broth of a marine bacterium Lentibacter algarum ZXM100T isolated from the seawater in the coastal region of Qingdao and purified by Q Sepharose Fast Flow ion-exchange chromatography and Superdex 75 gel-permeation chromatography. Its physiochemical properties and primary structural characters were investigated by chemical analysis together with high performance liquid chromatography (HPLC), high performance gel permeation chromatography (HPGPC) and gas chromatography and mass spectrometry (GC-MS). The results show that the total sugar content of the exoploysaccharide La0.1-1 was about 66% with an average molecular weight at 12.0 kDa. La0.1-1 is mainly composed of Gal, Man, GlcN at the ratio of 1.35:1.1:1.0. Results of GC-MS and NMR demonstrate that the exopolysaccharide La0.1-1 mainly exists with the beta configuration. The primary linkage styles are --> 2)-Manp(1 --> and --> 3)-Galp(1 --> with a small amount of --> 4)-Galp(--> 1 and --> 4)-Manp(1 --> linkages. The linkage mode of GlcN is --> 4)GlcN(1 --> and terminal linkage. The exopolysaccharide has mainly a linear sructure with a few branches linked to 0-6 of --> 2)-Manp(1 --> and 0-4 or 0-6 of --> 3)-Galp(1 -->. 1D-NMR data also revealed that La0.1-1 is substituted by certain acetyl; the acetyl is mainly linked to N-2 of GlcN. The exopolysaccharides of the bacterium of Lentibacter genus is reported for the first time, and an exopolysaccharide with novel structure was obtained, which enriched marine polysaccharide resources.
Chromatography, Gel ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; Gas Chromatography-Mass Spectrometry ; Magnetic Resonance Spectroscopy ; Molecular Weight ; Polysaccharides ; chemistry ; isolation & purification ; Rhodobacteraceae ; chemistry ; Seawater ; microbiology

Objective To provide the reference for the identification of unknown fentanyl analogues by studying the characteristic ions and main fragmentation pathways of fentanyl analogues in the modes of collision induced dissociation （CID） and electron ionization （EI）. Methods Nine fentanyl analogues （2, 2'-difluorofentanyl, acetyl fentanyl, fentanyl, butyl fentanyl, valeryl fentanyl, acryloyl fentanyl, furan fentanyl, 4-fluorine isobutyl fentanyl, carfentanyl） were selected and analyzed with ultra-high performance liquid chromatography-quadrupole time-of-flight-mass spectrometry （UHPLC-QTOF-MS） and gas chromatography-mass spectrometry （GC-MS）. The mass spectrum obtained was analyzed. The CID and EI fragmentation routes of fentanyl analogues were speculated. Results The CID and EI fragmentation pathways were highly similar. In the CID mode, characteristic ions were formed by the carbon-nitrogen bond cleavage between the piperidine ring and the N-phenyl-amide moiety, within the piperidine ring, and between the phenethyl and piperidine ring. While in the EI mode, dissociation of the piperidine ring, as well as cleavage between the piperidine ring and the phenethyl were the main fragmentation pathways. Conclusion This study summarizes the main fragmentation pathways and characteristic ions of fentanyl analogues in the CID and EI modes, which is useful for forensic laboratories to identify and structural analyze fentanyl type new psychoactive substance in practical work.
Chemistry Techniques, Analytical/methods* ; Chromatography, High Pressure Liquid ; Fentanyl/analysis* ; Gas Chromatography-Mass Spectrometry ; Humans ; Mass Spectrometry

This study examined the chemical composition of A. blasiliensis and the chemical structural properties of an immuno-stimulating polysaccharide. The amino acids, free sugars, and organic acids by HPLC and fatty acids by GC were analyzed. The immuno-stimulating substance from A. blasiliensis was extracted with hot water and purified by ethanol precipitation. It underwent ion exchange chromatography on DEAE-cellulose and gel filtration on Toyopearl HW 65F. Through GP-HPLC, the substance was found to be homogeneous. Its chemical structure was determined by 13C-NMR. Fatty acids, organic acids, and sugar alcohol composition consisted exclusively of linoleic acid, fumaric acid and mannitol, respectively. The amino acids were mainly glutamic acid, glycine, and arginine. By 13C-NMR analysis, the immuno-stimulating substance was identified as beta-(1-->3) (1-->6)-glucan, composed of a backbone with (1-->3)-linked D-glucopyranosyl residues branching a (1-->6)-linked D-glucopyranosyl residue. The beta-glucan from A. blasiliensis showed pronounced immuno-stimulating activity on the antibody-production ability of B-lymphocytes by the hemolytic suspension assay. In these results, A. blasiliensis was estimated to have potent pharmacological properties and potential nutritional values.
Agaricus ; Amino Acids ; Arginine ; B-Lymphocytes ; Carbohydrates ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; DEAE-Cellulose ; Ethanol ; Fatty Acids ; Fumarates ; Glutamic Acid ; Glycine ; Linoleic Acid ; Mannitol ; Nutritive Value ; Water

