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Chromatographic Behavior (chromatographic + behavior)
Selected AbstractsPeptides of human gingival crevicular fluid determined by HPLC-ESI-MSEUROPEAN JOURNAL OF ORAL SCIENCES, Issue 6 2005Elisabetta Pisano The acidic-soluble protein content of human gingival crevicular fluid was analyzed by reverse-phase high-performance liquid chromatography (RP-HPLC), and the eluent deriving from the chromatography separation was directly introduced into an ion-trap mass spectrometer through electrospray ionization (ESI-IT MS). By this technique the molecular weight of peptides/proteins was determined with a precision of ,,1/10,000 amu. On the basis of the chromatographic behavior and the knowledge of the molecular mass value, some peptides and proteins soluble in acidic solution were unambiguously recognized. Besides high quantities of human serum albumin, , -defensins 1,4 and minor amounts of cystatin A, statherin, basic PB salivary peptide and other unidentified components were detected. The presence of , -defensins in gingival crevicular fluid is in agreement with their relevant contribution to protein composition deriving from granulocyte secretions. Other peptides and proteins abundant in human saliva, such as proline-rich proteins (PRPs) and histatins, were not detected in gingival crevicular fluid. Further investigations will be necessary to establish the origin of statherin and PB salivary peptide in gingival crevicular fluid. [source] Micellar and aqueous-organic liquid chromatography using sub-2,,m packings for fast separation of natural phenolic compoundsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 13 2010Jun Cao Abstract The objective of the present work was to investigate the chromatographic behavior of natural phenolic compounds in micellar and aqueous-organic LC using a short column packed with 1.8,,m particles. Firstly, the effect of ACN and SDS on elution strength and selectivity was examined by isocratic submicellar (0,30% ACN/5% 1-butanol/1,6,mM SDS) and micellar (0,30% ACN/5% 1-butanol/40,60,mM SDS) systems. The varied concentrations of two modifiers in the mobile phases revealed different eluting power. Then, the application of organic modifier gradient was discussed in both submicellar and micellar LC using mobile phases of 4,mM SDS/5% 1-butanol or 50,mM SDS/5% 1-butanol containing ACN gradient from 0 to 30%, respectively. For micellar system, the separation was found to be better in gradient than isocratic elution. Additionally, the sensitivity of aqueous-organic LC was examined. The mobile phase was a mixture of ACN and water employing gradient elution at a flow rate of 0.5,mL/min, with analysis time below 9,min. It was found that separation efficiency was significantly better compared with micellar LC. Besides, the aqueous-organic LC has been applied to separation of various phenolic compounds in Yangwei granule or Radix Astragali samples. [source] Application of hydrophilic interaction chromatography retention coefficients for predicting peptide elution with TFA and methanesulfonic acid ion-pairing reagentsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 6-7 2010Chad E. Wujcik Abstract Hydrophilic retention coefficients for 17 peptides were calculated based on retention coefficients previously published for TSKgel silica-60 and were compared with the experimental elution profile on a Waters Atlantis HILIC silica column using TFA and methanesulfonic acid (MSA) as ion-pairing reagents. Relative peptide retention could be accurately determined with both counter-ions. Peptide retention and chromatographic behavior were influenced by the percent acid modifier used with increases in both retention and peak symmetry observed at increasing modifier concentrations. The enhancement of net peptide polarity through MSA pairing shifted retention out by nearly five-fold for the earliest eluting peptide, compared with TFA. Despite improvements in retention and efficiency (Neff) for MSA over TFA, a consistent reduction in calculated selectivity (,) was observed. This result is believed to be attributed to the stronger polar contribution of MSA masking and diminishing the underlying influence of the amino acid residues of each associated peptide. Finally, post-column infusion of propionic acid and acetic acid was evaluated for their potential to recover signal intensity for TFA and MSA counter-ions for LC-ESI-MS applications. Acetic acid generally yielded more substantial signal improvements over propionic acid on the TFA system while minimal benefits and some further reductions were noted with MSA. [source] Evaluation of mobile phase, ion pairing, and temperature influence on an HILIC-MS/MS method for L -arginine and its dimethylated derivatives detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 13 2008Giuseppe Paglia Abstract Asymmetric NG,,NG -dimethylarginine (ADMA) increases in diseases such as renal failure, diabetes mellitus, and hypercholesterolemia. The feasibility and utility of a hydrophilic interaction chromatography (HILIC) method for the separation of free L -arginine (Arg), ADMA, and symmetric NG,,NG, -dimethylarginine (SDMA) on a typical silica column were explored and the impact of some experimental parameters on the chromatographic behavior of these analytes was investigated. The effect of water and TFA content in mobile phase and of column temperature was investigated during the development of a fast and simple HILIC-MS/MS method that might be suitable for the quantification of free Arg, ADMA, and SDMA in plasma for routine analysis. Our results show that a good compromise between efficiency and peak shape with acceptable retention and total chromatographic run time is achieved using an ACN/water (90:10) mobile phase with TFA% as additive ranging from 0.015 to 0.025% and column temperature ranging from 25 to 30°C. [source] Preparation and characterization of C16 monolithic columns for capillary electrochromatographyJOURNAL OF SEPARATION SCIENCE, JSS, Issue 3 2005Kai Zhang Abstract A series of methacrylamide-based C16 monolithic columns were prepared and characterized to determine how their porous structural properties and chromatographic behavior are affected by the percentages of functional monomer, base monomer, and cross-linker in the polymerization solution. Baseline separation of 6 neutral compounds can be readily obtained in an optimized column. Furthermore, the effects of organic additive in the mobile phase, operating voltages, and temperature on retention behaviors and separation efficiencies were also studied. The separation