Acetic Acid ; analysis ; Chromatography, Ion Exchange ; Workplace

PURPOSE: To elucidate the anti-oxidant effect of extract fractions from Xanthium strumarium L. on lens protein by crosslinking assay. METHODS: [(1 4)C] N-formyl-lysine was synthesized and purified by ion exchange chromatography. The crosslinking activities of extract fractions(Xan Crude, Xan CHCl3, Xan EtAc and Xan H2O) to lens protein were determined by incorporation with [(14)C] N-formyl-lysine. RESULTS: It was observed that Xan Crude, Xan CHCl3, and Xan EtAc extracted from Xanthium strumarium L. showed approximately 10% of antioxidant effect whereas Xan H2O showed no effect by crosslinking assay. CONCLUSIONS: This study showed that the crosslinking assay described in this study can be developed as a potential tool to screen the anti-oxidant effect rapidly and accurately compared to MTT assay. The result was compared to MTT assay using Human Lens epithelial cell line.
Antioxidants ; Cataract ; Chromatography, Ion Exchange ; Epithelial Cells ; Humans ; Xanthium*

Chromatography, Ion Exchange ; Hemoglobins, Abnormal ; analysis ; Humans ; Male

Chromatography, Ion Exchange ; Hemoglobins, Abnormal ; analysis ; Humans ; Male

Air ; analysis ; Anions ; analysis ; Chromatography, Ion Exchange ; Ions ; analysis ; Workplace

OBJECTIVETo develop a method for determination of mandelic acid (MA) and phenylglyoxylic acid (PGA) in urine by reagent-free ion chromatography.

METHODSIon chromatography was performed on an AS19 column with a gradient elution solution containing 10-35 mmoL/L KOH at a flow rate of 1.00 ml/min, and MA and PGA were detected at ultraviolet wavelengths of 225 nm and 254 nm, respectively. The samples were diluted 10 times with purified water, then purified on a silver column to remove high concentrations of chloride ion, and injected after being filtered through a 0.2-µm m filter membrane.

RESULTSThe recoveries of standard addition of MA and PGA were 96.5% and 99.3%, respectively, with both relative standard deviations less than 5.0%. Good linear relationships were noted in the range of 1.0-100.0 mg/L for both MA and PGA (r >0.9995). The detection limits of MA and PGA were 0.02 mg/L and 0.05 mg/L, respectively; the minimum detectable concentrations of MA and PGA were 0.2 mg/L and 0.5 mg/L (when the sampling amount was 5.0 ml and diluted to 50.0 ml with water, and the injection volume was 300 µL).

CONCLUSIONSThis method is fast, convenient, and highly sensitive and selective. It can be used for the analysis of MA and PGA in the urine of styrene-exposed workers.

Chromatography, Ion Exchange ; Glyoxylates ; urine ; Humans ; Mandelic Acids ; urine ; Styrene

A bromoperoxidase from Gracilaria lemaneiformis was purified to homogeneity using a multi-step process of ammonium sulfate precipitation (AS), dialysis, and DEAE-cellulose 52 anion exchange chromatography. The bromoperoxidase activity was unstable or undetectable in crude extract solution. However, it became stable with electrophoretic purity after this multiple purification process. The anion exchange chromatography purification was a critical step in the purification process, which effectively eliminated the phycobiliprotein and smucilaginous polysaccharides. The purified bromoperoxidase was a monomeric enzyme with the relative molecular masses of 66 kD as determined by denaturing and native gradient gel electrophoresis. The optimal pH for bromoination was 6.0 and bromoperoxidase activity was stable as stored at a broad pH range of 3.0-9.0. Of a range of compounds tested, only vanadium enhanced bromoperoxidase activity. Kinetic studies for the bromination of monochlorodimedone (MCD) showed that the Km values of Br- and H2O2 are 53.5 micromol/L, 38 micromol/L respectively.
Chromatography, Ion Exchange ; methods ; Enzyme Stability ; Gracilaria ; enzymology ; Hydrogen-Ion Concentration ; Kinetics ; Peroxidases ; isolation & purification ; metabolism