Two new azaphilone derivatives containing 1,3-dioxolane moiety, penidioxolanes A (1) and B (2), were isolated from marine-derived fungus Penicillium sp. KCB12C078, together with four known compounds (3-6) by chemical investigation. Compounds 1 - 6 were isolated by combination of silica gel, ODS column chromatography and preparative HPLC. Their structures were determined by analysis of spectroscopic data including 1D-, 2D-NMR, and MS techniques. The isolates were evaluated against cancer cell growth inhibition effects and antimicrobial activity.
Chromatography ; Chromatography, High Pressure Liquid ; Fungi ; Penicillium* ; Silica Gel

OBJECTIVES: This study was conducted in order to understand the cellular events associated with silica-induced pathogenesis of the rat lung. METHODS: Silicosis was induced by an intratracheal instillation of 50 mg of silica (SiO2, 0.15 - 10 micrometer) suspended in 500 microliter of a sterile saline solution in Sprague-Dawley rats weighing 200g. Silicotic nodules were excised from the rat lungs 4 weeks after silica instillation, then boiled for 4 days at 110 degrees in solution containing 2% SDS, 10 M urea and 40 mM DTT. The insoluble cellular encapsulates were electrophoresed on 4-12 % gradient SDS-PAGE, and the amino acid composition was analyzed. Affinity chromatographies of the homogenate supernatants of the control lung, silicotic nodule, and normal rat plasma were performed using rabbit IgG, anti-rat, cross-linked protein from the silicotic nodule. The amounts of N epsilon-(gamma-glutamyl) lysine cross-linked in the control lungs and silicotic nodules were determined using HPLC analysis. RESULTS: The remaining cross-linked protein was insoluble in the 10 M urea and 40 mM sulfhydryl reagents even under prolonged boiling conditions. The encapsulate revealed the retention of silica particles within the protein whose amino acid composition showed a high percentage of alanine, leucine and glycine. A 46 KDa protein was identified as a cross-linked protein in the silicotic nodule by affinity chromatography. The level of N epsilon-(gamma-glutamyl) lysine dipeptide in the nodule digest was prominently increased compared with that in the control lung. CONCLUSIONS: Transglutaminase (TGase)-catalyzed cross-linking appears to be involved in the silicotic nodule formation, and the 46 KDa protein may be cross-linked to itself and other extracellular matrix proteins during fibrosis and the formation of eventually insoluble nodule.
Alanine ; Animals ; Chromatography ; Chromatography, Affinity ; Chromatography, High Pressure Liquid ; Electrophoresis, Polyacrylamide Gel ; Extracellular Matrix Proteins ; Fibrosis ; Glycine ; Immunoglobulin G ; Leucine ; Lung* ; Lysine ; Plasma ; Rats* ; Rats, Sprague-Dawley ; Silicon Dioxide ; Silicosis ; Sodium Chloride ; Sulfhydryl Reagents ; Urea

Human cytomegalovirus (HCMV)-specific monoclonal antibody, SCMVM 34, recognizes early antigen confined to the nucleus of HCMV-infected cells. This study was performed to identify the antigen reactive to SCMVM 34 with purification and amino acid sequencing. The nuclear and cytoskeletal fraction of HCMV-infected cells was subjected to 0.4 M NaCl extraction, DEAE-Sephacel ion exchange chromatography, DNA-cellulose chromatography and SDS-PAGE. The molecular weight of the reactive proteins was 52 kD, 40 kD and 34 kD. The modified or blocked amino termini of 52 kD and 40 kD showed resistance to Edman degradation. The internal peptide fragments were isolated by tryptic digeytion and reverse-phase HPLC. The internal amino acid sequence analysis of the peptides from HPLC profile revealed that the antigens recognized by SCMVM 34 was ppUIA4.
Chromatography ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; Cytomegalovirus* ; Electrophoresis, Polyacrylamide Gel ; Humans* ; Molecular Weight ; Peptide Fragments ; Peptides ; Sequence Analysis, Protein