mechanism is also discussed. High column efficiency and good reproducibility indicate that the monolithic columns hold considerable promise. [source] Study of influence of additives of tyloxapol on the chromatographic characteristics of the model compounds: the comparative characterization of micellar mobile phases of tyloxapol and Triton X-100BIOMEDICAL CHROMATOGRAPHY, Issue 7 2009Marina Rukhadze Abstract The chromatographic behavior of model compounds of biomedical significance (organic acids, amino acids, drugs) was investigated using mobile phases modified with tyloxapol. The influence of factors such as concentration of tyloxapol, content of organic modifier and pH of mobile phase on the retention factor of solutes was studied. The results were compared with the data obtained by elution with mobile phases containing Triton X-100 additives, since units of Triton X-100 are repeated in the structure of tyloxapol. Divergence in chromatographic behavior of model compounds was explained by the difference in physico-chemical characteristics (microviscosity, polarity, critical micelle concentration, shape of micelles, etc.) of tyloxapol and Triton X-100 micelles. Copyright © 2009 John Wiley & Sons, Ltd. [source] Protein loading, elution, and resolution behavior in a novel device that integrates ultrafiltration and chromatographic separationBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2003Xiao-Ping Dai Abstract Hollow fiber membranes and chromatographic resin beads are commonly employed in a variety of bioseparation processes. A new class of integrated separation devices is being studied in which the shell side of a hollow fiber device is filled with adsorbents/chromatographic resin beads. Such devices and the corresponding separation methods integrate feed broth clarification by the microfiltration/ultrafiltration membrane with bioproduct purification by the shell-side resin beads either as an adsorbent or as beads in elution chromatography. A mathematical model has been developed for the prediction of the chromatographic behavior of such an integrated device. Simulations have been done to study the effects of axial dispersion, feed flow rate, water permeation rate, fiber packing density, and void fraction. Numerical solutions were obtained by solving the governing equations. This model can reasonably describe the concentration profiles as well as the breakthrough and elution behaviors in the integrated device. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 125,139, 2003. [source] Comparative HPLC enantioseparation of ferrocenylalcohols on two cellulose-based chiral stationary phasesCHIRALITY, Issue 5 2007Angela Patti Abstract The direct HPLC enantiomeric separation of several ferrocenylalcohols on the commercially available Chiralcel OD and Chiralcel OJ columns has been evaluated in normal-phase mode. Almost all the compounds were resolved on one or both chiral stationary phases (CSPs) with separation factor (,) ranging from 1.06 to 2.88 while the resolution (Rs) varied from 0.63 to 12.70 In the separation of the ,-ferrocenylalcohols 1a,e and the phenyl analogues 2a,e, which were all resolved except 1c, a similar trend in the retention behavior for the two series of alcohols was evidenced and the selectivity was roughly complementary on the two investigated CSP. For three ferrocenylacohols, chosen as model compounds, the influence of the mobile phase composition and temperature on the enantioseparation were investigated and additional information on the chiral recognition mechanism were deduced from the chromatographic behavior of their acetylderivatives. Chirality, 2007. © 2007 Wiley-Liss, Inc. [source] New approach for rapid detection of known hemoglobin variants using LC-MS/MS combined with a peptide database,JOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 3 2007F. Basilico Abstract The identification of hemoglobin (Hb) variants is usually performed by means of different analytical steps and methodologies. Phenotypic methods, such as gel electrophoresis and high performance liquid chromatography, are used to detect the different electrophoretic or chromatographic behaviors of hemoglobin variants in comparison to HbA0 used as a control. These data often need to be combined with mass spectrometry analyses of intact globins and their tryptic peptide mixtures. As an alternative to a ,step-by-step' procedure, we have developed a ,single step' approach for the identification of Hb variants present in biological samples. This is based on the µHPLC-ESI-MS/MS analysis of the peptide mixture generated by a tryptic digestion of diluted Hb samples and an in-house new database containing solely the variant tryptic peptide of known human Hb variants. The experimental results (full MS and MS/MS spectra) are correlated with theoretical mass spectra generated from our in-house-built variant peptide database (Hbp) using the SEQUEST algorithm. Simple preparation of samples and an automated identification of the variant peptide are the main characteristics of this approach, making it an attractive method for the detection of Hb variants at the routine clinical level. We have analyzed 16 different samples, each containing a different known variant of hemoglobin. Copyright © 2006 John Wiley & Sons, Ltd. [source] Simultaneous Determination of Epinephrine, Noradrenaline and Dopamine in Human Serum Samples by High Performance Liquid Chromatography with Chemiluminescence DetectionCHINESE JOURNAL OF CHEMISTRY, Issue 7 2007Fu-Nan Chen Abstract A simple, rapid and accurate high performance liquid chromatographic (HPLC) technique coupled with chemiluminescence (CL) detection was developed for the simultaneous determination of epinephrine (E), noradrenaline (NA) and dopamine (DA). It was based on the analyte enhancement effect on the CL reaction between luminol and potassium ferricyanide. The effects of various parameters, such as potassium ferricyanide concentration, luminol concentration, pH value and component of the mobile phase on chromatographic behaviors of the analytes (E, NA and DA) were investigated. The separation was carried out on C18 column using the mobile phase of 0.01 mol/L potassium hydrogen phthalate solution and methanol (92:8, V/V). Under the optimum conditions, E, NA and DA showed good linear relationships in the range of 1×10,8,5×10,6, 5.0×10,9,1.0×10,6 and 5.0×10,9,1.0×10,6 g/mL respectively. The detection limits for E, NA and DA were 4.0×10,9, 1.0×10,9 and 8.0×10,10 g/mL. The proposed method has been applied successfully to the analysis of E, NA and DA in human serum samples. [source] | ||||||||||