A new configuration integrated ion exchange effect system with both electro-migration and electrochemical reaction in a single cell was developed to effectively retrieve metal ions from simulated wastewater using ion exchange resins without additive chemicals. By simply assembling cation exchange resins and anion exchange resins separated by homogeneous membranes, we found that the system will always be acidic in the concentrate compartment so that ion exchange resins could be in-situ regenerated without hydroxide precipitation. Such a realizable design will be really suitable for wastewater purification.
Absorption ; Chromatography, Ion Exchange ; methods ; Electrochemistry ; methods ; Ion Exchange Resins ; chemistry ; Membranes, Artificial ; Nickel ; chemistry ; isolation & purification ; Ultrafiltration ; methods ; Water Pollutants ; isolation & purification ; Water Purification ; methods

AIM: To prepare high-purity ginseng total saponins from a water decoction of Chinese ginseng root. METHOD: Total saponins were efficiently purified by dynamic anion-cation exchange following the removal of hydrophilic impurities by macroporous resin D101. For quality control, ultrahigh-performance liquid chromatography with a charged aerosol detector (CAD) was applied to quantify marker components. The total saponin content was estimated by a colorimetric method using a vanillin-vitriol system and CAD response. RESULTS: D201, which consisted of a cross-linked polystyrene matrix and -N(+)(CH3)3 functional groups, was the best of the four anion exchange resins tested. However, no significant difference in cation exchange ability was observed between D001 (strong acid) and D113 (weak acid), although they have different functional groups and matrices. After purification in combination with D101, D201, and D113, the estimated contents of total saponins were 107% and 90% according to the colorimetric method and CAD response, respectively. The total amount of representative ginsenosides Re, Rd, Rg1, and compound K was approximately 22% based on ultrahigh-performance liquid chromatography-CAD quantitative analysis. CONCLUSION These findings suggest that an ion exchange resin, combined with macroporous adsorption resin separation, is a promising and feasible purification procedure for neutral natural polar components.
Adsorption ; Chromatography, Ion Exchange ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ion Exchange Resins ; chemistry ; Panax ; chemistry ; Plant Roots ; chemistry ; Porosity ; Saponins ; chemistry ; isolation & purification

OBJECTIVETo optimize the isolation and purification conditions for Hap(s) protein of non-typeable Haemophilus influenzae.

METHODSHap(s) protein was purified by ammonium sulfate precipitation, dialysis desalting and Hitrap weak cation exchange columns of CM Sepharose Fast Flow. The condition of the elution was optimized for pH and ionic strength, the absorbance at 280 nm of the elution samples were detected, and the targeted protein band in the collected samples was observed by SDS-PAGE electrophoresis.

RESULTSThe Hitrap ion exchange column was eluted with buffer 1, which resulted in a baseline distribution of absorbance at 280 nm. Buffer 2 elution of the column resulted in the presence of peak absorbance with trails, which was identified to be constituted by some low molecular weight bands by subsequent SDS-PAGE. In serial column elution with buffer 3 with different ionic strength, a peak absorbance was observed with the ionic strength of 100 mmol/L NaCl, and SDS-PAGE confirmed that the peak was generated by the target protein. No obvious peaks or bands in SDS-PAGE occurred with the other ionic strengths.

CONCLUSIONThe pH of the buffer only affect the elution of the irrelevant proteins rather than the Hap(s) protein, and elution with the buffer containing 100 mmol/L NaCl can be optimal for eluting the Hap(s) protein.

Bacterial Proteins ; isolation & purification ; Buffers ; Chromatography, Ion Exchange ; methods ; Electrophoresis, Polyacrylamide Gel ; Haemophilus influenzae ; metabolism ; Hydrogen-Ion Concentration ; Osmolar Concentration