OBJECTIVE: : To analyze experimental factors affecting recovery of puerarin in microdialysis. METHODS: : Puerarin concentration in microdialysate samples was determined by high performance liquid chromatography. The methods of direct dialysis, retrodialysis and the zero-net flux were used to calculate recovery, respectively. The effects of perfusate composition, the analyte concentration, perfusate flow rate, medium temperature and stir rates of the dialysis medium on recovery were investigated. RESULTS: : There were significant differences in the recovery values among direct dialysis, retrodialysis and zero-net flux methods. The recovery for 0.9% NaCl solution, Ringer's solution, PBS and anticoagulant dextrose solution as perfusate fluid were (71.25±2.36)%,(73.48±1.41)%,(68.50±2.43)% and (74.98±1.16)%, respectively. The composition of perfusate fluid had significant influence on the recovery(<0.01). At the same flow rate, recovery was independent of the analyte concentration. At the same concentration, the recovery was decrease with the increasing flow rate in an exponential relationship. The recovery increased with the raising temperature and stir rate of the dialysis medium, and the recovery remained stable when the stir rate reached above 200 rpm. CONCLUSIONS : A study method for recovery of puerarin in microdialysis has been established, and the recovery of puerarin is affected by calculating methods, perfusate fluids, flow rate, medium temperature and stir rate, but not affected by analyte concentrations.
Chemistry Techniques, Analytical ; methods ; Chromatography, High Pressure Liquid ; Isoflavones ; isolation & purification ; Microdialysis

Biological desulfurization is a process in which sulfur compounds are removed from gas and oil using microorganisms. It is a simple process that has mild operating conditions, high desulfurization efficiency, low energy consumption and less environmental pollution. However, there is still a lack of simple and efficient analytical methods for quantitatively analyzing the sulfur compounds in the biological desulfurization process. In order to solve this problem, the analytical method for the simultaneous determination of sulfite, thiosulfate and sulfide in biological desulfurization solutions by pre-column fluorescence derivation using high performance liquid chromatography (HPLC) was developed. The standard curves of sulfur species in this analytical method had good linear relationships with correlation coefficients of 0.999 5, 0.999 7, and 0.999 7 for sulfite, thiosulfate and sulfide, respectively. The detection limits of these sulfur compounds were 0.000 6, 0.000 7 and 0.001 1 μmol/L; the range of recovery rates were 98.17 to 101.9%, 100.9 to 102.6%, and 101.1 to 104.2%; which had good repeatability and stability. The analytical method was simple, efficient and accurate, and could be used to simultaneously determine the sulfur compounds in different biological desulfurization systems.
Chemistry Techniques, Analytical/methods* ; Chromatography, High Pressure Liquid ; Sulfur Compounds/analysis*

Oxaliplatin-loaded nanostuctured lipid carriers (OP-NLC) were prepared by ultrasonic emulsification method. And its optimal prescription was selected by orthogonal design. The laser light scattering technique, zeta potential analyzer, TEM, DSC, XRD and HPLC were employed to study the physicochemical parameters of OP-NLC, which displayed in terms of particle size, zeta potential, crystalline, drug loading and encapsulation efficiency. The results showed that OP-NLC had an average diameter of (111 +/- 20) nm, zeta potential of (-27.4 +/- 13.1) mV, encapsulation efficiency of (77.4 +/- 2.5) % and drug content of (0.8 +/- 1.5) mg mL(-1). TEM, DSC and XRD indicated that OP-NLC was spherical and the drug was dispersed as nanoparticles by means of non-crystalline. The in vitro release test showed that the drug could be sustained-released from NLC in buffer solution (pH 4.5) after a burst release in initial phase.
Antineoplastic Agents ; administration & dosage ; chemistry ; Calorimetry, Differential Scanning ; Chromatography, High Pressure Liquid ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Drug Compounding ; Lipids ; chemistry ; Microscopy, Electron, Transmission ; Nanoparticles ; Organoplatinum Compounds ; administration & dosage ; chemistry ; Particle Size

To optimize the preparation method of the complex of dihydroartemisinin (DHA) included by hydroxypropyl-beta-cyclodextrin (HP-beta-CD), the molar ratio of DHA and HP-beta-CD, inclusion temperature and inclusion time were optimized by the orthogonal design method with the inclusion drug yield and drug loading as the evaluation indexes. The IR spectrum, DSC and PXRD analyses were employed to characterize the complex and the molecular simulation was processed to investigate the tendency of complex formation. The optimized molar ratio of DHA and HP-beta-CD was 1 : 5, and the optimized preparation was performed under 50 degrees C for 1 h. The IR spectrum, DSC and PXRD analyses indicated the formation of the complex. The low binding free energy and the high solvent accessible surface obtained by molecular simulation showed that DHA could be included by HP-beta-CD and its solubility could be improved significantly. In conclusion, the optimized conditions for the preparation of DHA-HP-beta-CD complex provide a theoretical and experimental basis for further scale-up research.
2-Hydroxypropyl-beta-cyclodextrin ; Artemisinins ; administration & dosage ; chemistry ; Calorimetry, Differential Scanning ; Chromatography, High Pressure Liquid ; Drug Compounding ; methods ; Solubility ; Spectroscopy, Fourier Transform Infrared ; Surface Properties ; Temperature ; Time Factors ; X-Ray Diffraction ; beta-Cyclodextrins ; administration & dosage ; chemistry